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 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.

Objective To investigate the effect and potential molecular mechanism of PTGS2 on the prognosis of colon cancer patients.Methods The transcriptomic and proteomic data of pan-cancer were collected from TCGA,HPA,UALCAN and other databases,and the expression pattern and prognostic value of PTGS2 were analyzed by combining the clinical data such as staging,histology,survival time and so on.Based on GSEA,the biological functions which were significantly activated in patients with high expression of PTGS2 were iden-tified and the colon cancer cell line SW480 was used as an example for in vitro validation.PTGS2 over-expressing cell strains were constructed,and the effect on cell proliferation was determined by CCK8 method.Different concen-trations of H2O2 were used to form gradient oxidative stress,and the changes in cell antioxidant capacity were detected.The regulatory mechanism was preliminarily verified by Western blot.Results The transcription and expression of PTGS2 were found to be significantly up-regulated in colon cancer patients(P<0.05),and the increased expression of PTGS2 was associated with an increased mortality risk(P<0.05).Data analysis and in vitro experiments showed that over-expression of PTGS2 may promote the proliferation of colon cancer cells by activating the mTOR pathway.The antioxidant effect of cells was regulated by up-regulating oxidative stress regulatory proteins SOD2 and NRF2.Conclusions PTGS2 is a potential risk factor for colon cancer and its over-expression promotes cell proliferation,enhances cell antioxidant effect and is associated with poor progno-sis of colon cancer patients.

Objective To evaluate the long-term efficacy of tumor necrosis factor-α(TNF-α)inhibitor(TNFi)in the treatment of ankylosing spondylitis(AS)complicated with osteoporosis(OP)and the impact on bone metabolism,bone density,and inflammatory factors.Methods The data of 158 patients with AS and OP,who were admitted to Department of Rheumatology and Immunology of The First Affiliated Hospital of Naval Medical University(Second Military Medical University)from Jan.1,2010 to Dec.31,2017,were retrospectively collected.The patients were divided into bisphosphonate group(n=54),TNFi group(n=58),and TNFi+bisphosphonate group(n=46)according to the treatment methods.All patients were treated with calcium combined with calcitriol as the background treatment.After 5 years of treatment,Bath ankylosing spondylitis disease activity index(BASDAI)and Bath ankylosing spondylitis functional index(BASFI)scores were evaluated,and inflammatory indexes,bone metabolism markers,and bone mineral density were detected.Results After 5 years of treatment,the BASDAI and BASFI scores,erythrocyte sedimentation rate(ESR),C reactive protein(CRP),TNF-α,and interleukin-17A of the TNFi+bisphosphonate group and TNFi group were significantly lower than those before treatment(all P＜0.05);in the bisphosphonate group only ESR and CRP were significantly lower than those before treatment(both P＜0.05),and the other inflammatory indexes and BASDAI and BASFI scores showed no significant changes(all P＞0.05).The bone mineral density of the 3 groups after 5 years of treatment was significantly higher than that before treatment(all P＜0.05),and the bone mineral density of the TNFi+bisphosphonate group was significantly higher than that of the other 2 groups(both P＜0.05).After 5 years of treatment,the levels of parathyroid hormone(PTH),procollagen type 1 N-terminal propeptide(P1NP)and β-C-terminal telopeptide of type Ⅰ collagen(β-CTX)in the TNFi+bisphosphonate group and bisphosphonate group were significantly decreased compared with those before treatment(all P＜0.05),while the levels of N-terminal midfragment of osteocalcin(N-MID)and 25-hydroxy-vitamin D(25VitD)were significantly increased(all P＜0.05);in the TNFi group only PTH and P1NP levels were significantly decreased(both P＜0.05),while β-CTX,N-MID and 25VitD levels showed no significant differences(all P＞0.05).Conclusion Long-term use of TNFi in patients with AS and OP can effectively reduce disease activity,improve physical function,decrease the level of inflammatory factors,alleviate abnormal bone metabolism,and increase bone mineral density;and the combined use of TNFi and bisphosphonates has better efficacy.

