bluuebunny/crossref_metadata_2025 huggingface dataset:
The legal and organizational principles of attestation of candidates to patent attorneys
Authors: Kulinich Olha , Pototskyi Mykola
Published: March 2023

Keywords: intellectual property, attestation, representation, legal status, patent attorneys

   The article is devoted to the study of the organizational and legal principles of attestation of candidates to patent attorneys in Ukraine. The article analyses formation and development of the institute of patent attorneys in the context of obtaining the legal status of a patent attorney and adducesperiodization of the stages of development of the attestation procedure from 1994 to2022. The possibility is substantiated of conditional distinguishing of four stages.   The first stage of formation of legal bases of the institute of patent attorneys in Ukraine (1994−1996) is connected with creation of legal bases of functioning of the institute, definition of standard and requirements for obtaining the legal status of a patent attorney, introduction of procedure of attestation of candidates to patent attorneys, formation of community of patent attorneys.     The second stage of the development of the institute of patent attorneys conditionallylasted from 1996 to 2016 and is conditionally characterized as a period of strong development of the patent attorneys community, in particular by regular attestation, the creation of the first professional association of patent attorneys and their active assistance in the development of intellectual property protection.

    The third stage, which has been ongoing from 2017 to the present, is actually characterized by the suspension of attestation and the block of the possibility of obtaining the legal status of a patent attorney.      It has been established that the result of the reform of the system of legal protection of intellectual property, in particular the liquidation of the State Intellectual Property Service and the subordination of the relevant sphere of intellectual property to the competence of the Ministry of Economy, was the suspension of the attestation procedure. The exclusion from participation in the attestation procedure of the educationalinstitution, based on which attestation and transfer of the relevant functions to the subordinate enterprise took place, partially complicated the continuation of the attestation procedure. The expulsion from participation of the educational institution in the attestation procedure, because of which the attestation took place and transfer of the relevant functions to the subordinate enterprise, partially complicated the continuation of the attestation procedure.    The attention has been drawn to the absence of the corresponding order of the Ministry of Economy on the approval of composition of the Attestation and the Appeal Commission. The authors have described the prospects for the development of the attestation procedure at the fourth stage, which can tentatively begin in 2022.
NALAR FIKIH MAZHAB SYAFI’I DALAM KASUS PERNIKAHAN WANITA HAMIL DAN STATUS ANAK YANG DILAHIRKAN
Authors: Rohmad Azis Muhammad Abdulloh
Published: March 2023

This article is the result of library research. This study aims to: track the shafi'iyah school of thought in determining the legal status of pregnant women marriage. This article discusses the reasoning or way of thinking of the Shafi'iyah school in determining the legal status of a pregnant woman's marriage and the status of the child she gives birth to. The claim of this research is that the approach taken by the syafi'iyah school in determining the legal status of pregnant women's marriages uses a textual approach. The source of the law used is the Hadith of the Prophet Muhammad which stipulates that a wife who is still in her husband's marriage contract, every child born to her, even though it is the result of adultery, is considered by law to be her husband's child. The results of this study conclude that a child born before a legal marriage, is considered an adulterous child, cannot be attributed to a father, and each of them cannot be an heir and neither can inherit. If the marriage is carried out to cover the disgrace, if it is carried out before six months of gestation, the status of the baby is considered a legal child according to Islamic law.
Pulsed electromagnetic fields as a promising therapy for glucocorticoid-induced osteoporosis
Authors: Zhang Tianxiao , Zhao Zhiliang , Wang Tiantian
Published: March 2023

Glucocorticoid-induced osteoporosis (GIOP) is considered the third type of osteoporosis and is accompanied by high morbidity and mortality. Long-term usage of glucocorticoids (GCs) causes worsened bone quality and low bone mass via their effects on bone cells. Currently, there are various clinical pharmacological treatments to regulate bone mass and skeletal health. Pulsed electromagnetic fields (PEMFs) are applied to treat patients suffering from delayed fracture healing and non-unions. PEMFs may be considered a potential and side-effect-free therapy for GIOP. PEMFs inhibit osteoclastogenesis, stimulate osteoblastogenesis, and affect the activity of bone marrow mesenchymal stem cells (BMSCs), osteocytes and blood vessels, ultimately leading to the retention of bone mass and strength. However, the underlying signaling pathways via which PEMFs influence GIOP remain unclear. This review attempts to summarize the underlying cellular mechanisms of GIOP. Furthermore, recent advances showing that PEMFs affect bone cells are discussed. Finally, we discuss the possibility of using PEMFs as therapy for GIOP.
Hyaluronic acid-based bioink improves the differentiation and network formation of neural progenitor cells
Authors: Pereira Inês , Lopez-Martinez Maria J. , Villasante Aranzazu , Introna Clelia , Tornero Daniel , Canals Josep M. , Samitier Josep
Published: March 2023

Introduction: Three-dimensional (3D) bioprinting is a promising technique for the development of neuronal in vitro models because it controls the deposition of materials and cells. Finding a biomaterial that supports neural differentiation in vitro while ensuring compatibility with the technique of 3D bioprinting of a self-standing construct is a challenge.

Methods: In this study, gelatin methacryloyl (GelMA), methacrylated alginate (AlgMA), and hyaluronic acid (HA) were examined by exploiting their biocompatibility and tunable mechanical properties to resemble the extracellular matrix (ECM) and to create a suitable material for printing neural progenitor cells (NPCs), supporting their long-term differentiation. NPCs were printed and differentiated for up to 15 days, and cell viability and neuronal differentiation markers were assessed throughout the culture.

