Amounts of tanshenoside Ⅰ in cultivated Ludangshen of different years of cultivation history was determined by TLC densitometry in comparison with that in wildly grown Ludangshen and cultivated Taidangshen. Results showed that tanshenoside Ⅰ in cultivated Ludangshen decreases with increased years of cultivation history while that in cultivated Taidangsihen is slightly lower than that of Ludangshen of the same years of cultivation. Tanshenosde Ⅰ in wild Taidangshen is also lower than that in cultivated Taidangshen

Aim To further study the molecular mecha-nism of the herbs with hot nature on the regulational action on TRPV1 channel based on the 7900 Real-time PCR instrument. Methods 7900 PCR instrument was applied to detect the intracellular flurescence of TRPV1 channel in the dorsal root ganglions ( DRG ) neurons and the effect on the TRPV1 ’ s thermo-sensational functions of the selected 11 ingredients from hot herbs was explored. Results TRPV1 channels could be ac-tivated by gradually elevated temperature; the activa-tion process could be blocked by the TRPV1 specific blocking agent capsaizepine. Most of the ingredients from hot-nature herbs had the potential to up-regualate TRPV1 channel function. Conclusions The estab-lished TRPV1 channel detection system based on PCR instrument is suitable for the analysis of regulational functions of drugs on the heat-activated TRPV1 chan-nel;the functions of hot herbs may be related to the up-regualtional effects of its active ingredients on the TRPV1 channel which will further up-regulate energy metabolism.

In order to meet the requirements of cultivating the practical abilities and creativities of students who receive higher education, we initiated the reformation of education in the microbiology experiment teaching methods, implementing a system for module-based education, carefully monitoring every link in teaching, combining the encouragement and strict requirements together, adopting a proper way of assessment. It is proven that the implementation of the educational reformation mobilizes the interests of students and enhances the comprehensive qualities of students, which accomplishes the purposes of teaching.

Objective To evaluate the effect of exogenous hydrogen sulfide (H2 S) on myocardial damage induced by renal ischemia-reperfusion (I/R) in rats.Methods Thirty pathogen-free male Wistar rats,weighing 250-300 g,were randomized into 3 groups (n =10 each) using a random number table:sham operation group (S group); myocardial damage induced by renal I/R group (I/R group); NaHS group.Renal I/R was induced by occlusion of the left kidney for 120 min followed by 60 min reperfusion in anesthetized rats.In NaHS group,NaHS 100 μg/kg was injected intraperitoneally at 20 min before reperfusion.At the end of reperfusion,the rats were sacrificed and the hearts were obtained for determination of malondialdehyde (MDA) content and glutathione peroxidase (GSH-Px) activity in myocardial tissues and for microscopic examination.Results Compared with S group,MDA content was significantly increased,and GSH-Px activity was decreased in I/R and NaHS groups.Compared with I/R group,MDA content was significantly decreased,and GSH-Px activity was increased in NaHS group.The pathological changes of myocardium were significantly attenuated in NaHS group as compared with I/R group.Conclusion Exogenous H2 S can alleviate myocardial damage induced by renal I/R in rats.

Objective To investigate the effects of dietary consumption of monounsaturated fatty acid (MUFA) on the control of type 2 diabetes mellitus. Methods One hundred and eighty-five patients with type 2 diabetes mellitus from a community in Shanghai were randomly divided into MUFA intervention group (MUFA group,n=125) and control group (n=60). The patients in MUFA group consumed tea oil enriched in MUFA for 3 months,while those in control group consumed normal oil. The serum glucose (fasting plasma glucose and 2-hour postprandial glucose),fasting insulin and blood lipid were examined before intervention and three months after intervention,and the parameters were compared within groups and between groups. Results The serum fasting glucose,fasting insulin,total cholesterol and low density lipoprotein-cholesterol 3 months after intervention were significantly lower than those before intervention in MUFA group (P0.05),while the serum fasting glucose,total cholesterol and low density lipoprotein-cholesterol in MUFA group were significantly lower than those in control group 3 months after intervention (P0.05). Conclusion Dietary consumption of MUFA can improve the glucose and lipid metabolism in patients with type 2 diabetes mellitus.

