1.Inhibition of Oxidative Stress of Sciatic Nerve in Diabetic Rats by Tangbikang Granules Regulating AMPK/PGC-1α/SIRT3 Signaling Pathway
Gang LIU ; Yaqi ZHANG ; Lingling QIN ; Chengfei ZHANG ; Qiue ZHANG ; Huizhong BAI ; Yi ZHAO ; Tonghua LIU ; Xiaohong MU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(6):75-82
		                        		
		                        			
		                        			ObjectiveTo investigate the effect of Tangbikang granules on oxidative stress of sciatic nerve in diabetic rats by regulating adenylate activated protein kinase/peroxisome proliferator-activated receptor γ coactivator-1α/mitochondrial Sirtuins 3 (AMPK/PGC-1α/SIRT3) signaling pathway. MethodThe spontaneous obesity type 2 diabetes model was established using ZDF rats. After modeling, they were randomly divided into high, medium, and low dose Tangbikang granule groups (2.5, 1.25, 0.625 g·kg-1·d-1) and lipoic acid group (0.026 8 g·kg-1·d-1), and the normal group was set up. The rats were administered continuously for 12 weeks after modeling. The blood glucose of rats was detected before intervention and at 4, 8, 12 weeks after intervention. At the 12th week, motor nerve conduction velocity (MNCV), sensory nerve conduction velocity (SNCV), nerve blood flow velocity, mechanical pain threshold, and thermal pain threshold were detected. The sciatic nerve was taken for hematoxylin-eosin (HE) staining to observe the tissue morphology. The ultrastructure of the sciatic nerve was observed by transmission electron microscope. The expression levels of superoxide dismutase (SOD), malondialdehyde (MDA), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in sciatic nerve were determined by enzyme-related immunosorbent assay (ELISA). The mRNA expressions of AMPKα, AMPKβ, PGC-1α, and SIRT3 in sciatic nerve were determined by real-time polymerase chain reaction (Real-time PCR). ResultCompared with the normal group, fasting blood glucose in the model group was increased at each time point (P<0.01). The mechanical pain threshold was decreased (P<0.05), and the incubation time of the hot plate was extended (P<0.01). MNCV, SNCV, and nerve blood flow velocity decreased (P<0.05). The expression level of SOD was decreased (P<0.01). The expression levels of MDA, IL-1β, and TNF-α were increased (P<0.01). The mRNA expression levels of AMPKα, AMPKβ, PGC-1α, and SIRT3 were decreased (P<0.01). The structure of sciatic nerve fibers in the model group was loose, and the arrangement was disordered. The demyelination change was obvious. Compared with the model group, the fasting blood glucose of rats in the high dose Tangbikang granule group was decreased after the intervention of eight weeks and 12 weeks (P<0.01). The mechanical pain threshold increased (P<0.05). The incubation time of the hot plate was shortened (P<0.01). MNCV, SNCV, and Flux increased (P<0.05). The expression level of SOD was increased (P<0.01). The expression levels of MDA, IL-1β, and TNF-α were decreased (P<0.01). The mRNA expression levels of AMPKα, AMPKβ, PGC-1α, and SIRT3 were increased (P<0.01). The sciatic nerve fibers in the high-dose Tangbikang granule group were tighter and more neatly arranged, with only a few demyelinating changes. The high, medium, and low dose Tangbikang granule groups showed a significant dose-effect trend. ConclusionTangbikang granules may improve sciatic nerve function in diabetic rats by regulating AMPK/PGC-1α/SIRT3 signaling pathway partly to inhibit oxidative stress. 
		                        		
		                        		
		                        		
		                        	
2.Relationship between lumbar spine development and hip development in children with spastic cerebral palsy
Xinwei ZUO ; Gang LIU ; Huizhong BAI ; Lin XU ; Yi ZHAO ; Jingpei REN ; Chuanyu HU ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(8):1247-1252
		                        		