Results and Discussion: This composite biomaterial presented the desired physical properties to mimic the ECM of the brain with high water intake, low stiffness, and slow degradation while allowing the printing of defined structures. The viability rates were maintained at approximately 80% at all time points. However, the levels of β-III tubulin marker increased over time, demonstrating the compatibility of this biomaterial with neuronal cell culture and differentiation. Furthermore, these cells showed increased maturation with corresponding functional properties, which was also demonstrated by the formation of a neuronal network that was observed by recording spontaneous activity via Ca2+ imaging.
Biomechanical study of internal fixation methods for femoral neck fractures based on Pauwels angle
Authors: Wang Fuyang , Liu Yuchen , Huo Yi , Wang Ziming , Zhang Jinge , Xu Minghao , Ma Kaiming , Wang Linbao , Lu Yongtao , Cheng Liangliang , Zhao Dewei
Published: March 2023

Objective: To select the most appropriate internal fixation method based on the Pauwels angle, in order to provide a new concept for clinical accurate treatment of femoral neck fractures (FNFs).

Methods: FNFs models of Pauwels 30°; 40°; 50°; 60° were created respectively. For Pauwels ≤ 50°, 1, 2 and 3 Cannulated Compression Screws (CCS) and Porous Tantalum Screws (PTS) were used to fix the fracture for the models. For Pauwels 60°, 3CCS and Medial Buttress Plate (MBP) combined with 1, 2 and 3CCS were used to fix the fracture. Based on the results of the finite element (FE) analysis, the biomechanical properties of each model were compared by analyzing and evaluating the following four parameters: maximal stress of the bone (MBS), maximal stress of the implants (MIS), maximal displacement of bone (MBD), interfragmentary motion (IFM).

Results: At Pauwels 30°, the larger parameters were found in 1CCS, which was 94.8 MPa (MBS), 307.7 MPa (MIS), 0.86 mm (MBD) and 0.36 mm (IFM). In 2CCS group, the parameters were 86.1 MPa (MBS), 254.4 MPa (MIS), 0.73 mm (MBD) and 0.27 mm (IFM), which were similar to those of PTS. At Pauwels 40°; 50°, with the increase of the number of used CCS, accordingly, the parameters decreased. Particularly, the MIS (Pauwels 50°) of 1CCS was 1,195.3 MPa, but the other were less than the yield range of the materials. At Pauwels 60°, the MBS of 3CCS group was 128.6 Mpa, which had the risk of failure. In 2CCS + MBP group, the parameters were 124.2 MPa (MBS), 602.5 MPa (MIS), 0.75 mm (MBD) and 0.48 mm (IFM), The model stability was significantly enhanced after adding MBP.

Conclusion: Pauwels type Ⅰ (<30°) fractures can reduce the number of CCS, and PTS is an appropriate alternative treatment. For Pauwels type Ⅱ fractures (30°∼50°), the 3CCS fixation method is still recommended. For Pauwels type Ⅲ fractures (>50°), it is recommended to add MBP to the medial femoral neck and combine with 2CCS to establish a satisfactory fracture healing environment.
Annexin A protein family: Focusing on the occurrence, progression and treatment of cancer
Authors: Zhang Huhu , Zhang Zhe , Guo Tingting , Chen Guang , Liu Guoxiang , Song Qinghang , Li Guichun , Xu Fenghua , Dong Xiaolei , Yang Fanghao , Cao Can , Zhong Di , Li Shuang , Li Ya , Wang Mengjun , Li Bing , Yang Lina
Published: March 2023

The annexin A (ANXA) protein family is a well-known tissue-specific multigene family that encodes Ca2+ phospholipid-binding proteins. A considerable amount of literature is available on the abnormal expression of ANXA proteins in various malignant diseases, including cancer, atherosclerosis and diabetes. As critical regulatory molecules in cancer, ANXA proteins play an essential role in cancer progression, proliferation, invasion and metastasis. Recent studies about their structure, biological properties and functions in different types of cancers are briefly summarised in this review. We further discuss the use of ANXA as new class of targets in the clinical diagnosis and treatment of cancer.
Environmentally sustainable implementations of two-dimensional nanomaterials
Authors: Shams Mehnaz , Mansukhani Nikhita , Hersam Mark C. , Bouchard Dermont , Chowdhury Indranil
Published: March 2023

Rapid advancement in nanotechnology has led to the development of a myriad of useful nanomaterials that have novel characteristics resulting from their small size and engineered properties. In particular, two-dimensional (2D) materials have become a major focus in material science and chemistry research worldwide with substantial efforts centered on their synthesis, property characterization, and technological, and environmental applications. Environmental applications of these nanomaterials include but are not limited to adsorbents for wastewater and drinking water treatment, membranes for desalination, and coating materials for filtration. However, it is also important to address the environmental interactions and implications of these nanomaterials in order to develop strategies that minimize their environmental and public health risks. Towards this end, this review covers the most recent literature on the environmental implementations of emerging 2D nanomaterials, thereby providing insights into the future of this fast-evolving field including strategies for ensuring sustainable development of 2D nanomaterials.
Chemical, biological and in silico assessment of date (P. dactylifera L.) fruits grown in Ha’il region
Authors: Alsukaibi Abdulmohsen Khalaf Dhahi , Alenezi Khalaf M. , Haque Ashanul , Ahmad Irfan , Saeed Mohd , Verma Mahima , Ansari Irfan Ahmad , Hsieh Ming-Fa
Published: March 2023

Background: Dates palm (Phoenix dactylifera L.) fruits are among the most widely used fruits in the Middle East and African nations. Numerous researchers confirmed the presence of phytochemicals in P. dactylifera L. fruit and its by-products with broad-ranging biological activities.

