1.Application value of detection on stathmin, p16 and Ki-67 in the cervical intractable cases
Hongyuan ZHOU ; Yanli ZOU ; Junju ZHUO ; Bofang PENG ; Hong LI
Cancer Research and Clinic 2016;28(7):459-463
Objective To investigate the clinical application value of stathmin, p16 and Ki-67 in the cervical intractable cases. Methods Immunohistochemical method was used to detect the expressions of stathmin, p16 and Ki-67 in surgical specimens of 288 cervical intractable cases, including 30 cases of cervical benign changes, 70 cases of cervical intraepithelial neoplasia (CIN)Ⅰ, 78 cases of CINⅡ, 85 cases of CINⅢand 25 cases of squamous cell carcinoma (SCC, as control group). The application value of stathmin, p16 and Ki-67 in the cervical cases were analyzed. Results The positive expression rates of Ki-67 of cervical benign changes and CINⅠwere 20.0 % (6/30) and 54.3 % (38/70) (χ2 = 3.29, P> 0.05). The expression rates of Ki-67 in CINⅡ, CINⅢ and SCC were all 100.0 %, and compared with the cervix benign changes, the differences were statistically significant (χ2= 112, P< 0.05). The expression rates of p16 in cervical benign changes and CINⅠwere 6.7 % (2/30) and 91.4 % (64/70), and there was significantly statistical difference (χ2=50.64, P<0.05). However, the expression rates of p16 in CINⅡ, CINⅢand SCC were all 100.0%, and compared with the cervix benign changes, the differences were statistically significant (χ2= 7.18, P< 0.01). The expression rates of stathmin in cervical benign changes, CINⅠ, CIN Ⅱ, CINⅢ and SCC were 3.3 %(1/30), 5.7 % (4/70), 23.1 % (18/78), 77.0 % (67/87) and 100.0 % (25/25), respectively, and there was no statistic difference in cervical benign changes, CINⅠand CINⅡ (χ2=0.68, P>0.05), but the expression rates in CINⅢ and SCC were higher than those in cervical benign change, CINⅠand CIN Ⅱ(P< 0.01). The positive expressions of stathmin, p16 and Ki-67 in each group of CIN were positively correlated (r= 0.412, P< 0.05). Conclusions Combined detection of p16 and Ki-67 can assist in the differential diagnosis of cervical intractable cases, and provide objective indicators for the classification and accurate diagnosis of CIN. Combined detection of p16 and stathmin may help to identify high-grade, low-grade CIN and cervix benign changes for the reduction of over-treatment.
2.Effect of Zuogui Jiangtang Qinggan Prescription on Glucose and Lipid Metabolism in MKR Mice with Type 2 Diabetes Mellitus and Non-alcoholic Fatty Liver Disease
Yiyang WANG ; Junju ZOU ; Yuanyuan LU ; Qin XIANG ; Rong YU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(10):102-109
ObjectiveTo investigate the mechanism of Zuogui Jiangtang Qinggan prescription (ZJQP) in improving glucose and lipid metabolism in loss of skeletalmuscle-specific insulin-like growth factor-1 receptor function (MKR) mice with type 2 diabetes mellitus (T2DM) and non-alcoholic fatty liver disease (NAFLD). MethodNAFLD was induced by high-fat diet feeding for 8 weeks in MKR mice, which were randomly divided into model group, metformin group (0.067 g·kg-1), and ZJQP high and low-dose groups(14.8, 7.4 g·kg-1). Ten FVB mice of the same age were used as the normal group. After 8 weeks of drug treatment, the oral glucose tolerance test (OGTT) was performed, the serum was taken to detect triacylglycerol (TG) and total cholesterol (TC), and the wet weight of the mouse liver was weighed. Haematoxylin-eosin (HE) staining and oil red O staining were performed to assess histopathology of liver. The mRNA expression and protein expression of Fork head box protein O1 (FoxO1), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6Pase), and apolipoprotein C3 (ApoC-Ⅲ) in liver tissues were detected by real-time fluorescent quantitative PCR (Real-time PCR) and Western blot, respectively. ResultAs compared with the normal group, the levels of fasting blood glucose, liver index, serum TG, TC, and OGTT of mice in the model group increased significantly (P<0.01). As compared with model group, the fasting blood glucose and liver index of the mice in the metformin group and the ZJQP group decreased significantly (P<0.01), the serum levels of TG and TC in the high-dose ZJQP group decreased significantly (P<0.05,P<0.01), and the OGTT of mice in the metformin group and the high-dose ZJQP group improved (P<0.05). In histopathology, as compared with the normal group, mice in the model group showed decreased lipid droplets and vacuoles in hepatocytes, and their volumes became larger. Compared with the model group, the ZJQP group and metformin group showed that the lipid droplets in liver tissues were reduced, the vacuoles in liver cells were reduced, and the volume was smaller. At the molecular level, as compared with the normal group, the mRNA and protein levels of FoxO1, PEPCK, G6Pase, and ApoC-Ⅲ in liver tissues of mice in the model group were significantly up-regulated (P<0.01). As compared with the model group, the mRNA and protein levels of FoxO1, PEPCK, G6Pase, and ApoC-Ⅲ in the ZJQP group was significantly decreased (P<0.01). ConclusionZJQP can improve the glucose and lipid metabolism of T2DM with NAFLD and repair the pathological damage of liver, which may be through regulating the expression of FoxO1, PEPCK, G6Pase, ApoC-Ⅲ-related proteins in liver tissues to achieve the effects of regulating lipid, lowering glucose, and delaying hepatic steatosis.
