This paper is to report the exploration of the activation of Rho/ROCK signal pathway in 5-HT-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs) and the inhibitory effect of m-Nis on this pathway. PASMCs were cultured with the explant technique. MTT assay was used to explore the proliferation of PASMCs after 5-HT treated for different time and the intervening effect of m-Nis. RT-PCR and Western blot were used respectively to explore the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 in 5-HT-treated PASMCs and intervening effect of m-Nis. The results of MTT assay suggested that 5-HT (1 µmol · L(-1)) treatment for 12-72 h significantly induced the proliferation of rat PASMCs (P<0.05 or P < 0.01), which were inhibited by m-Nis (1 x 10(-5), 1 x 10(-6), l x 10(-7), 1 x10(-8) mol · L(-1)) in dose-dependent manners (P < 0.05 or P < 0.01). Similarly, the mRNA expression of RhoA, ROCK1 and the protein expression of p-MYPT1 were also inhibited by m-Nis in different degrees (P < 0.05 or P < 0.01). Thus, the results of this study suggested that Rho/ROCK pathway played an important role in 5-HT-induced proliferation of rat PASMCs, m-Nis inhibited 5-HT-induced proliferation obviously, which may be related to the blockage of Rho/ROCK signal pathway.
Animals ; Cell Proliferation ; drug effects ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Nisoldipine ; pharmacology ; Protein Phosphatase 1 ; metabolism ; Pulmonary Artery ; cytology ; Rats ; Serotonin ; pharmacology ; Signal Transduction ; rho-Associated Kinases ; metabolism ; rhoA GTP-Binding Protein ; metabolism

This study established an HPLC fingerprint of Tibetan medicine Shaji Gao from different habitats and lay a foundation for Shaji Gao varieties identification and preparation process. The chromatographic condition was as follow: Agilent zorbax SB-C18 (4.6 mm x 250 mm, 5 μm) eluted with the mobile phases of acetonitrile and 0.4% phosphoric acid water in gradient mode. The flow rate was 1.0 mL x min(-1), and the detection wavelength was set at 360 nm. The fingerprints of 15 batches Shaji Gao were carried out by similarity comparation, 7 chromatographic peaks were extracted as the common peaks of fingerprint, 3 peaks were identified, which were quercetin, kaempferol and isorhamnetin. The similarity degrees of 14 batches of samples were above 0.9 and 1 batch of samples was below 0.9. This is the first established fingerprint of Shaji Gao by using HPLC. This method has good precision, stability and repeatability that it could provide basis for quality control and evaluation of Shaji Gao.
Chromatography, High Pressure Liquid ; methods ; Medicine, Tibetan Traditional ; Quality Control

Objective To analyze the clinical characteristics of pneumomediastinum in patients with dermatomyositis and polymyositis for demonstrating its pathogenesis and for predicting its prognosis. Methods The clinical records of 96 patients with PM/DM were reviewed, focusing on for perdicting its pneumomediastinum. Five patients with pneumomediastinum are described in detail. Case reports of pneumomediastinum in PM/DM in English publications are reviewed. Results Five DM cases complicated by pneumomediastinum all had lung infections. Twenty-nine cases (including our five cases) of DM/PM with pneumomediastinum have taken methylprednisolone, four cases alive, and six died. Nine cases have taken CsA,seven cases alive and two died. Conclusion The infections was strongly suspected as being responsible for the pneumomediastinum. Methylprednisolone has poor effect. CsA can be an effective therapeutic agent in PM/DM.

0.05).At different time points after ischemia-reperfusion,the expression of cytochrome C and activation of caspase-3 were lower in the transgen mice than that in the wild type rats.Conclusions Under standard condition,overexpression of bcl-xl could significantly reduce the infarct area and improve neurological function in transgene mice than those in the wild type rats.The effect of overexpression of bcl-xl might be realized through inhibiting the apoptosis of neuron,and the mechanism might be that the overexpression of bcl-xl inhibit the release of cytochrome C and the activation of caspase-3.

The internship is the transition period of a medico becoming a doctor,the cultivation of clinical work ability of interns is a comprehensive ability cultivation which includes the foundation theories' consolidation and use,the practical operative train- ing,the cultivation of clinical thought ability and communication between doctors and patients,etc.To educate pediatrics intern has its characteristics.

OBJECTIVETo investigate the effect of Gli1 gene silencing by RNA interference (RNAi) on proliferation of K562 cells and its mechanisms.

METHODSThe small interference RNA (siRNA) was synthesized in vitro. K562 cells were transfected with Gli1 siRNA by the way of lipofection (lipofectamine 2000). Non-specific siRNA transfected cells were used as control. Transfection efficiencies of different siRNA concentrations were detected by flow cytometry and the best siRNA concentration was selected. The silencing effect of siRNA was demonstrated by real time PCR and Westem blot analysis. Cell proliferation was measured by MTT method, cell cycle by PI assay, c-myc and p21 mRNA level was detected by real time PCR analysis.