Objectives: In the present work, phytochemical and biological assessments of two different cultivars of date fruit (ShishiM1 and MajdoolM2 grown in the Ha’il region of Saudi Arabia) have been carried out.

Methods: Date fruits were extracted and analyzed by gas chromatography-mass spectrometry (GS-MS),liquid chromatography-mass spectrometry (LC-MS) and Fourier-transform infrared spectroscopy (FT-IR)techniques. The lyophilized methanolic extracts were analyzed for their in-vitro antiproliferative andcytotoxicity against colon cancer (HCT116) cell line. To identify the possible constituents responsible for the bioactivity, in-silico molecular docking and molecular dynamics (MD) simulation studies were carried out.

Results: Both cultivars exhibited in-vitro anticancer activity (IC50 = 591.3 μg/mL and 449.9 μg/mL for M1 and M2, respectively) against colon cancer HCT-116 cells. The computational analysis results indicated procyanidin B2 and luteolin-7-O-rutinoside as the active constituents.

Conclusion: Based on these results, we conclude that these cultivars could be a valuable source for developing health promoter phytochemicals, leading to the development of the Ha’il region, Saudi Arabia.
Mycobacterium tuberculosis-macrophage interaction: Molecular updates
Authors: Bo Haotian , Moure Ulrich Aymard Ekomi , Yang Yuanmiao , Pan Jun , Li Li , Wang Miao , Ke Xiaoxue , Cui Hongjuan
Published: March 2023

Mycobacterium tuberculosis (Mtb), the causative agent of Tuberculosis (TB), remains a pathogen of great interest on a global scale. This airborne pathogen affects the lungs, where it interacts with macrophages. Acidic pH, oxidative and nitrosative stressors, and food restrictions make the macrophage’s internal milieu unfriendly to foreign bodies. Mtb subverts the host immune system and causes infection due to its genetic arsenal and secreted effector proteins. In vivo and in vitro research have examined Mtb-host macrophage interaction. This interaction is a crucial stage in Mtb infection because lung macrophages are the first immune cells Mtb encounters in the host. This review summarizes Mtb effectors that interact with macrophages. It also examines how macrophages control and eliminate Mtb and how Mtb manipulates macrophage defense mechanisms for its own survival. Understanding these mechanisms is crucial for TB prevention, diagnosis, and treatment.
Computed tomography–based COVID–19 triage through a deep neural network using mask–weighted global average pooling
Authors: Zhang Hong-Tao , Sun Ze-Yu , Zhou Juan , Gao Shen , Dong Jing-Hui , Liu Yuan , Bai Xu , Ma Jin-Lin , Li Ming , Li Guang , Cai Jian-Ming , Sheng Fu-Geng
Published: March 2023

Background

There is an urgent need to find an effective and accurate method for triaging coronavirus disease 2019 (COVID-19) patients from millions or billions of people. Therefore, this study aimed to develop a novel deep-learning approach for COVID-19 triage based on chest computed tomography (CT) images, including normal, pneumonia, and COVID-19 cases.

Methods

A total of 2,809 chest CT scans (1,105 COVID-19, 854 normal, and 850 non-3COVID-19 pneumonia cases) were acquired for this study and classified into the training set (n = 2,329) and test set (n = 480). A U-net-based convolutional neural network was used for lung segmentation, and a mask-weighted global average pooling (GAP) method was proposed for the deep neural network to improve the performance of COVID-19 classification between COVID-19 and normal or common pneumonia cases.

Results

The results for lung segmentation reached a dice value of 96.5% on 30 independent CT scans. The performance of the mask-weighted GAP method achieved the COVID-19 triage with a sensitivity of 96.5% and specificity of 87.8% using the testing dataset. The mask-weighted GAP method demonstrated 0.9% and 2% improvements in sensitivity and specificity, respectively, compared with the normal GAP. In addition, fusion images between the CT images and the highlighted area from the deep learning model using the Grad-CAM method, indicating the lesion region detected using the deep learning method, were drawn and could also be confirmed by radiologists.

Conclusions

This study proposed a mask-weighted GAP-based deep learning method and obtained promising results for COVID-19 triage based on chest CT images. Furthermore, it can be considered a convenient tool to assist doctors in diagnosing COVID-19.
Enhancing urinary tract infection diagnosis for negative culture patients with metagenomic next-generation sequencing (mNGS)
Authors: Jia Kaipeng , Huang Shiwang , Shen Chong , Li Hongjun , Zhang Zhe , Wang Lei , Zhao Gangjian , Wu Zhouliang , Lin Yuda , Xia Han , Tang Mingze , Yang Huifen , Hu Hailong
Published: March 2023

Background

Metagenomic next-generation sequencing (mNGS) is a promising technology that allows unbiased pathogen detection and is increasingly being used for clinical diagnoses. However, its application in urinary tract infection (UTI) is still scarce.

Methods

The medical records of 33 patients with suspected UTI who were admitted to the Second Hospital of Tianjin Medical University from March 2021 to July 2022 and received urine mNGS were retrospectively analyzed. The performance of mNGS and conventional urine culture in diagnosing infection and identifying causative organisms was compared, and the treatment effects were evaluated in terms of changes in urinalyses and urinary symptoms.