		                        			
		                        			BACKGROUND:Most scholars now believe that children with cerebral palsy who have severe spinal deformities in early childhood(<15 years of age)may have a higher risk of progression of spinal deformities,which may result from imbalances in movement due to pelvic tilt,pain,etc. OBJECTIVE:To investigate the relationship between lumbar spine development and hip joint development in children with spastic cerebral palsy. METHODS:A retrospective analysis was performed in 102 children with spastic cerebral palsy admitted at Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine from January 2014 to December 2021.All admitted children had X-rays of the pelvic position and the lumbar lateral position.Anteroposterior X-ray of the pelvis was performed to measure femoral head migration percentage,central edge angle,neck-shaft angle,and acetabular index.The sagittal Cobb angle,sacral slope,arch-top distance,and lumbar lordosis index were measured by the lateral X-ray of the lumbar spine.Correlation of the two sets of indicators was further analyzed.All children were divided into normal group,risk group,hip subluxation group and total hip dislocation group according to their femoral head migration percentage,and the differences in lumbar spine indexes between groups were evaluated. RESULTS AND CONCLUSION:Pearson correlation analysis showed that the femoral head migration percentage was moderately positively correlated with sagittal Cobb angle and arch-top distance,and weakly positively correlated with lumbar lordosis index;the central edge angle was moderately negatively correlated with the arch-top distance and weakly negatively correlated with the sagittal Cobb angle;the neck-shaft angle was weakly positively correlated or not correlated with the sagittal Cobb angle and lumbar lordosis index;and the acetabular index was weakly positively correlated with the sagittal Cobb angle and arch-top distance.No statistically significant correlation was found between the remaining indicators.According to the femoral head migration percentage,the children were divided into four groups,including 25 cases in the normal group,41 cases in the risk group,27 cases in the hip subluxation group,and 9 cases in the total hip dislocation group.The sagittal Cobb angle was significantly increased in the risk group,the hip subluxation group and the total hip dislocation group compared with the normal group,showing an increasing trend group by group,and there were significant differences between groups(P<0.05).Compared with the normal group,the lumbar lordosis index in the risk group and the hip subluxation group increased significantly,and there were significant differences between groups(P<0.05).There was an increase trend in the lumbar lordosis index of the total hip dislocation group compared with the normal group.Compared with the normal group,the arch-top distance in the hip subluxation group and the total hip dislocation group increased significantly(P<0.05),and there was a stepwise increasing trend.There was no significant difference in sacral slope between groups.To conclude,the development of the lumbar spine in children with cerebral palsy is closely related to the development of the pelvic hip joint,and the most obvious relationship is between lumbar lordosis and hip dislocation.
		                        		
		                        		
		                        		
		                        	
3.Effects of Shujin Jiannao Prescription on cell apoptosis in rats with hypoxic-ischemic brain injury
Yu JIANG ; Lin XU ; Yalin ZHAO ; Gang LIU ; Yaqi ZHANG ; Huizhong BAI ; Jingpei REN ; Jie ZENG ; Xiaohong MU
Chinese Journal of Tissue Engineering Research 2024;28(28):4477-4483
		                        		
		                        			
		                        			BACKGROUND:Perinatal hypoxic-ischemic brain injury is one of the most common causes of cerebral palsy.Shujin Jiannao Prescription is an experienced formula for treating cerebral palsy and improving blood supply to the brain developed by the Dongzhimen Hospital,Beijing University of Chinese Medicine. OBJECTIVE:To explore the possible mechanism of Shujin Jiannao Prescription in treating hypoxic-ischemic cerebral palsy. METHODS:Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into six groups.There were 12 rats in each of the control and model groups as well as 10 animals in each of the minocycline group,and the low-,medium-,and high-dose groups of Shujin Jiannao Prescription.The neonatal rat ischemic-hypoxic cerebral palsy model was established in all groups except for the control group.After successful modeling,rats in each drug group were respectively gavaged with minocycline and Shujin Jiannao Prescription at a dose of 4,8,and 16 g/kg per day for 1 week.Body mass of rats was measured and behavioral changes were detected before and after drug administration.Hematoxylin-eosin staining was used to observe the histomorphology of hippocampal CA1 region of rat brain tissue,and immunohistochemistry and western blot were used to detect the expression levels of Bcl-2,Bax,and Caspase-3 in the brain tissue of rats. RESULTS AND CONCLUSION:Compared with the model group,medium-and high-dose Shujin Jiannao Prescription significantly increased the body mass of rats(P<0.05).Compared with the model group,minocycline effectively prolonged the suspension time of ischemic-hypoxic cerebral palsy rats(P<0.05),while medium-and high-dose Shujin Jiannao Prescription significantly prolonged the suspension time,shortened the inclined plane test time,and increased the Longa score of rats(P<0.05).The pathological results showed that after drug intervention,only a small number of neuronal cells in the brain tissue of rats were necrotic,the cells were more neatly arranged,the cell structure was more complete,and only part of the cell nuclei became smaller.Compared with the model group,minocycline and medium-and high-dose Shujin Jiannao Prescription reduced the expression of Bax Caspase-3(P<0.05),medium-and high-dose Shujin Jiannao Prescription increased the expression of Bcl-2(P<0.05),and Bcl-2/Bax protein expression was increased in minocycline and three Shujin Jiannao Prescription groups(P<0.05).In addition,the protein expression was increased in a dose-dependent manner after intervention with Shujin Jiannao Prescription,and there was no significant difference between the minocycline and three Shujin Jiannao Prescription groups(P>0.05).To conclude,the mechanism by which Shujin Jiannao Prescription treats ischemic-hypoxic cerebral palsy in rats may be to enhance the expression of anti-apoptotic protein Bcl-2,inhibit the expression of pro-apoptotic protein Bax,and reduce the expression of Caspase-3,ultimately inhibiting the apoptosis of hippocampal neuronal cells in rats with cerebral palsy.Within a certain range,the higher dose of Shujin Jiannao Prescription indicates the better therapeutic effect,and the high-dose Shujin Jiannao Prescription is as effective as minocycline.
		                        		
