AIM:To evaluate the efficacy of inflix-imab in the maintenance of Perianal fstulizing Crohn's disease.METHODS:The clinical data of 24 patients with perianal fistula Crohn's disease(PFCD)treated with infliximab(IFX)in the Depart-ment of Anorectal Surgery of our hospital from No-vember 2020 to October 2023 were retrospectively collected.The clinical efficacy and safety were eval-uated by observing the clinical characteristics for 54 weeks.The fistula remission rate,clinical remis-sion rate,and endoscopic remission rate of pa-tients were calculated.The changes of laboratory indexes before and after treatment were recorded.Logistic regression was used to analyze the related factors of fistula remission.All adverse reactions oc-curred during IFX treatment were recorded.RE-SULTS:After 54 weeks of IFX treatment,the fistula remission rate,clinical remission rate,and endo-scopic remission rate were 37.5％,45.83％,and 33.33％,respectively.Fistula response at 14 weeks of treatment(OR=19.419,95％CI:1.267-297.559,P=0.033)was predictive factors for fistula remission at 54 weeks of treatment.The inflammatory index-es and nutritional indexes were significantly im-proved compared with those before treatment(P＜0.01).The scores of PDAI,CDAI and SES-CD were significantly different from those before treatment(P＜0.01).Five patients(20.83％)had adverse reac-tions,and the symptoms disappeared or improved after symptomatic treatment,and no patient had serious adverse reactions.CONCLUSION:IFX can ef-fectively promote the closure of PFCD fistula,im-prove the chronic inflammatory reaction of intesti-nal mucosa,alleviate clinical symptoms,and im-prove the quality of life of patients.IFX is effective and safe for PFCD maintenance treatment.

AIM:To evaluate the efficacy of inflix-imab in the maintenance of Perianal fstulizing Crohn's disease.METHODS:The clinical data of 24 patients with perianal fistula Crohn's disease(PFCD)treated with infliximab(IFX)in the Depart-ment of Anorectal Surgery of our hospital from No-vember 2020 to October 2023 were retrospectively collected.The clinical efficacy and safety were eval-uated by observing the clinical characteristics for 54 weeks.The fistula remission rate,clinical remis-sion rate,and endoscopic remission rate of pa-tients were calculated.The changes of laboratory indexes before and after treatment were recorded.Logistic regression was used to analyze the related factors of fistula remission.All adverse reactions oc-curred during IFX treatment were recorded.RE-SULTS:After 54 weeks of IFX treatment,the fistula remission rate,clinical remission rate,and endo-scopic remission rate were 37.5％,45.83％,and 33.33％,respectively.Fistula response at 14 weeks of treatment(OR=19.419,95％CI:1.267-297.559,P=0.033)was predictive factors for fistula remission at 54 weeks of treatment.The inflammatory index-es and nutritional indexes were significantly im-proved compared with those before treatment(P＜0.01).The scores of PDAI,CDAI and SES-CD were significantly different from those before treatment(P＜0.01).Five patients(20.83％)had adverse reac-tions,and the symptoms disappeared or improved after symptomatic treatment,and no patient had serious adverse reactions.CONCLUSION:IFX can ef-fectively promote the closure of PFCD fistula,im-prove the chronic inflammatory reaction of intesti-nal mucosa,alleviate clinical symptoms,and im-prove the quality of life of patients.IFX is effective and safe for PFCD maintenance treatment.

Drugs have both therapeutic and toxic side effects. How to quickly determine the toxicity of the test substance is very important for drug development. In vitro cytotoxicity testing compensates for the shortcomings of using animal models for toxicity evaluation. Its role in toxicity evaluation is increasingly important. The development of computer technology and in-depth research in proteomics, genomics, and metabolomics provide a method for in vitro cytotoxicity evaluation towards a faster and more accurate direction. This paper reviews the commonly used cells, evaluation indicators, and detection techniques for in vitro cytotoxicity evaluation, in order to provide some reference for related research.

AIM: To investigate the effect of Qiteng Xiaozhuo granule on inflammation indexes in rats with chronic glomerulonephritis (CGN) through miR-339-5p. METHODS: CGN model was established by adriamycin (ADR) injection into tail vein of rats. Qiteng Xiaozhu granules (21.6, 10.8, 5.4 g/kg) of different doses were intragastric-fed for 30 days. Pathological changes in kidney tissues of CGN rats were observed by HE staining and electron microscopy. The contents of IL-1β, IL-6, IL-10 and TNF-α in serum and kidney tissue of CGN rats were detected by qRT-PCR, ELISA and Western blot. Relative expressions of miR-339-5p, Syk and p-Syk proteins were detected by Western blot, the relative expressions of miR-339-5p mRNA and Syk mRNA were detected by qRT-PCR. RESULTS: There were thickening of basement membrane and glomerular atrophy in the model group. Compared with the normal group, the expression of miR-339-5p and IL-10 in the model group was significantly down-regulated, while the expression of IL-1β, IL-6, TNF-α and Syk was significantly up-regulated. Qiteng Xiaozhuo granule group could significantly reduce the protein and mRNA expression levels of IL-1β, IL-6, TNF-α, Syk in kidney tissue.CONCLUSION: Qiteng Xiaozhuo granule may down-regulate the expression of inflammatory factors through miR-339-5p in the treatment of inflammatory symptoms in CGN rats.

