Hibiscus sabdariffa calyx (HS) water decoction extract is a commonly consumed beverage with various pharmacological properties. This systematic review examines the possible effect of HS intake on immune mediators. The Scopus and PUBMED databases were searched for all human and animal studies that investigated the effect of HS administration on immune related biomarkers. For each of the immune biomarkers, the mean, standard deviation and number of subjects were extracted for both the HS treated and untreated group. These values were used in the computation of standardized mean difference (SMD). Statistical analysis and forest plot were done with R statistical software (version 3.6.1). Twenty seven (27) studies met the eligibility criteria. Twenty two (22) of the studies were used for the meta-analysis which included a total of 1211 subjects. The meta-analysis showed that HS administration significantly lowered the levels of TNF-α , IL-6 , IL-1β , Edema formation , Monocyte Chemoattractant Protein -1  and Angiotensin converting enzyme cascade . The levels of IL-10 , Interleukin 8 , iNOS  and C- Reactive Protein , were not significantly changed by HS administration. Some of the results had high statistical heterogeneity. HS may be promising in the management of disorders involving hyperactive immune system or chronic inflammation. Hibiscus sabdariffa (HS) is a shrub that belongs to the Malvaceae family  .Hibiscus sabdariffa” OR roselle). The title and abstract of all the resultant search results were then screened for eligibility.Online databases (Pubmed and Scopus) were systematically searched by two researchers reviewing independently (UF and NC) to find articles that documented the impact of HS on immune biomarkers. Date restriction was not applied in the search. The search was conducted between March 20, 2020, and June 5, 2020. The databases were searched with the keywords  or other species of Hibiscus were excluded.The following were inputted in the inclusion criteria; (1) original articles published in the English language. (2) Human and animal studies that used HS as an intervention agent in a healthy or disease model. (3) Studies that reported any immune or inflammatory related biomarkers such as Interleukins, Chemokines, Toll-like receptors (TLRs), Prostaglandins, Tumor Necrosis Factor-α (TNF-α), NF-κB, Monocyte chemoattractant protein-1 (MCP-1/CCL2), Inducible Nitric Oxide synthase (iNOS), components of the cyclooxygenase and lipoxygenases pathways, components of the ACE cascade , ACE), inflammatory endpoints such as edema formation, and inflammatory diseases, etc. (4) Studies done using any extract from the calyces of HS. (5) Only studies that included both a control and an HS-intervention group were considered. Data obtained from Duplicate records were deleted, after which two authors  reviewed the collated articles to ascertain their suitability for inclusion. Where the two authors had different opinions on the inclusion of an article, the other researchers  voted and a consensus was reached. Titles and abstracts of articles were first screened, and publications that did not meet the written criteria were removed. Five independent researchers  assessed the full texts of the remaining articles for eligibility. For each of the eligible studies, relevant data were extracted independently by each of the reviewers , into a predesigned data extraction table which consisted of the first author’s last name, year of study, experimental model, HS formulation, dosage/route of administration, analytical method, the inflammatory biomarker investigated, tissue investigated, study sample size and summary of major findings. The data extraction table generated by each reviewer was subsequently compared and collated.Missing or incomplete data was estimated with Cochrane’s review manager software (RevMan 5.3). Statistics and meta-analysis were done with R statistical software (version 3.6.1). A random effect model was assumed and meta-analysis was computed using the DerSimonian-Laird tau estimator. The forest plot was plotted with the “meta-forest” statistical package using R –35. The www.arulerforwindows.com.The forest plot is essential for analyzing the pooled estimate of the mean differences between the control and the HS intervention group across different studies. The overall standardized mean difference and the pooled 95% confidence interval were presented for each biomarker analyzed. A forest plot was carried out on a particular immune-related biomarker only if more than one study reported complete data for that particular biomarker. The effect of HS on the levels of inflammatory biomarkers across healthy and diseased subgroups of the experimental models, as well as across human and rodent models were examined using subgroup analysis. Raw data for mean and standard deviation (SD) were extracted from chart images with the aid of a digital pixel ruler downloaded from For studies that reported standard error of mean (SEM) values, the SD was calculated using SD = (SEM∗√n).In estimating the pooled effect of HS administration on ACE signaling, data from different members of the pathways were combined in one analysis. For example the Ang II, ACE1, and Ang I receptors were all analyzed under ACE signaling. In studies where more than one dose of the HS extract was given, the dose that elicited the most effect was reported and used for the analyses. Some of the studies recorded some missing data. Such missing data (when possible), was estimated using RevMan 5.3. In some cases, the published data were not reported in forms that can be subjected to analysis with other data. Such data were transformed appropriately before analysis.The total search hits from both databases were 3718 papers (1943 from Scopus and 1775 from PubMed). A total of 78 relevant articles were downloaded after screening the title and abstract of the search results against the eligibility criteria. Duplicates articles (n=19) were removed and the remaining unique articles (n=59) were subjected to full-text screening. One article was excluded on account of being inaccessible. Twenty seven  articlesAdministration of HS calyces extract significantly decreased the levels of different pro-inflammatory biomarkers such as TNF-α , 40–43, HS administration lowered the MCP-1 levels when compared to untreated groups in the studies examined. The pooled SMD showed a significant decrease in MCP-1 Figure 2The meta-analysis 11 showeWe also attempted to compare the immunosuppressive potential of HS in different animal disease models using TNF-α, IL-6, IL-1β, MCP-1, and ACE as biomarkers  of its flavonoids and anthocyanin contents are rapidly absorbed and metabolized before appearing in the blood plasma as flavonoid conjugates and hippuric acid respectively , 51. AntAs discussed previously, the presence of different bioactive agents in extracts of HS, suggests the possible involvement of several mechanisms in the suppression of immune and inflammatory responses. Suggested mechanisms include the suppression of cellular stress, inhibition of enzymes that can trigger inflammatory reactions, and down-regulation of pro-inflammatory genes.An important mechanism involved in the anti-inflammatory activities of HS is its ability to suppress the generation of oxidative stress and cellular damage in cells. Several pathological conditions are associated with oxidative stress. Increased oxidative stress can cause cellular damages leading to the release of stress factors such as heat shock protein, reactive oxygen species (ROS), necrotic, and apoptotic factors, etc. These factors are capable of activating TLR proteins in immune cells leading to downstream expression of several inflammatory genes . Oxidati2O2)-stimulated rat dermal fibroblast , which functions in an opposing manner to both increase and decrease vasoconstriction respectively . ACE1 caACE2, a homologue and regulatory arm of ACE 1, diverts some of the Ang II generated by ACE1 into the production of a potent vasodilatory growth inhibitory peptide, angiotensin , which aInhibition of components of the angiotensin converting enzyme (ACE) cascades by HS extract have been reported in both human and animal studies , 25. TheHS has also been reported to inhibit lipoxygenase and cyclooxygenase enzyme systems , 59. ThiDown-regulation of pro-inflammatory genes and factors is another potential mechanism involved in HS anti-inflammatory activities. Different in-vivo and in-vitro studies have demonstrated the ability of HS or known components of HS to downregulate the expression of pro-inflammatory genes such as NF-κB, TNF-α, interleukins, iNOS , 31, 44.Microarray gene profiling studies by Chou et al. , showed Treatment with HS extract has been reported to down-regulate the activity of NF-κB (p65) subunit in different animal and cell line models , 18, 31.The results from the meta-analysis suggest that HS administration suppressed the levels of IL-1β across different experimental models Figure 6The result from the meta-analysis indicates that HS supplementation significantly suppressed the levels of TNF-α across the different studies Figure 8Evidence from this meta-analysis shows that HS treatment can significantly suppress the levels of MCP-1 Figure 2The meta-analysis showed that HS administration significantly suppressed the cellular levels of IL-6 in different disease models Figure 6IL-10 is an anti-inflammatory cytokine produced by virtually all immune cells. Its main function is to prevent immunopathology during inflammatory responses . The resInducible nitric oxide synthase (iNOS) synthesizes nitric oxide a regulator of immune responses. Overexpression of iNOS and the consequent over production of nitric oxide can lead to cellular injury including DNA damage . FindingThe suggested mechanism of HS anti-inflammatory activities is outlined in Components of HS extracts can suppress TLR mediated expression of inflammatory mediators by reducing the levels of DAMP molecules  and suppressing the expression of TLR. HS extract also suppresses the levels of MCP-1, TNF-α, IL-1β, IL-6 resulting in a reduction in the overall inflammatory/immune responses. HS extract can inhibit ACE1 activity as well as reduce the levels of ACE1 and Ang II proteins hereby suppressing downstream activation of AT1 receptor-mediated expression of inflammatory cytokines. HS can also interfere with components of NF-κB cascade thereby further suppressing the expression of inflammatory genes.The mechanism involved in the anti-inflammatory effect of HS extract in different disease models is multifaceted and involves the interaction of different constituents of HS with different cellular targets. It appears that the mechanism that predominates depends on the causative factors of each disease model. Diseases linked to dysfunctional oxidative stress most likely could involve the antioxidant capacity enhancement potentials of HS to restore normal antioxidant levels, bringing about the cessation of the observed inflammation. In pathological conditions caused by localized inflammation, the MCP-1, COX2, and lipoxygenase suppressing properties of HS might come into play to reduce inflammation. Reduced expression of MCP-1 will suppress the infiltration of circulating monocytes into inflammatory sites, reducing the extent of the innate immune response at such sites. The ability to inhibit ACE suppresses the possibility of having an enhanced Ang II/AT1 receptor-mediated downstream oxidative dysfunction and inflammatory response. A combination of all these effects could be beneficial for suppressing immune responses in conditions of chronic systemic inflammatory disorders.Apart from anti-inflammatory effects, other beneficial health effects have also been observed following HS supplementation. Using a combination of transcriptomics and metabolomics, Beltrán-Debón et al.  showed tThis systematic review is affected by the various limitations that are characteristics of systematic reviews. Some of such limitations include the fact that only published data that were identified using the search strategies and databases specified in our method section were accounted for in the data synthesis. The possible omission of other relevant studies either because they were not published or because they not were indexed in the databases we consulted cannot be ruled out. The data used in this review were obtained from in-vivo  studies in a limited disease model. Furthermore, there were