		                        		
		                        		
		                        	
4.A Meta-analysis in efficacy and safety of selective posterior rhizotomy for patients with cerebral palsy at different grades of the Gross Motor Function Classification System
Yu JIANG ; Gang LIU ; Luyao HUO ; Huizhong BAI ; Jingpei REN ; Yi ZHAO ; Chuanyu HU ; Lin XU ; Xiaohong MU
Journal of Clinical Medicine in Practice 2024;28(19):60-67
		                        		
		                        			
		                        			Objective To investigate the differences in efficacy and safety in the treatment of patients with cerebral palsy at different grades of the Gross Motor Function Classification System (GMFCS) by selective posterior rhizotomy (SPR). Methods Relevant literatures on SPR treatment for cerebral palsy were retrieved from Pubmed, Embase, Web of Science, China Biology Medicine disc, China National Knowledge Infrastructure (CNKI), Wanfang Database, and VIP Database. Clinical trials on SPR treatment for cerebral palsy were included for Meta-analysis. At least two researchers independently screened the literatures, extracted data, and assessed the quality of the literatures. Data analysis was performed by Review Manager 5.4 software. Results A total of 2, 726 literatures were retrieved, and 8 literatures were finally included after screening. The results of the Meta-analysis showed that the gross motor function and self-care ability of patients with cerebral palsy at all GMFCS grades improved significantly after surgery, and muscle tone decreased significantly after surgery (
		                        		
		                        	
5.Epithelial-mesenchymal phenotypes analysis of circulating tumor cells with dual-antibody for the prognosis assessment in hepatocellular carcinoma
Huizhong ZHANG ; Fazhuang FANG ; Xiaodong ZHANG ; Junjie LIU ; Jianming FANG ; Jingchao TANG ; Haiyang LI ; Yibiao ZHANG
Chinese Journal of Hepatobiliary Surgery 2024;30(11):830-835
		                        		
