Objective To investigate the effect of TanshinoneⅡA (TSN) on the cell hypertrophy induced by angiotensinⅡ(AngⅡ) in the primary culture of neonatal rat cardiomyocytes. Method The effect of TSN on cardiomyocyte was evaluated by the 3-[4, 5-dimethylthiazol-2-yl] -3, 5-diphenylformazan (MTF) assay. As the index of eardiomyocyte hypertrophy, protein synthesis rate was measured by H-Leucine incorporation and the cell size was determined by phase contrast microscope. The proto-oncogene c-los mRNA and c-jun mRNA expression was assessed using reverse transcription polymerase chain reaction (RT-PCR). Results Exposure of the myocytes to TSN (5～80 mmol/L) for 24hours produced no cytotoxicity. Protein synthesis rate and proto-oneogene c-fos and c-Jun mRNA expression of eardinmyoeytes increased significantly after AngⅡtreatment, and TSN inhibited these effect of AngⅡ.Conclusions TSN can prevent the hypertrophy of eardiomyocytes induced by AngⅡ, which be attributable relate to the decreased expression of proto-oncogene c-los and c-jun mRNA by TSN.

BACKGROUND:Among the factors causing vascular endothelial cell injury,angiotensin Ⅱ(Ang Ⅱ) caused by renin-angiotensin system (RAS), especially by local RAS, plays an important patho-physiological role.OBJECTIVE: To observe the effect of tanshinone ⅡA on the vascular endothelial cells secreting nitric oxide (NO) and endothelial nitric oxide synthase (eNOS) gene expression as well as intracellular Ca2+ level induced by Ang Ⅱ, and investigate the protective effect of tanshinone ⅡA on vascular endothelial cells.DESIGN: Controlled observation experiment.SETTING: Department of Emergency, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: This experiment was carried out in the Experimental Center for Basic Medicine, Tongji Medical College,Huazhong University of Science and Technology from March 2006 to October 2006. Porcine aorta used in this experiment was provided by the Department of Pathophysiology of Tongji Medical College.METHODS: Nitric acid deoxidization method and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the effects of Ang Ⅱ of different concentrations (10-8 to 10-6 mol/L) on endothelial cells secreting NO and eNOS mRNA expression in cultured porcine aortic endothelial cells at different time points (1,6 and 24 hours) separately, then 50 mg/L tanshinone Ⅱ A was respectively added at different time point (0, 6 hours) when Ang Ⅱ was at 10-6 mol/L, and changes in NO production and eNOS gene expression were detected respectively at 1, 6 and 24 hours. Intracellular Ca2+ level was also detected with laser scanning confocal microscopy.MAIN OUTCOME MEASURES: ① NO content. ②eNOS mRNA expression. ③Intracellular Ca2+ level.RESULTS: ① NO production and eNOS mRNA expression were decreased with increase of Ang Ⅱ concentration and prolongation of time (P ＜ 0.01). ② NO production and eNOS mRNA expression in each tanshinone ⅡA-treated group were significantly higher than those in the Ang Ⅱ group. At 1 and 6 hours of tanshinone ⅡA treatment, production of NO and eNOS mRNA expression in the Ang Ⅱ + tanshinone ⅡA group were significantly higher than those in the Ang Ⅱ 6 hours + tanshinone ⅡA group (P ＜ 0.05); There were no significant differences in NO production and eNOS mRNA expression between two groups at 24 hours (P ＞ 0.05). ③Intracellular Ca2+ level in the Ang Ⅱ group was significantly higher than that of control group (P ＜ 0.01), and intracellular Ca2+ level in the tanshinone ⅡA + Ang Ⅱ was significantly lower than that in the Ang Ⅱ group (P ＜ 0.05).CONCLUSION: Tanshinone Ⅱ A has a protective effect on vascular endothelial cells and their functions by suppressing the inhibition of Ang Ⅱ on NO level and eNOS gene expression in cultured porcine aortic endothelial cells.

