Regulatory effect of physiological tensile stress on differentiation of ATDC5 chondrocytes through Nell-1/Ihh signaling pathway / 吉林大学学报(医学版)

Objective:To discuss the regulatory effect of physiological tensile stress on the differentiation of chondrocytes,and to clarify the associated signaling pathway mechanism.Methods:The ATDC5 chondrocytes were cultured in vitro and subjected to physiological tensile stress by four-point bending cell mechanical loading device.Initially,the cells were divided into control group and tensile stress group(2 000 μstrain/2 h group),and further divided into different stress magnitudes(1 000,2 000,and 3 000 μstrain)for 2 h,and 2 000 μstrain for different duration time(1,2,and 4 h)groups;the cells without tensile stress were used as control group.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of type Ⅱ collagen(Col-Ⅱ),type Ⅹ collagen(Col-Ⅹ),aggregated proteoglycom(Aggrecan),sex-determining region Y-box protein 9(SOX9),vascular endothelial growth factor(VEGF),proliferating cell nuclear antigen(PCNA),Nel-like molecule tyep 1(Nell-1),Runt-related transcription factor 2(Runx2),Indian hedgehog(Ihh),patched homolog 1(Ptch-1),GLI family zinc finger protein 1(Gli-1),and hedgehog interacting protein 1(Hhip-1)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of Nell-1,Runx2,and Ihh proteins in the cells in various groups.The ATDC5 cells were divided into control group,cyclopamine group,tensile stress group,and cyclopamine + tensile stress group.RT-qPCR method was used to detect the expression levels of Nell-1,Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of Nell-1 and Ihh proteins in the cells in various groups.Results:Compared with control group,the expression levels of Col-Ⅱ,Col-Ⅹ,Aggrecan,SOX9,VEGF,and PCNA mRNA in the cells in 2 000 μstrain/2 h group were significantly increased(P<0.01);after treated with 2 000 μstrain tensile stress for different duration time(1,2,and 4 h)or different tensile stresses(1 000,2 000,and 3 000 μstrain)for 2 h,compared with control group,the expression levels of Runx2 mRNA in the cells in other groups were increased with the prolongation of time or the increasing of tensile stress(P<0.01),and the expression levels of Nell-1,Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA were gradually increased(P<0.01),the expression levels reached the peaking at 2 000 μstrain/2 h,and then decreased but remained significantly higher than that in control group(P<0.01).The Western blotting results showed that the expression levels of Nell-1,Runx2,and Ihh proteins in the cells were consistent with the change trend of mRNA expression levels.After pre-treated with cyclopamine,compared with control group,the expression levels of Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA in the cells in cyclopamine group were significantly decreased(P<0.01),and the expression levels of Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA in the cells in tensile stress and cyclopamine+tensile stress groups were significantly increased(P<0.01);compared with cyclopamine group,the expression levels of Nell-1,Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA in the cells in cyclopamine+tensile stress group were significantly increased(P<0.01);compared with tensile stress group,the expression levels of Ihh,Ptch-1,Gli-1,and Hhip-1 mRNA in the cells in cyclopamine + tensile stress group were significantly decreased(P<0.01).Compared with control group,the expression level of Ihh protein in the cells in cyclopamine group was significantly decreased(P<0.01),but there was no significant difference in expression level of Nell-1 protein in the cells between control group and cyclopamine group(P>0.05),while the expression levels of Nell-1 and Ihh proteins in the cells in tensile stress group and cyclopamine + tensile stress group were significantly increased(P<0.01);compared with cyclopamine group,the expression levels of Nell-1 and Ihh proteins in the cells in tensile stress group and cyclopamine + tensile stress group were significantly increased(P<0.01);compared with tensile stress group,in the expression levels of Nell-1 and Ihh proteins in the cells in cyclopamine + tensile stress group had no significant differences(P>0.05).Conclusion:After stimulated with physiological tensile stress,Nell-1 can activate the Ihh signaling pathway upstream,and regulate the differentiation of the ATDC5 chondrocytes.

Effects of Xianglian Huazhuo Decoction on Wnt1, GSK-3β and β-catenin in chronic atrophic gastritis rats / 国际中医中药杂志

Objective:To investigate the mechanism of Xianglian Huazhuo Decoction in treating chronic atrophic gastritis (CAG) rats by detecting the protein and gene expressions of Wnt1, GSK-3β, β-catenin in Wnt signaling pathway.Methods:Totally 60 Wistar rats were randomly divided into blank group and model group. The CAG model was established by MNNG free drink combined with sodium salicylate gavage combined with abnormal hunger and satiety. After model establishment, the rats were randomly divided into model group, Xianglian Huazhuo Group and folic acid group. Folic acid group received the folic acid water solution 1.3 mg/kg for gavage; the Xianglian Huazhuo Group received Xianglian Huazhuo Decoction 15.5 g/kg for gavage; the blank group and the model group received the same volume of purified water for gavage. After 8 weeks of intervention, the general state and the atrophic degree of gastric mucosa were observed. The expressions of Wnt1, GSK-3β and β-catenin were detected by immunohistochemical method, and the mRNA expression of β-catenin expression was detected by real-time quantitative polymerase chain reaction PCR.Results:The general condition of rats in Xianglian Huazhuo group was improved, the atrophy of gastric mucosa was improved, and the effect was better than that in folic acid group. Compared with the model group, the positive area ratio of Wnt1 decreased in Xianglian Huazhuo group ( P<0.05), and the positive area ratio of β-catenin decreased in folic acid group and Xianglian Huazhuo group ( P<0.05 or P<0.01), the positive area ratio of GSK-3β increased ( P<0.05), and the β-catenin mRNA level of Xianglian Huazhuo group decreased ( P<0.05). Conciusion 　Xianglian Huazhuo Decoction may increase GSK-3β by downregulating abnormal expression of Wnt1, promoting β-catenin to degrade normally, thereby blocking the abnormal activation of the Wnt signaling pathway, delaying or reversing CAG carcinogenesis to a certain extent, and exerting therapeutic effects.