It has been revealed that hypoxia is dynamic in hypertrophic scars; therefore, we considered that it may have different effects on hypoxia-inducible factor-1α (HIF-1α) and p53 expression. Herein, we aimed to confirm the presence of a teeterboard-like conversion between HIF-1α and p53, which is correlated with scar formation and regression. Thus, we obtained samples of normal skin and hypertrophic scars to identify the differences in HIF-1α and autophagy using immunohistochemistry and transmission electron microscopy. In addition, we used moderate hypoxia in vitro to simulate the proliferative scar, and silenced HIF-1α or p53 gene expression or triggered overexpression to investigate the changes of HIF-1α and p53 expression, autophagy, apoptosis, and cell proliferation under this condition. HIF-1α, p53, and autophagy-related proteins were assayed using western blotting and immunofluorescence, whereas apoptosis was detected using flow cytometry analysis, and cell proliferation was detected using cell counting kit-8 (CCK-8) and 5-bromo-2'-deoxyuridine (BrdU) staining. Furthermore, immunoprecipitation was performed to verify the binding of HIF-1α and p53 to transcription cofactor p300. Our results demonstrated that, in scar tissue, HIF-1α expression increased in parallel with autophagosome formation. Under hypoxia, HIF-1α expression and autophagy were upregulated, whereas p53 expression and apoptosis were downregulated in vitro. HIF-1α knockdown downregulated autophagy, proliferation, and p300-bound HIF-1α, and upregulated p53 expression, apoptosis, and p300-bound p53. Meanwhile, p53 knockdown induced the opposite effects and enhanced HIF-1α, whereas p53 overexpression resulted in the same effects and reduced HIF-1α. Our results suggest a teeterboard-like conversion between HIF-1α and p53, which is linked with scar hyperplasia and regression.
Apoptosis ; Autophagy ; Cell Hypoxia ; Fibroblasts/metabolism* ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Tumor Suppressor Protein p53/metabolism*

OBJECTIVE: To investigate the effect of miR-324-5p on the proliferation of rat glomerular mesangial (HBZY-1) cells and the role of Syk/Ras/c-fos signaling pathway in mediating this effect. METHODS: HBZY-1 cells cultured in vitro were transiently transfected with miR-324-5p mimics or miR-324-5p-mimics-NC followed by treatment with lipopolysaccharide (LPS). MTT assay was used to detect the proliferation activity of HBZY-1 cells, and RT-qPCR was used to detect the expressions of miR-324-5p and the mRNA expressions of Syk, Ras, MEK1/2, ERK1/2 and c-fos mRNA. The protein expressions of p-Syk, Ras, p-MEK1/2, p-ERK1/2 and c-Fos were detected by Western blotting and immunofluorescence assay. RESULTS: MTT assay showed that exposure to LPS significantly enhanced the proliferative activity of HBZY-1 cells. Compared with the cells treated with LPS and LPS + mimics NC, the cells transfected with miR-324-5p mimics prior to LPS exposure exhibited significantly lowered proliferative activity. Transfection with miR-324-5p mimics significantly lowered the mRNA expressions of Syk, Ras, MEK1/2, ERK1/2 and c-fos and the protein expressions of p-Syk, Ras, MEK1/2, ERK1/2 and c-Fos ( CONCLUSIONS miR-324-5p can inhibit the proliferation of rat chronic glomerulonephritis cells induced by LPS by inhibiting Syk/Ras/c-fos signaling pathway and may potentially serve as a diagnostic indicator and a therapeutic target for chronic glomerulonephritis.
Animals ; Cell Proliferation ; Lipopolysaccharides ; Mesangial Cells ; MicroRNAs/genetics* ; Proto-Oncogene Proteins c-fos ; Rats ; Receptor Protein-Tyrosine Kinases ; Signal Transduction ; ras Proteins