Objective:To investigate the effect of 3D-printed customized flanged cup in hip revision with severe acetabular bone defects.Methods:Since February 2017, 10 cases of 3D-printed customized flanged cups were used in hip revision with severe acetabular bone defects, including 2 cases of Paprosky type IIIA and 8 cases of Paprosky type IIIB. There were 5 males and 5 females, mean age 73.6±8.1 yrs (range, 62-87 yrs), 5 left and 5 right cases. The preoperative thin-layer CT scan was preformed to reverse reconstruct digital pelvis. Five cases of one-piece flanged cups and 5 cases of decomposed flanged cups, including 3 cases of composite one-piece cups were designed by computer. The surgery was performed strictly according to the plan.Postoperative follow-up was performed to evaluate the Harris score. Operation time,intraoperative bleeding and other complications such as vascular and nerve injury, postoperative infection, and dislocation were counted. Pelvic X-ray was used to assess the height and horizontal position of the center of rotation and the stability of the prosthesis.Results:The surgical procedure was successful, with an average operative time of 147.9±48.3 min (range, 96-212 min) and an average intraoperative bleeding of 730.4±262.6 ml (range, 500-1 300 ml). The mean time of final follow-up was 40.8±18.7 months (range, 16-70 months) after surgery. At the last follow-up, the average Harris score was 83.80±6.73, with 4 cases excellent, 5 cases good, and 1 case fair. The excellent and good rate was 90%. The last Harris score was significantly higher than that before operation 28.60±8.40 ( t=16.84, P<0.001). The height of affected hip joint rotation center decreased from 46.24±7.74 mm before operation to 15.54±2.54 mm after operation with significant difference ( t=14.61, P<0.001). It was slightly higher than the opposite side (13.81±1.48 mm), which had no significant difference ( t=1.83, P=0.100). The horizontal distance of affected hip joint rotation center increased from 33.79±5.27 mm before operation to 40.53±4.50 mm after operation with significant difference ( t=3.62, P=0.006). It had no significant difference ( t=1.28, P=0.232) compared with the opposite side (38.54±3.46 mm). All incisions were healed in one stage without infection, vascular or nerve injury. During the following-up, all prostheses were in satisfied position without loosening, dislocation or screw breaks. Conclusion:Digitally assisted 3D-printed flanged cups can be used in hip revision with severe acetabular bone defect. It can not only improve hip joint function, but also restore the acetabular rotation center and the prosthesis stability, which can achieve good early and mid-term effect.

The tumor microenvironment of pancreatic carcinoma is a dense matrix environment with excessive fibrosis containing pancreatic stellate cells, cancer-associated fibroblasts, immune cells, and extracellular matrix, which not only creates an environment to promote the growth and invasion of tumors, but also makes them resistant to chemotherapy and other antitumor drugs. Intensive fibrosis reaction in the matrix and changes of tumors in immune environment are considered the main reasons for treatment failure in pancreatic cancer management. This article reviews the recent research advances in the tumor microenvironment of pancreatic cancer, summarizes its composition and pathogenesis and the targeted therapies for matrix and immune cells, and analyzes the importance of tumor microenvironment in the development and progression of pancreatic cancer and its impact on targeted therapy.

To investigate the therapeutic effects of oral osthole on streptozotocin (STZ)-induced type 1 diabetes mellitus(T1DM) mice and explore its internal mechanism. METHODS: The diabetes model induced by STZ was established. Mice were randomly divided into control group, STZ model group, STZ+osthole group (20 mg/kg). Body weight, blood glucose, urine protein, blood urea nitrogen and creatinine were observed to detect renal function. The degree of renal tissue damage was detected by H&E staining and PAS staining, and the degree of renal fibrosis was detected by Sirius Red staining. CD68 and F4/80 immunofluorescence staining was used to observe the infiltration of macrophages in kidney tissue. The mRNA expressive levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) in renal tissue were detected by RT-qPCR. The protein expressive levels of phospho-NF-κB p65, NF-κB p65, IκBα, phospho-IκBα, phospho-p38 and p38 were detected by Western blot in renal tissue. RESULTS: Compared with the STZ model group, the levels of urinary protein, blood urea nitrogen, creatinine were significantly decreased after osthole treatment (P<0.05 or P<0.01). The renal structure disorder, mesangial matrix area, collagen fiber accumulation, and macrophage infiltration were significantly improved (P<0.05 or P<0.01). The expression of mRNA of pro-inflammatory cytokines TNF-α and IL-6 were significantly decreased (P<0.05 or P<0.001). The expression of phospho-NF-κB p65, phospho-IκBα and phospho-p38 were significantly down-regulated (P<0.05 or P<0.01), while the protein expression level of NF-κB p65, IκBα was up-regulated (P<0.05). CONCLUSION: Osthole has a protective effect on kidney injury caused by diabetes and inhibits NF-κB and p38/MAPK signaling pathway.

Objective:To discuss the relationship between heart rate (HR) and heart rate differences (HRD) at different time points in head-up tilt test (HUTT) and the occurrence of postural tachycardia syndrome(POTS) in children and adolescents.Methods:A total of 217 children and adolescents diagnosed as POTS, who complained of syncope or presyncope, were chosen as POTS group (aged 6 to 16 years). During the same period, 73 healthy children and adolescents with matching gender and age were selected as control group.All subjects underwent HUTT from October 2000 to November 2019.Get HR (HR0, HR5, HR10) in baseline, HUTT 5 min and 10 min, HRD (HRD5, HRD10) of HR in HUTT 5 min and 10 min minus HR in baseline.Results:(1) HR5, HR10, HRD5, and HRD10 were higher in the POTS group than those in the control group( P<0.05). (2) Univariate Logistic regression: There was a correlation between HR5, HR10, HRD5, HRD10 and the risk of POTS( P<0.01). (3) Multivariable Logistic regression: For each additional unit of HRD5 and HRD10, the risk of POTS increased by 27% ( OR=1.27, 95% CI1.16 to 1.36) and 28% ( OR=1.28, 95% CI1.20 to 1.38). Conclusion:HR and HRD are related with the occurrence of POTS in children and adolescents, but HR and HRD at different time points of HUTT play the little role on the effect size of the occurrence of POTS.