Study on the mechanism of Tianshui Dichang Decoction in the treatment of ulcerative colitis based on network pharmacology and experimental verification / 国际中医中药杂志

Objective:To explore the potential mechanism of Tianshui Dichang Decoction in the treatment of ulcerative colitis through network pharmacology and experimental verification.Methods:The active components and targets of Tianshui Dichang Decoction were screened by TCMSP. The related targets of ulcerative colitis were screened by OMIM, GeneCard and TTD databases, and the effective component targets of Tianshui Dichang Decoction were intersected with the potential targets of ulcerative colitis. The PPI network was constructed by STRING database to screen the core targets, and the "Chinese materia medica-disease-active components-target" network was constructed by Cytoscape 3.8.0 software. GO function and KEGG pathway enrichment analysis were carried out using Metascape database. 48 mice were divided into control group, model group, mesalazine group (0.3 g/kg) and Tianshui Dichang Decoction low-, medium-, and high-dosage groups (7.5,15 and 30 g/kg) according to random number table method, with 8 mice in each group. Except the control group, the ulcerative colitis model was established in other groups. After 7 days of intervention with corresponding drugs, the disease activity index (DAI) was scored, the pathological changes of colon were observed by HE staining, and the expressions of IL-6, STAT3mRNA and protein in colon tissue were detected by PCR and Western blot methods.Results:Totally 127 active components in Tianshui Dichang Decoction and 560 targets of ulcerative colitis were obtained. 89 intersecting targets of Tianshui Dichang Decoction and ulcerative colitis were obtained, and the core targets included IL6, TNF, IL1B, AKT1, TP53, VEGFA, JUN, PTGS2, CXCL8, CCL2, STAT3, MMP9 and so on. Oxidative stress response, lipopolysaccharide metabolism, bacterial response, signal transduction and other biological processes were mainly involved, mainly through the cancer pathway, IL17, TNF, MAPK and other signal pathways to play a role in the treatment of ulcerative colitis. The results of experimental verification showed that the DAI score, the expressions of IL-6 and STAT3 protein in colon tissue of Tianshui Dichang Decoction medium- and high-dosage groups.decreased ( P<0.05). The levels of IL-6 and STAT3 mRNA in colon tissue decreased in the Tianshui Dichang Decoction low-, medium- and high-dosage groups.groups ( P<0.05). Conclusion:Tianshui Dichang Decoction has a certain therapeutic effect on UC through component-multitarget-signal pathway, and its mechanism is related to regulating IL-6/STAT3 signal pathway and inhibiting intestinal mucosal inflammation.

Causes and intervention measures of hypoglycemia in elderly patients after gastrointestinal polypectomy / 中华老年医学杂志

Objective:To analyze the causes of hypoglycemia in elderly patients after resection of gastrointestinal polyps.Methods:We retrospectively analyzed the data of 120 elderly patients who underwent gastrointestinal polypectomy in our hospital from November 2019 to November 2020, and analyzed the reasons for postoperative hypoglycemia.Results:(1)There were significant differences between the hypoglycemic group and the non-hypoglycemic group in age, diabetes, preoperative fluid replacement, preoperative fasting time, preoperative waiting time, preoperative emotional instability, and preoperative night sleep time(all P<0.05). (2)Logistic regression analysis showed that postoperative hypoglycemia was associated with age >70 years( OR=4.266, 95% CI: 1.120-16.162); Diabetes mellitus( OR=4.094, 95% CI: 1.311-12.773); No fluid was taken before surgery( OR=10.923, 95% CI=2.882-41.386); Preoperative fasting time ≥10 h( OR=9.042, 95% CI: 2.571-31.645); Preoperative wait time ≥12 h( OR=7.035, 95% CI: 1.855-26.651); Preoperative emotional instability( OR=4.25, 95% CI: 1.263-14.370); The preoperative night sleep time <6 h( OR=5.952, 95% CI: 1.760-19.992)was closely related(all P<0.05). Conclusions:There are many reasons for the occurrence of hypoglycemia in elderly patients after gastrointestinal polyp removal.Therefore, evaluation work should be done and preventive measures should be taken to reduce the occurrence of hypoglycemia and ensure the effectiveness and safety of treatment for patients.

Research advances in the v-raf murine sarcoma viral oncogene homolog gene mutation in ameloblastoma / 中华口腔医学杂志

V-raf murine sarcoma viral oncogene homolog B1 (BRAF) is a pro-oncogene, which is one member of the RAF family. Mutated BRAF is found in approximately 8% of human tumors. BRAF gene mutations lead to continuous activation of the mitogen-activatd protein kinase (MAPK) pathway, which resulting in abnormal cell proliferation and tumorigenesis. In recent years, recurrent MAPK signaling mutations were identified in ameloblastoma, among which BRAF-V600E is the most prominent type. This provides new strategies for the targeted treatment of ameloblastoma. This paper reviewed the latest advances in BRAF gene mutation associated with ameloblastoma and its potential clinical significance.