		                        			
		                        			Objective:To explore the feasibility of circulating tumor cell (CTC) dual-antibody enrichment and dual-antibody detection for epithelial-mesenchymal phenotypes in patients with hepatocellular carcinoma (HCC), and investigate the clinical diagnostic value of CTC typing in evaluating postoperative recurrence and prognosis of HCC.Methods:Of 89 HCC patients who underwent surgical treatment in Zhejiang Jinhua Guangfu Tumor Hospital from March 2020 to January 2024 were enrolled into this study, including 73 males and 16 females, aged (64.4±9.5) years old. The peripheral blood samples of patients were collected before operation. Epithelial CTC, mesenchymal CTC and hybrid epithelial-mesenchymal CTC in blood samples of patients with HCC were enriched and detected by EpCAM/CSV double capture antibodies and PanCK/CSV double detection antibodies. Kaplan-Meier analysis was employed to assess recurrence-free survival (RFS) and overall survival (OS) rates. Univariate and multivariate Cox regression were used to analyze the effects of different types of CTC on postoperative RFS and OS.Results:The detection rates of total CTC, epithelial CTC, mesenchymal CTC and hybrid epithelial-mesenchymal CTC were 92.1% (82/89), 64.0% (57/89), 62.9% (56/89) and 55.1% (49/89), respectively. Multivariate Cox regression analysis showed that HCC patients with more mesenchymal CTC ( HR=2.408, 95% CI: 1.580-3.668) and hybrid epithelial-mesenchymal CTC ( HR=1.840, 95% CI: 1.004-3.371) in peripheral blood had higher postoperative recurrence risk (both P<0.05). Univariate Cox regression showed patients with more total CTC ( HR=1.426, 95% CI: 1.040-1.954, P=0.028) was associated with survival. Conclusion:The technique of epithelial-mesenchymal phenotypes analysis of circulating tumor cells based on dual-antibody capture and dual-antibody detection is feasible. The number of mesenchymal CTC and hybrid epithelial-mesenchymal CTC before operation is the influencing factor of postoperative recurrence in patients with hepatocellular carcinoma.
		                        		
		                        		
		                        		
		                        	
6.Hypolipidemic effect of flavonoids in hyperlipidaemic rats and study of PPARαpathway in blue prickly head flavonoids
Ruining LIU ; Jianbing ZHANG ; Huizhong ZHANG ; Han WANG ; Puguo HAO ; Yu GUO ; Yu WANG ; Hongxia ZHAO
Chinese Journal of Veterinary Science 2024;44(10):2243-2250
		                        		
		                        			
		                        			Hyperlipidaemic rats were randomly divided into a model group,a total flavonoids from echinops latifolius tausch(TFET)high,medium and low dose group,and a positive control group;meanwhile,healthy rats were selected as a blank control group.The rats in each group were dosed with the corresponding concentrations of TFET,simvastatin and distilled water for 45 consecutive days,and were tested for relevant lipid biochemical indexes,antioxidant indexes and PPARα path-way-related gene expression.The results showed that high-dose TFET could reduce the concentra-tions of TC,TG and LDL-C and increase the concentration of HDL-C very significantly;medium-dose TFET could reduce the concentrations of TC and TG very significantly and reduce the con-centration of LDL-C significantly;and low-dose TFET could reduce the concentrations of TC and LDL-C significantly.High,medium and low doses of TFET can extremely significantly reduce in-crease SOD activity;high and medium doses of TFET can extremely significantly reduce MDA content;high dose of TFET can extremely significantly increase T-AOC activity.The high dose of TFET could extremely significantly increase the expression of PPARα,CYP7A1 and CPT-1 genes in rat liver;the medium dose of TFET could extremely significantly increase the expression of CYP7A1 and CPT-1 genes,and could significantly increase the expression of PPARα gene;the low dose of TFET could extremely significantly increase the expression of CYP7A1 gene,and signifi-cantly increase the CPT-1 gene expression.The results suggest that TFET has antioxidant and lip-id-lowering effects on hyperlipidaemic rats,and its mechanism may be related to the activation of PPARα and its downstream related genes to promote fatty acid β-oxidation.
		                        		
		                        		
		                        		
		                        	
7.Interaction analysis of mismatch repair protein and adverse clinicopathological features on prognosis of colon cancer
Kexuan LI ; Fuqiang ZHAO ; Qingbin WU ; Junling ZHANG ; Shuangling LUO ; Shidong HU ; Bin WU ; Heli LI ; Guole LIN ; Huizhong QIU ; Junyang LU ; Lai XU ; Zheng WANG ; Xiaohui DU ; Liang KANG ; Xin WANG ; Ziqiang WANG ; Qian LIU ; Yi XIAO
Chinese Journal of Digestive Surgery 2024;23(6):826-835
		                        		