RESULTSTransfection efficiency of siRNA was increased in a dose-dependent manner when siRNA concentration was below 200 pmol, and the highest transfection efficiency reached (80.11 ± 5.63)%. Both the mRNA and protein level of Gli1 was down-regulated in Gli1 specific siRNA group, the mRNA level was (52.60 ± 3.57)% of that of control group after 24 h (t = 20.33, P < 0.01) and the protein level was (79.31 ± 5.58)% of that of control group after 48 h (t = 6.54, P < 0.01). The cell proliferation rate in Gli1 siRNA group was (94.41 ± 3.58)% (t = 2.40, P = 0.05) and (90.22 ± 3.34)% (t = 4.37, P < 0.01) of that of control group after 24 h and 48 h, respectively. G(2)/M cell cycle arrest was observed, the mRNA level of c-myc was down-regulated while p21 was up-regulated in Gli1 siRNA group after 24 h and 48 h (P < 0.05).

CONCLUSIONSTargeted silencing of Gli1 gene by RNAi inhibits the proliferation of K562 cells, which acts through the down-regulation of c-myc and up-regulation of p21 expression.

Cell Proliferation ; Gene Silencing ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transcription Factors ; genetics ; Transfection ; Zinc Finger Protein GLI1

Recently, novel Psidium meroterpenoids were reported in the guava leaves. According to careful analysis of the spectral data of literatures, the spectroscopic characteristics and biosynthetic pathway of Psidium meroterpenoids were summarized in this paper. The results showed that Psidium meroterpenoids had distinct spectroscopic features and reasonable biosynthetic routines, however the number order of carbon atoms was not consistent in the reported literatures. It was concluded that Psidium meroterpenoids were the characteristic chemical constituents of Psidium guajava Linn.
Magnetic Resonance Spectroscopy ; Plant Leaves ; chemistry ; Psidium ; chemistry ; Spectrum Analysis ; Terpenes ; chemistry

Objective To retrospectively evaluate the safety ,technical success rate and long‐term efficacy of the hepatic mul‐tiple cavernous hemangioma with super selective arterial cheoembolization .Methods 6 cases multiple hepatic cavernous hemangio‐ma by clinical diagnosed between 2004-2011 years in our hospital ,Through arterial super selective and completely filling cheoem‐bolization by Pingyang mycin lipiodol emulsion(PYM‐Lip) ,To assess the long‐term efficacy .by multi slice spiral CT enhanced scan‐ning and carry on relevant statistics processing in postoperative 6 ,12 ,36months .Results 26 lesions were embolismed in 6 cases multiple hepatic cavernous hemangioma ,Among the number of successful embolization were 15 of 1 cases ,2 of 4 cases ,3 of 1 cases , respectively .26 lesions was decreased with different degrees ,which the diameter of lesions were reduced with embolismed by CT enhanced scanning in postoperative 6 ,12 ,36months and diameter reduced> 50% ,diameter reduced≤50% ,lesions disappear was 38% (10/26) ,54% (14/26) ,8% (2/26) ,62% (16/26) ,23% (6/26) ,15% (4/26) ,69% (18/26) ,12% (3/26) ,19% (5/26) .Technical operation success rate 100% ,not serious complications occurred .There are statistically significant differences in the size of lesions before and after operation(P<0 .01) .Conclusion The technique success rate was high ,minimally invasive ,the complications was less ,the curative efficacy was obvious by transcatheter arterial super selective cheoembolizaton with hepatic multiple cavernous he ‐mangioma .

To investigate cytotoxic secondary metabolites of Micrococcus sp. R21, an actinomycete isolated from a deep-sea sediment (-6 310 m; 142 degrees 19. 9' E, 10 degrees 54. 6' N) of the Western Pacific Ocean, column chromatography was introduced over silica gel, ODS, and Sephadex LH-20. As a result, eight compounds were obtained. By mainly detailed analysis of the NMR data, their structures were elucidated as cyclo(4-hydroxy-L-Pro-L-leu) (1), cyclo(L-Pro-L-Gly) (2), cyclo( L-Pro-L-Ala) (3), cyclo( D-Pro-L-Leu) (4), N-β-acetyltryptamine (5), 2-hydroxybenzoic acid (6), and phenylacetic acid (7). Compound 1 exhibited weak cytotoxic activity against RAW264. 7 cells with IC50 value of 9.1 μmol x L(-1).
Animals ; Biological Factors ; chemistry ; isolation & purification ; metabolism ; pharmacology ; Cell Survival ; drug effects ; Macrophages ; cytology ; drug effects ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Mice ; Micrococcus ; chemistry ; genetics ; isolation & purification ; metabolism ; Molecular Structure ; Phylogeny ; RAW 264.7 Cells ; Seawater ; microbiology ; Secondary Metabolism

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics