1.Quantitative evaluation of the degrees of qualitative syndromes commonly encountered in patients with coronary heart disease.
Dongtao LI ; Jie LI ; Jian WANG ; Fuyu LI ; Jingxiu ZHU ; Meizeng ZHANG ; Junyan LI ; Yanlai XU ; Lingbo WEI ; Wenyan JI ; Rongqin JIANG ; Xuefa LIU
Journal of Integrative Medicine 2010;8(8):750-6
Objective: To establish a quantitative model for evaluating the degree of traditional Chinese medicine (TCM) syndromes often seen in patients with coronary heart disease (CHD). Methods: Medical literature concerning clinical investigation of TCM syndromes of CHD was collected and organized, and the "Hall for Workshop of Metasynthetic Engineering" expert symposium method was applied. First, the 100 millimeter scaling was used for combining with scoring on degree of symptoms to establish a quantitative criterion for classification of symptom degree in CHD patients, and the model was established by using comprehensive analytic hierarchy process as the mathematical tool to estimate the weight of the criterion for evaluating qualitative syndromes in various layers by specialists. Then the model was verified in clinical practice and the outcomes were compared with fuzzy evaluation from the specialists. Results: A total of 287 clinical observation forms on CHD cases were collected, and 167 forms were available after excluding any irregular forms. The results showed that basic coincidence rate between the outcomes derived from specialists and those from the model was 68.26% (114/167), and part coincidence rate was 88.62%(148/167). Conclusion: This model, with good rationality and feasibility, has a high coincidence rate with fuzzy evaluation from specialists, and can be promoted in clinical practice. It is a good quantitative model for evaluating the degree of TCM syndromes of CHD.
2.Dynamic expression of protein tyrosine phosphatase SHP2 in liver tissue of rats with carbon tetrachloride-induced liver fibrosis
Lisen HAO ; Jie SONG ; Rongpeng WU ; Penglei ZHANG ; Jingxiu JI ; Meiyu JIANG ; Yanbo MO ; Jing WANG
Chinese Journal of Hepatology 2021;29(9):844-848
Objective:To investigate the dynamic expression of protein tyrosine phosphatase SHP2 in liver tissue of rats with carbon tetrachloride (CCl 4)-induced liver fibrosis. Methods:Rat liver fibrosis model was established by intraperitoneal injection of CCl 4. Rat liver tissue histopathological changes were detected by HE and Masson-trichrome staining. Immunohistochemical staining, Western blot and real-time fluorescent quantitative PCR were used to detect SHP2 protein and mRNA expression in rat liver tissue. One-way analysis of variance was used for the comparison of means between multiple groups, and the LSD test was used for further inter-group comparison. Results:CCl 4-induced rat liver fibrosis model was successfully constructed, and with the extension of modeling time, the degree of liver fibrosis in rats were aggravated gradually. Immunohistochemical staining results showed that SHP2 was mainly expressed in the cytoplasm of rat liver tissues. With the aggravation of liver fibrosis, the number of cells with positive expression of SHP2 was aggravated gradually ( P < 0.05). Western blot and real-time fluorescent quantitative PCR results showed that the expressions of SHP2 protein and mRNA in rat fibrotic liver tissues at different times in week 2, 4, 6, and 8 were higher in modeling than control group ( P < 0.05), and was aggravated gradually with the liver fibrosis aggravation ( P < 0.05). Conclusion:The expression of SHP2 protein and mRNA in the liver tissue of rats with CCl 4-induced liver fibrosis increased gradually with the degree of liver fibrosis, and the degree of increase was consistent with the degree of liver fibrosis.
3.Effect of adenovirus-mediated shRNA down-regulates SHP2 expression on the apoptosis of human hepatic stellate cells LX-2
Lisen HAO ; Zongyuan ZHAN ; Jie SONG ; Xiaojia MIAO ; Yu HE ; Meiyu JIANG ; Jingxiu JI ; Yanbo MO
Chinese Journal of Hepatology 2023;31(12):1313-1317
Objective:To investigate the effect of adenovirus-mediated short hairpin RNA (shRNA) downregulating SH2 domain-containing protein tyrosine phosphatase 2 (SHP2) on the apoptosis of human hepatic stellate cells LX-2 cultured in vitro.Methods:The recombinant adenovirus Ad-shRNA/SHP2 carrying shRNA targeted SHP2 and expressing green fluorescent protein (GFP), and the empty control virus Ad-GFP expressing GFP were transfected into LX-2 cells cultured in vitro. Real-time fluorescence quantitative PCR was used to detect SHP2 mRNA expression in LX-2 cells. Western blot was used to detect the protein expressions of SHP2, Bax, and Bcl-2 in LX-2 cells. TUNEL and annexin-V/propidium iodide dual-labeled flow cytometry were used to detect apoptosis in LX-2 cells. Experimental group: (1) Control group: LX-2 cells were transfected with DMEM instead of adenovirus; (2) Ad-GFP group: transfected with empty virus Ad-GFP; (3) Ad-shRNA/SHP2 group: transfected with recombinant adenovirus Ad-shRNA/SHP2. The means between multiple groups were compared using a one-way ANOVA and the LSD test was used for inter group comparisons.Results:shRNA-targeted SHP2 significantly down-regulated the expression of SHP2 protein and mRNA in LX-2 cells ( P < 0.05). The TUNEL and annexin-V/propidium iodide dual-labeled flow cytometry results showed that the apoptosis rate of LX-2 cells in the Ad-shRNA/SHP2 group (12.755%±1.606%, 19.340%±2.505%) ( P < 0.05) was significantly higher compared to the control group (3.077%±0.731%, 9.438%±0.804%) and the Ad-GFP group (3.250%±0.851%, 8.893%±1.982%), with no statistically significant difference between the control group and the Ad-GFP group ( P > 0.05). Western blot analysis of Bax and Bcl-2 protein expression in LX-2 cells of each group revealed that the Bax protein expression was significantly higher in the Ad shRNA/SHP2 group (2.493 ± 0.203) ( P < 0.05) compared to the control group and Ad-GFP group (1.989 ± 0.147, 1.999 ± 0.162), with no statistically significant difference between the control group and the Ad-GFP group ( P > 0.05), while the Bcl-2 protein was significantly decreased in the Ad-shRNA/SHP2 group (1.042±0.148) compared with the control group and the Ad-GFP group (1.707±0.146, 1.521±0.142), with no statistically significant difference between the control group and the Ad-GFP group ( P > 0.05). Conclusions:SHP2 expression down-regulation induces apoptosis of human hepatic stellate cells LX-2 in vitro by reducing Bcl-2/Bax.