High price and poor stability of both crocin-1 and crocin-2 reference substance have become obstacles to HPLC assay of Croci Stigma. A new method based on reference extract was proposed. In this study, the reference extract was prepared from gardenia yellow which is cheap and easy to get The content of crocin-1 and crocin-2 in reference extract was determined and factors affecting stability of reference extract were investigated. Twelve batches of Croci Stigma were analyzed with reference extract and reference substance respectively. The results showed no difference. The presented method is feasible for quality control of Croci Stigma and reference extract is suitable to replace reference substances in assay.
Carotenoids ; analysis ; Chromatography, High Pressure Liquid ; standards ; Crocus ; chemistry ; Drugs, Chinese Herbal ; analysis ; Quality Control ; Reference Standards

Objective To systematically review the effectiveness and safety of statins for chronic obstructive pulmonary diseases (COPD) combining with pulmonary hypertension (PH).Methods The electronic searches in databases of PubMed,EMbase,the Cochrane Library,Web of Science,CBM,CNKI,VIP and Wanfang Data were conducted from the date of their establishment to January 2016 and the references of the include studies were also retrieved for collecting randomized controlled trials (RCTs) or quasi-RCTs on statins treating COPD combining with PH.Two researchers independenlty screened the literature according to the inclusion and exclusion criteria,extracted the data,assessed the quality of the included studies by adopting the Cochrane collaboration' s tool for assessing risk of bias,and performed Meta-analysis by using RevMan 5.3 software.Results A total of 24 studies involving 1 587 cases were included.The results of Meta-analysis showed that:compared with the control group,simvastatin significantly improved FEV1 [MD =0.23,95％ CI:0.16-0.31,P ＜ 0.000 01],FEV1 ％ [MD =6.73,95％ CI:1.34-12.12,P =0.01],FVC [MD =0.39,95％ CI:0.34-0.45,P ＜ 0.000 01],6 minutes walk distance (6MWD)· [MD=59.09,95％CI:54.24-63.93,P ＜0.000 01] and decreased mPAP [MD=6.73,95％ CI:1.34-12.12,P =0.01],SPAP [MD =-4.53,95 ％ CI =-8.87--0.19,P =0.04].Atorvastatin significantly improved FEV1 [MD =6.22,95 ％ CI:2.51-9.93,P =0.001] and 6 MWD [MD =24.10,95 ％ CI:12.98-35.23,P ＜ 0.000 1] and decreased sPAP [MD =-6.44,95％CI:-7.95--4.93,P＜0.00001] andmPAP [MD=-3.51,95％CI:-5.81--1.22,P=0.003].But no significant difference was found in the improvement of FEV1,FVC or FEV1/FVC.Fluvastatin significantly decreased sPAP [MD=-5.89,95％ CI:-6.99--4.79,P ＜0.000 01].There was a significant decrease in the Borg dyspnoea score in statins group [MD =-3.37,95％ CI:-4.61--2.14,P ＜ 0.000 01] as compared with the controls.In addition,the incidence of adverse drug reactions (ADRs) was similar between statins and the control group.Conclusion Current evidence suggests that statins may decrease pulmonary hypertension in patients with COPD combining with PH.However,high-quality clinical trials with large sample size are needed to verify whether the improvement of pulmonary function,6MWD and Borg dyspnoea score are the class effect or the incidence of ADRs is disparate among different statins.

BACKGROUNDDiabetic myocardiopathy is characterized by myocardial interstitial fibrosis and cardiac dysfunction. Statins were found to exert protective effects on cardiovascular disease by suppressing activation of small G proteins, independently of their lipid-lowering effect. The study investigated the effect of fluvastatin on myocardial interstitial fibrosis, cardiac function and mechanism of its action in diabetic rats.

METHODSTwenty-four male SD rats were randomly assigned to 3 groups: control rats (n = 8), streptozotocin (STZ)-induced diabetic rats (n = 8), and diabetic rats treated with fluvastatin (administered fluvastatin orally, 10 mg/kg body weight per day, n = 8). Twelve weeks later, miniature cardiac catheter was inserted into the left ventricle to conduct hemodynamic examination. Then myocardium tissues were collected, collagen content was detected by picro-sirius red staining, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of connective tissue growth factor (CTGF), and Western blotting was used to detect the protein expression of CTGF. Rho activity was determined by pull-down assay.