Objective To detect the serum level of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF),a significant metabolite offish oil,in subjects with normal glucose tolerance (NGT) in local communities,and to investigate the association of CMPF with fatty acid metabolism.Methods A total of 272 NGT participants from screening for diabetes in Shanghai in 2013 were enrolled.Anthropometric measurements,biochemical evaluation,and questionnaire interview were performed for all the participants.The participants were divided into normal weight group [body mass index (BMI) ≤23.9 kg/m2,n =143] and overweight/obesity group (BMI ≥ 24 kg/m2,n =129).The serum CMPF concentrations were determined using an enzyme-linked immunosorbent assay.Results Serum CMPF level in overweight/obesity group was lower than that in normal weight group [96.50 (46.11,169.56) μmol/L vs 153.20 (83.16,282.97) μmol/L,P＜0.05].The serum CMPF level was negatively correlated with BMI (r =-0.256,P＜0.01),triglycerides (r =-0.175,P =0.004),and free fatty acid (r =-0.126,P =0.041) according to bivariate correlation analyses.A multivariate stepwise linear regression analysis showed that the serum CMPF level was independently associated with BMI,triglycerides,free fatty acid,and HbA1C.A logistic regression analysis showed that the CMPF was a protective factor against obesity (OR =0.324,95％ CI 0.158,0.664).Conclusion Serum CMPF level is reduced in overweight/obese subjects.CMPF is beneficial to lipid metabolism.

The method of " quantitative analysis of multi-components by single marker(QAMS)" combined with fingerprint often used in the quality control of traditional Chinese medicine (TCM) and its preparation.This study reviewed and analyzed the data.It showed that " QAMS" can be used in the determination of multiple indicators in some traditional Chinese medicine and Chinese herbal compound preparations.This is a supplementary method for the determination of content, combined with the fingerprint to mutual benefit, quality evaluation of TCM is more comprehensive and scientific.

AIM To observe the effects of Wuwei Wentong Chubi Capsules (Cinnamomi Ramulus,Poria,Epimedii Folium,etc.) on autophagy proteins of Beclin-1 and LC3-Ⅱ in adjuvant-induced arthritis rats and to explore the possible mechanism of action.METHODS Sixty SD rats were randomized into six groups:normal group,model group,Wuwei Wentong Chubi Capsules (0.8,1.6,3.2 g/kg) groups and tripterygium glycosides (TPT,40 mg/kg) group.In addition to the normal group,adjuvant arthritis (AA) was induced with Freund's complete adjuvant.From the 2th day after injection of FCA,Wuwei Wentong Chubi Capsules with different doses and TPT were given by gavage once a day for 12 days.At the end of the experiment,ankle-joint samples were taken to examine the degree of AA by HE.Beclin-1 and LC3-Ⅱ mRNA were determined by real-time fluorescent quantitative PCR.Meanwhile,the protein expression levels of Beclin-1 and LC3-Ⅱ were determined by immunofluorescence histochemical staining and Western blot.RESULTS As compared with the model group,Wuwei Wentong Chubi Capsules (1.60,3.20 g/kg) not only significantly reduced histopathological injuries,but also effectively up-regulated mRNA and protein expressions of Beclin-1 and LC3-Ⅱ.CONCLUSION Wuwei Wentong Chubi Capsules has a therapeutic action on AA in rats,which might be partly associated with promoting autophagy,decreasing excessive proliferation of synovial cells,leading to the reduction of damage to articular cartilage.

AIM To observe the effects of Huangdi Anxiao Capsules (Puerariae lobatae Radix,Eriobotryae Folium,Notoginseng Radix et Rhizoma,etc.) on blood glucose and insulin resistance in GK rats with type 2 diabetes mellitus (T2DM) and its possible mechanism of action.METHODS GK rats with T2DM included in the experiment were randomly divided into model group,rosiglitazone (1.44 mg/kg) group,Shenqi Jiangtang Granules (1.08 g/kg) group,Huangdi Anxiao Capsules high,medium and low (12,6,3 g/kg) group and normal control group of Wistar rats.After six weeks of consecutive administration,fasting blood glucose (FBG),serum insulin (FINS),fasting plasma glucose (FPG),and insulin resistance index (HOMA-IR) were measured in all groups.Serum biochemical indexes of (TG,TC,HDL-C,LDL-C),glycosylated hemoglobin (HbA1c),glucagon-like peptide (GLP-1) and insulin-like growth factor (IGF-1) were measured.The pancreatic tissue was obtained by routine paraffin embedding and HE staining to observe the pathological changes.RESULTS Compared with the normal group,FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1c of the model group were significantly higher (P ＜0.01),and GLP-1 and IGF-1 expressions were markedly decreased (P ＜ 0.01).Compared with the model group,the levels of FBG,FPG,FINS,HOMA-IR,TG,TC,HDL-C,LDL-C and HbA1 c in Huangdi Anxiao Capsules high-,medium-and low-dose group were significantly decreased (P ＜ 0.05,P ＜0.01);GLP-1 and IGF-1 expressions were markedly increased (P ＜0.05,P ＜0.01),compared with the model group,the structure changes of pancreatic tissue in Huangdi Anxiao Capsules high-,medium-and low-dose groups largely reduced.CONCLUSION Huangdi Anxiao Capsules can reduce GK rats fasting blood glucose,insulin resistance,and its mechanism may be related to promoting the emergence and proliferation of pancreatic islet cells,improving the function of islet beta cells,and increasing insulin secretion.