few studies per disease model and in some cases, conclusions were drawn using models from just two studies. Only two disease models were reported for humans (i.e hypertensive and metabolic syndrome), as such, the possible impact of the human disease model on HS efficacy could not be reliably ascertained from this study. The small number of studies (per each biomarker) made it impossible to identify the possible causes of the heterogeneity observed in some of the pooled estimates. The number of studies used to generate the pooled estimate might not be sufficient to entirely reflect the real situation. Finally, most of the data used for this systematic review and meta-analysis were obtained from studies that exposed experimental models to chronic or sub-chronic levels of HS. The instantaneous efficacy of HS administration could not be thus ascertained from this systematic review.This systematic review and meta-analysis revealed that HS extracts possess immunosuppressive capabilities. This finding suggests that HS extracts might be beneficial in the treatment/management of pathological conditions associated with a hyperactive immune system. Further studies should verify the efficacy of HS in suppressing immune responses in autoimmune or chronic inflammatory disease conditions.The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.FU conducted the search, participated in data extraction, ran the statistical and forest plot analyses, designed the graphic abstract, and contributed to the introduction and discussion sections. JU and GB wrote the introduction and materials and methods and undertook data extraction. BE-E participated in data extraction, contributed to the discussion section, and collated the work. NC searched the databases and participated in data extraction. OO mentored the authors, reviewed the draft manuscript, and certified the final manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


San-Huang-Yi-Shen capsule (SHYS) has been used in the treatment of diabetic kidney disease (DKD) in clinics. However, the mechanism of SHYS on DKD remains unclear. In this study, we used a high-fat diet combined with streptozocin (STZ) injection to establish a rat model of DKD, and different doses of SHYS were given by oral gavage to determine the therapeutic effects of SHYS on DKD. Then, we studied the effects of SHYS on PINK1/Parkin-mediated mitophagy and the activation of NLRP3 inflammasome to study the possible mechanisms of SHYS on DKD. Our result showed that SHYS could alleviate DKD through reducing the body weight loss, decreasing the levels of fasting blood glucose (FBG), and improving the renal function, insulin resistance (IR), and inhibiting inflammatory response and oxidative stress in the kidney. Moreover, transmission electron microscopy showed SHYS treatment improved the morphology of mitochondria in the kidney. In addition, western blot and immunoflourescence staining showed that SHYS treatment induced the PINK1/Parkin-mediated mitophagy and inhibited the activation of NLRP3 signaling pathway. In conclusion, our study demonstrated the therapeutic effects of SHYS on DKD. Additionally, our results indicated that SHYS promoted PINK1/Parkin-mediated mitophagy and inhibited NLRP3 inflammasome activation to improve mitochondrial injury and inflammatory responses. Diabetic kidney disease (DKD), one of the leading diabetic complications, is a chronic structural and functional kidney disorder . Early Dα axis in renal tissues, thereby regulating mitochondrial biosynthesis and alleviating renal impairment in a mouse DKD model  was given by intraperitoneal injection. Animals were fasted for 12 h before the injection but were allowed to drink water. After 72 h, blood was collected from the tail vein to check if blood glucose was ≥16.7 mmol/L, which was used to confirm that the diabetic rat model was successfully constructed. Diabetic rats were housed for another 1 week. After 1 week, the rats were housed in metabolic cages and we performed 24 h of urine protein quantitation. Blood glucose level ≥ 16.7 mmol/L and urine protein ≥20 mg/24 h were used as criteria for the successful induction of the DKD animal model [DKD rats were divided into the model group, the SHYS low-dose group, and the SHYS high-dose group, with 10 rats per group. The control group and the model group were given an equal volume of distilled water via intragastric administration. Meanwhile, 0.81 g/kg and 1.62 g/kg of SHYS were given via intragastric administration for 8 continuous weeks to the SHYS low-dose group and the SHYS high-dose group, respectively. The dosages of SHYS were calculated based on human–rat body surface area conversion. During the study, rats were given ad libitum access to water and a standard diet. Fasting blood glucose (FBG) and body weight was measured every two weeks during SHYS treatment.After 8 weeks of SHYS intervention, metabolic cages were used to collect urine output from the groups, which was then centrifuged at 3000 rpm for 10 minutes. The urine protein content was measured according to the manufacturer's instructions.Besides, sodium pentobarbital (50 mg/kg) was administered by intraperitoneal injection to anesthetize the rat, and inner canthus blood was collected. The blood was centrifuged at 3,000 rpm for 15 minutes to collect serum. A fully automatic biochemical analyzer measured serum Cr and BUN levels in the various groups.μL of normal saline and homogenized on ice. Then, the homogenized renal tissue mixture was centrifuged at 3,000 rpm for 15 min and the supernatant was collected to obtain the tissue homogenate. The activities of SOD and GSH-Px and the level of MDA were investigated following the established protocol of test kits. The total protein contents in tissue homogenates were quantified using BCA protein concentration test kits.In addition, 100 mg of renal tissues were weighed and immersed into 900 β, IL-6 and TNF-α) in tissue homogenates were measured using ELISA based on the established protocol of the kits. The IR was evaluated based on homeostatic model assessment of IR (HOMA-IR) using the following formula: HOMA − IR = FBG × FINS/22.5. Besides, the total protein content in tissue homogenates were quantified using BCA protein concentration test kits.The fasting insulin (FINS) level in serum and the levels of proinflammatory cytokines (IL-1μm sections were cut and used for the hematoxylin and eosin (HE), Masson, and Sirius red staining. Pathological changes in rat renal tissues were observed under a microscope. The pathological changes in HE staining were evaluated based on the HE staining score as described previously [Renal tissues from the groups were fixed with a 10% formalin solution for 24 h, washed with water for 20 minutes, followed by dehydration using an alcohol gradient, xylene cleared, and then paraffin-embedded. Five eviously  , LAMP2 (1 : 500), Parkin (1 : 500), and VDAC1 (1 : 500) were incubated at 4°C overnight. The corresponding secondary antibodies were added after washing with PBS three times, followed by DAPI counterstaining for 10 minutes. The sections were then washed with PBS three times and sealed with an antiquenching agent. Images were taken using an immunofluorescence microscope. Image Pro Plus 6.0 was used to quantitate the fluorescence intensity, total area, and mean optical density. The positive area was calculated based on the ratio between mean optical density and total area.Renal tissues were fixed with formalin solution, embedded in paraffin, and cut into 3 μg sample was added to a 10% SDS-polyacrylamide gel for electrophoresis. The target proteins resolved in the gel were transferred to methanol-activated PVDF membranes. Membranes were blocked in 5% skim milk at room temperature for 2 hours before the antibodies, anti-VDAC1 , TOM20 , COXIV , LC3 , p62 , PINK1 (1 : 500), Parkin , NLRP3 , IL-1β , caspase-1 , ASC (1 : 500), IL-18 (1 : 500), and β-actin , were added and incubated at 4°C overnight. On Day 2, 1 × TBST buffer was used to wash the membrane three times for 10 minutes each. The corresponding secondary antibodies were added, and the membrane was incubated at room temperature for 2 h. 1 × TBST buffer was used to wash the membrane three times, 5 minutes each time. The enhanced chemiluminescence method was used to visualize the bands, and β-actin was the internal control. A gel imaging system was used for imaging. Image J software was used to analyze the gray values of each band to calculate the relative protein expression.Rat renal tissues were lysed on ice for 30 minutes with RIPA lysis buffer. Samples were centrifuged at 12,000 rpm at 4°C for 10 minutes, and the supernatant was transferred to fresh EP tubes. BCA protein concentration test kit was used to measure the protein concentration in the supernatant. A 20 t-test was used for comparison. One-way ANOVA followed by Tukey's post-hoc test was used for multigroup comparison of means. A difference of P < 0.05 was considered to be statistically significant.The SPSS Statistics 20.0 statistical software was used to analyze the experimental results. Quantitative data are expressed as mean ± standard deviation, and a P < 0.01, respectively). Low-dose and high-dose of SHYS treatment reduced the body-weight loss  and decreased the FBG level  in DKD model rats  whereas the Cr level was decreased in SHYS low-dose and SHYS high-dose groups  and the level of BUN was lower in the SHYS high-dose group (P < 0.05) compared with the model group , low-dose and high-dose of SHYS treatment lowered the HOMA-IR in rats with DKD  compared with the control group  in renal tissue homogenate were detected using ELISA to evaluate the anti-inflammatory effects of SHYS on DKD model rats. Compared to the control group, the levels of IL-1β, IL-6, and TNF-α were significantly increased in the model group . High-dose of SHYS treatment decreased the levels of IL-1β, IL-6, and TNF-α in the DKD model rats . The activity of SOD was increased in SHYS high-dose group compared to the model group (P < 0.05). Besides, low-dose and high dose of SHYS treatment decreased the MDA level  and increased the GSH-Px  activity in the DKD model rats , whereas low-dose and high-dose of SHYS treatment decreased the proportion of fragmented mitochondria  in rats with DKD . The protein levels of VDAC1 , TOM20 , and COXIV  were lower in the SHYS low-dose and SHYS high-dose groups as compared with those in the model group . Compared with the model group, low-dose and high-dose of SHYS treatment increased the protein levels of PINK1  and Parkin  in the kidney  was decreased and the protein level of p62 (P < 0.05) was increased in the model group compared with the control group. The protein level of LC3-II was higher in the SHYS low-dose and SHYS high-dose groups as compared to the model group  and the protein level of p62 was lower in the SHYS high-dose group compared with the model group  in the kidney were increased in DKD model rats compared to the rats in the control group . The coexpression of Parkin and VDAC1 , ASC (P < 0.05), cleaved caspase-1 (P < 0.01), mature IL-1β (P < 0.05), and mature IL-18 (P < 0.01) were increased in the model group. The protein levels of NLRP3, ASC, and mature IL-1β were lower in the SHYS high-dose group as compared to the model group . The protein levels of cleaved caspase-1  and mature IL-18  were lower in the SHYS high-dose group as compared to the model group  to Ub substrate or mitochondrial outer membrane proteins. The ubiquitin chains formed can be selectively recognized by the autophagy receptor p62 . p62 is β, and IL-18 expression in DKD rat renal tissues. Under physiological conditions, NLRP3 within cells is in an autoinhibitory state. Hyperglycemia can cause NLRP3 to recruit ASC and bind to pro-caspase-1 to form an inflammasome and activate caspase-1. Activated caspase-1 can cleave pro-IL-1β and pro-IL-18 to produce active IL-1β and IL-18, respectively, releasing from cells and worsening renal tissue inflammation.A recent study found that blockade of PINK1/Parkin-mediated mitophagy can induce NLRP3 inflammasome activation, and the activation of PINK1/Parkin-mediated mitophagy can block NLRP3 inflammasome activation . Our resIn summary, our study demonstrated the therapeutic effects of SHYS on DKD. Additionally, our results indicated that SHYS promotes PINK1/Parkin-mediated mitophagy in renal tissues and inhibits NLRP3 inflammasome activation to improve mitochondrial injury and inflammatory responses.


