Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice.

Objective To evaluate the prevention and control effectiveness of four major chronic diseases in Youyang County, and find the weak link of prevention and control, and to provide theoretical support for improving prevention and control strategies. Methods Based on the death data of permanent residents from 2012 to 2020 extracted from the cause-of-death registration and reporting system of Youyang County, a statistical analysis was conducted using SPSS19.0. The annual percentage change (APC) was tested by t-test. Results From 2012 to 2020, the mortality rate of and the standardized mortality rate of the four major chronic diseases and the premature mortality rate of diabetes in males showed an increasing trend (APC was 3.05%, 1.82% and 27.12%, respectively, P < 0.05). The mortality rate of the four chronic diseases in females increased (APC was 2.53%, P < 0.05), while the proportion of premature death of the four chronic diseases and the probability of premature death of cardiovascular and cerebrovascular diseases in females decreased (APC was -2.37%, -5.73%, P < 0.05). The standardized mortality rate and premature death rate of the four major chronic diseases were higher in males than those in females. The mortality rate of the four major chronic diseases and the premature death rate of diabetes in the whole population were on the rise (APC was 2.84% and 12.86%, P < 0.05). It was expected that the early death probability of the four major chronic diseases in Youyang County would be 12.65% in 2030, higher than the target value of 12.59% of ^“Healthy China 2030^”. Conclusion The future focus of Youyang County is to prevent and control malignant tumors and diabetes, especially to strengthen the prevention and control of male diabetes.

As a severe clinical manifestation of nonalcoholic fatty liver disease， nonalcoholic steatohepatitis （NASH） is characterized by lipid deposition and inflammatory damage in the liver. At present， clinical medications for NASH are still in the exploratory phase， and it is urgent to make progress. Recent studies have shown that the pathogenesis of NASH is associated with circadian rhythm disorders in the liver， with the specific manifestation of dysregulated expression of liver clock genes such as BMAL1， which increases hepatic lipogenesis， reduces fatty acid oxidation， and activates pro-inflammatory factors. Therefore， improving circadian rhythm of the liver and regulating the expression of liver clock genes are feasible strategies for the prevention and treatment of NASH. Currently， some medications for NASH via activating the proteins encoded by clock genes have been applied in animal experiments， for example， the REVERB full-agonist SR9009 can inhibit the development of liver inflammation， which confirms the possibility of NASH treatment by targeting the proteins encoded by clock genes. This article summarizes the role of hepatic clock genes in regulating lipid metabolism and the development and progression of inflammation in the liver and elaborates on the recent advances in medications targeting clock genes and the proteins encoded by clock genes， in order to provide new targets for the treatment of NASH.

Objective To comprehend the present condition of sustaining within familial pulmonary rehabilitation for COPD patients,alongside exploring the factors that impact this continuity.Methods A convenience sampling method was used to select 392 COPD patients from outpatient clinics of 2 tertiary-level A hospitals and 1 community health service center in Huaihua in Hunan Province from December 2023 to February 2024 as survey respondents.Patients were surveyed using the general information questionnaire,the COPD Assessment Test,the Modified Medical Research Council Dyspnea Scale,the Exercise Benefits/Barriers Scale,and the Brief 2-Way Social Support Scale.Results We sent out 392 questionnaires and collected 362 valid ones,with an effective response rate of 92.47％.The percentage of COPD patients who never performed home pulmonary rehabilitation was 56.08％,and the maintenance time of health behaviors in home pulmonary rehabilitation was 17.40％for 1-3 months,7.18％for 4-6 months,4.70％for 7-12 months,and 14.64％for more than 1 year.The results of multiple linear regression analysis showed that exercise place,pulmonary rehabilitation health education,exercise habits before having COPD,perceived level of exercise behavior,and two-way social support were influential factors affecting the maintenance of health behaviors in pulmonary rehabilitation in the families of COPD patients,which explained 38.5％of the variability.Conclusion Patients with COPD have a low level of maintenance of health behaviors in home pulmonary rehabilitation,and health education on exercise rehabilitation for COPD patients and caregivers should be strengthened to enhance patients'level of exercise perception and ability to utilize social support.