ObjectiveTo investigate the features of liver injury and related influencing factors in adolescents and adults with infectious mononucleosis (IM). MethodsA retrospective analysis was performed to investigate the features of liver injury in adolescents and adults with IM who were hospitalized in Peking University First Hospital from January 2005 to December 31 2018, and the patients were divided into subgroups based on age, Epstein-Barr virus (EBV) DNA level, and presence or absence of jaundice or infection with cytomegalovirus or hepatitis E virus (HEV). The t-test was used for comparison of continuous data meeting analytical conditions between two groups, and the Mann-Whitney U test was used for comparison of continuous data which did not meet analytical conditions between two groups; the chi-square test was used for comparison of categorical data between two groups, and the Fisher’s exact test was used for comparison of categorical data which did not meet the analytical conditions of the chi-square test. A logistic regression analysis was used for multivariate analysis. ResultsA total of 274 patients were enrolled, with 154 male patients (56.2%) and 120 female patients (43.8%), and the mean age of onset was 22.3±67 years. The incidence rate of liver injury ［defined as alanine aminotransferase (ALT) ＞50 U/L and/or aspartate aminotransferase (AST)＞40 U/L］ was 97.4% (267/274), and that of jaundice was 27.6% (74/268). The patients, aged ≥20 years, tended to have a higher level of gamma-glutamyl transpeptidase (GGT) (Z=2.070, P=0.038). Serum EBV DNA was measured for 167 patients, among whom 90 had positive results and 77 had negative results. The positive serum EBV DNA group had significantly higher levels of GGT (Z=3.005, P=0.003) and lactate dehydrogenase (Z=2.162, P=0.031) than the negative serum EBV DNA group. The patients with cytomegalovirus infection tended to have a higher level of alkaline phosphatase (Z=2.351, P=0.019), and the patients with HEV infection presented with a higher level of GGT (Z=1.988, P=0.047). AST (odds ratio ［OR］=1.006, 95% confidence interval ［CI］: 1.002-1.010, P=0.005) and ALP (OR=1.012, 95%CI: 1.005-1.020, P=0.001) were independent risk factors for jaundice. ConclusionThere is a high incidence rate of liver injury in adolescents and adults with IM, and the patients with an older age or positive serum EBV DNA tend to have more severe liver injury.

Objective:To analyze the effectiveness of antiviral therapy on adolescents and adults with infectious mononucleosis (IM).Methods:The clinical data of patients aged≥16 years old with IM who were hospitalized in Peking University First Hospital from January 1, 2005 to December 31, 2018 were analyzed retrospectively, and the patients were divided into antiviral treatment group and non-antiviral treatment group. The duration of hospitalization day, fever duration, ratio of lymphocytes and duration for normalization of Epstein-Barr virus (EBV) markers were compared between the two groups through single factor and propensity score matching analysis. Statistical analysis was conducted by independent sample t test, Mann-Whitney U test, chi-square test or Fisher exact probability method. Results:A total of 274 cases were enrolled and 176 cases (64.23%) were divided into antiviral treatment group and 98 cases (35.77%) into non-antiviral treatment group. The proportion of male (56.25%(99/176) vs 56.12%(55/98)), age (21.0(18.0, 26.0) years old vs 21.0(18.0, 27.0) years old), the ratio of fever (98.30%(173/176) vs 93.88%(92/98)), sore throat (90.34%(159/176) vs 88.78%(87/98)), lymphocyte ratio (0.648(0.568, 0.707) vs 0.663(0.581, 0.711)), atypical lymphocyte ratio (0.150(0.100, 0.235) vs 0.135(0.060, 0.250)) and serum EBV DNA level (2.71(2.70, 3.47) lg copies/mL vs 2.70(2.70, 3.28) lg copies/mL) were comparable between two groups at admission, and the differences were all not statistically significant(all P>0.05). The durations of hospitalization and fever in antiviral treatment group were 14.0(10.0, 18.0) d and (14.91±7.24) d, respectively, which were both significantly longer than those in non-antiviral treatment group (11.0(7.0, 15.0) d and (9.95±5.67) d, respectively). The differences were both statistically significant ( Z=-3.294 and t=-5.035, respectively, both P<0.01). Twenty-six patients each in the antiviral treatment group and non-antiviral treatment group were included in the propensity score matching assessment. The fever days of the two groups were 15.0(10.0, 18.0) d and 7.5(5.0, 12.5) d, respectively, and the hospitalization days were (15.4±5.5) d and (12.0±5.7) d, respectively. The differences were both statistically significant ( Z=-3.781 and t=-2.187, respectively, both P<0.05). However, there were no significant differences in the time required for the ratio of lymphocytes returning to normal, the time required for the ratio of atypical lymphocytes decreasing to <0.100, and the time required for serum EBV DNA becoming negative(all P>0.05). Conclusion:The antiviral treatment could not improve the prognosis of adolescent and adult IM patients.