In this paper, a deep learning algorithm was proposed to ensure the voice call quality of the cellular communication networks. This proposed model was consecutively monitoring the voice data packets and ensuring the proper message between the transmitter and receiver. The phone sends its unique identification code to the station. The telephone and station maintain a constant radio connection and exchange packets from time to time. The phone can communicate with the station via analog protocol (NMT-450) or digital . Cellular networks may have base stations of different standards, which allow you to improve network performance and improve its coverage. Cellular networks are different operators connected to each other, as well as a fixed telephone network that allows subscribers of one operator to another to make calls from mobile phones to landlines and from landlines to mobiles. The simulation is conducted in Matlab against different performance metrics, that is, related to the quality of service metric. The results of the simulation show that the proposed method has a higher QoS rate than the existing method over an average of 97.35%. The first cellular networks were developed using analog first generation 1G) standards. The most common are NMT and AMPS. Usually, next to the standard name, they write the frequency in megahertz, followed by the frequency range for basic station communication with mobile phones. For example, the basic stations on NMT-450 networks communicate with cell phones on the 450 MHz bandwidth. Working simultaneously on analog standards to ensure that multiple mobile phones in a cell, as well as the base stations of different cells, used multiple frequency multiplexing , which would work in one of the conditions of scarcity of free frequencies G standar. This enCDMA (code segment multi-access) makes extensive use of sophisticated methods of separating radio air between different mobile phones. Also, no matter how many different phones are in one cell, and no matter how many base stations are in the neighborhood, each mobile phone uses the CDMA 2000 1x standard of 1.25 MHz to receive and transmit the entire frequency band (channel) of relatively large width . To distLater, with the advancement of mobile phones and the development of computerization, technologies for GSM networks such as computer data transfer and Internet access were introduced (cm Internet) . The firThe most common data transfer technology is GPRS . GPRS allows multiple mobile phones to use dedicated time intervals simultaneously, using different algorithms for different quality of communication with BS and different workloads of BS . Each phInitially, these technologies were used on mobile phones to access the Internet using personal computers, and only later, did further development of mobile phones provide direct Internet access from mobile phones. To obtain information on a mobile phone, WAP technology , which has relatively small requirements for technical specifications , pages cHowever, with the advancement of mobile phones, things changed quickly. First, the need for an intermediate server has disappeared—now the browsers of modern mobile phones do their work independently. Second, the specialized language WML is replaced by the XHTML standard, which differs from the language commonly used on the Internet . It is oThe authors devised a deep learning algorithm was proposed to ensure the voice call quality of the cellular communication networks.The deep learning model consecutively monitors the voice data packets and ensures the proper message between the transmitter and receiver.The main contribution of the paper includes:Carvalho et al.  discusseCells that use the same radio channel group can avoid interruptions if they are spaced apart. In this case, frequency reuse is observed. Cheng et al.  expresseAccording to Hsu et al. , the covHussien and Sadi  explaineEzaki et al.  discusseFrom the existing models, it is found that most of the communications are not analyzed at the micro-level and even the micro-level , dynamic channel assignment algorithm (DCAA), integrated structured cabling system (ISCS), and fuzzy probabilistic based semi-Markov model (FPSMM)) solutions provide a computational burden to the entire system. This leads to increased computational cost and communication cost in estimating the solution related to voice call QoS.The UMTS standard  is based on W-CDMA technology , somewhat compatible with GSM. The speed of data reception and transmission reaches 1920 kilobits per second.1xEV technology  for CDMA2000 networks. Data reception speeds reach 3.1 megabits per second, and transfer speeds reach 1.8 megabits per second.Technologies like TD-SCMA, HSDPA, and HSUPA. It allows you to reach even higher speeds. Until 2006, W-CDMA technologies mostly provided HSDPA support. TD-SCMA is in development.The data transfer rates on second-generation networks are not sufficient to carry out many new functions of mobile communications, especially high-quality video in real-time (videophone), modern optical computer games on the Internet, and others. New standards and protocols have been developed to ensure the required speed:In this way, modern technologies such as mobile communication are not mobile phone technologies like global communication technologies. Microcells are commonly used in densely populated areas. Due to their small range, microcells are less susceptible to transfer degradation effects such as reflection and signal delays. A macrocell integrates with a group of microcells. The microcell serves slow-moving mobile devices and the macrocell serves fast-moving devices. The mobile device can determine the speed of its movement. This makes it possible to reduce the number of hops from one cell to another and adjust the location data.N = the number of complete dual cellular channels available in the cluster; A = the number of channels in the cell; and D = the number of cells in a cluster.When the distance between the mobile device and the base station of the microcell is small, the transition mechanism from one cell to another can be changed. When planning systems using hexagonal cells, the base station transmitters are located at the center of the cell, at the edge of the cell, or at the top of the cell. Cells with a transmitter at the center typically use omnidirectional antennas, and cells with transmitters at the edge or apex typically use sector directional antennas. Consider a system with a fixed number of full-duplex channels in an area. Each service area is split up into groups called “clusters.” Each cluster gets a group of channels that are spread out among the cells of the cluster in different ways.M = total number of channels in a given region; A = the number of channels in the cell; D = the number of cells in a cluster; and k = the number of clusters in a given zone.If the cluster is duplicated within the specified service area, then the total number of full-duplex channels is :From the exposures, we can see that the total number of channels in the cellular telephone system is directly proportional to the number of back clusters in a given service area. If the cluster size decreases but the cell size does not change, more clusters will be needed to cover a given service area, and the total number of channels in the system will increase. Due to the absence of neighboring cells in a small service area , the number of subscribers, who can use the same frequencies (channels) simultaneously depends on the total number of cells in the area. The average number of such subscribers is four, but in densely populated areas, the number is much higher.With the planned increase in cellular transport, the increased demand for service is met by reducing the size of the cell, which divides it into several cells, each with its own base station. If the cells are not very small, efficient cell division allows the system to handle more calls. If the cell diameter is less than 460 m, the base stations of neighboring cells will affect each other. The criterion for determining the relationship between frequency reuse and cluster size is the cellular structure as the subscriber density increases.S = the number of cells in the S-cluster; z and x = non-negative integers ;Execution: direct transfer of control from one base station to another;Shutdown: unwanted network resources are released and available to other mobile devices.The transfer process from one cell to another, i.e., when the mobile device moves from the base station 1 to base station 2, has four main steps shown in Q–the cellular utilization point, R–the range cellular cluster, t–the quantity of signal, uee) was compared with the existing optimal call admission control (OCAC), dynamic channel assignment algorithm (DCAA), integrated structured cabling system (ISCS), and fuzzy probabilistic based semi-Markov model (FPSMM). The entire simulation is conducted in a Matlab environment that runs on a high-end computing system with 8 GB of RAM and an i5 computing processor.The moving vehicle's location is found by the automatic telephone transmission, while the call is sent to the communication channel by the circuit controller. In the cutoff region of the channel location area, all the existing models achieved 63.05%, 71.32%, 63.36%, and 87.24% of channel allocations. But the proposed model achieved 92.08% of the channel allocation. This is possible because the proposed model already predicted the upcoming signal location. Hence, this achieves a high allocation rate.When the vehicle leaves the area of the remote base station, the driver cannot use cellular communications. If the call is made on the way to the boundary of the zone, the signal will weaken and eventually disappear completely. Where the cutoff region of the base station management for all the existing models was achieved at 64.90%, 65.48%, 58.43%, and 79.86% of base station management. But the proposed model achieved 93.88% of base station management. This is possible because the proposed model was already predicted for the base station. It is useful for the users to periodically modify the base stations while traveling. Hence, this achieves high base station management. In most communication networks, the transmitter transmits the higher data rate messages through the air medium. This was always at a higher level because, based on the modulation and filter quality, this may be affected. So the data rate from the transmitter was always huge. From the receiver end, the same data rate was received, and then the demodulation process was performed with the help of different filters. Where the cutoff region of the data transfer rate for all the existing models was achieved, 56.23%, 68.98%, 57.61%, and 79.87% of transfer communication between the transmitter and receiver. But the proposed model achieved 92.36% of the channel allocation. This is possible because the proposed model already found the shortest path between two endpoints. Hence, this achieves a high data transfer rate.In general, the communication mediums delay the process. The delay from the transmitter is called the transmission delay, and the delay from the receiving medium is called the receiver delay. So the speed of the transmission and receiving medium are always high for cellular communication. In the cutoff region of the communication speed, all the existing models achieved 63.09%, 64.79%, 54.57%, and 75.54% of channel allocations. But the proposed model achieved 96.70% of communication speed. This is possible because the proposed model already predicted the upcoming destination. Hence, this achieves a high speed.In general, the quality of service in cellular communication is very essential because this is the parameter used to evaluate the service range of the cellular base station. These cellular communication instruments are performed well to increase the quality of service of the network. A modern MTA can be automatically switched to Dictaphone mode by a signal or a given program without its owner's approval. It is not true that every MDA records the owner's speech and voice and then sends the information, but such an opportunity is technically provided in every modern MDA. The proposed LWDLA was compared with the existing OCAC, DCAA, ISCS, and FPSMM. If an action takes place during a show in a theater, it is almost obvious that the gun will explode by the end of the show. So, in this case, MTA has the ability to record and transfer information, and this factor should be taken into account when using your mobile phone. The MTA receives information from the station closest to the cell. In it, information spreads in the air. The MTA interacts with the station at bursts of digital pulse signals called “time intervals.” The duration of a service contact session can range from fractions of a second to several seconds. In the future, the solutions can be improved by increasing the rate of QoS of voice calls or HD voice calling using artificial intelligence approaches.


