Patent Description:
Inflammation is a very common and important basic pathological process, and a common and frequently-occurring disease of most organs and trauma infections on the body surface. Inflammation can be caused by infection (such as pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, etc.). It can also be non-infectious inflammation, which is usually closely related to the immunity of the body (such as allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, etc.). Meanwhile, inflammation is also one of the main predisposing factors of the onset of cancer (such as lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, etc.). Also, studies have shown that some inflammatory factors are also related to metabolic diseases (such as diabetes, gout, etc.). Under normal circumstances, inflammation is beneficial and is an automatic defense response of the body. But sometimes inflammation is also harmful, for example, attacks on the own tissues of the body, inflammation that occurs in hyaline tissues, and the like.

The clinical manifestations of inflammation include redness, swelling, fever, pain, dysfunction, etc., while the biochemical indicators of inflammation usually refer to high expression of inflammatory factors which are involved in the inflammation process and mediate the inflammatory response, for example, IL-1beta, IL-<NUM> and TNF-alpha. At present, the prevention and treatment of inflammation is still mainly based on western medicine, but there are also many traditional Chinese medicine products that have certain anti-inflammatory effects, but all have their own unavoidable shortcomings. RNA therapeutics and their use for the treatment of inflammation is disclosed inter alia in <CIT>, <CIT>, <NPL>), and <NPL>). Therefore, it is still necessary to find novel anti-inflammatory treatment measures.

The present invention provides the following as defined in the claims:.

Any small RNAs other than BZL-sRNA-<NUM> having SEQ ID NO: <NUM> are comparative examples and do not form part of the present invention.

The present invention relates to a small RNA having the sequence of SEQ ID NO: <NUM> for use in the treatment of an IL-1beta, IL-<NUM> or/and TNF-alpha related disease as specified in claim <NUM>, wherein administration of the small RNA rescues cell death caused by an infection with a virus, preferably with H5N1. Generally, siRNA, miRNA and other non-coding small RNAs are indiscriminately referred to as small RNAs (sRNAs). Unless otherwise specified, the term "small RNA (sRNA)" herein refers to non-coding small RNAs including siRNA and miRNA. The "small" in the term does not limit the RNA to a specific size.

The terms "including", "comprising" and "containing" mean that in addition to the listed characteristic elements, there may be other additional characteristic elements. In particular, it can also consist of only the listed characteristic elements.

The term "subject" refers to, for example, a subject who suffers from inflammation and/or H5N1 infection and needs treatment or who has signs of potential inflammation development and/or who is susceptible to H5N1 infection and in need of prevention.

Sequence "identity" is used herein to describe the relativity between two amino acid sequences or between two nucleotide sequences. As used herein, the sequence identity between two deoxyribonucleotide sequences is determined by using the Needleman-Wunsch algorithm (Needleman and Wunsch, <NUM>) implemented in the Needle program of the EMBOSS package (<NPL>) (preferably version <NUM>. <NUM> or later versions). The parameters used are a gap open penalty of <NUM>, a gap extension penalty of <NUM>, and the EDNAFULL substitution matrix (EMBOSS version of NCBI NUC4. The Needle output (obtained with the -nobrief option) marked as "the longest identity" is used as the percentage identity and calculated as follows:<MAT>.

As used herein, the term "stringent conditions" refers to, for example. hybridization in <NUM>×SSC at <NUM>, followed by washing several times in <NUM>×SSC at <NUM>, for a total of approximately <NUM> hour. The term "stringent hybridization conditions" used herein can also refer to hybridization in <NUM> sodium phosphate, <NUM>% SDS pH <NUM>, <NUM> EDTA and <NUM>% BSA at <NUM> for <NUM> hours, followed by washing twice in <NUM>×SSC and <NUM>% SDS at <NUM>. Those skilled in the art can determine the stringent conditions according to the specific sequence.

The term "cell survival rate" is also called cell viability. Cell survival rate can be calculated by using the MTS detection kit according to the manufacturer's instructions.

The term "IL-1beta, IL-<NUM> or/and TNF-alpha related disease" refers to the diseases characterized by an increase of the expression level of IL-1beta, IL-<NUM> or/and TNF-alphaselected from the group consisting of pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, allergic dermatitis, multiple sclerosis, systemic lupus erythematosus, lung cancer, gastric cancer, colorectal cancer, liver cancer, cervical cancer, breast cancer, leukemia, diabetes, gout, etc..

The small RNA having the sequence of SEQ ID NO: <NUM> was found to reduce or down-regulate the expression level of IL-1beta, IL-<NUM> or/and TNF-alpha in vivo or in vitro, thus rescuing cell death caused by H5N1 infection in a subject. The small RNA sequences used in the experiment are as shown in Table <NUM> below (any small RNAs other than BZL-sRNA-<NUM> having SEQ ID NO:<NUM> are comparative examples and do not form part of the present invention).

As used herein, the concentration of the aforementioned small RNA employed is <NUM>. The synthetic small RNAs may be used to test their ability to inhibit any one or more pathway(s) or gene(s) listed in Table <NUM>, or used to reduce or down-regulate the expression level of IL-1beta, IL-<NUM> or/and TNF-alpha in vitro or in vivo, and/or to improve cell survival rate and/or to treat or prevent IL-1beta, IL-<NUM> or/and TNF-alpha related diseases and/or H5N1 infection in a subject.

It should be understood that those skilled in the art can prepare a nucleic acid construct encoding the small RNA of the present invention, and can introduce the such nucleic acid construct into suitable expression vectors. The expression vector expressing the small RNA of the present invention can be directly introduced into a cell, provided that the expression vector can be expressed in the cell. For example, see US Patent <CIT>.

In addition, those skilled in the art can prepare constructs for expressing small RNA in cells, for example retroviral constructs, and through transfecting the packaging cell line with the constructs also produce recombinant retroviral particlest. For example, see US Patent <CIT>.

Those skilled in the art can introduce the cells containing expression vectors or constructs into a cell n. Alternatively, those skilled in the art can isolate the small RNAs from cells by conventional techniques. Therefore, the present invention encompasses methods for expressing small RNAs, which include the steps of cultivating the cells under suitable conditions and recovering small RNAs.

The small RNA claimed is BZL-sRNA-<NUM> having the sequence as shown nin SEQ ID NO: <NUM>. The inventors found that BZL-sRNA-<NUM> has a very favorable effect in inhibiting TNF-alpha, IL-1beta or IL-<NUM> protein or the respective mRNAs thereof. Therefore, the inventors selected BZL-sRNA-<NUM> as the basic small RNA, and combined it with other small RNAs to prepare the small RNA mixtures described in Table <NUM>.

The siRNA mixtures are prepared by mixing <NUM> of BZL-sRNA-<NUM> and <NUM> each of other small RNAs at a volume ratio of <NUM>:<NUM>. In one embodiment, the molar concentration of BZL-sRNA-<NUM> and the other small RNAs is <NUM>:<NUM>. In one embodiment, the molar concentration of total small RNA in the small RNA mixture is <NUM>. In the Figures, the mixtures are indicated by the symbol MIX.

The small RNA BZL-sRNA-<NUM> having SEQ ID NO:<NUM> can be used to treat IL-<NUM> related diseases. IL-<NUM> related diseases include (diseases other than those specified in claim <NUM> are comparative examples and do not form part of the present invention):.

The BZL-sRNA-<NUM> may be used in combination with one or more of the following:.

In order to treat inflammatory diseases, BZL-sRNA-<NUM> may be used in combination with one or more of the following agent(s): for example, non-steroidal anti-inflammatory agent(s) (hereinafter referred to as NSAIDs), including non-selective cyclooxygenase COX-<NUM>/COX-<NUM> inhibitors, regardless of topical or systemic application (for example piroxicam, diclofenac, propionic acids, such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates, such as mefenamic acid, indomethacin, sulindac, azapropazone, pyrazolones, for example phenylbutazone, salicylates, for example aspirin); selective COX-<NUM> inhibitors (for example, meloxicam, celecoxib, rofecoxib, valdecoxib, lumarocoxib, parecoxib and etoricoxib); cyclooxygenase inhibiting nitric oxide donators (CINODs); glucocorticoids (whether through local, oral, intramuscular, intravenous or intraarticular routes); methotrexate, leflunomide; hydroxychloroquine, d-penicillamine, auranofin or other parenteral or oral gold products; analgesics; diacerein; intraarticular treatments, for example hyaluronic acid derivatives; and nutritional additives, for example glucosamine.

BZL-sRNA-<NUM> may also be used in combination with existing therapeutic agents for the treatment of cancer.

Suitable agents that can be used in combination include: (i) anti-proliferative/anti-tumor agent(s) and combinations thereof used in medical oncology, for example gleevec (imatinib mesylate), alkylating agents (for example, cisplatin, carboplatin, cyclophosphamide, chlormethine, melphalan, chlorambucil, busulfan and nitrosourea); antimetabolites (for example, antifolates, such as fluoropyrimidines, such as <NUM>-fluorouracil and tegafur, raltitrexed, methotrexate, cytarabine, hydroxyurea, gemcitabine and paclitaxel; antitumor antibiotics (for example, anthracycline antibiotics, such as adriamycin, bleomycin, doxorubicin, daunorubicin, epirubicin, idarubicin, mitomycin-C, dactinomycin and mithramycin); anti-(mitosis) division agents (for example, vinblastines, such as vincristine, vinblastine, vindesine and vinorelbine and taxanes, such as paclitaxel and taxotere); and topoisomerase inhibitors (for example, etoposides, such as etoposide and teniposide, amsacrine, topotecan and camptothecin);.