Results

In the detection of bacteria and fungi, mNGS detected at least one pathogen in 29 (87.9%) cases, including 19 (57.6%) with positive mNGS but negative culture results and 10 (30.3%) with both mNGS and culture positive results. The remaining 4 (12.1%) patients were negative by both tests. Overall, mNGS performed better than culture (87.9% vs. 30.3%, P < 0.001). Within the 10 double-positive patients, mNGS matched culture results exactly in 5 cases, partially in 4 cases, and not at all in 1 case. In addition, mNGS detected a broader pathogen spectrum, detecting 26 species compared to only 5 species found in culture. The most abundant bacteria detected by mNGS was Escherichia coli, detected in 9 (27.2%) patients. All anaerobic bacteria, Mycobacterium Tuberculosis and all mixed pathogens were detected by mNGS. The final clinical diagnosis of UTI was made in 25 cases, and the sensitivity of mNGS was significantly higher than culture (100.0% vs 40.0%; P < 0.001) when using the diagnosis as a reference standard; the positive predictive value, negative predictive value and specificity were 86.2%, 100% and 50.0%, respectively. Importantly, targeted antibiotic therapy based on mNGS resulted in significant improvement in urinalyses and urinary symptoms in patients.

Conclusions

mNGS is a technology that has shown clear advantages over culture, particularly in the context of mixed infections and UTIs that are difficult to diagnose and treat. It helps to improve the detection of pathogens, guide changes in treatment strategies, and is an effective complement to urine culture.
The role of red blood cell distribution width in predicting coronary artery lesions in pediatric patients with kawasaki disease
Authors: Cai Jianghui , Tang Mi , Shuai Shuping , Zhang Rui , Zhang Hongxi , Yang Yanfeng , Wu MengJun , Liang Hua , Xing Shasha
Published: March 2023

Background

Recent studies have shown that red blood cell distribution width (RDW) has emerged as a novel predictor of cardiovascular diseases. We aim to investigate the association between RDW and the risk of coronary artery lesions (CALs) in pediatric patients with Kawasaki disease (KD).

Methods

KD patients were classified as the CALs group (patients with CALs) and non-CALs group (patients without CALs). Differences among the groups were analyzed by Mann-Whitney U-test and Chi-square analysis. The independent risk factors of CALs were identified by multivariate logistic regression analysis, followed by receiver operating characteristic (ROC) curve analysis to calculate the optimal cut-off value.

Results

The red blood cell distribution width (RDW) and C-reactive protein were significantly higher in the CALs group than those in the non-CALs group (p < 0.01). Multivariate logistic regression analysis revealed that RDW (OR = 5.2, 95% CI, 4.064 to 6.654) was independent risk factors of CALs in KD patients (p < 0.01). The subgroup analysis also confirmed that the high level of RDW was an independent risk factor for the development of CALs in patients with complete and incomplete KD. The ROC analysis showed the optimal cut-off value of RDW for predicting CALs was >13.86%, with a sensitivity of 75.79% and specificity of 92.81% (AUC = 0.869, 95% CI = 0.844–0.892; p < 0.0001).

Conclusions

RDW is an independent predictor with high sensitivity and specificity to predict CALs in KD patients. The elevation in RDW level (>13.86%) may be used as novel biomarkers for early predicting CALs in KD patients during the acute phase.
Causal association between sleep traits and the risk of coronary artery disease in patients with diabetes
Authors: Tian Mengyun , Ma Hongchuang , Shen Jiaxi , Hu Teng , Cui Hanbin , Huangfu Ning
Published: March 2023

Background and aims

The association between sleep traits and coronary artery disease (CAD) in patients with diabetes has been reported in previous observational studies. However, whether these potential relationships are causal remains unclear. We aim to assess the causal relationship between sleep traits and CAD in diabetic.

Methods

Genetic instrumental variables associated with five sleep-related traits (insomnia, sleep duration, ease of getting up, morningness and snoring) were extracted from corresponding genome-wide association studies (GWAS). The associations of genetic variants with CAD were based on 15,666 individuals with diabetes (3,968 CAD cases and 11,696 controls). The primary analysis was derived using the inverse variance weighting method. Further sensitivity analysis was conducted to confirm the robustness and consistency of the main results.

Results

Genetic liability to insomnia was significantly related to the increased risk of CAD in individuals with diabetes [odds ratio (OR): 1.163; 95% CI: 1.072–1.254; p = 0.001]. Suggestive evidence was found for the borderline associations between both sleep duration (OR: 0.629; 95% CI: 0.380–1.042, p = 0.072) and snoring (OR: 1.010, 95% CI: 1.000–1.020, p = 0.050) with CAD risk. However, no consistent evidence was found for the association between ease of getting up and morningness with the risk of CAD in diabetic. Similar results can be verified in most sensitivity analyses.

Conclusions

We provide consistent evidence for the causal effect of insomnia on the increased risk of CAD in individuals with diabetes. The management of sleep health should be emphasized to prevent CAD in diabetic patients.
Circulating insulin-like growth factor-1 and risk of lung diseases: A Mendelian randomization analysis
Authors: Jiang Sujing , Xu Zhiyong , Shi Yueli , Liang Sibei , Jiang Xinyuan , Xiao Mingshu , Wang Kai , Ding Liren
Published: March 2023

Background

Insulin-like growth factor-1 (IGF-1) display a vital role in in the pathogenesis of lung diseases, however, the relationship between circulating IGF-1 and lung disease remains unclear.