Objective:To investigate the effect and potential mechanism of exosomal miR-21 derived from Schwann cell(SC)in the repairment of sciatic nerve injury(SNI). Method:SC was infected with negative control lentivirus and lentivirus that expression of miR-21,and its exosome was collected and identified respectively.The SNI rat model was established by nerve stump anasto-mosis.After surgery,the rats were randomly divided into the model group(n=10),the SC-derived exosome group(n=10)and the SC-derived exosome group(n=10)that high expression of miR-21.The SC-derived exosomes group and SC-derived exosome group that high expression of miR-21 were treated with local injec-tion of exosome collected in vitro.After 3 weeks,the functional recovery was detected by behavioral experi-ment.The nerve regeneration was evaluated by immunofluorescence staining.RT-qPCR was used to detect the expression of miR-21 in serum exosome and miR-21 and SPRY2 in sciatic nerve.The targeting interaction be-tween miR-21 and SPRY2 was verified by dual-luciferase reporter gene experiment. Result:Compared with the model group,the functional recovery and regeneration of sciatic nerve were signifi-cantly improved in the other two groups.The expression of miR-21 was significantly increased,while the ex-pression of SPRY2 was significantly decreased.The improvement in the SC-derived exosome group with high expression of miR-21 were more significantly than those in the SC-derived exosome group. Conclusion:Exosomal miR-21 derived from SC can significantly promote the repair of sciatic nerve injury by targeting SPRY2.

Objective To summarize the current research status and cutting-edge trends of the off-label drug use in China,with a view to providing reference for researchers in this field.Methods CNKI and SinoMed databases were searched to collect research of the off-label drug use in China,and used Microsoft Excel 2021,the R software Bibliometric,and VOSviewer 1.6.18 to visualize the time and trend of publication,province,issuing authors and units,journals,keywords,and topic evolution of the included studies.Results 1 475 papers were included in the research.A total of 2 808 authors from 31 provinces,cities and regions had conducted relevant studies on over-the-counter medication,with an overall increasing trend in the number of publications.Among them,Guangdong province published the most studies related to this field,the Straits Pharmacy Journal and China Pharmacy published the most studies in this field.Proprietary Chinese medicines,antimicrobials,antitumor drugs,and other drugs were the research hotspots.In addition,the patients in pediatrics,outpatient emergency,obstetrics and gynecology,psychiatry and other departments as a special sick population,the clinical use of medication exists in the overspecification situation was also a future research trend.Conclusion At present,research in this field focuses more on OLDU for special populations,special diseases,special drugs,etc.In the future,researchers should conduct evidence-based evaluation of drugs on the basis of more high-quality evidence in order to seek the best evidence for guiding the clinical use of medication.At the same time,drug administration and medical institutions should also develop standardized management policies and systems to promote the rational and safe use of medication in healthcare institutions.

ObjectiveTo explore the distribution of traditional Chinese medicine （TCM） syndromes of alopecia areata （AA）， and to provide reference for TCM clinical syndrome differentiation and classification of AA. MethodsAA patients who visited the specialized hairiness clinic of Beijing China-Japan Friendship Hospital were included. A questionnaire was developed including general information of the patients， history of hair loss （onset time， triggers and exacerbating factors， disease progression）， current symptoms （symptoms and signs）， medical history， personal history， family history， and hair microscopy examination results. The factor analysis and cluster analysis were used to determine the syndrome elements and to summarize the syndrome types. ResultsA total of 600 patients with AA were included， including 218 males （36.33%） and 382 females （63.67%）. Totally， 128 patients （21.33%） had a family history of hair loss， and 326 patients （54.33%） had a previous related underlying disease. The leading triggering and exacerbating factors of AA were tension and anxiety， accounting for 335 cases （55.83%） and 285 cases （47.50%）， respectively. The top 10 symptoms involved among patients were scalp oil， anxiety， irritability， dreaminess， fatigue， itching， tension， weakness and dandruff. The factor analysis showed that the factor rotation converged after 9 iterations， and finally obtained 12 common factors and 34 variables， with a cumulative contribution rate of 58.59%. In terms of disease location of AA， the main syndrome elements were liver， spleen and kidney， and the disease nature syndrome elements were mainly dampness-heat， qi stagnation， yin deficiency， qi deficiency， and blood deficiency. The clustering analysis of the 12 common factors showed that TCM syndromes could be summarized into four categories： internal retention of damp-heat， liver-kidney deficiency， qi and blood deficiency， and liver constraint and spleen deficiency. There were significant differences in the distribution of TCM syndromes in patients of different ages and genders （P＜0.001）. ConclusionThe main disease location of AA is in the liver， spleen， and kidney， with the liver being the key. The disease mechanism of AA is a deficiency-excess complex， initially manifested as excess and later becoming deficiency. The TCM syndromes mainly include four types which are internal retention of damp-heat， liver-kidney deficiency， qi and blood deficiency， and liver constraint and spleen deficiency.