IL-1beta plays a key role in the pathology related to a variety of diseases involving immune and inflammatory elements. The small RNA BZL-sRNA-<NUM> having SEQ ID NO: <NUM> can be used to treat IL-1beta related diseases. These diseases include(diseases other than those specified in claim <NUM> are comparative examples and do not form part of the present invention): acquired immunodeficiency syndrome; acquired immunodeficiency-related diseases; acquired pernicious anemia; acute coronary syndrome; acute and chronic pain (different forms of pain); acute idiopathic polyneuropathy inflammation; acute immune diseases related to organ transplantation; acute or chronic immune diseases related to organ transplantation; acute inflammatory demyelinating polyneurotic neuropathy; acute ischemia; acute liver disease; acute rheumatic fever; acute transverse myelitis; Addison's disease; adult (acute) respiratory distress syndrome; adult Still's disease; alcohol-induced cirrhosis; alcohol-induced liver injury; allergic disease; allergy; alopecia; alopecia areata; Alzheimer's disease; allergic reaction; ankylosing spondylitis; ankylosing spondylitis-related lung disease; antiphospholipid antibody syndrome; aplastic anemia; arteriosclerosis; arthropathy; asthma; atherosclerotic disease/ arteriosclerosis; atherosclerosis; atopic allergy; atopic eczema; atopic dermatitis; atrophy autoimmune hypothyroidism; autoimmune bullous disease; autoimmune dermatitis; autoimmune diabetes; autoimmune disorders related to streptococcal infection; autoimmune enteropathy; autoimmune hemolytic anemia; autoimmune hepatitis; autoimmune hearing loss; autoimmune lymphoproliferative syndrome (ALPS); autoimmune-mediated hypo- glycemia; autoimmune myocarditis; autoimmune neutropenia; autoimmune premature ovarian failure; autoimmune thrombocytopenia (AITP); autoimmune thyroid disease; autoimmune uveitis; bronchiolitis obliterans; Behcet's disease; blepharitis; bronchiectasis; bullous pemphigoid; cachexia; cardiovascular disease; catastrophic antiphospholipid syndrome; celiac disease; cervical joint stiffness; chlamydia; choleosatatis; chronic active hepatitis; chronic eosinophilic pneumonia; chronic fatigue syndrome; chronic immune diseases related to organ transplantation; chronic ischemia; chronic liver disease; chronic mucocutaneous candidiasis; cicatricial pemphigoid; clinical isolation syndrome with risk of multiple sclerosis (CIS); common immunodeficiencies (common variable hypogammaglobulinemia); connective tissue disease-related interstitial lung disease; conjunctivitis; Coombs positive hemolytic anemia; childhood-onset psychosis; chronic obstructive pulmonary disease (COPD); Crohn's disease; cryptogenic autoimmune hepatitis; cryptogenic fibrotic alveolitis; dacryocystitis; depression; dermatitis scleroderma; dermatomyositis; dermatomyositis/polymyositis-related lung disease; diabetic retinopathy; diabetes; dilated cardiomyopathy; discoid lupus erythematosus; disc herniation; disc prolapse; disseminated intravascular coagulation; drug-induced hepatitis; drug-induced interstitial lung disease; drug-induced immune hemolytic anemia; endocarditis; endometriosis; endophthalmitis; enteropathic synovitis; episcleitis; erythema multiforme; severe erythema multiforme; female infertility; fibrosis; fibrotic lung disease; gestational pemphigoid; giant cell arteritis (GCA); glomerulonephritis; goiter autoimmune hypothyroidism (Hashimoto's disease); Goodpasture's syndrome; gouty arthritis; graft-versus-host disease (GVHD); Grave's disease; group B streptococcus (BGS) infection; Guillain-Barre syndrome (GBS); hemosiderinosis-related lung disease; hay fever; heart failure; hemolytic anemia; Henoch-Schonlein purpura; hepatitis B; hepatitis C; Hughes syndrome; Huntington's disease; hyperthyroidism; hypoparathyroidism; idiopathic leukopenia; idiopathic thrombocytopenia; idiopathic Parkinson's disease; idiopathic interstitial pneumonia; idiocratic liver disease; IgE mediated allergies; immune hemolytic anemia; inclusion body myositis; infectious diseases; infectious ophthalmic diseases; inflammatory bowel disease; inflammatory demyelinating disease; inflammatory heart disease; inflammatory nephropathy; insulin dependent diabetes; interstitial pneumonia; IPF/UIP; iritis; juvenile chronic arthritis; juvenile pernicious anemia; juvenile rheumatoid arthritis (JRA); Kawasaki's disease; keratitis; keratoconjunctivitis sicca; Kussmaul disease or Kussmaul-Meier disease; Landry's paralysis; Langerhans cell histiocytosis; linear IgA disease; livedo reticularis; Lyme arthritis; lymphocytic infiltrating lung disease; macular degeneration; male idiopathic infertility or NOS; malignant tumor; microvasculitis of the kidney; microscopic polyangiitis; mixed connective tissue disease-related lung disease; Morbus Bechterev; motor neuron disease; mucosal pemphigoid; multiple sclerosis (all subtypes: primary progressive, secondary progressive, relapsing remitting, etc.); multiple organ failure; myalgia encephalitis/Royal Free disease; myasthenia gravis; myelodysplastic syndrome; myocardial infarction; myocarditis; nephrotic syndrome; nerve root disorder; neuropathy; non-alcoholic steatohepatitis; non-A, non-B hepatitis; optic neuritis; organ transplant rejection; osteoarthritis; osteolysis; ovarian cancer; ovarian failure; pancreatitis; parasitic disease; Parkinson's disease; pauciarticular JRA; pemphigoid; pemphigus foliaceus; pemphigus vulgaris; peripheral arterial occlusive disease (PAOD); peripheral vascular disease (PVD); peripheral artery disease (PAD); phacolytic uveitis; phlebitis; polyarteritis nodosa (or nodular epiarteritis); polychondritis; polymyalgia rheumatica; poliosis; polyarticular JRA; polyendocrine deficiency syndrome; polymyositis; polyglandular type I and polyglandular type II deficiency; polymyalgia rheumatica (PMR); post-infection interstitial lung disease; post-inflammatory interstitial lung disease; postpump syndrome; premature ovarian failure; primary biliary cirrhosis; primary mucinous edema; primary Parkinson's syndrome; primary sclerosing cholangitis; primary sclerosing hepatitis; primary vasculitis; prostate and rectal cancer and hematopoietic malignancies (leukemia and lymphoma); prostatitis; psoriasis; psoriasis type <NUM>; psoriasis type <NUM>; psoriatic arthritis; psoriatic arthropathy; pulmonary hypertension secondary to connective tissue disease; pulmonary manifestations of polyarteritis nodosa; pure red cell aplasia; primary adrenal insufficiency; radiation fibrosis; reactive arthritis; Reiter's disease; recurrent neuromyelitis optica; renal disease NOS; restenosis; rheumatoid arthritis; rheumatoid arthritis-related interstitial lung disease; rheumatic heart disease; SAPHO (synovitis, acne, pustulosis, hyperostosis and osteitis); sarcoidosis; schizophrenia; Schmidt's syndrome; scleroderma; secondary amyloid-dosis; shock lung; scleritis; sciatica; secondary adrenal insufficiency; sepsis syndrome; septic arthritis; septic shock; seronegative arthropathy; silicone-related connective tissue disease; Sjogren's disease-related lung disease Sjogren's syndrome; Sneddon-Wilkinson skin disease; sperm autoimmunity; spondyloarthropathy; spondylitis ankylopoietica; Stevens- Johnson syndrome (SJS); Still's disease; stroke; sympathetic ophthalmia; systemic inflammatory response syndrome; systemic lupus erythematosus; systemic lupus erythematosus-related lung disease; systemic scleroderma; systemic scleroderma-related interstitial lung disease; Takayasu's disease/arteritis; temporal arteritis; Th2 type and Th1 type mediated diseases; thyroiditis; Toxic shock syndrome; toxoplasma retinitis; toxic epidermal necrolysis; transverse myelitis; TRAPS (tumor necrosis factor receptor type I (TNFR) associated periodic syndrome); type B insulin resistance with acanthosis nigricans; type <NUM> allergy; type <NUM> autoimmune hepatitis (traditional autoimmune or lupus-like hepatitis); type <NUM> autoimmune hepatitis (anti-LKM antibody hepatitis); type II diabetes; ulcerative colitis arthropathy; ulcer colitis; urticaria; usual interstitial pneumonia (UIP); uveitis; vasculitis disseminated lung disease; vasculitis; vernal conjunctivitis; viral retinitis; vitiligo; Vogt-Koyanagi-Harada syndrome (VKH syndrome); Wegener's granulomatosis; wet macular degeneration; wound healing; Yersinia and Salmonella associated arthropathy.

The the small RNA BZL-sRNA-<NUM> may be used in combination withat least one additional agent listed below. The combination may also include more than one additional agent, for example, <NUM> or <NUM> additional agents.

Exemplary combinations include the small RNA BZL-sRNA-<NUM> and non-steroidal anti-inflammatory drugs (NSAIDS), for example ibuprofen. Other exemplary combinations include BZL-sRNA-<NUM> and corticosteroids, including prednisolone. Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for rheumatoid arthritis include: cytokine suppressive anti-inflammatory drugs (CSAIDs); antibodies or antagonists against other human cytokines or growth factors, for example, TNF, LT, IL-1α, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL- <NUM>, IL-<NUM>, interferon, EMAP-II, GM-CSF, FGF and PDGF. BZL-sRNA-<NUM> can be combined with antibodies against cell surface molecules or ligands thereof including CD154 (gp39 or CD40L), said cell surface molecules are for example CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, CD80 (B7. <NUM>), CD86 (B7. <NUM>), CD90 and CTLA.

Exemplary therapeutic agents used in combination with BZL-sRNA-<NUM> can interfere at different points in the autoimmune and subsequent inflammatory cascades, for example TNF antagonists, such as chimeric, humanized or human TNF antibodies, D2E7 (<CIT>), CA2 (REMICADEa), CDP <NUM>, and soluble p55 or p75 TNF receptors, derivatives thereof (p75TNFR1gG (ENBRELa) or p55TNFR1gG (Lenercept), and TNF-alpha converting enzyme (TACE) inhibitors, and other IL-<NUM> inhibitors (interleukin-<NUM> converting enzyme inhibitors, IL-1RA, etc.). Other reagents used in combination with BZL-sRNA-<NUM> include interleukin <NUM>, reagents that act in parallel with IL-1a function, depend on IL-1a function, or is consistent with IL-1a function, for example IL-<NUM> antagonists (for example IL-<NUM> binding proteins for example antibodies or soluble IL-<NUM> receptors, or antigen-binding fragments thereof). Additional reagents used in combination with BZL-sRNA-<NUM> include non-exhaustive anti-CD4 inhibitors, costimulatory pathway CD80 (B7. <NUM>) or CD86 (B7. <NUM>) antagonists, including antibodies, soluble receptors, antagonistic ligands or antigen-binding fragments thereof.

BZL-sRNA-<NUM> can also be combined with agents for the treatment of rheumatoid arthritis, for example methotrexate, <NUM>-MP, azathioprine sulfasalazine, mesalazine, olsalazine chloroquinine/hydroxychloroquine, penicillamine, gold sodium thiomalate (intramuscular and oral), azathioprine, colchicine, corticosteroids (oral, inhalation and local injection), beta2 adrenergic receptor agonists (salbutamol, terbutaline and salmeterol), xanthine (theophylline and aminophylline), cromoglycates, nadocromil, ketotifen, ipratropium and oxitropium, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs for example ibuprofen, corticosteroids for example prednisolone, phosphodiesterase inhibitors, adenosine agonists, anticoagulants, complement inhibitors, adrenergics, reagents that interfere with the sinaling through pro-inflammatory cytokines for example TNF-alpha or IL-<NUM> (for example IRAK, NIK, IKK, p38 and MAP kinase inhibitors), IL-1beta converting enzyme inhibitors, TNF-alpha converting enzyme (TACE) inhibitors, T cell signaling inhibitors for example kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, azathioprine, <NUM>-mercaptopurine, angio- tensin converting enzyme inhibitors, soluble cytokine receptors and derivatives thereof (for example, soluble p55 or p75 TNF receptors and derivatives p75TNFRIgG (ENBRELTM and p55TNFRIgG (Lenercept)), sIL-1RI, sIL-1RII, sIL-6R), anti-inflammatory cytokines (for example, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM> and TGFbeta), celecoxib, folic acid, hydroxychloroquine sulfate, profencoxib, etanercept, infliximab, naproxen, valdecoxib, sulfasalazine, methylprednisolone, meloxicam, methylprednisolone acetate, gold sodium thiomalate, aspirin, triamcinolone, dextropropoxyphene naphthalenesulfonate/paracetamol, folates, naproxen, voltarin, piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone hydrochloride, dihydrocodeinone bitartrate/paracetamol, diclofenac sodium/misoprostol, fentanyl, anakinra, human recombinant, tramadol hydrochloride, salsalate, sulindac, cyanocobalamin/fa/ pyridoxine, paracetamol, sodium alendronate, prednisolone, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulfate (glucosamine sulf)/chondroitin, amitriptyline hydrochloride, sulfadiazine, oxycodone hydrochloride/paracetamol, olopatidine hydrochloride, misoprostol, sodium methoxy naphthyl propionate, omeprazole, cyclophosphamide, rituximab, IL-<NUM> TRAP, MRA, CTLA4-IG, IL-<NUM> BP, anti-IL-<NUM>, anti-IL15, BIRB-<NUM>, SCIO-<NUM>, VX-<NUM>, AMG-<NUM>, VX-<NUM>, roflumilast, IC-<NUM>, CDC-<NUM> and mesopram.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for inflammatory bowel disease include: budesonide; epidermal growth factors, corticosteroids, cyclosporine, sulfasalazine, aminosalicylates, <NUM>-mercapto-purine, azathioprine, metronidazole, lipoxygenase inhibitors, mesalazine, olsalazine, balsala-zide, antioxidants, thromboxane inhibitors, IL-<NUM> receptor antagonists, anti-IL-1beta monoclonal antibodies, anti-IL-<NUM> monoclonal antibodies, growth factors, elastase inhibitors, pyridyl-imidazole compounds, antibodies or antagonists against other human cytokines or growth factors for example TNF, LT, IL-1beta, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, EMAP-II, GM-CSF, FGF and PDGF. BZL-sRNA-<NUM> can be combined with antibodies against cell surface molecules and ligands thereof, said cell surface molecules are for example CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69 and CD90. BZL-sRNA-<NUM> can also be combined with reagents for example methotrexate, cyclosporine, FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs such as ibuprofen, corticosteroids such as prednisolone, phosphodiesterase inhibitors, adenosine agonists, anticoagulants, complement inhibitors, adrenergic agent(s), reagents that interfere with the signaling via pro-inflammatory cytokines for example TNF-alpha or IL-<NUM> (e.g., IRAK, NIK, IKK, p38 or MAP kinase inhibitors), IL-1beta converting enzyme inhibitors, TNF-alpha converting enzyme inhibitors, T cell signaling inhibitors for example kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, azathioprine, <NUM>-mercaptopurine, angiotensin-converting enzyme inhibitors, soluble cytokine receptors and derivatives thereof (e.g., soluble p55 or p75 TNF receptors, sIL-1RI, sIL-1RII, sIL-6R) and anti-inflammatory cytokines (e.g., IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, TGFbeta).