Methods

Single nucleotide polymorphisms (SNPs) associated with the serum levels of IGF-1 and the outcomes data of lung diseases including asthma, chronic obstructive pulmonary disease (COPD), lung cancer and idiopathic pulmonary fibrosis (IPF) were screened from the public genome-wide association studies (GWAS). Two-sample Mendelian randomization (MR) analysis was then performed to assess the independent impact of IGF-1 exposure on these lung diseases.

Results

Totally, 416 SNPs related to circulating IGF-1 levels among 358,072 participants in UK Biobank. According to a primary casual effects model with MR analyses by the inverse variance weighted (IVW) method, the circulating IGF-1 was demonstrated a significantly related with the risk of asthma (OR, 0.992; 95% CI, 0.985-0.999, P=0.0324), while circulating IGF-1 showed no significant correlation with CODP (OR, 1.000; 95% CI, 0.999-1.001, P=0.758), lung cancer (OR, 0.979, 95% CI, 0.849-1.129, P=0.773), as well as IPIGFF (OR, 1.100, 95% CI, 0.794-1.525, P=0.568).

Conclusion

The present study demonstrated that circulating IGF-1 may be causally related to lower risk of asthma.
Quantitative proteomics identified a novel invasion biomarker associated with EMT in pituitary adenomas
Authors: Zhang Yu , Li Lei , Ma Xin , Liu Chenan , Liu Gemingtian , Bie Zhixu , Yang Zhijun , Liu Pinan
Published: March 2023

Background

Complete resection of invasive pituitary adenoma is usually difficult, resulting in a high recurrence rate. Therefore, it is needed to find potential diagnostic markers and therapeutic targets for invasive pituitary adenoma.

Methods

We collected samples from patients with invasive and non-invasive pituitary adenomas from Beijing Tiantan Hospital for protein extraction and quantitative analysis. We identified differential proteins (DEPs) by differential analysis of the two groups. The intersection of differential proteins related to invasion and epithelial-mesenchymal transition (EMT) in the GeneCards database was identified as EMT-DEPs. The protein network of EMT-DEPs was analyzed using the STRING database and Cytoscape software, and the hub EMT-DEPs were obtained by the MCC algorithm of the cytoHubba plugin. Correlation analysis was used to obtain the interpairing proteins among EMT-DEPs, and core EMT-DEPs were identified based on the number of paired proteins. The Venn program was used to identify the intersection of hub EMT-DEPs and core EMT-DEPs as key EMT-DEPs. Finally, a series of analyses plus experiments were used to verify the correlation of the target protein with invasion and EMT in pituitary adenoma.

Results

Quantitative comparison of proteins between invasive and non-invasive pituitary adenomas indicated 833 differential proteins. The overlaps of EMT-related proteins and differential proteins consisted of 46 EMT-DEPs. There were 6 intersections between the hub EMT-DEPs and core EMT-DEPs. Using quantitative protein data and GSE169498 chip, we found that solute carrier family 2 member 1 (SLC2A1) was our target protein. SLC2A1 was significantly correlated with the invasiveness of pituitary adenoma, and the ROC curve was satisfactory. The functions and pathways of SLC2A1 and paired protein enrichment were closely linked to the EMT. Consistently, SLC2A1 expression was significantly and positively correlated with the expression of classical markers of EMT. The final experiment revealed that SLC2A1 was significantly upregulated in invasive pituitary adenoma.

Conclusion

SLC2A1 is significantly upregulated in invasive pituitary adenoma with satisfactory predictive value. It may regulate EMT. It may be a potential diagnostic marker for invasive pituitary adenoma.
Prognostic biomarker GSTK1 in head and neck squamous cell carcinoma and its correlation with immune infiltration and DNA methylation
Authors: Feng Yao , Zhou Ying-Hui , Zhao Jie , Su Xiao-Lin , Chen Ning-Xin , Zhao Ya-Qiong , Ye Qin , Hu Jing , Ou-Yang Ze-Yue , Zhong Meng-Mei , Yang Yi-Fan , Han Peng-Ju , Guo Yue , Feng Yun-Zhi
Published: March 2023

Background: Glutathione S-transferase kappa 1 (GSTK1) is critical in sarcoma and breast cancer (BRCA) development. However, the clinical significance of GSTK1 in head and neck squamous cell carcinoma (HNSC) remains unclear. This study is the first investigation into the role of GSTK1 in HNSC.

Methods: All original data were downloaded from the Cancer Genome Atlas (TCGA) dataset and verified by R Base Package 4.2.0. The expression of GSTK1 in various cancers was explored with TIMER and TCGA databases. Prognostic value of GSTK1 was analyzed via survival module of Kaplan-Meier plotter and Human Protein Atlas database and Cox regression analysis. The association between GSTK1 and clinical features was evaluated by Wilcoxon signed-rank test and logistic regression analysis. The relationship between GSTK1 and immune infiltration and methylation level was further explored. The expression of GSTK1 and its correlation with immune cell infiltration was verified by Immunohistochemical staining (IHC).