		                        			
		                        			Objective:To investigate the interactive effect of mismatch repair (MMR) protein status and adverse clinicopathological features on prognosis of stage Ⅰ-Ⅲ colon cancer.Methods:The retrospective cohort study was conducted. The clinicopathological data of 1 650 patients with colon cancer of stage Ⅰ-Ⅲ who were admitted to 7 hospitals in China from January 2016 to December 2017 were collected. There were 963 males and 687 females, aged 62(53,71)years. Patients were classified as 230 cases of MMR deficiency (dMMR) and 1 420 cases of MMR proficiency (pMMR) based on their MMR protein status. Observation indicators: (1) comparison of clinicopathological characteristics between patients of different MMR protein status; (2) analysis of factors affecting the survival outcomes of patients of dMMR; (3) analysis of factors affecting the survival outcomes of patients of pMMR; (4) interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Comparison of ordinal data was conducted using the Mann-Whitney U test. The random forest interpolation method was used for missing values in data interpolation. Univariate analysis was conducted using the COX proportional risk regression model, and multivariate analysis was conducted using the COX stepwise regression with forward method. The coefficient of multiplication interaction effect was obtained using the interaction term coefficient of COX proportional risk regression model. Evaluation of additive interaction effects was conducted using the relative excess risk due to interaction ( RERI). Results:(1) Comparison of clinicopathological characteristics between patients of different MMR protein status. There were significant differences in age, T staging, the number of lymph node harvest, the number of lymph node harvest <12, high grade tumor between patients of dMMR and pMMR ( P<0.05). (2) Analysis of factors affecting the survival outcomes of patients of dMMR. Results of multivariate analysis showed that T staging, N staging, the number of lymph node harvest <12 were independent factors affecting the disease-free survival (DFS) of colon cancer patients of dMMR ( hazard ratio=3.548, 2.589, 6.702, 95% confidence interval as 1.460-8.620, 1.064-6.301, 1.886-23.813, P<0.05). Age and N staging were independent factors affecting the overall survival (OS) of colon cancer patients of dMMR ( hazard ratio=1.073, 10.684, 95% confidence interval as 1.021-1.126, 2.311-49.404, P<0.05). (3) Analysis of factors affecting the survival outcomes of patients of pMMR. Results of multivariate analysis showed that age, T staging, N staging, vascular tumor thrombus were independent factors affecting the DFS of colon cancer patients of pMMR ( hazard ratio=1.018, 2.214, 2.598, 1.549, 95% confidence interval as 1.006-1.030, 1.618-3.030, 1.921-3.513, 1.118-2.147, P<0.05). Age, T staging, N staging, high grade tumor were independent factors affecting the OS of colon cancer patients of pMMR ( hazard ratio=1.036, 2.080, 2.591, 1.615, 95% confidence interval as 1.020-1.052, 1.407-3.075, 1.791-3.748, 1.114-2.341, P<0.05). (4) Interaction analysis of MMR and adverse clinicopathological features on survival outcomes. Results of interaction analysis showed that the multiplication interaction effect between the number of lymph node harvest <12 and MMR protein status was significant on DFS of colon cancer patients ( hazard ratio=3.923, 95% confidence interval as 1.057-14.555, P<0.05). The additive interaction effects between age and MMR protein status, between high grade tumor and MMR protein status were significant on OS of colon cancer patients ( RERI=-0.033, -1.304, 95% confidence interval as -0.049 to -0.018, -2.462 to -0.146). Conclusions:There is an interaction between the MMR protein status and the adverse clinicopathological features (the number of lymph node harvest <12, high grade tumor) on prognosis of colon cancer patients of stage Ⅰ-Ⅲ. In patients of dMMR, the number of lymph node harvest <12 has a stronger predictive effect on poor prognosis. In patients of pMMR, the high grade tumor has a stronger predictive effect on poor prognosis.
		                        		
		                        		
		                        		
		                        	
8.A national multi-center prospective study on the perioperative practice of enhanced recovery after surgery for choledochal cysts in children
Ming YUE ; Jiexiong FENG ; Yan′an LI ; Yuanmei LIU ; Zhigang GAO ; Qi CHEN ; Hongwei XI ; Qiang YIN ; Chengji ZHAO ; Yuzuo BAI ; Wanfu LI ; Libin ZHU ; Weibing TANG ; Hongqiang BIAN ; Huizhong NIU ; Zhiheng GUO ; Heying YANG
Chinese Journal of General Surgery 2024;39(11):827-832
		                        		