The model group and the drug group are given intraperitoneal injections of vitamins, and the model group and the drug group are given a high-fat diet. Rats in the low-dose group and high-dose group are given low-dose and high-dose Dahuang Zhechong pill lavage solution, respectively. Besides, the control group is given simvastatin solution by gavage, and intervention is performed once a day for 12 weeks. Ubiquitin (Ub) protein expression, ubiquitin activase (UBE1), nuclear factor-κB, nuclear inhibitory factor-κB (IκB) gene expression, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and serum tumor necrosis factor-α (TNF-α) are compared. The experimental result shows that Dahuang Zhechong pills can reduce inflammation and prevent and treat AS by blocking the activation of the UPP/NF-κB signaling pathway and can be used as a proteasome inhibitor in the clinical treatment of AS.In order to investigate the effects of different doses of Dahuang Zhechong pills on the ubiquitin proteasome pathway/nuclear factor- The ubiquitin proteasome pathway (UPP) is a pathway involved in the NF-κB activated signal pathway that plays an important role in regulating the expression of inflammatory factors. Therefore, it is of important reference value to clarify the change characteristics and mechanism of the UPP-NF-κB signaling pathway in AS for the optimization of subsequent clinical targeted therapy [With the change of people's lifestyle and the aggravation of population aging, the incidence rate of cardiovascular disease is increasing year by year. As one of the pathological bases of various cardiovascular and cerebrovascular diseases, the incidence rate of atherosclerosis (AS) has increased significantly. Effective prevention and treatment of AS has important guiding significance for disease control, improving quality of life and reducing the risk of other cardiovascular diseases . The cli therapy . At pres therapy . In rece therapy .κB signaling pathway and its mechanism, aiming to provide data support for subsequent application of paternal treatment in the clinical prevention and treatment of AS.Dahuang Zhechong pill is a prescription from the ancient book Synopsis of the Golden Chamber by Zhang Zhongjing, which can play a good role in removing blood stasis, generating new vitality, and nourishing deficiency. Previous studies have confirmed that Dahuang Zhechong pills can inhibit the activity of and factor signaling pathway, reduce inflammation, and play a certain role in anti-AS, but the specific mechanism has not been clarified in relevant studies. Therefore, this study further explored the effect of different doses of paternal treatment on the UPP-NF-This paper is organized as follows: α is one of the polytropic inflammatory factors in AS, which can activate cyclic adenosine monophosphate (cAMP) before AS [α is reversely upregulated and activated to promote the capture of a large number of granulocytes in the blood stream, thus inducing a vicious cycle of calcification and inflammation mutually promoting [Oxidative stress, high glucose, and inflammation play an important role in the formation of atherosclerotic calcification. TNF-efore AS . The cAMromoting .κB signaling pathway in several ways. For example, the degradation of NF-κB precursor protein requires UPP to generate p50 and p52. The NF-κB signaling pathway can be activated only after ubiquitination and degradation of IκB protein, leading to the high expression of related inflammatory factors and genes. Therefore, inhibition of ubiquitination and degradation of IκB can indirectly inhibit the activation and transcription of the NF-κB signaling pathway, thereby inhibiting inflammatory response and improving arteriosclerosis [As a nonlysosomal protein degradation pathway, UPP is widely present in eukaryotic cells and has a certain dependence on adenosine triphosphate (ATP), participating in and playing a decisive role in the activation of nuclear factor signaling pathways . UPP regclerosis .κB promotes the inflammatory signaling pathway and increases IL-1β and macrophage content, thereby increasing serum IL-6 and TNF-α concentration, aggravating inflammatory response, and aggravating AS. Inhibition of the NF-κB signaling pathway can reverse the downregulation of contractile protein in some smooth muscle cells and inhibit cell proliferation and secretion. NF-κB may be a key signaling pathway node in the phenotypic transformation of smooth muscle cells and participates in the occurrence and development of AS by regulating the phenotypic transformation of vascular smooth muscle cells [κB was highly activated. Activation of NF-κB and high expression of inflammatory genes were important factors leading to the occurrence and development of AS. In addition, the UPP pathway was highly activated in the aorta tissues of AS rats. It indicates that inhibition of IκB degradation and activation of the NF-κB signaling pathway play an important role in improving AS [. As a statin, simvastatin can inhibit cholesterol synthesis and promote LDL metabolism by upregulating LDL receptors on the cell surface, reducing TC and LDL, and thus achieving anti-AS effects [Scutellaria baicalensis, peach kernel, almond, Rehmannia glutinosa, grubs, Tabanidae, insects, leeks, peony, dried lacquer, and licorice. In terms of pharmacology, they can reduce the plaque area of AS, inhibit the proliferation of smooth muscle cells and collagen fiber proliferation, and then inhibit intima thickening and foam cell formation. Clinical studies showed that Dahuang Zhechong pills can play an anti-inflammatory role and inhibit the expression of TNF-α and NF-κB in the AS lesion area. The insects and leeches in the prescription can, respectively, play the roles in reducing blood viscosity, antiplatelet aggregation, and blood lipid [κB in rats with atherosclerosis.NF-le cells , 11. Abnle cells . Yang etle cells  showed toving AS . As a st effects . Dahuangod lipid . It is i58-week-old male clean grade Wistar rats were purchased from the Animal Experiment Center of Hebei Medical University for the experiment, weighing 180–200 g, with Certificate No. 1305143 and License No. SCXK(Ji)2016-1-004. After adaptive feeding for a week, the rats are divided into 5 groups, including the normal group, model set, matched group, LDG, and HDG, with 10 rats in each group.Before the start of the experimental rats, in addition to the normal group, the rest of the group by intraperitoneal injection of rats are given vitamin D360 unit/kg, the experimental group; after the start of normal rats using normal feed, the rest of the group of rats were with 10.2% lard propylthiouracil, 3% cholesterol, 0.5% sodium cholic acid, 5% sugar, and high-fat feed, continuous feeding for 12 weeks.−1·d−1 distilled water intragastric administration. The low-dose group and high-dose group are given 0.7 g·kg−1·d−1 and 1.4 g·kg−1·d−1 Dahuang Zhechong pills, liquid intragastric administration, respectively; the control group is given 5 mg·kg−1·d−1 simvastatin liquid intragastric administration, once at 9 am every day. The intervention lasted for 12 weeks.At the beginning of modeling, rats in each group are given corresponding drugs by gavage intervention. The normal group and the model group are given 10 ml·kgThe expression level of Ub protein is detected by immunohistochemistry. The rat thoracic aorta tissue of 1-2 cm is prepared into wax blocks, and the slices are dewaxed into water. The antigen is repaired by microwave for 10 min and washed repeatedly with phosphate buffered saline (PBS) for 3 times. After 20 min, goat blood is added, cleaned, and sealed for 30 min in a constant temperature environment of 37°C. After washing, rabbit anti-ub antibody (1 : 200) is added and stayed overnight at 4°C. After washing with PBS, the secondary antibody working solution is dropped, respectively, and incubated for 30 min in a warm box. After dropping horseradish peroxidase-labeled chain enzyme ovalbumin, the reaction is continued to be incubated for 30 min, and the reaction is terminated by DAB. Known Ub-positive is used as a positive control, and PBS is used as a negative control instead of the primary antibody. Two experienced pathologists independently read and interpreted the radiographs. Five high magnification fields (×400) are randomly selected. The percentage of Ub-positive cells is calculated according to the brownish yellow particles in the cytoplasm and nucleus, of which 26%–50% is 1 point, 51%–75% is 2 points, and more than 75% is 3 points. The staining intensity score is 0 for no color, 1 for light brown or yellow mark, 2 for tan, and 3 for tan. Add the two score values, and ≥ 3 is positive and < 3 is negative. Pathologists who read these films selected typical pigmentation sites, excluded background pigmentation and nonspecific pigmentation, and then performed micrographs. Image Pro Plus software is used for gray scale scanning of photos. The gray value is proportional to the protein expression intensity.μL reverse transcription reaction system is composed of 3.0 μg total RNA, 0.5 μg oligo dT, 1.3 μl dNTP, 40U RNasin, 200U MMLV-RT, 5 μL5 × loading buffer, and appropriate DEPC water. After kept in a waterbath at 37°C for 1 hour, the cDNA is inactivated at 70°C for 5 min. The cDNA is stored at−20°C. The 50 μL PCR amplification reaction system is composed of 5 μL cDNA, 1 μL dNTP, 5 μL10 × loading buffer, 20 μmol primers, 25 μL MgCl, 1 μL Taq enzyme, and DEPC water. UBE1 mRNA, NF-κB mRNA, and IκB mRNA are pre-denatured at 50°C for 2 min, pre-denatured at 95°C for 10 min, and then annealed at 60°C for 60 seconds. The operation is repeated 40 times, with U6 as an internal reference. 