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for Crohn's disease include: TNF antagonists, for example anti-TNF antibodies, D2E7 (<CIT>; HUMIRA®), CA2 (REMICADE®), CDP <NUM>, TNFR-Ig constructs, (p75TNFRIgG (ENBREL®) and p55TNFRIgG (Lenercept)) inhibitors and PDE4 inhibitors. BZL-sRNA-<NUM> can be combined with corticosteroids, for example budesonide and dexamethasone. BZL-sRNA-<NUM> can also be combined with reagents for example sulfasalazine, <NUM>-aminosalicylic acid and olsalazine, and reagents that interfere with the synthesis or action of pro-inflammatory cytokines (for example IL-<NUM>) (for example IL-1beta converting enzyme inhibitors and IL-1RA). Small RNAs can also be used together with T cell signaling inhibitors, for example, tyrosine kinase inhibitor <NUM>-mercaptopurine. BZL-sRNA-<NUM> can be combined with IL-<NUM>. BZL-sRNA-<NUM> can be combined with the following reagents: mesalazine, prednisone, azathioprine, mercaptopurine, infliximab, methylprednisolone sodium succinate, diphenoxylate/atropine sulfate, loperamide hydro- chloride, methotrexate, omeprazole, folates, ciprofloxacin/glucose-injection, dihydrocodeinone bitartrate/paracetamol, tetracycline hydrochloride, fluocinolone, metronidazole, thimerosal/boric acid, cholestyramine/sucrose, ciprofloxacin hydrochloride, hyoscyamine sulfate, pethidine hydrochloride, midazolam hydrochloride, oxycodone hydrochloride/paracetamol, promethazine hydrochloride, sodium phosphate, sulfamethoxazole/trimethoprim, celecoxib, polycarbophil, propoxyphene napsylate, hydrocortisone, multivitamins, balsalazide disodium, codeine phosphate/paracetamol, colesevelam hydrochloride (colesevelam hcl), cyanocobalamin, folic acid, levofloxacin, methylprednisolone, natalizumab and interferon γ.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for multiple sclerosis include: corticosteroids, prednisolone, methylprednisolone, azathioprine, cyclophosphamide, cyclosporin, methotrexate, <NUM>-amino-pyridine, tizanidine, interferon-β1a (AVONEX®, Biogen), interferon-β1b (BETASERON®, Chiron/Berlex), interferon α-n3 (Interferon Sciences/Fujimoto), interferon-α (Alfa Wassermann/J&J), interferon beta1A-IF (Serono/Inhale Therapeutics), pegylated interferon (peginterferon) α2b (Enzon/Schering-Plough), copolymer <NUM> (Cop-<NUM>, COPAXONE®; Teva Pharmaceutical Industries, Inc. ), hyperbaric oxygen, intravenous immunoglobulin, cladribine, antibodies, antagonists or inhibitors against other human cytokines or growth factors and receptors thereof, for example, TNF, LT, IL-1beta, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, IL-1A, IL-<NUM>, IL-<NUM>, IL-<NUM>, EMAP-II, GM-CSF, FGF and PDGF. BZL-sRNA-<NUM> can be combined with antibodies against cell surface molecules or ligands thereof, said cell surface molecules are for example CD2, CD3, CD4, CD8, CD19, CD20, CD25, CD28, CD30, CD40, CD45, CD69, CD80, CD86 and CD90. BZL-sRNA-<NUM> can also be combined with reagents for example FK506, rapamycin, mycophenolate mofetil, leflunomide, NSAIDs for example ibuprofen, phosphodiesterase inhibitors, adenosine agonists, anticoagulants, complement inhibitors, adrenergic agent(s), reagents that interfere with the signaling via pro-inflammatory cytokines for example TNF-alpha or IL-<NUM> (for example IRAK, NIK, IKK, p38 or MAP kinase inhibitors), IL-1b converting enzyme inhibitors, TACE inhibitors, T cell signaling inhibitors for example kinase inhibitors, metalloproteinase inhibitors, sulfasalazine, azathioprine, <NUM>-mercaptopurine, angiotensin-converting enzyme inhibitors, soluble cytokine receptors and derivatives thereof (for example soluble p55 or p75 TNF receptors, sIL-1RI, sIL-1RII, sIL-6R), anti-inflammatory cytokines (for example IL-<NUM>, IL-<NUM>, IL-<NUM> and TGFbeta), COPAXONE®, and caspase inhibitors for example caspase-<NUM> inhibitors.

BZL-sRNA-<NUM> can also be combined with reagents for example alemtuzumab, dronabinol, Unimed, daclizumab, mitoxantrone, xaliproden hydrochloride, <NUM>-aminopyridine, glatiramer acetate, natalizumab, sinnabidol, a-immunokine NNSO3, ABR-<NUM>, AnergiX. MS, chemokine receptor antagonists, BBR-<NUM>, calagualine, CPI-<NUM>, LEM (liposome encapsulated mitoxantrone), THC. CBD (cannabinoid agonist), MBP-<NUM>, mesopram (PDE4 inhibitor), MNA-<NUM>, anti-IL-<NUM> receptor antibodies, neurovax, pirfenidone alltrap <NUM> (RDP-<NUM>), sTNF-R1, talampanel, teriflunomide, TGF-beta2, tiplimotide, VLA-<NUM> antagonist (e.g., TR-<NUM>, VLA4 ultrahaler, antegran-ELAN/Biogen), interferon γ antagonists and IL-<NUM> agonists.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of angina include: aspirin, nitroglycerin, isosorbide mononitrate, metoprolol succinate, atenolol, metoprolol tar- trate, amlodipine besylate, diltiazem hydrochloride, isosorbide dinitrate, clopidogrel disulfate, nifedipine, atorvastatin calcium, potassium chloride, furosemide, simvastatin, verapamil hydrochloride, digoxin, propranolol hydrochloride, carvedilol, lisinopril, spironolactone, hydrochlorothiazide, enalapril maleate, nadolol, ramipril, enoxaparin sodium, heparin sodium, valsartan, sotalol hydrochloride, fenofibrate, ezetimibe, bumetanide, losartan potassium, lisinopril/hydrochlorothiazide, felodipine, captopril and bisoprolol fumarate.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of ankylosing spondylitis include: ibuprofen, voltaren and misoprostol, naproxen, meloxicam, indometacin, voltaren, celecoxib, lofencoxib, sulfasalazine, methotrexate, azathioprine, minocycline, prednisone, etanercept and infliximab.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of asthma include: salbutamol, salmeterol/fluticasone, montelukast sodium, fluticasone propionate, budesonide, prednisone, salmeterol xinafoate, levalbuterol hydrochloride, salbutamol sulfate/ipratropium, prednisolone sodium phosphate, triamcinolone, beclomethasone dipropionate, ipratropium bromide, azithromycin, pirbuterol acetate, prednisolone, anhydrous theophylline, methyl- prednisolone sodium succinate, clarithromycin, zafirlukast, formoterol fumarate, influenza virus vaccine, methylprednisolone, amoxicillin trihydrate, flunisolide, allergy injection, cromolyn sodium, fexonadine hydrochloride, flunisolide/menthol, amoxicillin/clavulanate potassium, levofloxacin, inhaler auxiliary device, guaifenesin, dexamethasone sodium phosphate, moxifloxacin hydrochloride, doxycycline hydrochloride, guaifenesin/d-methylmorphan, p-ephedrine/cod/chlorphenir, gatifloxacin, cetirizine hydrochloride, mometasone furoate, salmeterol xinafoate, benzonatate, cefalexin, dihydrocodeinone/chlorpheniramine, cetirizine hydrochloride/pseudoephedrine, phenylephrine/promethazine, codeine/promethazine, cef-prozil, dexamethasone, guaifenesin/ pseudoephedrine, chlorpheniramine/dihydrocodeinone, nedocromil sodium, terbutaline sulfate, epinephrine, methylprednisolone and m-hydroxy-isoproterenol sulfate.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of COPD include: salbutamol sulfate/ipratropium, ipratropium bromide, salmeterol/fluticasone, salbutamol, salmeterol xinafoate, fluticasone propionate, prednisone, anhydrous theophylline, methylprednisolone sodium succinate, montelukast sodium, budesonide, formoterol fumarate, triamcinolone, levofloxacin, guaifenesin, azithromycin, beclomethasone dipropionate, levalbuterol hydrochloride, funisolide, ceftriaxone sodium, amoxicillin trihydrate, gatifloxacin, zafirlukast, amoxicillin/clavulanate potassium, flunisolide/ menthol, chlorpheniramine/dihydro- codeinone, m-hydroxy-isoproterenol sulfate, methylprednisolone, mometasone furoate, p-ephedrine/cod/chlorpheniramine, pirbuterol acetate, p-ephedrine/loratadine, terbutaline sulfate, tiotropium bromide, (R,R)-formoterol, TgAAT, cilomilast and roflumilast.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of HCV include: interferon-α-2a, interferon-α-2b, interferon-α con1, interferon-α-n1, pegylated interferon-α-2a, pegylated interferon-α-2b, ribavirin, peginterferon α-2b+ribavirin, ursodeoxycholic acid, glycyrrhizic acid, thymalfasin, Maxamine, VX-<NUM> and any compound used to treat HCV by interfering with the following targets: HCV polymerase, HCV protease, HCV helicase, HCV IRES (internal ribosome entry site).

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of idiopathic pulmonary fibrosis include: prednisone, azathioprine, salbutamol, colchicine, salbutamol sulfate, digoxin, γ- interferon, methylprednisolone sodium succinate (sod succ), lorazepam, furosemide, lisinopril, nitroglycerin, spironolactone, cyclophosphamide, ipratropium bromide, actinomycetes D, alteplase, fluticasone propionate, levofloxacin, m-hydroxy-isoproterenol sulfate, morphine sulfate, oxycodone hydrochloride, potassium chloride, triamcinolone, anhydrous tacrolimus, calcium, interferon-α, methotrexate, mycophenolate mofetil and interferon-γ-1b.

E examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of myocardial infarction include: aspirin, nitroglycerin, metoprolol tartrate, enoxaparin sodium, heparin sodium, clopidogrel bisulfate, carvedilol, atenolol, morphine sulfate, metoprolol succinate, warfarin sodium, lisinopril, isosorbide mononitrate, digoxin, furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate, torsemide, reteplase, losartan potassium, quinapril hydrochloride/mag carb, bumetanide, alteplase, enalaprilat, amiodarone hydrochloride, tirofiban hydrochloride monohydrate, diltiazem hydrochloride, captopril, irbesartan, valsartan, propranolole hydrochloride, fosinopril sodium, lidocaine hydrochloride, eptifibatide, cefazolin sodium, atropine sulfate, aminocaproic acid, spironolactone, interferon, sotalol hydrochloride, potassium chloride, docusate sodium, dobutamine hydrochloride, alprazolam, pravastatin sodium, atorvastatin calcium, midazole hydrochloride, pethidine hydrochloride, isosorbide dinitrate, epinephrine, dopamine hydrochloride, bivalirudin, rosuvastatin, ezetimibe/ simvastatin, avasimibe and cariporide.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of psoriasis include: calcipotriene, clobetasol propionate, triamcinolone, halobatasol proionate, tazorotene, methotrexate, fluocinolone, enhanced betamethasone dipropionate, fluocinolone acetate, Acitretin, tar shampoo, betamethasone valerate, mometasone furoate, ketoconazole, pramocaine/fluocinolone, hydrocortisone valerate, fluoxycortisone, urea, betamethasone, clobetasol propionate/emollient (emoll), fluticasone propionate, azithromycin, hydrocortisone, moisturizing formula, folic acid, desonide, pimecrolimus, coal tar, diflorasone diacetate, etanercept folate, lactic acid, <NUM>-methoxypsoralen, hc/bismuth subgal/znox/resor, methyl- prednisolone acetate, prednisone, sunscreen, clofloxasone, salicylic acid, anthralin, clocotrolone, coal extract, coal tar/salicylic acid, coal tar/salicylic acid/sulfur, desoximetasone, diazepam, emollient, fluocinolone/ emollient, mineral oil/castor oil/na lact, mineral oil/peanut oil, petroleum/isopropyl myristate, psoralen, salicylic acid, saponificated/tribromsalan, thimerosal/boric acid, celecoxib, infliximab, cyclosporine, alacepril, efalizumab, tacrolimus, pimecrolimus, PUVA, UVB and salicylazosulfapyridine.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of psoriatic arthritis include: methotrexate, etanercept, lofencoxib, celecoxib, folic acid, sulfasalazine, naproxen, leflunomide, methylprednisolone acetate, indomethacin, hydroxychloroquine sulfate, predni- sone, sulindac, enhanced betamethasone dipropionate, infliximab, methotrexate, folates, triamcinolone, voltarin, dimethyl sulfoxide, piroxicam, diclofenac sodium, ketoprofen, meloxicam, methylprednisolone, naproxen, tolmentin sodium, calcipotriol, cyclosporine, diclofenac sodium/misoprostol, fluocinolone, glucosamine sulfate, gold sodium thiomalate, dihydrocodeinone bitartrate/paracetamol, ibuprofen, risedronate sodium, sulfadiazine, thioguanine, valdecoxib, alacepril and efalizumab.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of restenosis include: sirolimus, paclitaxel, everolimus, tacrolimus, ABT- <NUM> and paracetamol.