		                        			
		                        			Objective:To evaluate the safety and efficacy of enhanced recovery after surgery(ERAS) in the perioperative period of congenital choledochal cysts in children.Methods:This is a multicenter prospective randomized controlled study. The clinical data of 273 pediatric congenital choledochal cysts(CCC) patients who underwent surgery at 14 medical centers with complete follow-up data were collected through the medical data analysis platform. Among them, 123 cases in ERAS group were managed perioperatively in strict accordance with ERAS mode, and 150 cases in conventional group were managed according to traditional mode. The length of hospital stay,time to first farting, time to complete feeding, the incidence of complications, cost and readmission rate within 30 days,stress indexes and liver function were compared between the two groups.Results:Compared with the conventional group, median time to start farting (2.0 d vs. 3.0 d, P<0.001), median time to complete feeding (5.0 d vs. 7.0 d, P<0.001), median postoperative hospitalization time (6.0 d vs. 9.0 d, P<0.001),the median total length of stay(13.0 d vs. 15.0 d, P<0.001) were shorter,the median hospitalization cost (37,000 yuan vs.43,000 yuan P<0.001) was lower, and stress indexes recovered quickly. The incidence of postoperative hospital stay and readimission rate within 30 d were not statistically different between the two groups. Conclusion:It is safe and feasible to implement ERAS for children with CCC in the perioperative period, which can reduce stress response, speed up recovery,and save medical costs.
		                        		
		                        		
		                        		
		                        	
9.Effect of Tangbikang Granules on PI3K Pathway in Type 2 Diabetic Mellitus Rats with Non-alcoholic Fatty Liver Disease
Yaqi ZHANG ; Lingling QIN ; Chengfei ZHANG ; Qiue ZHANG ; Huizhong BAI ; Gang LIU ; Xinwei ZUO ; Yi ZHAO ; Tonghua LIU ; Xiaohong MU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(9):71-80
		                        		
		                        			
		                        			ObjectiveThis study aims to investigate the therapeutic effect of Tangbikang granules(TBK) on type 2 diabetes mellitus (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD) and to elucidate the underlying mechanism. MethodT2DM and NAFLD were induced in ZDF rats, which were then respectively treated (ig) with low-dose (0.625 g·kg-1), medium-dose (1.25 g·kg-1), and high-dose (2.5 g·kg-1) TBK for 12 weeks. Fasting blood glucose (FBG) and body mass were recorded every 4 weeks during the treatment. One week before sampling, the feed intake of rats was detected, and after 12 h night fasting, oral glucose tolerance test (OGTT) was performed. The area under the curve (AUC) was used to evaluate glucose tolerance, and the homeostatic model assessment for insulin resistance (HOMA-IR) was calculated. Blood in abdominal aorta and liver were collected for determination of blood glucose and lipid metabolism indexes: Fasting serum insulin (FINS), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), and nonesterified fatty acids (NEFA). The liver was weighed to calculate the liver index, and the liver tissue morphology was observed and analyzed based on hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining. The protein levels of insulin receptor substrate (IRS), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and phosphorylated IRS and Akt were detected by Western blotting. All data were analyzed by SPSS 20.0. ResultThe feed intake of the model group was higher than that in the normal group (P<0.01), and the feed intake the administration groups was lower than that in the model group (P<0.05, P<0.01). At the 8th and 12th week, the body mass in the model group was lower than that in the normal group (P<0.01). Compared with the model group, TBK reduced FBG in a concentration-dependent manner. The blood glucose level in OGTT and AUC in the model group were higher/larger than those in the normal group (P<0.01). The blood glucose value in OGTT was decreased in TBK groups and the metformin group compared with that in the model group, and AUC in the administration groups was significantly different from that in the model group (P<0.01). The serum level of FINS and HOMA-IR in the model group were higher than those in the normal group (P<0.01), and they were lower in the TBK groups than in the model group (P<0.01). Serum levels of TG, TC, HDL-C, NEFA (P<0.05, P<0.01), and LDL-C were higher in the model group than in the normal group. Serum levels of TG, TC, LDL-C, and NEFA in the TBK groups were lower than those in the model group, and the levels of TG, LDL-C, and NEFA in TBK groups were concentration-dependent (lowest levels in high-dose TBK group). Compared with the model group, high-dose TBK significantly increased the level of HDL-C (P<0.05). Liver index of the model group was higher than that in the normal group (P<0.01). The liver index of the administration groups showed a decreasing trend with no significant difference from that in the model group. As for the HE staining result of liver, the model group had unclear structure of liver lobule, enlarged cells of different sizes, and obvious steatosis of hepatocytes. TBK of all doses alleviated liver injury, particularly the high dose. For the PAS staining, compared with the normal group, the model group demonstrated significant fat vacuoles and significant reduction in purplish red glycogen granules in the cytoplasm. The staining results of high- and medium-dose groups of TBK were more similar to the normal group. Western blot was used to detect the protein expression of liver tissue. The expression of PI3K protein, p-IRS1/IRS1, and p-Akt/Akt in the model group were lower than those in the normal group (P<0.01), and they were higher in the high-dose TBK group than in the model group (P<0.01). ConclusionTBK exerts therapeutic effect on T2DM combined with NAFLD in ZDF rats by activating the typical PI3K signaling pathway. 
		                        		