RESULTSAfter 12 weeks, the left ventricular systolic pressure (LVSP) and maximum rate of left ventricular (LV) pressure rise and fall (+dP/dt max and -dP/dt max) were significantly lower and left ventricular end diastolic pressure (LVEDP) was higher in the diabetic rats than those in the control rats (P < 0.01). Moreover, in LV myocardial tissue of diabetic rats the collagen content, fibronectin, mRNA and protein expression of CTGF and the activity of RhoA were all significantly increased compared with the control rats (P < 0.01). Administration of fluvastain obviously improved the cardiac function of diabetic rats, attenuated fibronectin expression, mRNA and protein expression of CTGF and the activity of RhoA in LV myocardium of diabetic rats.

CONCLUSIONSFluvastatin attenuates cardiac dysfunction and myocardial interstitial fibrosis of diabetic rat by inhibiting activity of RhoA to down-regulate the overexpression of CTGF, and Rho/Rho-kinase pathway may be an important target in the treatment of diabetic cardiomyopathy.

Animals ; Anticholesteremic Agents ; therapeutic use ; Blood Glucose ; metabolism ; Blotting, Western ; Cholesterol ; blood ; Connective Tissue Growth Factor ; metabolism ; Fatty Acids, Monounsaturated ; therapeutic use ; Fibrosis ; blood ; drug therapy ; metabolism ; Hemodynamics ; drug effects ; Immunohistochemistry ; Indoles ; therapeutic use ; Male ; Myocardium ; metabolism ; pathology ; Random Allocation ; Rats ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .

AIM:R-spondin 2 (Rspo2), one member of R-spondin family which contains four secreted proteins , plays an important role in skeletal muscle development .However, the impact of Rspo2 on vascular smooth muscle cell ( SMC) differentiation is little known . This study aims at revealing the role and mechanism of Rspo 2 on SMC differentiation from embryonic stem cells (ESCs).METHODS:A well-established model for studying SMC differentiation from ESCs were used , in which mouse embryonic stem cells ( ES-D3) were seeded on collagen IV-coated flasks and cultured in differentiation medium (DM) for 2, 4, 6 and 8 days.Smooth muscle specific markers, includingα-smooth muscle actin (α-SMA), SM22 and smooth muscle myosin heavy chain (SM-MHC), were detected to in-sure the successful model by qRT-PCR and Western blot .After 3-day pre-differentiation, ESCs were treated with recombinant Rspo 2 protein, overexpression plasmid or shRNA plasmid for 96 h, and the mRNA and protein expression of smooth muscle markers was detected.To explore the role of Rspo2 on SMC differentiation in vivo, 3-day predifferentiated ESCs (106 in 50μLα-MEM) incubated with Rspo2-overexpression plasmid were mixed with 50 μL of Matrigel ( Becton Dickinson Labware ) and then subcutaneously injected into C57BL/6J mice.After 12 days, mice were sacrificed and the implants were harvested for immunofluorescence staining , qRT-PCR and Western blot.Furthermore, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation assay (ChIP) and lucif-erase reporter assay were performed to investigate the transcriptional activity of SMC differentiation related transcription factors , inclu-ding serum response factor (SRF), myocardin (MYO), myocyte-specific enhancer factor 2C (MEF-2C).Involvement of Rspo2 re-ceptor, leucine-rich repeat-containing, G-protein-coupled receptors (Lgr)4,5,6, and β-catenin pathway during Rspo2-induced MSC differentiation were also uncovered by overexpression or inhibition of the respective protein .RESULTS:Our results showed that Rspo 2 mRNA and protein expression was significantly and consistently increased during ESC differentiation towards SMCs .Recombinant Rs-po2 protein and enforced Rspo 2 expression in ESCs resulted in up-regulation of smooth muscle markers and transcription factors , while knockdown decreased the expression of these genes .Expectedly , Rspo2 overexpression also promotes SMC differentiation in vivo.
Mechanistically , our data showed that Rspo 2 could promote SRF binding to SM22 promoter region .Evidence also revealed that one of three Rspo2 receptors, LGR5, was up-regulated while the other two , LGR4 and LGR6, was down-regulated.Silencing of LGR5 inhibi-ted Rspo2-induced SMC differentiation, whereas knockdown of LGR4 had no impact.Finally, activation or inhibition of β-catenin could promote or inhibit SMC differentiation , respectively .CONCLUSION: Our findings demonstrate for the first time that Rspo 2 plays a positive role during smooth muscle cell differentiation from embryonic stem cells .We confirmed that Rspo 2 can up-regulate smooth muscle markers at transcription level .We also revealed Rspo promote smooth muscle cell differentiation through activation of LGR 5 re-ceptor and Wnt/β-catenin pathway .