Results: GSTK1 was lower in HNSC, BRCA, Lung squamous cell carcinoma, and Thyroid carcinoma than in para-carcinoma. Low GSTK1 expression was associated with worse overall survival in Bladder urothelial carcinoma, Kidney renal papillary cell carcinoma, BRCA, and HNSC. However, only in BRCA and HNSC, GSTK1 expression in tumors was lower than that in normal tissues. Cox regression analyses confirmed that GSKT1 was an independent prognostic factor of overall survival in HNSC patients. The decrease in GSTK1 expression in HNSC was significantly correlated with high T stage and smoker history. IHC showed that the expression level of GSTK1 in HNSC was lower than that in para-carcinoma. In addition, GSEA showed that three pathways related to immune infiltration were positively correlated, while two pathways related to DNA methylation were negatively correlated with expression of GSTK1. Further analysis showed that GSTK1 was moderately positively correlated with the infiltration level of T cells and Cytotoxic cells, which was further confirmed by IHC. The methylation level of GSTK1 was associated with prognosis in patients with HNSC.

Conclusion: Low GSTK1 expression may be a potential molecular marker for poor prognosis in HNSC and provide new insight for the development of diagnostic marker or therapeutic target.
N6-methyladenosine related gene expression signatures for predicting the overall survival and immune responses of patients with colorectal cancer
Authors: Yu Lili , Wang Lijuan , Sun Jing , Zhou Xuan , Hu Yeting , Hu Lidan , He Yazhou , Lin Chunqing , Chen Jie , Xu Xiaolin , Dunlop Malcolm G. , Theodoratou Evropi , Ding Kefeng , Li Xue
Published: March 2023

N6-methyladenosine (m6A) modification has been demonstrated to exhibit a crucial prognostic effect on colorectal cancer (CRC). Nonetheless, potential mechanism of m6A in survival rate and immunotherapeutic response remains unknown. Here we investigated the genes associated with m6A regulators and developed a risk score for predicting the overall survival (OS) of CRC patients. RNA-seq transcriptomic profiling data of COAD/READ samples were obtained from The Cancer Genome Atlas (TCGA) database. Absolute Shrinkage and Selection Operator (LASSO)- Cox regression analysis was conducted to identify the m6A-related gene expression signatures and the selected genes were inputted into stepwise regression to develop a prognostic risk score in TCGA, and its predictive performance of CRC survival was further validated in Gene Expression Omnibus (GEO) datasets. According to our results, the risk score comprising 18 m6A-related mRNAs was significantly associated with CRC survival in both TCGA and GEO datasets. And the stratified analysis also confirmed that high-risk score acted as a poor factor in different age, sex, T stage, and tumour, node, metastasis (TNM) stages. The m6A-related prognostic score in combination with clinical characteristics yielded time-dependent area under the receiver operating characteristic curve (AUCs) of 0.85 (95%CI: 0.79–0.91), 0.84 (95%CI: 0.79–0.90) and 0.80 (95%CI: 0.71–0.88) for the prediction of the 1-, 3-, 5-year OS of CRC in TCGA cohort. Furthermore, mutation of oncogenes occurred more frequently in the high-risk group and the composition of immune cells in tumour microenvironment (TME) was significantly distinct between the low- and high-risk groups. The low-risk group had a lower microsatellite instability (MSI) score, T-cell exclusion score and dysfunction score, implying that low-risk patients may have a better immunotherapy response than high-risk patients. In summary, a prognostic risk score derived from m6A-related gene expression signatures could serve as a potential prognostic predictor for CRC survival and indicator for predicting immunotherapy response in CRC patients.
Dual oxidase 1 is dispensable during Mycobacterium tuberculosis infection in mice
Authors: Gupta Tuhina , Sarr Demba , Fantone Kayla , Ashtiwi Nuha Milad , Sakamoto Kaori , Quinn Frederick D. , Rada Balázs
Published: March 2023

Introduction

Mycobacterium tuberculosis (Mtb) is the primary cause of human tuberculosis (TB) and is currently the second most common cause of death due to a singleinfectious agent. The first line of defense against airborne pathogens, including Mtb, is the respiratory epithelium. One of the innate defenses used by respiratory epithelial cells to prevent microbial infection is an oxidative antimicrobial system consisting of the proteins, lactoperoxidase (LPO) and Dual oxidase 1 (Duox1), the thiocyanate anion (SCN-) and hydrogen peroxide (H2O2), which together lead to the generation of antimicrobial hypothiocyanite (OSCN-) in the airway lumen. OSCN- kills bacteria and viruses in vitro, but the role of this Duox1-based system in bacterial infections in vivo remains largely unknown. The goal of this study was to assess whether Duox1 contributes to the immune response against the unique respiratory pathogen, Mtb.

Methods

Duox1-deficient (Duox1 KO) and wild-type (WT) mice were infected with Mtb aerosols and bacterial titers, lung pathology, cytokines and immune cell recruitment were assessed.

Results and discussion

Mtb titers in the lung, spleen and liver were not different 30 days after infection between WT and Duox1 KO mice. Duox1 did not affect lung histology assessed at days 0, 30, and 90 post-Mtb infection. Mtb-infected Duox1 KO animals exhibited enhanced production of certain cytokines and chemokines in the airway; however, this response was not associated with significantly higher numbers of macrophages or neutrophils in the lung. B cell numbers were lower, while apoptosis was higher in the pulmonary lesions of Mtb-infected Duox1 KO mice compared to infected WT animals. Taken together, these data demonstrate that while Duox1 might influence leukocyte recruitment to inflammatory cell aggregates, Duox1 is dispensable for the overall clinical course of Mtb lung infection in a mouse model.
Characteristics of humoral and cellular responses to coronavirus disease 2019 (COVID-19) inactivated vaccine in central China: A prospective, multicenter, longitudinal study
Authors: Yuan Youhua , Xu Junhong , Ma Bing , Chen Guohua , Wang Zhibin , Wang Shanmei , Jing Nan , Zhang Jiangfeng , Wang Baoya , Yan Wenjuan , Zhang Qi , Zhao Qiongrui , Li Yi
Published: March 2023

Introduction

In China, the long-term immunogenicity and adverse effects of inactivated vaccines produced by different or the same manufacturer remain unclear. Therefore, the objective of this study was to evaluate the cellular immune responses and neutralizing antibody kinetics of homologous and heterologous administrations of an inactivated coronavirus disease 2019 (COVID-19) vaccine 240 days after the second vaccination.