		                        		
		                        		
		                        	
10.Molecular Mechanism of Tangbikang Granules Against Diabetic Peripheral Neuropathy: Based on Network Pharmacology and Experimental Verification
Yaqi ZHANG ; Lingling QIN ; Huizhong BAI ; Chengfei ZHANG ; Qiue ZHANG ; Xinwei ZUO ; Shengyuan JIANG ; Yi ZHAO ; Tonghua LIU ; Xiaohong MU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(9):81-90
		                        		
		                        			
		                        			ObjectiveTo explore the mechanism of Tangbikang granules (TBK) against diabetic peripheral neuropathy (DPN) based on network pharmacology and in-vivo experiment. MethodThe active components in medicinals of TBK and their target genes were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The active components of the medicinals which are not included in TCMSP were searched from previous research. After the analysis of drug-likeness by SwissADME, the target genes of them were predicted with SwissTargetPrediction. DPN-related target genes were retrieved from GeneCards. The common targets of the disease and the prescription were the hub genes of TBK against DPN, which were uploaded to Metascape for Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. High-sugar and high-fat diet and low-dose streptozotocin (STZ, ip) were employed to induce diabetes in rats, and then the model rats were respectively treated with low-dose (0.625 g·kg-1), medium-dose (1.25 g·kg-1), and high-dose (2.5 g·kg-1) TBK for 12 weeks. Sensory nerve conduction velocity (SNCV) was evaluated. After hematoxylin and eosin (HE) staining, the sciatic nerve was observed under light microscope to examine the nerve damage. Real-time PCR was performed to detect the gene expression of adenosine monophosphate-activated protein kinase (AMPK) pathway-related targets in rat sciatic nerve, and Western blot to measure the protein expression of AMPK and phosphorylated (p)-AMPK in rat sciatic nerve. ResultThe main active components of TBK, such as quercetin, kaempferol, β-sitosterol, leech pteridine A, stigmasterol, and baicalein were screened out, mainly acting on interleukin-6 (IL-6), tumor necrosis factor (TNF), protein kinase B (Akt), JUN, and HSP90AA1 and signaling pathways such as AMPK, nuclear factor-κB (NF-κB), and Janus kinase/signal transducer and activator of transcription (JAK/STAT). Molecular docking results showed that β-sitosterol and stigmasterol had high binding affinity with IL-6, TNF, JUN, and HSP90AA1. As for the animal experiment, compared with the normal group, model group had low SNCV of sciatic nerve (P<0.01), disordered and loose myelinated nerve fibers with axonotmesis and demyelinization, low mRNA expression of AMPKα, AMPKβ, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), Sirtuin 3 (SirT3), mitochondrial transcription factor A (TFAM), and low p-AMPK/AMPK ratio in sciatic nerve (P<0.05, P<0.01). Compared with the model group, TBK of the three doses raised the SNCV (P<0.01), restored nerve morphology and nerve compactness, and increased the mRNA expression of AMPKα, AMPKβ, PGC-1α, SirT3, and TFAM (P<0.05, P<0.01). The ratio of p-AMPK/AMPK in the high-dose and medium-dose TBK groups was higher than that in the model group (P<0.01), while the protein expression in the low-dose TBK group was insignificantly different from that in the model group. ConclusionTBK exerts therapeutic effect on DPN through multiple pathways and targets. The mechanism is that it activates and regulates AMPK/PGC-1α/SirT3 signaling, which lays a basis for further study of TBK in the treatment of DPN. 
		                        		
		                        		
		                        		
		                        	
            

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