Whether Fanconi anemia (FA) heterozygotes are predisposed to bone marrow failure and hematologic neoplasm is a crucial but unsettled issue in cancer prevention and family consulting. We retrospectively analyzed rare possibly significant variations (PSVs) in the five most obligated FA genes, BRCA2, FANCA, FANCC, FANCD2, and FANCG, in 788 patients with aplastic anemia (AA) and hematologic malignancy. Sixty-eight variants were identified in 66 patients (8.38%). FANCA was the most frequently mutated gene (n = 29), followed by BRCA2 (n = 20). Compared with that of the ExAC East Asian dataset, the overall frequency of rare PSVs was higher in our cohort (P = 0.016). BRCA2 PSVs showed higher frequency in acute lymphocytic leukemia (P = 0.038), and FANCA PSVs were significantly enriched in AA and AML subgroups (P = 0.020; P = 0.008). FA-PSV-positive MDS/AML patients had a higher tumor mutation burden, higher rate of cytogenetic abnormalities, less epigenetic regulation, and fewer spliceosome gene mutations than those of FA-PSV-negative MDS/AML patients (P = 0.024, P = 0.029, P = 0.024, and P = 0.013). The overall PSV enrichment in our cohort suggests that heterozygous mutations of FA genes contribute to hematopoietic failure and leukemogenesis.
Anemia, Aplastic/genetics* ; Epigenesis, Genetic ; Fanconi Anemia/genetics* ; Germ Cells ; Hematologic Neoplasms/genetics* ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Retrospective Studies

Ulcerative colitis(UC)is a chronic inflammatory bowel disease caused by many factors including colonic inflammation and microbiota dysbiosis.Previous studies have indicated that celastrol(CSR)has strong anti-inflammatory and immune-inhibitory effects.Here,we investigated the effects of CSR on colonic inflammation and mucosal immunity in an experimental colitis model,and addressed the mechanism by which CSR exerts the protective effects.We characterized the ther-apeutic effects and the potential mechanism of CSR on treating UC using histological staining,intestinal permeability assay,cytokine assay,flow cytometry,fecal microbiota transplantation(FMT),16S rRNA sequencing,untargeted metabolomics,and cell differentiation.CSR administra-tion significantly ameliorated the dextran sodium sulfate(DSS)-induced colitis in mice,which was evidenced by the recovered body weight and colon length as well as the decreased disease activity index(DAI)score and intestinal permeability.Meanwhile,CSR down-regulated the production of pro-inflammatory cytokines and up-regulated the amount of anti-inflammatory mediators at both mRNA and protein levels,and improved the balances of Treg/Thl and Treg/Th1 7 to maintain the colonic immune homeostasis.Notably,all the therapeutic effects were exerted in a gut microbiota-dependent manner.Furthermore,CSR treatment increased the gut microbiota diversity and changed the compositions of the gut microbiota and metabolites,which is probably associated with the gut microbiota-mediated protective effects.In conclusion,this study provides the strong evidence that CSR may be a promising therapeutic drug for UC.

Objective:To observe the repairing effect of ozone water injection in the articular cavity for the treatment of knee osteoarthritis (KOA) on articular cartilage and to explore its repair mechanism.Methods:48 rats were randomly divided into fourgroups, the normal, model, normal saline and ozone water group, each group had 12 rats. The rats were injectied into the joint cavity with papain to establish a KOA model other than the normal group. After confirming the success of the model, the ozone water group and normal saline group was treated with ozone water and normal saline injection into the joint cavity once a week for a total of 3 treatments, the normal group and the model group are all raised routinely. Before and after the treatment, the ratknee joint behavioral score MG score was conducted; after the treatment articular cartilage surface gross score, hematoxylin and eosin (HE) staining and modified Mankin score of articular cartilage pathological changes was measured, and Western blot and Rt-PCR to measure the level of protein and mRNA expression of NF-κB p65, IKKβ and IκBα in articular cartilage tissues.Results:Compared with before the treatment, the rat knee joint behavioral score of the ozone water group was significantly lower (all P<0.05); after the treatment, the gross articular cartilage surface score and the modified Mankin score of the ozone water group were significantly reduced compared with the model and normal saline group (all P<0.05); Compared with the model and normal saline group, the protein and mRNA expression levels of NF-κB p65 and IKKβ in the ozone water group are significantly lower (all P<0.05), and the levels of IκBα are significantly higher (all P<0.05). Conclusion:Ozone water injection in the articular cavity can effectively repair damaged articular cartilage. The repair mechanism may be achieved by inhibiting the activation of NF-κB signaling pathway.