10 μL PCR product is extracted and electrophoretic is performed with 20 gl−1 agarose gel. The results are observed by the UV lamp. The absorbance ratio of UBE1 mRNA, NF-κB mRNA, and IκB mRNA is calculated by semiquantitative detection of the optical density scanner. All samples are repeated twice, and the average value is taken.Total RNA is extracted by a one-step method using the Invitrogen TRIzol kit. 25 α detection kit .After 12 weeks of administration, the head is cut off and 2 ml blood samples are collected by two test tubes, one of which is centrifuged at 800r/min for 10 min. Plasma is separated by a Roche automatic biochemical analyzer . Measured total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C). serum is separated by centrifugation at 3500 r/min for 10 min in another test tube, and serum is determined by enzyme linked immunosorbent assay (ELISA) in strict accordance with the TNF-For each rat aorta tissue, made paraffin section, dewaxing in normal operation to the water, with hematoxylin staining for 10 minutes, immersed in water ish, appropriate differentiation is accomplished with 1% volume fraction of hydrochloric acid alcohol and ishing in water, which lasts for 5 minutes with eosin stain, also ish in flowing water for five minutes. The 70%, 80%, 95%, and 100%gradient alcohols are dehydrated, according to the normal steps, and neutral gum is used to seal the tablets. Images are taken under an Eclipse TE 2000-S immunofluorescence microscope purchased from Japan, and morphological changes in rat aorta are observed.x ± s) and the t-test is adopted. P < 0.05 indicated that the data are statistically significant.The software that effectively processed the data in the study is SPSS 22.0, which was used to test the normality of the measurement data. The normal distribution of the measurement data is presented in the form of , as shown in ∗” means prompt comparison model set, P < 0.05.Compared with the model set, the normal group, the matched group, and the HDG had lower lipid levels, and there are statistical differences between the groups , as shown in Compared with the model set, TNF-P < 0.05), as shown in Compared with the model set, Ub protein in the normal group, matched group, and HDG is lower, and there is a statistical difference between groups , as shown in Table 4.Compared with the model set, the mRNA levels of UBE1, NF-κB in rats with AS are investigated. The experimental results demonstrate that Dahuang Zhechong pills can reduce the expression of inflammatory factors and improve AS by blocking the activation process of the UPP/NF-κB signaling pathway. It is recommended that high-dose Dahuang Zhechong pills be promoted as a proteasome inhibitor in the clinical treatment of AS [In this study, the effects of different doses of Dahuang Zhechong pills on the UPP-NF-nt of AS .


Guidelines for management and prevention of contrast media extravasation have not been updated recently. In view of emerging research and changing working practices, this review aims to inform update on the current guidelines.In this paper, we review the literature pertaining to the pathophysiology, diagnosis, risk factors and treatments of contrast media extravasation. A suggested protocol and guidelines are recommended based upon the available literature.• Risk of extravasation is dependent on scanning technique and patient risk factors.• Diagnosis is mostly clinical, and outcomes are mostly favourable.• Referral to surgery should be based on clinical severity rather than extravasated volume.The online version contains supplementary material available at 10.1007/s00330-021-08433-4. Contrast media extravasation (CMEX) is a complication where there is leakage of intravenously administered contrast agents (either iodine or gadolinium-based), into the surrounding soft-tissues . This caCMEXs are thought to be one of the most frequent adverse events in radiology but are much less studied than others such as contrast-associated acute kidney injury –10. WhilPrevious guidelines around CMEX from the Contrast Media Safety Committee (CMSC) of the European Society of Urogenital Radiology (ESUR) published in 2002 related to older contrast injection protocols . Since tAuthors prepared seven clinical questions in Patient–Intervention–Comparison–Outcome (PICO) format . A systeRisk of bias of each study included was graded according to National Institutes of Health (NIH) study quality assessment tools. The strength of recommendation of different risk factors, diagnosis, detection and management of CMEX were graded according to OCEBM Levels of Evidence Working Group “The Oxford 2011 Level of Evidence”  plus location of the cannula; more damage occurs with involvement of tight sub-fascial compartments compared to looser subcutaneous layers . An acutConsequently, much research has been based on preventing cannula dislodgement and exploring the impact of variables that increase risk of leakage from the puncture site.With CMEX contrast escapes and infiltrates the interstitium during injection. This is mostly a clinical diagnosis and routine use of imaging is not indicated for its detection.Non-physicians are usually first responders to CMEX, i.e. radiology technicians/radiographers and healthcare support workers. Hence, it is recommended that imaging departments follow a protocol which allows identification of at-risk patients, easy detection of CMEX, awareness of monitoring needs and effective management of extravasation by a wide range of staff . As an eWe summarise three main ways of detecting CMEX in Table Many vendors have developed Extravasation Detection Accessories (EDA) which are sensors that allow detection and interruption of automated injection in the event of an extravasation. Dykes et al. demonstrated the use of EDA resulted in smaller volumes of CMEX when they occurred compared to when no EDA was utilised  . HoweverOne quality improvement project and three case-series have investigated EDA use; each assessed a different device –27. Two Proper documentation of CMEX is crucial ; at the With severe extravasations, imaging documentation may be helpful . Plain rSix studies evaluated the influence of the physical properties of cannulae; none were prospective randomised trials. A pseudo-randomised trial compared between a fenestrated 20G and a non-fenestrated 18G cannula which showed no effect on CMEX rates or volume . FurtherCentral venous catheters (CVC—tunnelled or non-tunnelled), totally implantable vascular access devices, haemodialysis catheters and peripherally inserted central catheters (PICCs) are increasingly used for patients in critical care, on chemotherapy or long-term antibiotics. Of course these patients often require regular cross-sectional imaging with IV contrast. Power injector compatible versions of these have been shown to be safe , with a Prior to the advent of power-injectable CVCs, the Medical and Healthcare Products Regulatory Agency (MHRA) in the UK published recommendations regarding rates and volumes for CM injections. However, these have been withdrawn and current advice is to follow specific manufacturer guidance. Plumb and Murphy recommended a maximum flow rate of 2.0 ml/s for CVC and to use the distal lumen if a multi-lumen CVC is in place . A CT toSinan et al. did not find any significant difference in extravasation rates in patients receiving CM via power injection vs. manual injection . A studySix studies have assessed rate and volume impact on CMEX. A randomised controlled trial by Kok et al. demonstrated no significant difference in CMEX when assessing different flow rates  . Howeverp = 0.0005) [A non-randomised retrospective study found higher rates of extravasation in patients who had ultrasound-guided cannula insertion (3.6%) vs. standard insertion (0.3%). However, the numbers of patients in each group were drastically different—364 in the ultrasound-guided cannula inserted group and 896 in the standard cannula inserted group . The difference in results in this study is likely due to confounding variables; i.e. prior failure of standard insertion and with deeper veins there is potentially a shorter length of intravascular cannula hence greater potential for dislodgement upon injection . The typ 0.0005) , despiteThree main contrast media properties have been studied in relation to CMEX: osmolarity, charge and viscosity.2O) and cellular lysis which is thought to be a factor in the degree of tissue damage caused by extravasation. This has only been studied in animal models where old ionic CM  was used [Osmolarity—A relationship has been demonstrated between higher osmolarity CM  are well tolerated in humans , 50. On Viscosity—This is thought to influence the likelihood of extravasation occurring. A computational fluid dynamics study by Sakellariou et al. demonstrated the potential for increased incidence of extravasation with more viscous CM, especially in CT angiography when performed with smaller peripheral cannulas . A viscoOverall, this suggests that using CM with lower viscosity and/or warming CM and/or diluting/mixing CM with saline has a preventative role in CMEX.There is much less data on rates of CMEX for MRI; the incidence of CMEX is reported as approximately 0.06% with no serious complications described—likely due to low infusion rates and lower CM volumes compared to IBCM uses. Theoretically, the extravasation of GBCAs could lead to oedema, necrosis or haemorrhage, potentially exacerbated due to their ionicity and higher osmolarity when compared to IBCM, although this does not seem to be borne out in practice , 48.Based on a meta-analysis of 356,582 patients by Ding et al., females and older patients (> 60 years) are at greater risk of developing CMEX; however, gender and age have no impact on the volume of CM extravasation when it does occur . In-patiIt is suggested based on the risk factors discussed in Table The untreated sequelae of severe CMEX are as follows: increased intra-compartmental pressure (compartment syndrome), with subsequent risk of ischemia due to venous congestion and low arterial gradient causing disproportionate necrosis, severe neurovascular compromise or even limb loss. Treatments to avoid these serious complications can be divided into passive conservative measures and more active therapies Table .Table 5Studies assessing conservative methodologies to treat extravasation events are few and not of robust quality in terms of applicability for radiology, i.e. laboratory-based studies or investigations of extravasation of cytotoxic drugs rather than contrast media. Most recommendations are based on “good clinical practice”. The aim of conservative measures is to reduce the morbidity associated with CMEX. The