Examples of therapeutic agents with which BZL-sRNA-<NUM> can be combined for the treatment or prevention of sciatica include: dihydrocodeinone bitartrate/paracetamol, profencoxib, cyclobenzaprine hydrochloride, methylprednisolone, naproxen, ibuprofen, oxycodone hydrochloride/paracetamol, celecoxib, valdecoxib, methylprednisolone acetate, prednisone, codeine phosphate/paracetamol, tramadol hydrochloride/paracetamol, metaxalone, meloxicam, metopamol, lidocaine hydrochloride, diclofenac sodium, gabapentin, dexamethasone, cariprado, ketorolac tromethamine, indomethacin, paracetamol, diazepam, naproxen, oxycodone hydrochloride, tizanidine hydrochloride, sodium diclofenac/misoprostol, dextropropoxyphene naphthalenesulfonate/paracetamol, asa/oxycod/oxycodone, ibuprofen/ dihydrocodeinone bit, tramadol hydrochloride, etodolic acid, propoxyphene hydrochloride, amitriptyline hydrochloride, cariprado/codeine phosphate/asa, morphine sulfate, multivitamins, sodium methoxy naphthyl propionate, orphenadrine citrate and temazepam.

Examples of therapeutic agents in which BZL-sRNA-<NUM> can be combined for the treatment or prevention of systemic lupus erythematosus (SLE) include: NSAIDS, for example, voltaren, naproxen, ibuprofen, piroxicam and indometacin; COX2 inhibitors, for example, celecoxib, profencoxib and valdecoxib; antimalarial agent(s), for example, hydroxychloroquine; steroids, for example, prednisone, prednisolone, budesonide and dexamethasone; cytotoxins, for example, azathioprine, cyclophosphamide, mycophenolate mofetil and methotrexate; PDE4 inhibitors or purine synthesis inhibitors, for example CELLCEPT®. BZL-sRNA-<NUM> can also be combined with reagents for example sulfasalazine, <NUM>-aminosalicylic acid, olsalazine, imulan, and reagents that interfere with the synthesis, production or action of pro-inflammatory cytokines (for example IL-<NUM>) (for example caspase inhibitors, such as IL-1beta converting enzyme inhibitors and IL-1ra). BZL-sRNA-<NUM> can also be used together with T cell signaling inhibitors, for example tyrosine kinase inhibitors, or molecules that target T cell activation molecules, for example CTLA-<NUM>-IgG or anti-B7 family antibodies and anti-PD-<NUM> family antibodies. BZL-sRNA-<NUM> can be combined with IL-<NUM> or anti-cytokine antibodies, for example fonotolizumab (anti-IFNg antibody), or anti-receptor receptor antibodies, for example anti-IL-<NUM> receptor antibodies and antibodies against B cell surface molecules. BZL-sRNA-<NUM> can also be used together with the following reagents: LJP <NUM> (abetimus), reagents that deplete or inactivate B cells, for example rituximab (anti-CD20 antibody), lymphostat-B (anti-BlyS antibody), TNF antagonists, for example, anti-TNF antibodies, D2E7 (<CIT>, HUMIRA®), CA2 (REMICADE®), CDP <NUM>, TNFR-Ig constructs (p75TNFRIgG (ENBREL®) and p55TNFRIgG (Lenercept)).

TNF-alpha has proven pathophysiological effects in various human diseases, especially inflammatory disorders, immune and immune regulation disorders, infections that cause septic, endotoxic and cardiovascular shock, neurodegenerative diseases and malignant diseases. The small RNA BZL-sRNA-<NUM> having SEQ ID NO:<NUM> can be used to treat the diseases listed below (diseases other than those specified in claim <NUM> are comparative examples and do not form part of the present invention).

Autoimmune or chronic inflammation: general chronic inflammation and/or autoimmune state, general immune-mediated inflammatory disorders, inflammatory CNS diseases, inflammatory diseases affecting eyes, joints, skin, mucous membrane, central nervous system, gastrointestinal tract, urinary tract or lung, general uveitis state, retinitis, HLA-B27+ uveitis, Behcet's disease, dry eye syndrome, glaucoma, Sjgren syndrome, diabetes (including diabetic neuropathy), insulin resistance, general arthritis state, rheumatoid arthritis, osteoarthritis, reactive arthritis and Reiter's syndrome, juvenile arthritis, ankylosing spondylitis, multiple sclerosis, Guillain-Barre syndrome, myasthenia gravis, amyotrophic lateral sclerosis, sarcoidosis, glomerulonephritis, chronic kidney disease, cystitis, psoriasis (including psoriatic arthritis), hidradenitis suppurativa, panniculitis, pyoderma gangrenosum, SAPHO syndrome (synovitis, acne, pustulosis, hyperostosis and osteitis), acne, Sweet syndrome, pemphigus, Crohn's disease (including extraintestinal manifestations), ulcerative colitis, bronchial asthma, allergic pneumonia, general allergies, allergic rhinitis, allergic sinusitis, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, Wegener granulomatosis, Kawasaki syndrome, giant cell arteritis, Churg-Strauss vasculitis, polyarteritis nodosa, burns, graft-versus-host disease, host-versus-graft reaction, rejection after organ or bone marrow transplantation, general systemic or local vasculitis state, systemic and discoid lupus erythematosus, multiple myositis and dermatomyositis, scleroderma, preeclampsia, acute and chronic pancreatitis, viral hepatitis and alcoholic hepatitis. Acute inflammation and/or prevention of post-operative or post-traumatic inflammation and pain: prevention of general post-operative inflammation, eye surgery (for example cataract (eye lens replacement) or glaucoma surgery), joint surgery (including arthroscopic surgery), joint-related structures (for example ligament) surgery, oral and/or dental surgery, minimal interventional cardiovascular procedures (for example PTCA, atherectomy, stent placement), laparoscopic and/or endoscopic abdominal and gynecological procedures, endoscopic urology procedures (for example prostate surgery, ureteroscopy, cystoscopy and interstitial cystitis), general pre- and post-operative inflammation (prevention). Neuropathy and neurodegenerative diseases: Alzheimer disease, Parkinson's disease, Huntington's disease, Bell's palsy and Creutzfeld-Jakob disease. Cancer: cancer-related osteolysis, cancer-related inflammation, cancer-related pain, cancer-related cachexia and bone metastasis. Pain: acute and chronic forms of pain (regardless of being caused by the central or peripheral effects of TNF-alpha and regardless of being classified as inflammatory, noxious or neuropathic forms of pain), sciatica, low back pain, carpal tunnel syndrome, complex regional pain syndrome (CRPS), gout, post-herpetic neuralgia, fibromyalgia, local pain state, chronic pain syndrome due to metastatic tumor and dismenorrhea. Infection: bacterial, viral or fungal sepsis, tuberculosis, AIDS. Cardiovascular diseases: atherosclerosis, coronary artery disease, hypertension, dyslipidemia, cardiac insufficiency and chronic heart failure. In one embodiment, the TNF-alpha-related disease is spondyloarthropathy, lung-related disorders, coronary heart disease, metabolic disorders, anemia, pain, liver disorders, skin disorders, nail disorders, or vasculitis. In another embodiment, the TNF-alpha-related disease is age-related cachexia, Alzheimer's disease, cerebral edema, inflammatory brain injury, chronic fatigue syndrome, dermatomyositis, drug reaction, intraspinal and/or peripheral edema, family periodic fever, Felty's syndrome, fibrosis, glomerular nephropathy (for example glomerulonephritis after streptococcal infection or IgA nephropathy), prosthesis relaxation, microscopic polyangiitis, mixed connective tissue disorder, multiple myeloma, cancer and cachexia, multiple organ disorders, myelodysplastic syndrome, orchitism, osteolysis, pancreatitis including acute, chronic and pancreatic abscess, periodontal polymyositis, progressive renal failure, pseudogout, pyoderma gangraenosum, recurrent polychondritis, rheumatic heart disease, sarcoidosis, cholangitis sclerosus, stroke, thoracic-abdominal aortic aneurysm (TAAA) repair, TNF receptor-associated periodic syndrome (TRAPS), and yellow fever vaccination-related syndromes, inflammatory diseases associated with ears, chronic otitis or pediatric otitis. In another embodiment of the present invention, the TNF-alpha related disease is Crohn's disease-related disease, juvenile arthritis/Still's disease (JRA), uveitis, sciatica, prostatitis, endometrial ectopic, choroidal neovascularization, lupus, Sjogren's syndrome and wet macular degeneration.

Examples of therapeutic agent(s) with which BZL-sRNA-<NUM> can be used in combination include: non-steroidal anti-inflammatory drugs (NSAIDs); cytokine suppressive anti-inflammatory drugs (CSAIDs); CDP-<NUM>/BAY-<NUM> -<NUM> (humanized anti-small RNA; Celltech/Bayer); cA2/infliximab (chimeric anti-small RNA; Centocor); 75kd TNFR-IgG/etanercept (75kD TNF receptor-IgG fusion protein; Immunex (<NPL>); 55kd TNF-IgG (55kD TNF receptor-IgG fusion protein; Hoffmann-LaRoche); IDEC-CE <NUM>/SB <NUM> (non-depleted primatized anti-CD4 antibody); IDEC/SmithKline; DAB <NUM>-IL-<NUM> and/or DAB <NUM>-IL-<NUM> (IL-<NUM> fusion protein; Seragen); Anti-Tac (humanized anti-IL-2Rα; Protein Design Labs/Roche); IL-<NUM> (anti-inflammatory cytokine; DNAX/Schering); IL-<NUM> (SCH <NUM>; recombinant IL-<NUM>, anti-inflammatory cytokine; DNAX/Schering); IL-<NUM>; IL-<NUM> and/or IL-<NUM> agonists (e.g., agonist antibodies); IL-1RA (IL-<NUM> receptor antagonist; Synergen/Amgen); TNF-bp/s-TNF (soluble TNF binding protein); R973401 (phosphodiesterase type IV inhibitor); MK-<NUM> (COX-<NUM> inhibitor); iloprost; methotrexate; thalidomide and thalidomide-related agent(s) (for example, Celgen); leflunomide (anti-inflammatory and cytokine inhibitor); tranexamic acid (plasminogen activation inhibitor); T-<NUM> (cytokine inhibitor); prostaglandin El; tenidap (non-steroidal anti-inflammatory drug); naproxen (non-steroidal anti-inflammatory drug); mobic (non-steroidal anti-inflammatory drug); ibuprofen (non-steroidal anti-inflammatory drug) Drug); piroxicam (non-steroidal anti-inflammatory drug); diclofenac sodium (non-steroidal anti-inflammatory drug); indomethacin (non-steroidal anti-inflammatory drug); salicylazosulfapyridine; azathioprine ; ICE inhibitors (inhibitor of the enzyme interleukin-1beta converting enzyme); zap-<NUM> and/or Ick inhibitors (casein kinase inhibitor zap-<NUM> or Ick); VEGF inhibitors and/or VEGF-R inhibitors (vascular endothelial cell growth factor or vascular endothelial cell growth factor receptor; inhibitors of angiogenesis); corticosteroid anti-inflammatory agent(s) (for example, SB203580); TNF-converting enzyme inhibitors; anti-IL-<NUM> antibodies; anti-IL-<NUM> antibodies; interleukin-<NUM>; interleukin-<NUM>; interleukin-<NUM> inhibitors; gold; penicillin; chloroquine; hydroxychloroquine; chlorambucil; cyclophosphamide; cyclosporine; total lymphocyte irradiation method; antithymocyte globulin; anti-CD4 antibodies; CD5-toxin; orally administered peptides and collagen; disodium lobenzarit; cytokine regulatory agents (CRAs) HP <NUM> and HP <NUM> (Houghten Pharmaceuticals, Inc. ); ICAM-<NUM> anti-allergy phosphorothioate oligodeoxynucleotide (ISIS <NUM>; Isis Pharmaceuticals, Inc. ); soluble complement receptor <NUM> (TP10; T Cell Sciences, Inc. ); prednisone; occulin; glycosaminoglycan polysulfate; minocycline; anti-IL2R antibodies; fish and plant seed fatty acids; auranofin; phenylbutazone; meclofenamic acid; flufenamic acid; intravenous immunoglobulin; zileuton; mycophenolic acid (RS-<NUM>); tacrolimus (FK-<NUM>); sirolimus (rapamycin); amprilose (therafectin); cladribine (<NUM>-chlorodeoxyadenosine); azuridine; methotrexate; antiviral agents; and immunomodulators. Any of the above-mentioned agent(s) can be combined with the small RNAs of the present invention to treat TNF-α related diseases.

BZL-sRNA-<NUM> may becombined with one of the following agent(s) to treat rheumatoid arthritis: small molecule inhibitor KDR (ABT-<NUM>), small molecule inhibitor of Tie-<NUM>; methotrexate; prednisone; celecoxib; folic acid; hydroxy-chloroquine sulfate; rofecoxib; etanercept; infliximab; leflunomide; naproxen; valdecoxib; sulfapyridine; methylhydro-prednisolone; ibuprofen; meloxicam; methylprednisolone acetate; gold sodium thiomalate; aspirin; azathiopurine; triamcinolone acetate; propoxyphene naphthalenesulfonate/ paracetamol; folates; nabumetone; diclofenac sodium; piroxicam; etodolac; diclofenac sodium; oxaprazine; oxycodone hydrochloride; hydrocodone ditartrate/ paracetamol; diclofenac sodium/misoprote; fentanyl; anakinra, human recombinant; tramadol hydrochloride; salsalate; sulindac; vitamin B12/fa/vitamin B6; acetaminophen; arsendronate sodium; hydroprednisone; morphine sulfate; lidocaine hydrochloride; indomethacin; glucosamine sulfate/chondroitin; cyclosporine; amitriptyline hydrochloride; sulfadiazine; oxycodone hydrochloride/ acetaminophen; olopatidine hydrochloride; misoprote; sodium methoxy naphthyl propionate; omeprazole; mycophenolate mofetil; cyclophosphamide; rituximab; IL-<NUM> TRAP; MRA; CTLA4-IG; IL-8BP; ABT-<NUM>; ABT-<NUM> (anti-IL18); anti- IL15; BIRB-<NUM>; SCIO-<NUM>; VX-<NUM>; AMG-<NUM>; VX-<NUM>; roflumilast; IC-<NUM>; CDC-<NUM>; and mesopram. BZL-sRNA-<NUM> and the above-mentioned drug for the treatment of rheumatoid arthritis may also be used in combination for the treatment of TNF-alpha related diseases.

BZL-sRNA-<NUM> may becombined with one of the following agent(s) to treat TNF-alpha related diseases in which TNF-alpha activity is harmful: anti-IL12 antibody (ABT874); anti-IL18 antibody (ABT <NUM>); small molecule inhibitors of LCK; small molecule inhibitors of COT; anti-III antibodies; small molecule inhibitors of MK2; anti-CD19 antibodies; small molecule inhibitors of CXCR3; small molecule inhibitors of CCR5; small molecule inhibitors of CCR11; anti-E/L selectin antibodies; small molecule inhibitors of P2X7; small molecule inhibitors of IRAK-<NUM>; small molecule agonists of glucocorticoid receptor; anti-C5a receptor antibodies; small molecule inhibitors of C5a receptor; anti-CD32 antibodies; and CD32 as therapeutic proteins.

BZL-sRNA-<NUM> may be administered in combination with antibiotics and anti-infectives. The term "antibiotic" used herein refers to a chemical substance that inhibits the growth of microorganisms or kills microorganisms. The term includes antibiotics produced by microorganisms known in the art, as well as synthetic antibiotics (for example, analogs). Antibiotics includeclarithromycin (Biaxin), ciprofloxacin (Cipro), and metronidazole (Flagyl).

BZL-sRNA-<NUM> may also beadministered in combination with other therapeutic agent(s) for treating sciatica or pain. Examples of agent(s) that can be used to relieve or suppress the symptoms of sciatica or pain include hydrocodone ditartrate/paracetamol, rofecoxib, cyclobenzaprine hydrochloride, methylprednisone, naproxen, ibuprofen, oxycodone hydrochloride/acetaminophen, celecoxib, valdecoxib, methylprednisone acetate, prednisone, cocaine phosphate/paracetamol, tramadol hydrochloride/ acetaminophen, metaxalone, meloxicam, methocarbamol, lidocaine hydrochloride, diclofenac sodium, gabapentin, dexamethasone, carlipodor, ketorolac, indometacin, acetaminophen, diazepam, nabumetone, oxycodone hydrochloride, tizanidine hydrochloride, diclofenac sodium/misoprostol, propoxyfennaphthalene sulfonate/paracetamol, a small amount of ibuprofen/hydrocodone; tramadol hydrochloride, etodolic acid; propoxyphene hydrochloride, amitriptyline hydrochloride, carliprol/codeine phosphate, morphine sulfate, multivitamins, sodium methoxy naphthyl propionate, orphenadrine citrate and temazepam.

BZL-sRNA-<NUM> may be used in combination with hemodialysis to treat TNF-alpha related diseases.

BZL-sRNA-<NUM> may also be used in combination with agent(s) used to treat Crohn's disease or Crohn's disease-related diseases. Therapeutic agent(s) that can be used to treat Crohn's disease include mesalazine, prednisone, azathioprine, mercaptopurine, infliximab, budesonide, salicylazosulfapyridine, methylprednisolone, diphenoxylate, loperamide hydrochloride, methotrexate, folates, cipro-floxacin/glucose-injection, hydrocodone ditartrate, tetracycline hydrochloride, fluocinolone acetate, metronidazole, thimerosal/boric acid, cholestyramine/sucrose, ciprofloxacin hydro-chloride, hyoscyamine sulfate, dolantin hydrochloride, midazolam hydrochloride, oxycodone hydrochloride/acetaminophen, promethazine hydrochloride, sodium phosphate, sulfamethoxazole/trimethoprim, celecoxib, polyacrylic resin, propoxyphene napsylate, hydrocortisone, multivitamins, balsalazide disodium, cocaine phosphate/paracetamol, colesevelan hydrochlo-ride, vitamin B12, folic acid, levofloxacin, methylprednisolone, natalizumab and γ-interferon.

BZL-sRNA-<NUM> may beadministered in combination with other therapeutic agent(s) for treating asthma. Examples of agent(s) that can be used to reduce or suppress asthma symptoms include: salbutamol; salmeterol/fludesone; sodium; fludexone propionate; budesonide; prednisone; salmeterol xinafoate; levalbuterol hydrochloride; sulfate/ipratropium; prednisone sodium phosphate; triamcinolone acetonide; beclomethasone dipropionate; ipratropium bromide; azithromycin; pirbuterol acetate, prednisone, anhydrous theophylline, methylprednisolone, clarithromycin, zafirlukast, formoterol fumarate, influenza virus vaccine, methylprednisolone trihydrate, flunisolide, allergic allergy injection, cromolyn sodium, fexofenadinehydrochloride, flunisolide/menthol, amoxicillin/potassium clavulanate, levofloxacin, inhalation aid device, guaifenesin, dexamethasone sodium phosphate; moxifloxacin hydrochloride; hyclate; guaifenesin/dextro-methorphan; chlorpheniramine; gatifloxacin; cetirizine hydrochloride; mometasone furoate; salmeterol xinafoate; cough syrup; cephalexin; hydrocodone/chlorpheniramine; cetirizine hydrochloride/pseudoephedrine; phenylephedrine/promethazine; codeine/promethazine; cefprozil; dexamethasone; guaifenesin/pseudoephedrine; chlorpheniramine/ hydrocodone, nedocromil sodium, terbutaline sulfate, epinephrine, methylprednisone and orciprenaline sulfate.

BZL-sRNA-<NUM> may be administered in combination with other therapeutic agent(s) used for treating COPD. Examples of agent(s) that can be used to reduce or suppress COPD symptoms include: salbutamol sulfate/ipratropium; ipratropium bromide; salmeterol/fludexone; salbutamol; salmeterol xinafoate; fludexone propionate; prednisone; anhydrous theophylline; methylprednisolone sod succ; montelukast sodium; budesonide; formoterol fumarate; triamcinolone acetonide; levofloxacin; guaifenesin; azithromycin; beclomethasone; dipropionic acid; levalbuterol hydrochloride; flunisolide; sodium; trihydrates; gatifloxacin; zafirlukast; amoxicillin/clavulanate potassium; flunisolide/menthol; chlorpheniramine/hycodone; orciprenaline sulfate; methylprednisolone; furoates; ephedrine/cod/chlorpheniramine; pirbuterol hydrochloride; ephedrine/loratadine; terbutaline sulfate; tiotropium bromide; (R, R)-formoterol; TgAAT; cilomilast and roflumilast.

BZL-sRNA-<NUM> may be administered in combination with other therapeutic agent(s) used for treating IPF. Examples of agent(s) that can be used to reduce or suppress the symptoms of IPF include: prednisone; azathioprine; salbutanolamine; colchicine; sulfates; digoxin; γ-interferon; methylprednisolone sod succ; furosemide; lisinopril; nitroglycerin; spironolactone; cyclophosphamide; ipratropium bromide; actinomycin d; alteplase; fluticasone propionate; levofloxacin; oxinaline sulfate; morphine sulfate; oxycodone hydrochloride; potassium chloride; triamcinolone acetonide; anhydrous tacrolimus; calcium; α-interferon; methotrexate; mycophenolate mofetil.

BZL-sRNA-<NUM> may be administered in combination with other therapeutic agent(s) used for treating spondyloarthropathy. Examples of such agent(s) include: non-steroidal antiinflammatory drugs (NSAIDs), COX <NUM> inhibitors, including Celebrex, Vioxx; and Bextra, and etoricoxib. Physical therapy is also used to treat spondyloarthropathy, usually in combination with non-steroidal anti-inflammatory drugs.

BZL-sRNA-<NUM> may be administered in combination with other therapeutic agent(s) used for treating ankylosing spondylitis. Examples of agent(s) that can be used to reduce or suppress the symptoms of ankylosing spondylitis include: ibuprofen, diclofenac and misoprostol, naproxen, meloxicam, indomethacin, diclo-fenac, celecoxib, rofecoxib, salicylazosulfapyridine, prednisone, methotrexate, azathioprine, minocycline, prednisone, etanercept and infumab.

BZL-sRNA-<NUM> may be administered in combination with other therapeutic agent(s) used for treating psoriasis arthritis in patients. Examples of agent(s) that can be used to reduce or suppress the symptoms of arthritis in patients with psoriasis include: methotrexate; etanercept; rofecoxib; celecoxib; folic acid; salicylazosulfapyridine; naproxen; leflunomide; methylprednisolone acetate; indomethacin; hydroxychloroquine sulfate; sulindac; prednisone; betamethasone (diprospan); infliximab; methotrexate; folic acid; triamcinolone acetonide; diclofenac; dimethyl sulfoxide; piroxicam; diclofenac sodium; ketoprofen; meloxicam; prednisone; methylprednisolone; nabumetone; sodium tetrabenzoylpyrrole acetate; calcipotriene; cyclosporine; diclofenac; sodium/ misoprostol; fluocinolone acetate; glucosamine sulfate; gold sodium thiomalate; hydrocodone; ditartrate/paracetamol; ibuprofen; risedronate sodium; sulfadiazine; thioguanine; valdecoxib; alefacept; and efalizumab.

BZL-sRNA-<NUM> can be administered in combination with other therapeutic agent(s) used for treating restenosis. Examples of agent(s) that can be used to reduce or inhibit restenosis include: rapamycin, paclitaxel, everolimus, tacrolimus, ABT-<NUM>, and acetaminophen.