Methods

This prospective, multicenter, observational, longitudinal study involved 595 participants with a negative SARS-CoV-2 polymerase chain reaction result who were serologically tested and followed for 8 months after vaccination. Neutralizing antibodies, interferon-gamma (IFN-γ), interleukin (IL)-6, CD4+ T-lymphocyte, and B-lymphocyte counts were evaluated in serum samples after stimulation with 2 μg/mL SARS-CoV-2 spike protein for 16 h at follow-up intervals of 2 months.

Results

Most participants [582/595; 146 male participants, 449 female participants; mean age 35 (26–50 years)] rapidly developed neutralizing antibodies after two doses of the vaccine administered 3-weeks apart. The positive rate of neutralizing antibodies peaked at 97.7% at 60–90 days, decreased, and stabilized at 82.9% at 181–240 days post-vaccination. Lower antibody concentrations were correlated with older age, longer duration after vaccination, non-health care workers, mixed-manufacturer vaccinations, and intervals of less than 40 days between two doses of vaccination, whereas lower IFN-γ levels and B-lymphocyte counts were associated with older age, blood type A, and non-health care workers. A higher IL-6 level was associated with older age, mixed-manufacturer vaccinations, intervals of less than 40 days between two doses of vaccination, and medical staff. Adverse reactions were mild or moderate and self-limited, with no serious events reported.

Discussion

Two doses of the Chinese inactivated vaccine induced robust and rapid antibody expression and cellular immune responses. Boosting vaccination is considered important, as antibodies and cellular immune responses were reduced in susceptible populations.
Intra-articular delivery of AAV vectors encoding PD-L1 attenuates joint inflammation and tissue damage in a mouse model of rheumatoid arthritis
Authors: Li Wenjun , Sun Junjiang , Feng Susi Liu , Wang Feng , Miao Michael Z. , Wu Eveline Y. , Wallet Shannon , Loeser Richard , Li Chengwen
Published: March 2023

Objective

Rheumatoid arthritis (RA) is the most common form of autoimmune inflammatory arthritis. Intra-articular gene delivery to block proinflammatory cytokines has been studied in pre-clinical models and human clinical trials. It has been demonstrated that the level of programmed death-ligand 1 (PD-L1) is associated with rheumatoid arthritis (RA). This study examined the therapeutic role of PD-L1 by intra-articular delivery via adeno-associated virus (AAV) vectors in the mouse collagen-induced arthritis (CIA) model.

Methods

Mice were intra-articularly injected with AAV5 vectors encoding human PD-L1 on day 0 and immunized with bovine type II collagen to induce CIA simultaneously. On day 49 post AAV administration, joints were collected for histo-pathological and cytokine analysis. Additionally, the systemic impacts of intra-articular injection of AAV5/PD-L1 vectors were also studied. To study the therapeutic effect of PD-L1, AAV5/PD-L1 vectors were administered into the joints of RA mice on day 21.

Results

After administration of AAV5/PD-L1 vectors, strong PD-L1 expression was detected in AAV transduced joints. Joints treated with PD-L1 at the time of arthritis induction exhibited significantly less swelling and improved histopathological scores when compared to untreated joints. Additionally, the infiltration of T cells and macrophages was decreased in joints of CIA mice that received AAV5/PD-L1 vectors (P<0.05). The levels of pro-inflammatory cytokines, including IL-1, IL-6, IL-17 and TNFα, were lower in AAV5/PD-L1 treated than untreated joints (P<0.05). Furthermore, the administration of AAV5/PD-L1 vectors into the joints of CIA mice did not impact serum cytokine levels and the antibody titers to type II collagen. Biodistribution of AAV vectors after intra-articular injection showed undetectable AAV genomes in other tissues except for a low level in the liver. Similar to the results of AAV5/PD-L1 vector administration on day 0, decreased joint swelling and lower histopathological damage were observed in joints treated with AAV5/PD-L1 vectors on day 21.

Conclusion

The results from this study demonstrate that local AAV mediated PD-L1 gene delivery into the joints is able to prevent the development and block the progression of arthritis in CIA mice without impacting systemic immune responses. This study provides a novel strategy to effectively treat inflammatory joint diseases using local AAV gene therapy by interference with immune checkpoint pathways.
T regulatory cells metabolism: The influence on functional properties and treatment potential
Authors: Tomaszewicz Martyna , Ronowska Anna , Zieliński Maciej , Jankowska-Kulawy Agnieszka , Trzonkowski Piotr
Published: March 2023