evidence base for use of invasive treatments is limited with mostly retrospective studies, small case numbers, lack of control groups, data based on older CM or even not CMEX.Increased swelling or painIncreased rednessChange in sensation in the affected limbSkin ulceration or blisteringIt is important that clear instructions to the patient who has suffered CMEX are given regarding when to seek additional medical care if there are worsening symptoms. A patient information leaflet is recommended, available in the languages appropriate for the institution. Patients should be warned about red flag signs and symptoms which are as follows:In Table Compartment syndrome is the most serious among the consequences of CMEX. It is extremely rare with less than 12 cases reported in a recent literature review however must be suspected if patient complains of severe pain and/or neurovascular compromise . Signs wOverall, the decision to refer for surgical intervention should be a clinical one—i.e. based on red flag signs and symptoms, rather than arbitrary CMEX volumes.This review highlights the key up-to-date evidence pertaining to CMEX summarising the important risk factors and a systematic approach to management. Whilst this review has been all encompassing, there are some limitations. Heterogeneity of the studies included in the paper have made performing a meta-analysis tricky and often difficult to compare data across different types of studies. Most of the studies are retrospective and with rates of CMEX being generally low, there are inadequately statically powered studies. There have been technological changes in CT and MRI, especially with use of non-ionic CM, increased understanding of the risk factors and use of EDAs as well as systems which halt injection if problems are encountered during infusion, all of which inform update on the previous guidelines. However, there remain important areas where further research would be merited. There is no data available on patient experience of CMEX and this is an important impact to explore as the patient may refuse to have CM in the future. Long-term follow-up of patients after CMEX and prospective trials with CMEX interventions are also not well-researched. Important questions to ask are appropriate time interval when to re-scan after a CMEX, impact on workflow and cost implications.Supplementary file1 (DOCX 27.1 KB)Supplementary file2 (DOCX 27.3 KB)Supplementary file3 (DOCX 27.5 KB)Supplementary file4 (DOCX 90.8 KB)Supplementary file5 (DOCX 33.3 KB)Below is the link to the electronic supplementary material.


Significant evidence links white matter (WM) microstructural abnormalities to cognitive impairment in schizophrenia (SZ), but the relationship of these abnormalities with functional outcome remains unclear.n = 25; SZ-HCP-C2, n = 24) and patients with lower cognitive performance . Healthy controls (HC) were included in both cohorts . We compared fractional anisotropy (FA) of the whole-brain WM skeleton between the three groups  by a whole-brain exploratory approach and an atlas-defined WM regions-of-interest approach via tract-based spatial statistics. In addition, we explored whether FA values were associated with Global Assessment of Functioning (GAF) scores in the SZ groups.In two independent cohorts , patients with SZ were divided into two subgroups: patients with higher cognitive performance  was positively correlated with GAF score, in C2 the FA of the temporal part of the left IFOF was positively correlated with GAF score.We provide robust evidence for WM microstructural abnormalities in SZ. These abnormalities are more prominent in patients with low cognitive performance and are associated with the level of functioning.The online version contains supplementary material available at 10.1007/s00406-021-01363-8. Diagnostic and Statistical Manual of Mental Disorders (DSM-V) describes a broad range of severities of cognitive impairment in SZ, ranging from intact to severe [Schizophrenia (SZ) is a severe neuropsychiatric disorder that is associated with poor social –3 and oco severe . HoweverThe disconnection hypothesis in SZ has been put forward in the form of various disconnection theories, such as disconnection of fronto-temporal regions , 12, forTherefore, we performed a replication study that applied a whole-brain exploratory approach and atlas-defined WM regions-of-interest approach via tract-based spatial statistics (TBSS) in two different cohorts. In the present study, we aimed to robustly identify microstructural differences in WM in patients with SZ subdivided into groups with good and poor cognitive performance. Moreover, we aimed to investigate the relationship between regions-of-interest (ROIs) selected on the basis of exploratory findings and functional outcomes in SZ. We hypothesized that brain WM microstructure in SZ is more severely disturbed in patients with poor cognitive performance and poor functioning.International Statistical Classification of Diseases and Related Health Problems, 10th revision (ICD-10) [Participants in cohort (1) were recruited from the University Hospital, LMU Munich, Germany. This study was approved by the local ethics committee of the University Hospital, LMU Munich (project number: 17–13), and written informed consent was obtained from all participants. The patients were diagnosed by two independent, experienced psychiatrists using the criteria of the (ICD-10) . Individ(ICD-10) ) were re(ICD-10)  and addi(ICD-10)  and leve(ICD-10) . GAF sco(ICD-10) –40. Part(ICD-10) , 42.n = 25) and LCP group , and cohort (2) comprised 27 HC and 48 patients with SZ subdivided into an HCP group  and LCP group .The demographic and clinical characteristics of the study participants in cohorts 1) and (2) are shown in Table  and (2) We used neurocognitive instruments that are related to functional deficits in SZ , 30, 43.b values (b = 1000 s/mm2 and b = 0).All magnetic resonance imaging (MRI) examinations were performed with a 3.0 T MR scanner  with a standard 20-channel phased-array head coil. DTI was performed with 64 non-collinear diffusion-encoding directions and the following parameters: repetition time, 9600 ms; echo time, 95 ms; field of view, 244 mm; voxel size, 2.0 × 2.0 × 2.0 mm; slice thickness, 2.0 mm; 65 slices; and multiple diffusion weighting b values (b = 1000 s/mm2 and b = 0).All MRI examinations were performed with a 3.0 T MR scanner  with a standard 8-channel phased-array head coil. DTI was performed with 12 non-collinear diffusion-encoding directions and the following parameters: repetition time, 6500 ms; echo time, 96 ms; field of view, 256 mm; voxel size, 2.0 × 2.0 × 2.0 mm; slice thickness, 2.0 mm; 49 slices; and multiple diffusion weighting DTI data were processed with TBSS programs  in the Ft tests for continuous variables  and chi-square tests for categorical variables (sex distribution and hand preference), with a significance level of α < 0.05. Differences in demographic and clinical characteristics between the SZ-HCP and SZ-LCP groups in each cohort were analyzed with independent samples t tests for continuous variables  and chi-square tests for the number of patients taking medication, with a significance level of α < 0.05. Differences in demographic and clinical characteristics between the HC, SZ-HCP, and SZ-LCP groups in each cohort were analyzed by analysis of variance and Bonferroni’s post hoc test for continuous variables  and chi-square tests for categorical variables (sex distribution and hand preference), with a significance level of α < 0.05. Group differences in demographic and clinical characteristics between the SZ groups in cohorts (1) and (2) were analyzed by independent samples t tests for continuous variables (age and duration of school education) and chi-square tests for categorical variables (sex distribution and hand preference), with a significance level of α < 0.05. All statistical analyses were performed with IBM SPSS statistics 20.In each cohort, differences in demographic and clinical characteristics between the HC and SZ groups were analyzed by independent samples α < 0.05.Voxel-wise statistics of the skeletonized FA data were applied using randomize in FSL, version 6.0.0. The HC and SZ groups were compared by an analysis of covariance design, with age and sex as nuisance covariates. We randomly performed permutation-based testing with 5000 permutations and inference by threshold-free cluster enhancement (TFCE) with a threshold of less than 0.05. The mean FA values of the whole skeleton in the HC, SZ-HCP, and SZ-LCP groups were examined for differences by analysis of variance and Bonferroni’s post hoc test, with age and sex as covariates, with a significance level of t tests, with age and sex as covariates, with significance set at p < 0.00125. (= 0.05/40 WM tracts because 20 WM tracts were examined in each cohort). The mean FA values of 20 ROIs in the HC, SZ-HCP, and SZ-LCP groups were examined for differences by analysis of variance and Bonferroni’s post hoc test, with age and sex as covariates and significance set at p < 0.00125 (= 0.05/40 WM tracts because 20 WM tracts were examined in each cohort). In the voxels with a statistical difference in the mean FA values of 20 ROIs, voxel-wise multiple regression analyses were performed with TBSS to examine the relationship between FA values and demean GAF scores. We used 5000 permutations to calculate FA values using age and sex as covariates. Spearman's rank correlation test was carried out between the demeaned GAF scores and mean FA values of the voxels that were statistically significant in the voxel-wise multiple regression analysis.For atlas-based segmentation, all extracted skeletons were overlaid with the Johns Hopkins University DTI-based WM Atlas in FSL , 50. Difp < 0.01).Table p < 0.001). No significant differences were found between the SZ-HCP and SZ-LCP groups with regard to the duration of illness, PANSS subscales, GAF scores, chlorpromazine daily dose equivalents, and the number of patients taking an antidepressant or benzodiazepine.In cohort (2), no differences in age, sex, hand preference, or duration of school education were observed between the HC and SZ groups. Age, sex, and hand preference were not different between the HC, SZ-HCP, and SZ-LCP groups, but duration of school education was significantly lower in the SZ-LCP group than in the SZ-HCP group (p < 0.05).Further analyses of demographic and clinical characteristics between the SZ groups in cohorts (1) and (2) are shown in Supplementary Table 1. The SZ group in cohort (2) had significantly higher PANSS negative, general, and total scores than the SZ group in cohort (1) , the FA values in the SZ group were significantly lower than those in the HC group in the left temporal basal areas , the FA values in the SZ group were significantly lower than those in the HC group in widespread regions , FA values in the frontal part of the left IFOF were significantly associated with the demean GAF scores . Second, in the SZ group we found a significant positive relationship between cognition-related FA values in the fronto-temporal part of the left IFOF and the GAF score.Our results are consistent with previous findings which revealed that WM volumes were significantly smaller in patients with SZ with cognitive impairment than in healthy individuals but WM volumes in patients without cognitive impairment were not . MoreoveIn this study, we could replicate in two independent cohorts of patients with SZ, that especially patients with poor cognitive performance were affected by WM microstructural abnormalities. Moreover, cognition-related WM abnormalities, which were mainly found in fronto-temporal parts of the left IFOF, were related to general, social, and occupational functioning.It is worth emphasizing here that in our study FA was measured with different MRI scan parameters in two independent cohorts. A clinical application of structural MRI in clinical trials requires a certain robustness of readouts across different scanners and protocols, despite all biological and technical variability.The advantage of the current study is the replication of our findings in two independent cohorts and thus increases generalizability. Therefore, our results provide robust evidence that emphasizes the relevance of large reductions in FA as indicating a neurobiological mechanism of SZ in patients with severe cognitive impairment. Beyond the group comparison, the future goal is to find a framework for individual patients  to allowIn its atlas-based analysis, the current study found a significant positive relationship in both cohorts between cognition-related FA, mainly in the fronto-temporal part of the left IFOF, and demean GAF scores in patients. The IFOF is the longest associative bundle and connects the occipital cortex, superior parietal lobule, and temporal basal areas to the frontal lobe , 56. MorAn interesting finding from our analysis of the whole skeleton is that the FA values in widespread regions were significantly lower in the SZ group than in the HC group in cohort (2) but not in cohort (1). Another interesting finding from our atlas-based analysis is that we found a significant positive relation between the FA values and the demean GAF scores in the temporal part of the left IFOF in cohort (2) but in the frontal part of the left IFOF in cohort (1), i.e., the local regions in the left IFOF could not be replicated in both cohorts. This discrepancy between the independent cohorts might be explained, at least in part, by some factors affecting FA, such as differences in symptom severity for SZ , the useSome limitations of this study must be noted. First, the patients were taking a variety of pharmacological agents not only at the time of scanning, but prior to the scanning . Second, they were evenly divided with regard to the cognitive composite score and could not be classified according to unique score criteria , though previous findings show that approximately one quarter of schizophrenia have similar cognitive performance as healthy . Future The main strength of this study is the replication approach that used independent samples with different MRI parameters. The results provide a foundation for developing the neurobiological basis of cognitive function, which could serve as a functional proxy in SZ and other psychiatric disorders. Of particular importance will be the design of an approach to transform individual changes of the structural and functional connectome towards a clinical application. Here, cognition and functionality for psychiatric disorders will also play an important role across diagnoses.Supplementary file1 (DOCX 28 KB)Below is the link to the electronic supplementary material.


The identification of novel molecular systems with high fluorescence and significant non-linear optical (NLO) properties is a hot topic in the continuous search for new emissive probes. Here, the photobehavior of three two-arm bis[(dimethylamino)styryl]benzene derivatives, where the central benzene was replaced by pyridine, furan, or thiophene, was studied by stationary and time-resolved spectroscopic techniques with ns and fs resolution. The three molecules under investigation all showed positive fluorosolvatochromism, due to intramolecular charge-transfer (ICT) dynamics from the electron-donor dimethylamino groups, and significant fluorescence quantum yields, because of the population of a planar and emissive ICT state stabilized by intramolecular hydrogen-bond-like interactions. The NLO properties (hyperpolarizability coefficient and TPA cross-section) were also measured. The obtained results allowed the role of the central heteroaromatic ring to be disclosed. In particular, the introduction of the thiophene ring guarantees high fluorescent quantum yields irrespective of the polarity of the medium, and the largest hyperpolarizability coefficient because of the increased conjugation. An important and structure-dependent involvement of the triplet state was also highlighted, with the intersystem crossing being competitive with fluorescence, especially in the thiophene derivative, where the triplet was found to significantly sensitize molecular oxygen even in polar environment, leading to possible applications in photodynamic therapy. In our long-term research project on the push-pull behavior of stilbenoid compounds bearing donor–acceptor (D/A) groups linked by a conjugated bridge (D-π-A systems) ,3,4,5,6,These push-pull systems show large charge displacement during the excitation and the presence of intramolecular charge transfer (ICT) in the excited state particularly favored in polar solvents.Recently we have studied the photobehavior of distyrylbenzene analogs in which the central benzene was replaced by a heteroaromatic ring , and the lateral benzenes functionalized with strong electron-withdrawing nitro groups in the para position ,26.3 states able to generate strong spin-orbit coupling benzene derivatives  spectroscopic techniques coupled with theoretical calculations at the TD–DFT level was performed. The combined experimental and theoretical study allowed the competitive processes deactivating the excited electronic states and their mechanisms to be pointed out, highlighting the effect given by the central heteroaromatic ring . The spectral characterization of the three compounds in solvents of different polarity to derive their hyperpolarizability coefficient through a solvatochromic method is also reported, together with their two-photon absorption (TPA) cross-sections measured by the two-photon excited fluorescence technique.Particular attention was paid to the comparison of the photobehavior of the compounds investigated of the present study, with the symmetric dinitro-derivative analogs and the asymmetric ones that carry both the dimethylamino group on one edge and the nitro group on the opposite side, with the intent to investigate how the replacement of strong electron-withdrawing nitro substituents with strong electron-donor dimethylamino ones can tune the photobehavior and NLO properties in these symmetrical systems.−1 cm−1. The absorption spectrum shifted to the red at about 60 to 70 nm when replacing the π-deficient pyridine unit with the π-rich furan and thiophene rings, respectively, due to the increased degree of conjugation , an, an49], f DMA-QT . TD–DFT f DMA-QT  managed tals see  showed atals see , a consiCT, between the first excited singlet state and the ground state was determined. The Stokes shift recorded in EtOH was excluded from the plot for both DMA-QP and DMA-QF because of the intramolecular H-type bonds with the solvent affecting spectral positions beside solvatochromism. The ΔμCT values match the fluorosolvatochromic behavior of the DMA derivatives; the largest difference was found for the furan-derivative, followed by the pyridine- and thiophene-substituted compounds. This information was then manipulated using Oudar’s formula (Equation (2)) for the pyridine and furan derivatives and in the modified equation (Equation (3)) for the quadrupolar DMA-QT compound, to give the dynamic hyperpolarizability coefficients (βCT) in Tol  were found to be comparable to other NLO materials and their trend reproduces the increased conjugation when replacing the central pyridine with the furan and, particularly, with the thienyl unit, as found for other push-pull molecules benzene derivatives under investigation proved very informative about the role of the central heteroatoms in tuning the spectral, photophysical, and NLO properties of this molecular series. In fact, the π-rich furan and thiophene rings endow the system with an increased degree of conjugation, shifting the absorption spectra towards longer wavelengths as opposed to the absorption of DMA-QP, which instead features a π-deficient pyridine ring. Conjugation also plays a leading role in determining the entity of the hyperpolarizability coefficients, which were found to be large and comparable when dealing with DMA-QF and DMA-QT, due to the great electron-transport ability of furan and thiophene , and abomophores ,51, as wmophores ,57, analTPA were, however, modest and the TPE spectra did not overlap the OPE profiles. This finding could be explained by taking into account the conformational equilibria between compressed and elongated forms. On the one hand, the compressed and