BZL-sRNA-<NUM> can be administered in combination with other therapeutic agent(s) used for treating myocardial infarction. Examples of agent(s) that can be used to reduce or suppress myocardial infarction include: aspirin, nitroglycerin, metoprolol tartrate, enoxaparin sodium, heparin sodium, clopidogrel hydrosulfate, carvedilol, atenolol, morphine sulfate, metoprolol succinate, warfarin sodium, lisinopril, isosorbide mononitrate, digoxin, furosemide, simvastatin, ramipril, tenecteplase, enalapril maleate, torsemide, reteplase, losartan potassium, quinapril hydrochloride/mag carb, bumetanib, alteplase, enalaprilat, amiodarone hydrochloride, tirofiban hydrochloride m-hydrate, diltiazem hydrochloride, captopril, irbesartan tablets, valsartan, propranolol hydrochloride, fosinopril sodium, lidocaine hydrochloride, eptifibatide, cefazolin sodium, atropine sulfate, leucine, spironolactone, interferon, sotalol hydrochloride, potassium chloride, docusate sodium, dobutamine hydrochloride, alprazolam, pravastatin sodium, lipitor, midazolam hydrochloride, dolantin hydrochloride, isosorbide dinitrate, epinephrine, dopamine hydrochloride, bivalirudin, rosuvastatin, ezetimibe/simvastatin, avasimibe, abciximab and cariporide.

BZL-sRNA-<NUM> can be administered in combination with other therapeutic agent(s) used for treating angina. Examples of agent(s) that can be used to reduce or suppress angina include: aspirin; nitroglycerin; isosorbide mononitrate; metoprolol succinate; atenolol; metoprolol tartrate; alodipine sulfonate, dilitiazem hydropchloride, isosorbide dinitrate, clopidogrel hydrosulfate; nifedipine; lipitor; potassium chloride; furosemide; simvastatin; verapamil hydrochloride; digoxin; propranolol hydrochloride; carvedilol; lisinopril; sprionolactone; dihydrochlorothiazide; enalapril maleate; madolol; ramipril; enoxaparin sodium; heparin sodium; valsartan; sotalol hydrochloride; fenofibrate; ezetimibe; bumetanide; losartan potassium lisinopril/hydrochlorothiazide; felodipine; captopril; and bisoprolol fumarate.

BZL-sRNA-<NUM> can be administered in combination with agent(s) usually used for treating hepatitis C virus. Examples of such agent(s) include: interferon-α-2a, interferon-α-2b, interferon-α conl, interferon-α-nl, pegylated interferon-α-2a, pegylated interferon-α-2b, ribavirin, pegylated interferon-α-2b and ribavirin, andro-deoxycholic acid, glycyrrhizic acid, thymalfasin, maxamine and VX-<NUM>.

BZL-sRNA-<NUM> can beused in combination with corticosteroids, vitamin D analogs, and topical or oral retinoic acid, or a combination thereof, for the treatment of psoriasis. In addition, BZL-sRNA-<NUM> can be used in combination with one of the following agent(s) for the treatment of psoriasis: small molecule inhibitor of KDR (ABT-<NUM>), small molecule inhibitor of Tie-<NUM>, calcipotriene, clobetasol propionate, triamcino-lone acetonide, halobetasol propionate, tazorotene, methotrexate, fluocinolone acetate, fluocinolone, Acitretin, tar shampoo, betamethasone valerate, mometasone furoate, ketoconazole, pramocaine/fluocinolone, hydrocortisone valerate, fludrolone, urea, betamethasone, clobetasol propionate/emoll, fludiasone propionate, azithromycin, hydrocortisone, prescription for increasing moisture, folic acid, desonide, coal tar, diflurazone acetate, etanercept, folates, lactic acid, methoxsalin, hc/bismuth subgallate/znox/resor, methylprednisolone acetate, prednisone, sunscreen substances, salicylic acid, hascinonide, anthranol, clocortolone pivalate, coal extracts, coal tar/salicylic acid, coal tar/salicylic acid/sulfur, desoxy-methasone, diazepam, emollient, pimecrolimus emollient, fluocinolone acetate/emollient, mineral oil/castor oil /na lact, mineral oil/peanut oil, isopropyl petroleum myristate, psoralen, salicylic acid, saponificated/ tribromosalen, thimerosal/boric acid, celecoxib, infliximab, alefacept, efalizumab, tacrolimus, pimecrolimus, PUVA, UVB and salicylazosulfapyridine.

BZL-sRNA-<NUM> can be administered in combination with other therapeutic agent(s) for treating skin diseases. For example, BZL-sRNA-<NUM> can be combined with PUVA therapy. PUVA is a combination of psoralen (P) and long-wave ultraviolet rays, which is used to treat many different skin diseases. BZL-sRNA-<NUM> can also be combined with pimecrolimus. BZL-sRNA-<NUM> can also beused to treat psoriasis in combination with tacrolimus. Tacrolimus and BZL-sRNA-<NUM> may be administered in combination with methotrexate and/or cyclosporin. BZL-sRNA-<NUM> may also beadministered in combination with stimulated excimer laser therapy for the treatment of psoriasis.

Examples of other therapeutic agent(s) with which BZL-sRNA-<NUM> can be combined to treat skin diseases or nail diseases include UVA and UVB phototherapy. Further examples to be used in combination with BZL-sRNA-<NUM> include anti-IL-<NUM> and anti-IL-<NUM> therapeutic agent(s), including antibodies.

BZL-sRNA-<NUM> can beadministered in combination with other therapeutic agent(s) for the treatment of Behcet's disease. Therapeutic agent(s) for the treatment of Behcet's disease include: prednisone, cyclophosphamide (cytoxan), azathioprine (also referred to as imuran), methotrexate, timethoprim/sulfamethoxazole (also known as compound sulfamethoxazole tablets or TMP-SMZ) and folic acid.

The present invention is further illustrated below with reference to the examples. These embodiments are only illustrative.

ELISA (enzyme-linked immunosorbent assay) and RT-qPCR (real-time fluorescent quantitative PCR).

Main experimental instruments and equipment: <NUM> cell culture dishes, <NUM>-well cell culture plates, pipettors, pipettes, optical microscopes.

<NUM> THP-<NUM> cells (monocyte macrophages, purchased from the Cell Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured in RPMI <NUM> culture medium containing fetal bovine serum to the logarithmic growth phase. They were distributed into <NUM>-well plates with <NUM> medium/well, incubated overnight at <NUM> for subsequent experiments.

<NUM> The groups of the experiment were as follows: Blank group, i.e. empty group, referred to untreated cells. This group served as a blank control; LPS group: this group was treated as follows: <NUM>µl RNAimax was diluted with <NUM>µl Opti-MEM and added to the cells, which were stimulated with LPS. This group served as a negative control; NC group: the random nonsense sequence <NUM>' UUC UCC GAA CGU GUC ACG UTT-<NUM> (double-stranded, Genepharma) was added to the cells with the same concentration and the same transfection method as that of the experimental group, and the cells were stimulated with LPS. This group served as a negative control.

<NUM> The artificially synthesized small RNAs were transfected by using RNAimax at RNAimax <NUM>µl/<NUM>µl Opti-MEM, small RNA (<NUM>) <NUM>µl/<NUM>µl Opti-MEM. The above liquids were mixed and incubated for <NUM> minutes at room temperature and added to the cells.

<NUM> LPS was added for stimulation <NUM> hours after the transfection, and the final concentration of LPS was <NUM>µg/ml.

<NUM> The cell supernatant was collected <NUM> hours after LPS stimulation, and the concentration of the protease inhibitor added was <NUM>µl/ml.

<NUM> The expressions of the three factors IL-1beta/IL-<NUM>/TNF-alpha were detected by ELISA kit.

<NUM> The groups of the experiment were as follows: Blank group: empty group, referred to untreated cells. This group served as a blank control; LPS group: in this group, <NUM>µl RNAimax was diluted with <NUM>µl Opti-MEM and added to the cells which were subjected to LPS stimulation. This group served as a negative control; NC group: the random nonsense sequence <NUM>' UUC UCC GAA CGU GUC ACG UTT-<NUM> (Genepharma) was added to the cells with the same concentration and the same transfection method as that of the experimental group, and the cells were stimulated with LPS. This group served as a negative control.

<NUM> Nine hours after LPS stimulation, the cells were collected by centrifugation at <NUM> for <NUM> minutes.

<NUM> Total cell RNA was extracted by using Total RNA Rapid Extraction Kit according to the manufacturer's instructions.

<NUM> Reverse transcription of RNA into cDNA: reverse transcription of small RNA into cDNA was carried out using a reverse transcription kit (High-Capacity cDNA Reverse Transcription Kits, Applied Biosystems, cat. <NUM>) according to the manufacturer's instructions. The reverse transcription system was as follows: template RNA (<NUM> ng/µl) <NUM>µl, <NUM>× RT Buffer <NUM>µl, <NUM>× dNTP Mix (<NUM>) <NUM>µl, <NUM>× Random Primer (included in the kit) <NUM>µl, MultiScribe™ reverse transcriptase <NUM>µl, RNase inhibitor <NUM>µl , Nuclease-free H<NUM>O <NUM>µl. After transient centrifugation, the system was put into the PCR instrument for reaction, and the reaction conditions were as follows: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) the reaction was stopped at <NUM>. After the reaction, <NUM>µl RNase Free dH<NUM>O was added to make up the final volume to <NUM>µl.

<NUM> Quantitative PCR amplification reaction: the total volume of the qPCR reaction system was <NUM>µl, including: <NUM>µl <NUM>×SYBR Green Master Mix, <NUM>µl forward primer (<NUM>), <NUM>µl reverse primer (<NUM>), <NUM>µl cDNA obtained by reverse transcription and <NUM>µl RNase Free dH<NUM>O. A LightCycler <NUM> fluorescent quantitative PCR instrument was used and the PCR reaction conditions were: pre-denaturation for <NUM> minutes at <NUM>, then PCR amplification cycle: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; for a total of <NUM> cycles; finally <NUM> for <NUM> to cool down. The forward primers and reverse primers for the amplification reaction were all designed and synthesized by Beijing Tsingke Xinye Biological Technology Co. The primer sequences used are as follows:.

<NUM> Calculation of the relative expression by using the <NUM>-ΔΔCt method.

Main experimental instruments and equipment: <NUM> cell culture dishes, <NUM>-well cell culture plates, pipettors, pipette, optical microscopes, <NUM> centrifuge tubes, microplate reader, MTS detection kit (Promega, Celltiter <NUM> AQueous One Solution Cell Proliferation Assay, REF: G3581, LOT: <NUM>).

Main experimental reagents: Cell culture: F12 culture medium (Hyclone), FBS (Gibico); Model establishment and transfection: artificially synthesized small RNA, RNAimax, opti-MEM, H5N1 virus (A/Jilin/<NUM>/<NUM>); Cell survival rate: MTS cell viability detection kit.

<NUM> The function of artificially synthesized small RNAs derived from Chinese herbal medicine in resisting H5N1 infection and alleviating cell death was verified by applying an A549 cell model infected by the H5N1 strain derived from <NUM> Jilin (A/Jilin/<NUM>/<NUM>).

<NUM> A549 cells (human lung adenocarcinoma epithelial cells, purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) were cultured in <NUM> cell culture dishes (cultured in Ham's F12 nutrient medium (HyClone, Logan, UT, USA), distributed into <NUM>-well plates, <NUM>µl cell-containing culture medium per well).

<NUM> When the cells were observed to grow to <NUM>% confluence (about <NUM> hours) under an optical microscope, the artificially synthesized plant small RNAs were transfected by using a transfection reagent, transfection reagent <NUM>µl/ml, small RNA <NUM> nmol/ml.

<NUM> The cells were infected with H5N1 virus <NUM> hours after transfection, and the amount of challenge was <NUM>.

<NUM> The cell death status was detected by using the MTS kit <NUM> hours after challenge. The relevant reagents for MTS detection were mixed thoroughly according to: serum-free culture medium: solution A: solution B = <NUM>:<NUM>:<NUM>. The supernatant of the cells in the <NUM>-well plates were aspirated. MTS detection reagent mixture was added to the <NUM>-well plates at <NUM>µl/well and incubated in an oven at <NUM> for <NUM> (protected from light) after adding.

<NUM> The cell survival status was detected by using a microplate reader: the absorbance at <NUM> was detected three times per plate, and the result of the third time shall prevail.

<NUM> THP-<NUM> cells (monocyte macrophages, purchased from the Cell Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured to the logarithmic growth phase. They were distributed into <NUM>-well plates with <NUM> medium/well, incubated overnight at <NUM> for subsequent experiments.