CD4+CD25highFoxP3+ regulatory T cells (Tregs) constitute a small but substantial fraction of lymphocytes in the immune system. Tregs control inflammation associated with infections but also when it is improperly directed against its tissues or cells. The ability of Tregs to suppress (inhibit) the immune system is possible due to direct interactions with other cells but also in a paracrine fashion via the secretion of suppressive compounds. Today, attempts are made to use Tregs to treat autoimmune diseases, allergies, and rejection after bone marrow or organ transplantation. There is strong evidence that the metabolic program of Tregs is connected with the phenotype and function of these cells. A modulation towards a particular metabolic stage of Tregs may improve or weaken cells’ stability and function. This may be an essential tool to drive the immune system keeping it activated during infections or suppressed when autoimmunity occurs.
Deciphering comprehensive features of tumor microenvironment controlled by chromatin regulators to predict prognosis and guide therapies in uterine corpus endometrial carcinoma
Authors: Wu Qihui , Tian Ruotong , Liu Jiaxin , Ou Chunlin , Li Yimin , Fu Xiaodan
Published: March 2023

Background

Dysregulation of chromatin regulators (CRs) can perturb the tumor immune microenvironment, but the underlying mechanism remains unclear. We focused on uterine corpus endometrial carcinoma (UCEC) and used gene expression data from TCGA-UCEC to investigate this mechanism.

Methods

We used weighted gene co-expression network analysis (WGCNA) and consensus clustering algorithm to classify UCEC patients into Cluster_L and Cluster_H. TME-associated CRs were identified using WGCNA and differential gene expression analysis. A CR risk score (CRRS) was constructed using univariate Cox and LASSO-Cox regression analyses. A nomogram was developed based on CRRS and clinicopathologic factors to predict patients' prognosis.

Results

Lower CRRS was associated with lower grade, more benign molecular subtypes, and improved survival. Patients with low CRRS showed abundant immune infiltration, a higher mutation burden, fewer CNVs, and better response to immunotherapy. Moreover, low CRRS patients were more sensitive to 24 chemotherapeutic agents.

Conclusion

A comprehensive assessment of CRRS could identify immune activation and improve the efficacy of UCEC treatments.
Identification of lactylation related model to predict prognostic, tumor infiltrating immunocytes and response of immunotherapy in gastric cancer
Authors: Yang Hao , Zou Xiaoming , Yang Shifeng , Zhang Ange , Li Nana , Ma Zhen
Published: March 2023

Background

The epigenetic regulatory chemical lactate is a product of glycolysis. It can regulate gene expression through histone lactylation, thereby promoting tumor proliferation, metastasis, and immunosuppression.

Methods

In this study, a lactylation-related model for gastric cancer (GC) was constructed, and its relationships to prognosis, immune cell infiltration, and immunotherapy were investigated. By contrasting normal tissues and tumor tissues, four lactylation-related pathways that were substantially expressed in GC tissues were found in the GSEA database. Six lactylation-related genes were screened for bioinformatic analysis. The GC data sets from the TCGA and GEO databases were downloaded and integrated to perform cluster analysis, and the lactylation related model was constructed by secondary clustering.

Results

The fingding demonstrated that the lactylation score has a strong correlation with the overall survival rate from GC and the progression of GC. Mechanistic experiments showed that abundant immune cell infiltration (macrophages showed the highest degree of infiltration) and increased genetic instability are traits of high lactylation scores. Immune checkpoint inhibitors (ICIs) demonstrated a reduced response rate in GC with high lactylation scores. At the same time, tumors with high lactylation scores had high Tumor Immune Dysfunction and Exclusion scores, which means that they had a higher risk of immune evasion and dysfunction.

Discussion

These findings indicate that the lactylation score can be used to predict the malignant progression and immune evasion of GC. This model also can guide the treatment response to ICIs of GC. The constructed model of the lactate gene is also expected to become a potential therapeutic target for GC and diagnostic marker.
Fluorescence activated cell sorting and fermentation analysis to study rumen microbiome responses to administered live microbials and yeast cell wall derived prebiotics
Authors: Klassen Leeann , Reintjes Greta , Li Meiying , Jin Long , Amundsen Carolyn , Xing Xiaohui , Dridi Lharbi , Castagner Bastien , Alexander Trevor W. , Abbott D. Wade
Published: March 2023

Rapid dietary changes, such as switching from high-forage to high-grain diets, can modify the rumen microbiome and initiate gastrointestinal distress, such as bloating. In such cases, feed additives, including prebiotics and live microbials, can be used to mitigate these negative consequences. Bio-Mos® is a carbohydrate-based prebiotic derived from yeast cells that is reported to increase livestock performance. Here, the responses of rumen bacterial cells to Bio-Mos® were quantified, sorted by flow cytometry using fluorescently-labeled yeast mannan, and taxonomically characterized using fluorescence in situ hybridization and 16S rRNA sequencing. Further, to evaluate the effects of bovine-adapted Bacteroides thetaiotaomicron administration as a live microbial with and without Bio-Mos® supplementation, we analyzed microbial fermentation products, changes to carbohydrate profiles, and shifts in microbial composition of an in vitro rumen community. Bio-Mos® was shown to be an effective prebiotic that significantly altered microbial diversity, composition, and fermentation; while addition of B. thetaiotaomicron had no effect on community composition and resulted in fewer significant changes to microbial fermentation. When combined with Bio-Mos®, there were notable, although not significant, changes to major bacterial taxa, along with increased significant changes in fermentation end products. These data suggest a synergistic effect is elicited by combining Bio-Mos® and B. thetaiotaomicron. This protocol provides a new in vitro methodology that could be extended to evaluate prebiotics and probiotics in more complex artificial rumen systems and live animals.