bent configuration is favored because of intramolecular hydrogen-like bonds; on the other hand, the elongated form is expected to show a higher TPA cross-section because of the larger transition dipole moment associated with a straight configuration, as already reported for other systems [Conversely, the ICT character is fundamental for enhancing third order NLO properties, such as TPA abilities, with DMA-QF exhibiting higher TPA cross-sections relative to DMA-QT. The σ systems , thus coF = 0.20, 0.37, and 0.53 for DMA-QP, DMA-QF, and DMA-QT, respectively, vs. ΦF = 0.004, 0.10, and 0.21 for the corresponding dinitro compounds) [T = 0.43 and very significant production of singlet oxygen by sensitization from the triplet. These properties make the dimethylamino-thiophene derivative even more attractive than its nitro-substituted counterpart, where both fluorescence and triplet production were found to reduce with polarity [The presence of ICT dynamics for the three molecules was evidenced by the important fluorosolvatochromism, especially marked for DMA-QF, and the significant spectral evolution probed by fs-resolved ultrafast spectroscopy. However, the lack of excited-state lifetime quenching with solvent polarity, which was instead found to increase for DMA-QP and DMA-QF when passing from a scarcely to a highly polar medium because of the competition with intersystem crossing, points to a planar ICT state which remains significantly fluorescent. The emissive capability in a polar environment differed radically from what has been previously observed for asymmetric styrylbenzene analogs, whose highly dipolar push-pull A-π-D structure leads to the formation of an ICT state that becomes non-emissive upon increasing polarity . As for mpounds) , making polarity .DMA-QT, therefore, features a very interesting photobehavior in a polar environment, where its excited state deactivates entirely by fluorescence and intersystem crossing (followed by molecular oxygen sensitization) in an almost 50/50 ratio, making the molecule promising for applications as both an imaging probe and a PDT agent.The synthesis procedure of the DMA-QP compound has been already described in a previous paper , while tE,1′E)-furan-2,5-diylbis)bis.DMA-QF , p-dimethylaminobenzaldehyde , KOAc (1 eq), and acetic anhydride (3 eq) were added. In addition, a catalytic amount of I2 was added and the reaction mixture was left on reflux for 24 h. The reaction mixture was poured into the ice and saturated with aqueous NaOH. The precipitate was formed, filtered, and washed with water. After removal of the solvent, the residue was worked up with water and toluene and dried over MgSO4. The crude reaction product was chromatographed and the pure E,E-isomer of DMA-furan was obtained in the last fractions using petroleum ether–diethylether (5%) mixture as eluent.DMA-QF: yellow solid; isolated yield 21% (32 mg); fR = 0.35 (petroleum ether–diethylether (5%)); m.p. 119–122 °C; 1H NMR  δ/ppm of 7.37 , 6.93 , 6.72 , 6.68 , 6.09 , 2.97 ; 13C NMR  δ/ppm of 152.1 (s), 150.8 (s), 149.4 (s), 128.8 (2d), 125.4 (2d), 112.1 (d), 107.5 (d), 107.1 (d), 39.9 (2q); HRMS for C24H26N2O: M+calcd 358.2045; M+found 358.2049.E,1′E)-thiophene-2,5-diylbis)bis).DMA-QT  and azobisisobutyronitrile (0.1 eq) were added. The reaction mixture was heated on reflux and irradiated with a halogen lamp (75 W) overnight. After cooling down to RT, the reaction mixture was filtrated to remove succinimide and evaporated. After the removal of the solvent, an extraction with dichloromethane and water was carried out. The extract was dried and concentrated. The crude product was dissolved in benzene and triphenylphosphine (PPh3) in benzene was added. After stirring overnight at RT the precipitate was filtered off and used in the next step after drying. To a stirred solution of obtained phosphonium salt (0.45 mmol) and p-dimethylaminobenzaldehyde (0.45 mmol) in p.a. ethanol, sodium ethoxide was dropwise added . Stirring continued for 4 h at RT. After removal of the solvent, the residue was worked up with water and toluene and dried over MgSO4. The crude reaction product was chromatographed and the pure E,E-isomer of DMA-thiophene was obtained in the last fractions  by repeated column and thin-layer chromatography using petroleum ether–diethylether mixture as eluent (5%).DMA-QT: yellow solid; isolated yield 14% (25 mg); fR = 0.30 (petroleum ether–diethylether (5%)); m.p. 128–130 °C; 1H NMR  δ/ppm of 7.36 , 6.99 , 6.84 , 6.82 , 6.70 , 2.99 ; 13C NMR  δ/ppm of 146.8 (s), 141.8 (s), 137.9 (s), 127.4 (d), 127.2 (2d), 125.6 (2d), 124.8 (d), 112.5 (d), 40.4 (2q); HRMS for C24H26N2S: M+calcd 374.1817; M+found 374.1814.1H and 13C NMR spectra were recorded on a spectrometer at 600 MHz (3 using tetramethylsilane as a reference. High-resolution mass spectra (HRMS) were obtained on a matrix-assisted laser desorption/ionization time-of-flight MALDI-TOF/TOF mass spectrometer  equipped with Nd:YAG laser operating at 355 nm with a firing rate of 200 Hz in the positive ion reflector mode, described in detail in Ref. [The  600 MHz . All NMR in Ref. . A totalThe structures of the molecules under investigation are presented in F, experimental error ± 10%) of dilute solutions (A at λexc < 0.1) were obtained by exciting each sample at the relative maximum absorption wavelength by employing 2-(1-naphthyl)-5-phenyl-1,3,4-oxadiazole (α-NPD), 9,10-diphenylanthracene and tetracene (ΦF = 0.70 [Absorption spectra were recorded with a Cary 4E (Varian) spectrophotometer. Fluorescence emission and excitation spectra were detected using a FluoroMax-4P (HORIBA Scientific) spectrofluorimeter and analyzed by FluorEssence software with appropriate instrumental response correction files. The fluorescence quantum yields  were measured in air-equilibrated Tol and DMF solutions using the DMA derivatives as sensitizers. The 1O2 phosphorescence spectra were detected using a spectrofluorimeter FS5 (Edinburgh Instrument) equipped with an InGaAs detector. Phenalenone  was employed as a reference compound for comparison purposes [Singlet oxygen quantum yields  or HRS (hyper–Rayleigh scattering) techniques, with the advantages of simplicity and the application of conventional steady-state stationary spectroscopy only. As for the case of the EFISH method, the solvatochromic method provides the βCT dominant contribution, which corresponds to the βXXX component of the β tensor describing the CT transition and being referred to as the frequency of the exciting laser of EFISH allows for direct comparison and good agreement between the results of the two approaches.Even though approximated, the solvatochromic method managed to give a valid estimation of the second order hyperpolarizability coefficients of βn) expressed as CT = μE − μG) according to Equation (1):−1), a is the cavity radius within Onsager’s model, taken as 60% of the calculated diameter along the CT direction resulting from the optimized geometry [h is the Planck constant and c is the speed of light in a vacuum.This method is based on the results of the fluorosolvatochromic behavior. The dependence of the Stokes shift  as follows:d and thus ΔQCT = 4d2(ΔμCT)2. In this case, the quadrupole is considered as two opposite dipoles, separated by a distance d between their barycenter, sharing the central heteroatomic ring.In the case of DMA-QT, as also reported for other quadrupolar push-pull systems ,25, anotf) obtained by the integrated absorption band as −1) of the bathochromic CT transition, and 0 has been calculated considering Equation (4) [The dynamic hyperpolarizability coefficient was then derived through Oudar’s formula (3)βCT=β9∫ε(v)dv  or predi−3 J/cm2 of energy fluence and under the magic angle condition, stirring the solution in a 2 mm cuvette (0.5 < A < 1.0 at λexc = 400 nm) during the experiments to avoid photoproduct interferences. Photodegradation was checked by recording the absorption spectra before and after the time-resolved measurement, where no significant change was observed. The experimental data matrixes were first analyzed using the Surface Xplorer PRO (Ultrafast Systems) software, where it was possible to perform SVD of the 3D matrix to derive the principal components (spectra and kinetics). Successively, a global analysis using GloTarAn software was performed in order to obtain the lifetimes and the evolution-associated spectra (EAS) of the detected transient.The experimental setup for the femtosecond transient absorption and fluorescence up-conversion measurements have been widely described elsewhere ,24,27. B−1) laser flash photolysis (Edinburgh LP980) with a pump pulse centered at 355 nm , coupled with a PMT for signal detection. A pulsed xenon lamp was then used to probe the absorption properties of the produced excited states. The setup was calibrated using an optically matched solution of Benzophenone in AcCN (ΦT = 1.0 and εT = 5200 M−1 cm−1) [T) were measured by energy transfer experiments from dithienylketone  to the DMA-QP and DMA-QF compounds, and from DMA-QF and DMA-QT to the all-trans-α, ω-di(2-thienyl)octatetraene  [T = 0.66 [T = 30,000 M−1 cm−1 at λT = 630 nm) [T = 0.71 and εT = 45,500 M−1 cm−1 at λT = 422 nm) [T). All measurements were performed by bubbling the sample with pure N2.Triplet formation quantum yields, transient absorption, and triplet lifetimes were measured using nanosecond (pulse width 7 ns and laser energy < 1 mJ pulse−1 cm−1) . Triplet 465 nm) . The triT = 0.66  and εT = 630 nm)  and Anthsignal between 410 and 750 nm; idler between 820 and 2200 nm). The fluorescence intensity is detected by a PTM tube (Hamamatsu R2257-Y001), powered by a high-voltage power supply  and given in mV by an oscilloscope LeCroy-Wave Runner-LT322 . Solutions of known concentration (about 1 × 10−5 M) in DMF were prepared together with fluorescein in buffered water at pH 11 (σ = 26 GM at 930 nm) [The same Nd:YAG pump coupled with an optical parametric oscillator  was used to perform the two-photon excited fluorescence experiments. The OPO can be tuned to produce radiation in the 410–2200 nm range ( 930 nm)  as referQuantum mechanical calculations were performed by using the Gaussian 16 package . DFT wit