<NUM> The groups of the experiment were as follows: Blank group: empty group, referred to untreated cells. This group served as a blank control; LPS group: in this group, <NUM>µl RNAimax was diluted with <NUM>µl Opti-MEM and added to the cells which were subjected to LPS stimulation. This group served as a negative control; NC (native) group: the random nonsense sequence <NUM>' UUC UCC GAA CGU GUC ACG UTT-<NUM> (double-stranded, Genepharma) was added to the cells with the same concentration and the same transfection method as that of the experimental group, and the cells were stimulated with LPS. This group served as a negative control.

<NUM> The artificially synthesized small RNA mixtures (Table <NUM>) were transfected by using RNAimax. The volume ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (the initial concentrations of various small RNAs were all <NUM>), RNAimax <NUM>µl/<NUM>µl Opti-MEM, small RNA mixture (<NUM>) <NUM>µl/<NUM>µl Opti-MEM. The above liquids were mixed and incubated for <NUM> minutes at room temperature and added to the cells.

<NUM> The groups of the experiment were as follows: Blank group: empty group, referred to untreated cells. This group served as a blank control; LPS group: in this group, <NUM>µl RNAimax was diluted with <NUM>µl Opti-MEM and added to the cells which were subjected to LPS stimulation. This group served as a negative control; NC group: the random nonsense sequence <NUM>' UUC UCC GAA CGU GUC ACG UTT-<NUM> (double-stranded, Genepharma) was added to the cells with the same concentration and the same transfection method as that of the experimental group, and the cells were stimulated with LPS. This group served as a negative control.

<NUM> The artificially synthesized small RNA mixtures (Table <NUM>) were transfected by using RNAimax. The volume ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM>, RNAimax <NUM>µl/<NUM>µl Opti-MEM, small RNA mixture (<NUM>) <NUM>µl/<NUM>µl Opti-MEM. The above liquids were mixed and incubated for <NUM> minutes at room temperature and added to the cells.

<NUM> The cells were lysed by <NUM> TRI Reagent (sigma, T9424-<NUM>), centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM> and the precipitate was discard. Chloroform was added at the ratio of <NUM>µl/ml TRIzol, shaken and mixed throughly, and left at room temperature for <NUM>. The mixture was centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM>. The upper water phase was transferred to another centrifuge tube. The upper water phase was transferred to another new EP tube. Isopropanol was added at <NUM>/ml TRIzol, mixed well and left at room temperature for <NUM>-<NUM>. The mixture was centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM>. The supernatant was discarded, <NUM> of <NUM>% ethanol was added and the centrifuge tube was gently shaken to suspend the precipitate. The mixture was centrifuged at <NUM>, <NUM> for <NUM>. The supernatant was discarded to the greatest extent and the tube was dried at room temperature for <NUM>-<NUM>. The RNA sample was dissolved by using <NUM>µl DEPC-treated H<NUM>O.

<NUM> Reverse transcription of RNA into cDNA: reverse transcription of small RNA into cDNA was carried out using a reverse transcription kit (High-Capacity cDNA Reverse Transcription Kits, Applied Biosystems, cat. The reverse transcription system was as follows: template RNA (<NUM> ng/µl) <NUM>µl, <NUM>× RT Buffer <NUM>µl, <NUM>× dNTP Mix (<NUM>) <NUM>µl, <NUM>× Random Primer (included in the kit) <NUM>µl, MultiScribe™ reverse transcriptase <NUM>µl, RNase inhibitor <NUM>µl, Nuclease-free H<NUM>O <NUM>µl. After transient centrifugation, the system was put into the PCR instrument for reaction, and the reaction conditions were as follows: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) the reaction was stopped at <NUM>. After the reaction, <NUM>µl RNase Free dH<NUM>O was added to make up the final volume to <NUM>µl.

<NUM> Quantitative PCR amplification reaction: the total volume of the qPCR reaction system was <NUM>µl, including: <NUM>µl <NUM>×SYBR Green Master Mix, <NUM>µl forward primer (<NUM>), <NUM>µl reverse primer (<NUM>), <NUM>µl cDNA obtained by reverse transcription and <NUM>µl RNase Free dH<NUM>O. A LightCycler <NUM> fluorescent quantitative PCR instrument was used and the PCR reaction conditions were: pre-denaturation for <NUM> minutes at <NUM>, then PCR amplification cycle: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; for a total of <NUM> cycles; finally <NUM> for <NUM> to cool down. The forward primers and reverse primers for the amplification reaction were all designed and synthesized by Beijing Tsingke Xinye Biological Technology Co. The UBC gene was used as an internal reference gene. The primer sequences used are as follows:.

<NUM><NUM>-week-old male C57 mice weighing <NUM>-<NUM> were divided into <NUM> groups, one of which remained untreated during the entire experiment, i.e. the blank group.

<NUM> The mice were given a dose of <NUM> nmol/animal of BZL-sRNA-<NUM> or NC small RNA by gavage <NUM> days, <NUM> days and <NUM> day in advance, respectively, and the groups were BZL-sRNA-<NUM> or NC group (native group), respectively.

<NUM> After <NUM>% pentobarbital sodium anesthesia at <NUM>, the mice were tracheally injected with a dose of LPS (<NUM>/ml) <NUM>µl, at <NUM>µg/animal. Among them, the group only treated with LPSwas denoted as the LPS group.

<NUM> Nine hours after <NUM>% pentobarbital sodium anesthesia, alveolar lavage (<NUM>µl) was performed for <NUM> times, each time with <NUM>µl PBS pipetted repeatedly for <NUM> times.

<NUM> The obtained lavage fluid was centrifuged at <NUM> for <NUM>. The obtained exfoliated lung cells were lysed by <NUM> Trizol (Thermo), centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM> and the precipitate was discard. Chloroform was added at the ratio of <NUM>µl/ml TRIzol, shaken and mixed throughly, and left at room temperature for <NUM>. The mixture was centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM>. The upper water phase was transferred to another centrifuge tube. The upper water phase was transferred to another new EP tube. Isopropanol was added at <NUM>/ml TRIzol, mixed well and left at room temperature for <NUM>-<NUM>. The mixture was centrifuged at <NUM>,<NUM> rpm, <NUM> for <NUM>. The supernatant was discarded, <NUM> of <NUM>% ethanol was added and the centrifuge tube was gently shaken to suspend the precipitate. The mixture was centrifuged at <NUM>, <NUM> for <NUM>. The supernatant was discarded to the greatest extent and the tube was dried at room temperature for <NUM>-<NUM>. The RNA sample was dissolved by using <NUM>µl DEPC-treated H<NUM>O.

<NUM> Reverse transcription of RNA into cDNA: reverse transcription of small RNA into cDNA was carried out using a reverse transcription kit (High-Capacity cDNA Reverse Transcription Kits, Applied Biosystems, cat. The reverse transcription system was as follows: template RNA (<NUM> ng/µl) <NUM>µl, <NUM>× RT Buffer <NUM>µl, <NUM>× dNTP Mix (<NUM>) <NUM>µl, <NUM>× Random Primer (included in the kit) <NUM>µl, MultiScribe™ reverse transcriptase <NUM>µl, RNase inhibitor <NUM>µl , Nuclease-free H<NUM>O <NUM>µl. After transient centrifugation, the system was put into the PCR instrument for reaction, and the reaction conditions were as follows: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) the reaction was stopped at <NUM>. After the reaction, <NUM>µl RNase free dH<NUM>O was added to make up the final volume to <NUM>µl.

<NUM> Quantitative PCR amplification reaction: the total volume of the qPCR reaction system was <NUM>µl, including: <NUM>µl <NUM>×SYBR Green Master Mix, <NUM>µl forward primer (<NUM>), <NUM>µl reverse primer (<NUM>), <NUM>µl cDNA obtained by reverse transcription and <NUM>µl RNase Free dH<NUM>O. A LightCycler <NUM> fluorescent quantitative PCR instrument was used and the PCR reaction conditions were: pre-denaturation for <NUM> minutes at <NUM>, then PCR amplification cycle: (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; (<NUM>) <NUM>, <NUM>; for a total of <NUM> cycles; finally <NUM> for <NUM> to cool down. The forward primers and reverse primers for the amplification reaction were all designed and synthesized by Beijing Tsingke Xinye Biological Technology Co. The GAPDH gene was used as an internal reference gene. The primer sequences used are as follows:.

<NUM> The supernatant was centrifuged at <NUM> rpm for <NUM>, and the cell debris was removed. The expression of the factors was verified by detection with ELISA kits (DuoSet Mouse IL-1beta/IL-<NUM>/TNF-alpha, R&D, DY401/DY406/DY410).

<NUM> Preparation and extraction of RNA for sequencing.

<NUM> The groups of the experiment:_Blank group: empty group, referred to untreated cells. This group served as a blank control;_LPS group: in this group, <NUM>µl RNAimax was diluted with <NUM>µl Opti-MEM and added to the cells which were subjected to LPS stimulation. This group served as a negative control;_NC group: the random nonsense sequence <NUM>' UUC UCC GAA CGU GUC ACG UTT-<NUM> (Genepharma) was added to the cells with the same concentration and the same transfection method as that of the experimental group, and the cells were stimulated with LPS. This group served as a negative control.

<NUM> The artificially synthesized plant small RNAs were transfected by using RNAimax at RNAimax <NUM>µl/<NUM>µl Opti-MEM, small RNA (<NUM>) <NUM>µl/<NUM>µl Opti-MEM. The above liquids were mixed and incubated for <NUM> minutes at room temperature and added to the cells.

<NUM> LPS was added for stimulation <NUM> hours after the transfection, and the final concentration of LPS was <NUM>µg/ml.

<NUM> Nine hours after LPS stimulation, the cells were collected by centrifugation at <NUM> for <NUM> minutes.

<NUM> The cells were fully lysed by using <NUM> Trizol Reagent (sigma). <NUM>µl chloroform was added, mixed well and centrifuged at <NUM>, 13200rpm for <NUM>. <NUM>µl supernatant was taken and the same amount of isopropanol was added, mixed well and let stand at -<NUM> for <NUM>. The mixture was centrifuged at <NUM>, <NUM> rpm for <NUM>. The supernatant was discarded and the precipitate was washed twice with <NUM>% ethanol prepared with DEPC water. The precipitate was dried and dissolved with <NUM>µl DEPC water.

<NUM> The obtained RNA solution was sent to the company for sequencing.

<NUM> Uploading the sequencing data: A total of <NUM> sample data (including <NUM> NC) was uploaded to the <NUM>. <NUM> port <NUM> bioinformatics server using SSH protocol, using Xftp (version Xftp <NUM>) as the transfer tool and XShell (version XShell <NUM>) as the secure terminal simulation software on WINDOW10 platform.

<NUM> Preparation of database data and calculation of sequencing data: The next step was carried out after uploading the data. The hg19 version of the human genome of UCSC was downloaded and the library was built by using bowtie2 (version bowtie2 <NUM>. The annotation file that matched hg19 in the UCSC database was used as the annotation file. The sequenced <NUM> bp fragments were matched to the human genome file of the gene name of each annotated segment by using the shell script to run Tophat (version <NUM>. <NUM>) and cufflink (version <NUM>. <NUM>), and the statistics of the expression count of each gene were completed.

Tophat running parameters: average separation distance between each sequenced fragment pair - r: <NUM>; standard deviation of the separation distance --mate-std-dev: <NUM>; library-type (chain-specific): fr-secondstrand; number of the threads - p: <NUM>.

<NUM> Summary of the sequencing data results: The sequencing data sorted by Tophat was written into a new text by using python (version <NUM>. <NUM>) script.

<NUM> Statistics of differential genes of <NUM> sequencing result samples: The running script was written by using DEGseq of R (version <NUM>. The expressions of each sample and each gene were sorted out and compared with the expressions of each gene in the NC group to calculate the FC (fold change).

<NUM> Statistics and clustering of differentially down-regulated genes: The results of step <NUM> were processed by screening the genes (wherein the gene expression level FC was down-regulated by more than <NUM> fold in each sample as compared to the control group NC). These genes were uploaded to the Metacore database for analysis. The parameters were selected (ignore first line; species: homo sapiens; type: down regulate; p-value and FDR: no limit).

<NUM> Statistics of the number of differentially down-regulated genes: Statistics of <NUM> small RNA samples were performed by using the script in python (<NUM>. The expression levels of down-regulated genes (down-regulation level: fold change > <NUM>) was calculated, andthe genes that could be down-regulated by small RNAs in <NUM> samples were obtained.