Lipopeptides produced by Bacillus subtilis JA antagonized a broad spectrum of fungal pathogens. Crude lipopeptides were extracted with methanol from the precipitate which was obtained by adding 6 mol/L HCl to the cell-free culture broth.The crude extract was run on Diamonsil C_(18)column(5?m,250 mm?4.6 mm) in reverse phase HPLC system to purify the lipopeptides.Inhibitory ability and IC_(50)values of lipopeptides towards various microorganisms were determined by agar diffusion method.The results showed lipopeptides exhibited strong inhibitory activity against some important plant pathogenic fungi,including R.solani and F.oxysporum.The ability of B.subtilis JA to antagonize against the growth of the post-harvest pathogen -B,cinerea was tested in vitro.Spore germination of B.cinerea was strongly inhibited in the presence of JA cell suspension.Furthermore,B.subtilis JA can produce antifungal volatiles which strongly inhibited the spore germination and mycelial growth of B.cinerea.As a biocontrol agent,the synergic effect of lipopeptides and volatiles may play a major role in controlling the pathogens by B.subtilis JA.

Objective To study the intervention effect of tanshinone on electrophysiological abnormality of hypertrophic cardicoyte in order to illuminate the underlying mechanism of tanshinone in preventing the arrhythmia induced by myocardial hypertrophy. Method Twenty-week-rid SD rats (200～250 g) were divided into 4 groups (8 in each group) randomly. Of 4 groups, rats of three groups were operated on by a procedure of 'one kidney one clamp' to make renal artery constriction. The rest group served as sham operation group (control group). When the blood pressure increased,rats of operation groups were divided into tanshinone group, captopril group and hyper-trophic group. The effects of tanshinoe and captopril were observed and compared on the action potential duration (APD),L-type calcium current (ICa, L) and transient outward potassium current (Ito) density in cellular membrane of hypertrophic myocardium by using patch clamp and intra-cellular calcium survey technique. Results The blood pressure in operation groups was obviously higher than that in sham-operation group (P<0.01), but there was no difference between operation groups (P>0.05). The ratio of ventricle weight to body weight (VW/BW) was much higher in hypertrophic group than in control group (P<0.01), and it significantly decreased after interven-tion with tanshinone or captopril (P<0.01). Compared with hypertrophic group, tanshinone markedly shortened the prolongation of action potential duration (P<0.01), decreased membrane capacity and peak amplitude of ICa,L(P<0.01), but had no effect on the density of ICa,L. Tanshinone also significantly increased Ito current density and peak amplitude, which were completely different from hypertrophic group (P<0.05). There were similar results foundin captopril intervention. Conclusions Tanshinone could reduce calcium influx and resume the activity of ho ion channels, and thus shorten the first phase and the plateau phase of repolarization and decrease the prolongation of APD in hypertrophic cadiocyte. So tanshinone can prevent the onset of arrhythmia attributed to the myocardial hypertrophy.

Adolescent ; Adult ; Female ; Health Personnel ; Health Status ; Humans ; Middle Aged ; Occupational Health ; Young Adult

Eleven type 1 diabetic patients who received fixed regime of insulin Glargine were included in the study.The levels of fasting serum insulin were measured for each subject at 6:00 in three consecutive mornings.The variability of mean fasting serum insulin in each subject was 3.3%-41.5% (mean 15.4%).The variability did not correlate with the dose of Glargine statistically.

OBJECTIVETo observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells (MSCs) derived from bone marrow during osteogenic differentiation.

METHODSMSCs were induced to differentiate with MSC osteogenic differentiation medium for 7, 14, 21 and 28 days respectively. Alizarin Red staining was used to detect matrix mineralization. Expression of osteoblast-related genes, including osteocalcin, osteopontin, Runt-related transcription factor 2 (Runx2), alkaline phosphatase and collagen type 1, was assessed with quantitative reverse transcription-polymerase chain reaction.

