1.Establishment and application of a TaqMan real-time PCR assay for the detection of encephalomyo-carditis virus
Haixia ZHANG ; Ruofei FENG ; Dan WANG ; Jingjing FAN ; Jingying XIE ; Zhongren MA ; Yuping FENG
Chinese Journal of Microbiology and Immunology 2013;(9):706-710
Objective To establish a TaqMan real-time PCR assay for the detection of encephalo-myocarditis virus ( EMCV) .Methods Based on the conservative region of 3D gene of EMCV published in GenBank , a pair of primers and one TaqMan probe were designed and synthesized .Then a TaqMan real-time PCR assay was set up and the reactive system was optimized .The sensitivity and specificity of the assay was evaluated respectively .The TaqMan real-time PCR assay was then carried out to detect 98 randomly selected swine serum samples and the results were compared with those by using ELISA .Results The Ct value of the templates had a good linear relationship with the log starting quantity , with a correlation coefficient of 0.995.The TaqMan real-time PCR assay was only specific for EMCV and its sensitivity was 100 times higher than that of the ordinary PCR .The coincidence rate between the established assay and the ELISA assay was 98.0%in the detection of 98 blood samples.Conclusion The TaqMan real-time PCR assay for the detec-tion of EMCV was successfully established with advantages of high sensitivity and good specificity .It could be used for detection of EMCV and quantitative analysis .
2.Effect of QizhiJiangtang Capsule on insulin resistance in diabetic rats and its mechanism
Xiaotian ZHANG ; Yu CHEN ; Chunjiang YU ; Yuze YUAN ; Jingtong ZHENG ; Chao ZHANG ; Jingying SAI ; Chenxue SONG ; Jingshu XIE ; Fang WANG
Journal of Jilin University(Medicine Edition) 2014;(4):805-811
Objective To explore the effect of QizhiJiangtang Capsule on the insulin resistance (IR)in the diabetic rats,and to clarify the action mechanism.Methods The diabetes rat models were induced by high fat diet combined with STZ injection.The successful models of the rats were randomly divided into diabetes group (DM), ShenqiJiangtang Granule group (SQ)and high (QJH),middle-(QJM),low (QJL)doses of QizhiJiangtang Capsule groups;at the same time control group (NC)was established. The drug concentrations in high, middle and low-doses of QizhiJiangtang Capsules groups were 1.35, 0.68, and 0.34 g · kg-1 respectively;and the concentration of ShenqiJiangtang Granule was 0.27 g·kg-1.After the diabetic model was established successfully, the rats were treated for 8 weeks on the basis of drug dose.Then the levels of fasting blood glucose (FBG),fasting insulin (FINS),insulin resistance index (IRI)and biochemical indexes related to lipid metabolism of the rats were measured using blood glucose detector and automatic biochemistry analyser.The gene expression of insulin receptor substrate-1 (IRS-1),phosphatidyl inositol 3-kinase (PI3K),and glucose transporter 4 (GLUT4)in liver tissue were examined by Real Time PCR.The levels of tumor necrosis factorα(TNF-α)and adiponectin (ADPN)in serum were detected using ELISA.Results Compared with control group,the levels of FBG,FINS and IRI of the rats in diabetes group were significantly increased (P<0.05 or P<0.01 );the serum total cholesterol (TC), triglyceride (TG)and low density lipoprotein (LDL)levels were significantly increased (P<0.05 ), while the serum high-density lipoprotein (HDL)level was significantly decreased (P<0.05);the mRNA expression levels of IRS-1,PI3K and GLUT4 in liver tissue were decreased (P<0.05);the level of serum TNF-αwas increased (P<0.05),but the ADPN level was decreased (P<0.05).Compared with diabetes group,the FBG level and IRI of the rats in QizhiJiangtang Capsule and ShenqiJiangtang Granule groups were significantly decreased (P<0.01);the levels of FINS of the rats middle and high doses of in QizhiJiangtang Capsule groups and ShenqiJiangtang Granule group were significantly decreased (P<0.05);the levels of serum TC,TG and LDL of the rats in middle dose of QizhiJiangtang Capsule group and ShenqiJiangtang Granule group were significantly decreased (P<0.05 or P<0.01),but the HDL level was increased (P<0.05);the mRBA expression lvels of IRS-1,PI3K and GLUT4 inliver tissue were increased (P<0.05);the levels of serum TNF-αof the rats in middle dose of QizhiJiangtang Capsule group and Shenqijiangtang Granule group were significantly decreased (P<0.05),but the serum ADPN levels were increased (P<0.05 ). Conclusion QizhiJiangtang Capsule can significantly improve the IR in the diabetic rats,and the pharmacological mechanisms are related to adapting the blood lipid component and insulin signal transduction pathways.
3.Improvement effects of 3,5,6,7,8,3′,4′-heptamethoxyflavone of Fructus Aurantii on rats with damp blockage of the middle energizer
Wenhui GONG ; Yating XIE ; Li XIN ; Shihao YAN ; Beibei ZHAO ; Yuqing ZHENG ; Jingying GUO ; Jie SHANG ; Peng ZHENG ; Jinlian ZHANG
China Pharmacy 2024;35(7):819-824
OBJECTIVE To investigate the improvement effects of 3,5,6,7,8,3′,4′-heptamethoxyflavone (HMF) of Fructus Aurantii on rats with damp blockage of the middle energizer. METHODS The rats were randomly divided into normal group, model group, positive control group (Raceanisodamine tablet, 1 mg/kg), HMF low-dose, medium-dose and high-dose groups (0.3, 0.6, 0.9 mg/kg), with 7 rats in each group. Except for the normal group, the other groups were modeled by internal and external composite factors. After successful modeling, the rats in each group were given the corresponding drug or normal saline, once a day, for 14 days. The general behavioral states such as dietary intake, water intake and mental state of the rats were observed, and the fecal water content rate and saliva flow rate were measured. Hematoxylin-eosin (HE) staining was used to observe the pathological and morphology in gastric and small intestinal tissues of rats. The plasma content of aldosterone was detected, and the expression of aquaporins (AQP3) in the gastric tissue of rats was determined. RESULTS Compared with the normal group, the dietary intake and water intake of the model group rats were significantly decreased (P<0.01), the fecal water content rate, salivary flow rate, plasma content of aldosterone and the expression of AQP3 in gastric tissue were increased significantly (P<0.01). Gastric tissue injury invaded the mucosal muscle layer, resulting in mucosal muscle layer rupture; pathological and morphological changes such as small intestinal villous erosion and glandular structure destruction were observed in the small intestine. Compared with the model group, the dietary intake and water intake of rats were increased in HMF groups; fecal water content rate, salivary flow rate, plasma content of aldosterone, the expression of AQP3 in gastric tissue were decreased, most of the above differences were statistically significant (P<0.05 or P<0.01). The pathological and morphological changes in the gastric and small intestine tissues of rats had been improved to varying degrees. CONCLUSIONS HMF of Fructus Aurantii with dry property HMF could improve the symptoms of rats with damp blockage of middle energizer, the mechanism of which may be associated with reducing the content of plasma aldosterone and down-regulating the expression of gastric AQP3.