AIM:To investigate the changes of Wnt signaling pathway in catalpol-induced proliferation of rat bone marrow mesenchymal stem cells (BMSCs).METHODS:The BMSCs were isolated from SD rats , purified by differ-ential time adherent method and divided into control group and catalpol (1.0 mg/L) group.Flow cytometry was used to de-tect the proliferation index of BMSCs .The mRNA levels of Wnt3a, Wnt5a, Wnt11 and β-catenin was evaluated by real-time PCR.In addition, the protein expression level of β-catenin was determined by Western blotting .RESULTS:Prolife-ration index was increased from 8.90%±0.46% to 17.93%±1.68% after treatment with catalpol (P<0.01).Com-pared with control group , the mRNA expression of Wnt5a, Wnt11 andβ-catenin was all increased with catalpol treatment . No difference of Wnt3a mRNA expression between control group and catalpol group was observed .Meanwhile, the protein expression of β-catenin was increased in catalpol group compared with control group .CONCLUSION:Catalpol promotes BMSCs going into the cell cycle .Classical and non-classical Wnt signaling pathways are activated in this process .

Objective To construct microRNA (miRNA) expression vector targeting surviving,and to investigate its effect on transfected human colorectal carcinoma (HT-29) cell apoptosis and proliferation.Methods miRNA targeting survivin was synthesized and transfected HT-29 cells by lipofectin.HT-29 cells were cultured in the 6 orifices.The cultured cells were divided into control,liposome,negative control and positive control groups.Transient transfected cells were collected and the proliferation index and apoptosis rate of HT-29 cells were detected by flow cytometry.The expressions of survivin mRNA and protein were detected by RT-PCR and Western blot.Results The proliferation index and apoptosis rate of the positive control group were significantly higher compared with normal group,transfection group and mock-vehicle group (17.98％ ± 2.35％ vs 38.04％ ±2.11％ vs 36.73％ ±2.51％ vs 36.57％ ±3.05％; t =20.05,P＜0.01; t =18.75,P＜0.01; t=18.59,P＜0.01; 19.54％ ±1.74％ vs 3.13％ ±0.29％ vs 3.70％ ±0.44％ vs 3.61％ ± 0.50％ ; t =16.40,P ＜ 0.01 ; t =15.84,P ＜ 0.01 ; t =15.92,P ＜ 0.01).Survivin mRNA and protein expression levels were specifically suppressed in transfected HT-29 cells (t =0.68,P ＜0.01 ; t =0.58,P ＜ 0.01; t=0.61,P＜0.01;t=0.64,P＜0.01; t=0.62,P＜0.01;t=0.67,P＜0.01).Conclusion Survivin targeted silence can effectively decrease the expression of survivin mRNA and protein,induce colorectal carcinoma HT-29 cell apoptosis and inhibit cell proliferation.

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human fibrinogen gene, in order to achieve high level secretory expression in extracellular.Methods Expression plasmid,pGAPZαA-FGB-FGG-FGA-AOX1,was constructed by inserting the synthesized sequence encoding human fibrinogen(FGA, FGB,FGG) and then introduced into Pichia pastoris SMD1168H by electroporation.Transformants were availably screened by Zeocin resistance,the expression of recombinant protein was identified by SDS-PAGE and Western blot analysis, the protein yield was tested by ELISA assay.After ultrafiltration and purification, the biological activity of protein was detected.Results The crude yield of human fibrinogen in Pichia pastoris supernatant reached 15 mg/L in flask and the biological aggregation activity was determined.Conclusion The human fibrinogen gene was obtained and successfully expressed in Pichia pastoris and the active products were secreted into the medium.

AIM: To illuminate the effect of SWI/SNF-related, matrix-associated, actin-dependent regulator of chromatin, subfamily b, member 1 (SMARCB1) in early diagnosis and prognosis of hepatocellular carcinoma (HCC) by determining the clinical expression of SMARCB1 in HCC tissue and benign liver tissue.METHODS: The specific target gene SMARCB1 was selected from these genes by using The Cancer Genome Atlas (TCGA).SMARCB1 expression in HCC tissue and benign liver tissue was measured by immunohistochemistry.Further statistical analysis of TCGA was performed to illuminate the role of SMARCB1 on HCC occurrence and progression.RESULTS: Compared with the benign liver tissue, immunohistochemical staining showed that SMARCB1 expression was significantly up-regulated in the HCC tissue (P<0.01).In addition, SMARCB1 expression was significantly associated with advanced tumor stage (P<0.05).The relation between SMARCB1 expression at mRNA level and clinical prognosis was analyzed.The results indicated that high SMARCB1 expression was an independent prognostic factor for HCC (P<0.05).CONCLUSION: SMARCB1 may play a part as a carcinogenic gene in tumorigenesis.We can distinguish primary HCC samples from non-malignant samples according to its different clinical expression.High SMARCB1 expression probably predicts poor outcome in HCC patients.

Objective To study the intervention effect of Jianpi Zhidong Decoction on the expressions of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus of tic disorder mice.Methods According to the random number table method,60 male ICR mice were divided into the blank control group(n=15)and modeling group(n=45).The mice in the modeling group were intraperitoneally injected with iminodipropanitrile(350 mg/kg,once a day for 7 days)to establish a tic disorder model,and the mice in the blank control group were intraperitoneally injected with equal volume of normal saline.The stereotyped behavior score was calculated to evaluate the success of modeling.According to the random number table method,the mice in the modeling group were divided into the model group,tiapride group(29.5 mg/kg)and Jianpi Zhidong Decoction group(17.8 g/kg),with 15 mice in each group.The drug was administered by gavage for 4 weeks.After administration,stereotyped behavior score was calculated.The positive protein expression,protein expression,and mRNA expression of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus and hippocampus were determined by immunohistochemistry,Western blotting and real-time PCR.Results Compared with the blank control group,the stereotyped behavior score of mice in the model group was increased(P＜0.05).Compared with the model group,the stereotyped behavior scores of mice in the tiapride group and the Jianpi Zhidong Decoction group were decreased(P＜0.05).Compared with the blank control group,the average optical densities of Cx43 in the cortex,corpus striatum,hypothalamus,and hippocampus of mice in model group were decreased(P＜0.05).Compared with the model group,the average optical densities of Cx43 in the cortex and hippocampus in the tiapride group were increased,and the average optical densities of Cx43 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the tiapride group,the average optical density of Cx43 in the cortex in the Jianpi Zhidong Decoction group was increased(P＜0.05).Compared with the blank control group,the average optical densities of Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus of mice in model group were decreased(P＜0.05).Compared with the model group,the average optical densities of Cx30 in the corpus striatum and hypothalamus in the tiapride group were increased,and the average optical densities of Cx30 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the tiapride group,the average optical density of Cx30 in the hippocampus in the Jianpi Zhidong Decoction group was increased(P＜0.05).Compared with the blank control group,the protein expressions of Cx43 in the cortex,corpus striatum,and hippocampus of mice in the model group were decreased(P＜0.05).Compared with the model group,the protein expression of Cx43 in the cortex in the tiapride group was increased,and the protein expressions of Cx43 in the cortex,corpus striatum,and hippocampus in the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the protein expressions of Cx30 in the cortex and hippocampus of mice in the model group were decreased(P＜0.05).Compared with the model group,the protein expressions of Cx30 in the cortex and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the mRNA expressions of Cx43 in the cortex,corpus striatum and hippocampus in the model group were decreased(P＜0.05).Compared with the model group,the mRNA expressions of Cx43 in the cortex,corpus striatum,and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Compared with the blank control group,the mRNA expressions of Cx30 in the cortex and hippocampus in the model group were decreased(P＜0.01).Compared with the model group,the mRNA expressions of Cx30 in the cortex and hippocampus in the tiapride group and the Jianpi Zhidong Decoction group were increased(P＜0.05).Conclusion The decreased expression of Cx43 and Cx30 in the cortex,corpus striatum,hypothalamus,and hippocampus may be involved in the occurrence of stereotyped behavior in tic disorder mice.The intervention mechanism of Jianpi Zhidong Decoction may be related to the increased expression of Cx43 and Cx30 in various brain regions.

Objective:To evaluate the effect of applying high-fidelity simulation model in the teaching of lower urinary tract injury.Methods:A total of 50 junior medical students majoring in clinical medicine were enrolled. They were randomly assigned to the control group or the experimental group. The students in the control group were taught using the traditional method involving PPT presentation and videos on lower urinary tract anatomy, surgical basis of lower urinary tract injury, and operative methods for lower urinary tract injury. The students in the experimental group were taught using the simulation model for observation and related operation. After four class hours (40 minutes/class hour) of teaching, the students in the two groups were subjected to theoretical assessment and questionnaire survey. The differences between the two groups were compared and analyzed using the t and χ 2 tests in SPSS 26.0. Results:The total scores of theoretical assessment were (36.80±4.00) and (33.12±3.62) in the experimental group and the control group, respectively, and the difference was significant ( P<0.05). The results of the questionnaire showed that the self-evaluation of the experimental group was better than that of the control group in the mastery of theoretical knowledge, clinical operation, early treatment, operation process, and key steps, and the differences were statistically significant ( P<0.05). Conclusions:Compared with the traditional teaching method, the high-fidelity simulation model improves the efficiency of learning anatomy knowledge and operation process, and promotes the mastery of important and difficult knowledge in lower urinary tract injury. This method merits popularization.