RESULTSOn day 14 after induction of differentiation, cells were stained positively with Alizarin Red. The expression levels of these genes exhibited an upward trend as induction time was prolonged. Exposure to osteogenic differentiation medium less than 21 days did not significantly induce osteocalcin expression; osteocalcin expression levels in the differentiated cells induced for 21 and 28 days were 1.63 and 2.46 times as high as the undifferentiated cells respectively (all P<0.05). Stimulation with MSC osteogenic differentiation medium over 14 days significantly enhanced bone marrow-derived MSCs to express osteopontin and Runx2 genes (all P<0.05). Osteogenic differentiation medium could significantly induce the expressions of alkaline phosphatase and collagen type1 genes (all P<0.05). Their expressions reached the peak levels on day 21, which were increased more than 4- and 3-fold respectively.

CONCLUSIONHuman bone marrow-derived MSCs could exhibit the sequential expression pattern of osteoblast marker genes during osteogenic differentiation in vitro.

Alkaline Phosphatase ; genetics ; Cell Differentiation ; Cells, Cultured ; Collagen Type I ; genetics ; Core Binding Factor Alpha 1 Subunit ; genetics ; Genetic Markers ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Osteocalcin ; genetics ; Osteogenesis ; Transcriptome

AIM To establish an HPLC-ELSD method for the simultaneous content determination of gastrodin,parishin A,parishin B,ginsenosides Rg1,Re,Rf,Rb1 and Rd in Tiantai No.1 Tablets (Ginseng Radix et Rhizoma,Gastrodiae Rhizoma,Cistanches Herba,etc.).METHODS The analysis of methanol extract of this drug was performed on a 45 ℃C thermostatic Alltima C1scolumn (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrie-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient elution manner.RESULTS Eight constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 96.94％-97.93％ with the RSDs of 1.06％-2.48％.CONCLUSION This simple,accurate and reliable method can be used for the quality control of Tiantai No.1 Tablets.

Objective To investigate the role of white matter astrocytes and their specific protein S100A4 in sensory neurite outgrowth in vitro.Methods White matter astrocyte cultures expressing S100A4 were prepared. Dissociated adult dorsal root ganglion(DRG)cells were placed on the top of the astrocytes and co-cultured for 6,12, 18,24 hours.Small interfering S100A4 RNA was used to eliminate S100A4 expression.The growth of DRG cell neu- rites on S100A4-sileneed and S100A4-expressing astrocytes was compared.Results 12,18 and 24 hours after the co-culture with S100A4-expressing or S100A4-silenced astroeytes,neurite growth from the DRG cells was observed. Neurite outgrowth was significantly greater in S100A4 siRNA treated cultures compared to control siRNA treated white matter astrocyte cultures.Conclusion These findings suggest that white matter astroeytes are able to support axonal regeneration and,furthermore,that administration of small interfering S100A4 RNA provides strong additional support for axon regeneration.

Objective To investigate the correlation of pretreatment serum squamous cell carcinoma antigen(SCCAg)with the clinico-pathological features of squamous cell carcinoma of uterine cervix and its significance as a prognostic factor.Methods One hundred and fourteen patients of squamous cell carcinoma of the uterine cervix(Ⅰbl-Ⅱa),who underwent pretreatment serum SCCAg evaluation and long-term follow-up after treatment were selected for this study.Clinical data were used to investigate the correlation between SCCAg and clinico-pathological features and factors that influence prognosis through univariate and multivariate analysis.Results Univariate analysis showed elevation of SCCAg(using≤1.5 mg/L as the cut-off value)was correlated with tumor size,deep stromal invasion and pelvic node metastasis (P0.05).However,between no pelvic node metastasis+elevated SCCAg cases and no pelvic node metastasis+normal SCCAg cases,there was a significant difference in DFS (71.8% vs 98.0%,P=0.003),recurrence rate(33.3% vs 9.8%,P=0.006)and local recurrence (26.5% vs 2.1%,P=0.001).Conelusions The independent prognostic factors for Ⅰ bl-Ⅱa squamous cell carcinoma of uterine cervix include elevated pretreatment SCCAg and pelvic node metastasis. Patients with elevated pretreatment serum SCCAg and no metastasis to pelvic lymph node(s)are at significantly elevated risk of local recurrence,and therefore need individualized treatment to improve local control and long-term survival.