<NUM> Classification tabulation of <NUM> small RNA target genes, and the pathways or biological processes involved by the target genes: According to the pathways or biological processes to which the down-regulated genes of each sample belong, the data was divided to <NUM> categories and tabulated by referring to the PUBMED and KEGG databases, as well as the clustering results of the down-regulated genes (criteria: FC > <NUM>) for each sample in the Metacore database of <NUM> samples as described in <NUM>.

Any small RNAs other than BZL-sRNA-<NUM> having SEQ ID NO:<NUM> are comparative examples and do not form part of the present invention.

<FIG>: The expression of the inflammatory factor IL-1beta at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Scutellaria barbata (BZL) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Bupleurum (CHu) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Andrographis paniculata (CXL) and Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. In <FIG>, "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs shown in <FIG> had significantly higher effect in reducing the protein expression of IL-1beta than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> had the smallest value in ELISA of the inflammatory factor IL-1beta, indicating the best effect on inhibiting IL-1beta protein level among the small RNAs tested.

<FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Scutellaria barbata (BZL) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Bupleurum (CHu) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with Viola philippica (DDi), Scutellaria baicalensis (HQi), Lonicera japonica (JYH), Fructus forsythiae (LQi), and Prunella vulgaris (XKC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Andrographis paniculata (CXL) and Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. "*" means that an unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs in <FIG> had significantly higher effect in reducing the protein expression of IL-<NUM> than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> inhibited IL-<NUM> protein level very well.

<FIG>: The expression of the inflammatory factor TNF-alpha at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with Scutellaria barbata (BZL), Viola philippica (DDi), Scutellaria baicalensis (HQi), Fructus forsythiae (LQi) and Prunella vulgaris (XKC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Bupleurum (CHu) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at protein level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. "*" means that an unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs shown in <FIG> had significantly higher effect in reducing the protein expression of TNF-alpha than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> had the smallest value in ELISA of the inflammatory factor TNF-alpha, indicating the best effect on inhibiting TNF-alpha protein level among the small RNAs tested.

<FIG>: The expression of the inflammatory factor IL-1beta at mRNA level (relative expression level of IL-1beta compared to UBC) compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Scutellaria barbata (BZL) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Bupleurum (CHu), Fructus forsythiae (LQi) and Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-1beta at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs shown in <FIG> had significantly higher effect in reducing the mRNA expression of IL-1beta than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> had the smallest value in qPCR of the inflammatory factor IL-1beta, indicating the best effect on inhibiting IL-1beta mRNA level among the small RNAs tested.

<FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Scutellaria barbata (BZL) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Bupleurum (CHu) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the control group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Viola philippica (DDi), Scutellaria baicalensis (HQi), Lonicera japonica (JYH), Fructus forsythiae (LQi) and Prunella vulgaris (XKC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP -<NUM> cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Andrographis paniculata (CXL) and Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs shown in <FIG> had significantly higher effect in reducing the mRNA expression of IL-<NUM> than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> had relatively low value in qPCR of the inflammatory factor IL-<NUM>, indicating a relatively favorable effect on inhibiting IL-<NUM> mRNA level among the small RNAs tested.

<FIG>: The expression of the inflammatory factor TNF-alpha at mRNA level (relative expression level of TNF-alpha compared to UBC) compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Scutellaria barbata (BZL) and Bupleurum (CHu) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Viola philippica (DDi), Lonicera japonica (JYH), Fructus forsythiae (LQi) and Prunella vulgaris (XKC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Houttuynia cordata (YXC) small RNA <NUM> hours in advance, as specified in the figure. <FIG>: The expression of the inflammatory factor TNF-alpha at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the Taraxacum (PGY) small RNA <NUM> hours in advance, as specified in the figure. "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNAs shown in <FIG> had significantly higher effect in reducing the mRNA expression of TNF-alpha than the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. BZL-sRNA-<NUM> had the smallest value in qPCR of the inflammatory factor TNF-alpha, indicating the best effect on inhibiting TNF-alpha mRNA level among the small RNAs tested.

<FIG>: BZL: after H5N1 (<NUM>. I) infection, the rescue results of Scutellaria barbata (BZL) small RNA on cell death, as specified in the figure. <FIG>: CHu: after H5N1 (<NUM>. I) infection, the rescue results of the Bupleurum (CHu) small RNA on cell death, as specified in the figure. <FIG>: LQi/XKC: after H5N1 (<NUM>. I) infection, the rescue results of the Fructus forsythiae (LQi) / Prunella vulgaris (XKC) small RNA on cell death, as specified in the figure. <FIG>: XKC/YXC: after H5N1 (<NUM>. I) infection, the rescue results of the Prunella vulgaris (XKC) / Houttuynia cordata (YXC) small RNA on cell death, as specified in the figure. <FIG>: YXC: after H5N1 (<NUM>. I) infection, the rescue results of Houttuynia cordata (YXC) small RNA on cell death, as specified in the figure. In <FIG>, unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the rescuing effect on cell death caused by H5N1 infection. "*" represents P < <NUM> in unpaired t test, and "**" represents P < <NUM> in unpaired t test. As shown in <FIG>, the small RNAs as specified in the figure significantly improved the cell survival rate, showing a more obvious effect of rescuing cell death compared with the NC group. The values in <FIG> were all values obtained by normalization relative to the NC group. Among them, BZL-sRNA-<NUM> was very effective in rescuing cell death.

As mentioned above in "Functional experiment of artificially synthesized small RNA mixtures at protein level verified by using the THP-<NUM> cell model stimulated by LPS", the effects of the mixtures in Table <NUM> were verified.

<FIG>: The expression of the inflammatory factor IL-1beta at protein level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the small RNA mixtures <NUM> hours in advance, as specified in the figure. The mixing ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (v/v). "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNA mixtures shown in <FIG> had significantly higher effect in reducing IL-1beta protein level than the NC group, among which MIX20, <NUM>, <NUM>, <NUM> and <NUM> had significantly higher effect in reducing IL-1beta protein level than the BZL-sRNA-<NUM> group. The values in <FIG> were all values obtained by normalization relative to the NC group. For the mixture in the figure that was comparable effect in reducing IL-1beta protein level to that of BZL-sRNA-<NUM> group, as the molar concentration of BZL-sRNA-<NUM> small RNA in the mixture in the test cell liquid was much lower than that in the BZL-sRNA-<NUM> group, this indicated that the various small RNAs in the mixture also had a favorable synergistic effect in reducing IL-1beta protein level.

<FIG>: The expression of the inflammatory factor IL-<NUM> at protein level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the small RNA mixtures <NUM> hours in advance, as specified in the figure, wherein the mixing ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (v/v). "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The results showed that the small RNA mixtures shown in <FIG> had significantly higher effect in reducing IL-<NUM> protein level than the NC group, among which those in <FIG> (except MIX10), and MIX22, <NUM>-<NUM>, <NUM>-<NUM> and <NUM>-<NUM> in <FIG> had significantly higher effect in reducing IL-<NUM> protein level than the BZL-sRNA-<NUM> group. The values in <FIG> were all values obtained by normalization relative to the NC group. Mixture <NUM> was comparable to the BZL-sRNA-<NUM> group in reducing IL-1beta protein level. As the molar concentration of BZL-sRNA-<NUM> small RNA in Mixture <NUM> in the test cell liquid was much lower than that in the BZL-sRNA-<NUM> group, this indicated that the various small RNAs in Mixture <NUM> also had a favorable synergistic effect in reducing IL-<NUM> protein level.

<FIG>: The expression of the inflammatory factor IL-1beta at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the small RNA mixtures <NUM> hours in advance, as specified in the figure. The mixing ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (v/v). "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNA mixtures shown in <FIG> had significantly higher effect in reducing IL-1beta mRNA level than the NC group, among which MIX23, <NUM> and <NUM> had significantly higher effect in reducing IL-1beta mRNA level than the BZL-sRNA-<NUM> group. The values in <FIG> were all values obtained by normalization relative to the NC group. For the mixture that was comparable to the BZL-sRNA-<NUM> group in reducing IL-1beta mRNA level, as the molar concentration of BZL-sRNA-<NUM> small RNA in the mixture in the test cell liquid was much lower than that in the BZL-sRNA-<NUM> group, this indicated that the various small RNAs in the mixture also have a favorable synergistic effect in reducing IL-1beta mRNA level.

<FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the small RNA mixtures <NUM> hours in advance, as specified in the figure, the mixing ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (v/v). "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The results showed that the small RNA mixtures shown in <FIG> had significantly higher effect in reducing IL-<NUM> mRNA level than the NC group, among which those in <FIG> (except MIX10 and <NUM>), and MIX25-<NUM>, <NUM>, <NUM> and <NUM> in <FIG> had significantly higher effect in reducing IL-<NUM> mRNA level than the BZL-sRNA-<NUM> group. The values in <FIG> were all values obtained by normalization relative to the NC group. For the mixture that was comparable to the BZL-sRNA-<NUM> group in reducing IL-<NUM> mRNA level, as the molar concentration of BZL-sRNA-<NUM> small RNA in the mixture in the test cell liquid was much lower than that in the BZL-sRNA-<NUM> group, this indicated that the various small RNAs in the mixture had a favorable synergistic effect in reducing IL-<NUM> mRNA level.

<FIG>: The expression of the inflammatory factor TNF-alpha at mRNA level compared to the NC group, in the cell inflammation model after <NUM> hours of LPS stimulation, with the THP1 cells transfected with the small RNA mixtures <NUM> hours in advance, as specified in the figure, the mixing ratio of BZL-sRNA-<NUM> to other small RNAs was <NUM>:<NUM> (v/v). "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factors in in vitro experiments. The experimental results showed that the small RNA mixtures shown in <FIG> had significantly higher effect in reducing TNF-alpha mRNA level than the NC group, among which MIX32, <NUM>, <NUM> and <NUM> had significantly higher effect in reducing TNF-alpha mRNA level than the BZL-sRNA-<NUM> group. The values in <FIG> were all values obtained by normalization relative to the NC group. A mixture shows comparable effect in reducing TNF-alpha mRNA level to that of BZL-sRNA-<NUM> group. As the molar concentration of BZL-sRNA-<NUM> small RNA in the mixture in the test cell liquid was much lower than that in the BZL-sRNA-<NUM> group, this indicated that the various small RNAs in the mixture also had a favorable synergistic effect in reducing TNF-alpha mRNA level.

<FIG>: The expression of the inflammatory factor TNF-alpha at protein level compared to the NC group, in the alveolar lavage fluid of the animal inflammation model after <NUM> hours of LPS stimulation, wherein the mice were gavaged with small RNA, three days in advance. "*" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating that BZL-sRNA-<NUM> had the effect of inhibiting the expression of inflammatory factor TNF-alpha in in vivo experiments.

<FIG>: The expression of the inflammatory factor IL-<NUM> at mRNA level compared to the NC group, in the exfoliated lung cells of the animal inflammation model after <NUM> hours of LPS stimulation, wherein the mice were gavaged with small RNA, three days in advance. "**" means that unpaired t test P < <NUM> was considered statistically significant in statistical analysis, indicating the effect of inhibiting the expression of inflammatory factor IL-<NUM> in in vivo experiments.

Any small RNAs other than BZL-sRNA-<NUM> having SEQ ID NO:<NUM> are comparative examples and do not form part of the present invention.

Classification tabulation of <NUM> small RNA target genes and the pathways or biological processes they involved was performed as mentioned above.

Table <NUM>: Classification table of small RNA target genes and the pathways or biological processes they involved, obtained by calculation of transcriptome sequencing of <NUM> small RNAs selected from Table <NUM>. This table shows thatinflammatory factors and/or H5N1 infection were inhibited by small RNAs through: the mechanism of action, targets of action and pathways of action of chemokine related signaling pathways, JAK/STAT signal transduction, amino acid metabolism, mRNA activation and function, coenzyme metabolism, small nucleolar RNA, etc., suggesting that different small RNAs had the possibility of synergistic effects in related signaling pathways and target genes.

The genes in the following table correspond to the pathways in the corresponding rows in the above table.

Claim 1:
A small RNA for use in the treatment of an IL-1beta, IL-<NUM> and/or TNF-alpha related disease,
wherein the small RNA has a sequence shown in SEQ ID NO: <NUM>;
wherein administration of the small RNA rescues cell death caused by an infection with a virus; and
wherein the IL-1beta, IL-<NUM> and/or TNF-alpha related disease is selected from the group consisting of pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, diabetes, and gout.