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.

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

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.

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 prepare a single-epitope polyclonal antibody against mouse long peptidoglycan recognition protein (mPGRP-L).

METHODSB cell dominant epitopes of mPGRP-L predicted by bioinformatics were synthesized, and the immunogen was prepared by conjugation of the synthetic peptide and the carrier protein key-hole limpet hemocyanin (KLH) by MBS method. The single-epitope polyclonal antibody was obtained by immunizing rabbits with the KLH-peptide conjugate, purified by SPG affinity columns or antigenic peptide affinity columns, and identified by ELISA and Western blotting.

RESULTS AND CONCLUSIONA dominant epitope in N-terminal region of mPGRP-L, with amino acid sequence of NH(2)-(C)DPHSLSPELQALISEVAQHD-COOH, was chosen and synthesized. The titer of anti-serum of the rabbits immunized with the KLH-peptide conjugate was 1:256,000. The polyclonal antibody purified with SPG affinity columns and antigenic peptide affinity columns were named as mPGRP-Ln1 and mPGRP-Ln2, respectively. Western blotting demonstrated that the antibody mPGRP-Ln1 could recognize the recombined N-terminal fragment of mPGRP-Ln with a clear band at relative molecular mass of about 29,000, suggesting the successful preparation of the single-epitope polyclonal antibody against the N-terminal region of mPGRP-L.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; chemistry ; immunology ; isolation & purification ; Antibody Specificity ; immunology ; Blotting, Western ; Carrier Proteins ; chemistry ; immunology ; Enzyme-Linked Immunosorbent Assay ; Epitope Mapping ; Epitopes ; chemistry ; immunology ; Hemocyanins ; chemistry ; Immune Sera ; immunology ; Mice ; Molecular Sequence Data ; Protein Binding ; Rabbits

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

The differences of arrhythmias among distinct left ventricular geometric patterns in the patients with essential hypertension were studied. 179 patients with essential hypertension received 24 h dynamic ECG recording, ambulatory blood pressure monitoring, echocardiography examination, etc. According to the examinations, left ventricular geometric patterns and arrhythmias were identified. The comparison of morbidity of arrhythmias between the left ventricular remodeling group and the normal geometric pattern group was performed. The multiple stepwise regression analysis was carried out to identify the independent determinants of arrhythmias. After these predictors were controlled or adjusted, the severity of arrhythmias among different left ventricular geometric patterns was compared. It was found that the morbidity of atrial arrhythmia, ventricular arrhythmia and complex ventricular arrhythmias in the left ventricular remodeling group was significantly higher than in the normal geometric pattern group respectively. There were many independent factors influencing on arrhythmias in essential hypertension. Of all these factors, some indices of left ventricular anatomic structure, grade of hypertension, left atrial inner dimension, E/A, diastolic blood pressure load value at night and day average heart rate and so on were very important. After the above-mentioned factors were adjusted, the differences of the orders of arrhythmias between partial geometric patterns were reserved, which resulted from the differences of the geometric patterns. Many factors contributed to arrhythmias of essential hypertension, such as grade of hypertension, LVMI, LA, PWT and so on. The severity of arrhythmias was different in different left ventricular geometric patterns.

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.