3.Danggui Shaoyaosan Combined with Yinchenhaotang Regulates Lipid Metabolism to Ameliorate Type 2 Diabetes Mellitus Complicated with Metabolic Dysfunction-associated Steatotic Liver Disease
Yilin XU ; Liu LI ; Junju ZOU ; Hong LI ; Rong YU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):38-47
ObjectiveTo explore the regulatory effect and mechanism of Danggui Shaoyaosan combined with Yinchenhaotang on lipid metabolism in the mouse model of type 2 diabetes mellitus (T2DM) complicated with metabolic dysfunction-associated steatotic liver disease (MASLD) based on network pharmacology and animal experiments. MethodsTwenty-four MKR transgenic diabetic mice were randomly allocated into 4 groups: Model, low-dose (12.6 g·kg-1) Chinese medicine (concentrated decoction of Danggui Shaoyaosan combined with Yinchenhaotang), high-dose (25.2 g·kg-1) Chinese medicine, and Western medicine (metformin, 0.065 g·kg-1). Six FVB mice were used as the normal group. All groups were treated for 6 consecutive weeks. The mice in the drug treatment groups were administrated with corresponding agents by gavage, and those in the normal group and model group received the same volume of distilled water. Fasting blood glucose, body weight, liver weight, glucose tolerance, liver function indicators, blood lipid levels, and pathological changes in the liver were evaluated for each group. Network pharmacology was employed to analyze the targets and pathways of Danggui Shaoyaosan combined with Yinchenhaotang in the treatment of T2DM complicated with MASLD. Molecular biological techniques were used to verify the enriched key targets. ResultsCompared with the model group, each treatment group showed reduced fasting blood glucose, body weight, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and liver weight (P<0.01). The high-dose Chinese medicine group was superior to the low-dose group in reducing low-density lipoprotein (LDL), increasing high-density lipoprotein (HDL), and recovering glucose tolerance (AUC) and ALT (P<0.05), with the effect similar to that of the Western medicine group. Morphologically, Chinese medicine groups showed reduced lipid accumulation and alleviated pathological damage in the liver tissue, with the high-dose group demonstrating more significant changes. Network pharmacology results showed that Danggui Shaoyaosan combined with Yinchenhaotang may exert therapeutic effects through multiple targets such as fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), B-cell lymphoma-2 (Bcl-2), MYC oncogene (MYC), and interleukin-1β (IL-1β). Western blot showed that compared with the model group, the treatment groups demonstrated down-regulated protein levels of FAS and ACC (P<0.01) and up-regulated protein levels of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and UCP1 (P<0.01). Compared with the low-dose Chinese medicine group, the high-dose Chinese medicine group exhibited down-regulated protein levels of FAS and ACC and up-regulated protein levels of PGC-1α and UCP1 (P<0.05). ConclusionDanggui Shaoyaosan combined with Yinchenhaotang has the effect of ameliorating T2DM complicated with MASLD and can improve the liver lipid metabolism by up-regulating the protein levels of Fas and ACC and down-regulating the protein levels of PGC-1α and UCP1.