Objective: To determine the contents of paeoniflorin and glycyrrhizic acid in Gianggan Capsules by RP HPLC. Methods: The Chromatographic method was carried out on Kromasil C 18 column using acetonitrile phosphate buffer solution (pH=5) (20∶80) as a mobile phase. The detection wavelength was set at 230nm and 250nm. The flow rate was 1ml/min. The column temperature was 40℃. Results: The paeoniflorin and glycyrrhizic acid calibration curves were linear at the ranges of 0.48～4.80?g and 1.08～10.81?g(r=0.9999). The average recoveries were 100.5% with RSD 1.2% for paeoniflorin and 98.9% with RSD 0.9% for glycyrrhizic acid. Conclusion: This method is simple, reliable and provides a reference standard for the quality control of Gianggan Capsules.

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.

Objective To investigate autoantibody against endothelin receptor type A (ENRA-Ab) in patients with systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH).The possibility of autoantibody-mediated pathogenesis in the development of SLE-PAH has also been explored.Methods ENRA-Ab in the serum of SLE-PAH and controls were detected by using a human ETRA epitope peptide-based ELISA.The clinical relevance of ENRA-Ab in SLE-PAH was analyzed.Proliferation of vascular smooth muscle cells (SMCs) and permeability of endothelial cells in vitro under the stimulation of polyclonal ENRA-Ab IgG were assessed.The expressions of PAH-related markers, i.e., 5-HTT, PDGFR-b, VEGF-A and PDGF-B were measured by qPCR.The effect of ENRA-Ab in vivo was also determined in a suboptimaldose monocrotaline-induced model with the assessment of right ventricle hypertrophy index and pathology parameters.Independent t-test, Tukey-Kramer test of variance analysis and Pearson' s correlation analysis were used for statistical analysis.Results ENRA-Abs was presented in a higher occurrence in SLE-PAH (35/85,41％) compared with controls (0/60;0, 13/80, 16％).There was a significant correlation between ENRA-Ab and echocardiograph estimated pulmonary arterial systolic pressure (r=0.392, P=0.002) in SLE-PAH.ENRA-Ab could promote SMCs proliferation, disrupt endothelial barrier and up-regulate PAH-related markers expression,which could be blocked in the presence of ETR antagonist.ENRA-Ab aggravated right ventricle hypertrophy and vascular remodeling in vivo.Conclusion ENRA-Ab is a new biomarker, in SLE-PAH, which may mediate PAH development in SLE.

Objective To investigate the effectiveness and safety of minimal invasive surgery combined with ventriculoscope for the treatment of basal ganglia hemorrhage.Methods From June 2014 to June 2015 there were 57 patients with basal ganglia hemorrhage were selected into this study.These patients were signed into the minimal invasive surgery combined with ventriculoscope group and the craniotomy group according to the methods of surgery they received.And the clinical outcomes of the two groups were compared.Results Compared with the craniotomy group,patients in the minimal invasive surgery combined with ventriculoscope group got a significantly lower Glasgow coma scale at 7 days,14 days and 28 days after the operation (P =0.02,0.04,0.04);the hospital stays were significantly reduced in the minimal invasive surgery combined with ventriculoscope group [(21.45 ±5.67)d vs.(25.67 ±7.45)d,P =0.02];and the operation time were significantly reduced as well [(134.45 ±21.11)min vs.(178.65 ±45.32)min,P =0.000)].There was no significant difference in intra-cranial pressure,rate of hematoma clearance,rate of organ functional failure,rate of re-bleeding and mortality 28 days after operation (P >0.05).Conclusion The minimal invasive surgery combined with ventriculoscope is effective and safe for the treatment of basal ganglia hemorrhage,which is worthy of popularization.

Objective To investigate the anti-fibrosis effect of a 5-lipoxygenase (5-LO)specific inhibitor AA-861 on liver fibrosis. Methods Liver fibrosis was induced in male Sprague Dawley rats by intraperitoneal injection of carbon tetrachloride(CCI4).AA-861(0.2 mg/100 g/d)was administrated by intraperitoneal injection starting 2 days before the first dosage of CCI4 and rats were killed at weeks 2,4,and 6.Liver specimens were obtained from each animal and fixed with 4%formaldehyde for histological analysis. The Mrna expression of 5-LO,smooth muscle-alpha(α-SMA),Collagen-1,matrix metalloproteinase-2(MMP-2)and tissue inhibitors of metalloproteinase-1(TIMP-1),the protein expression of 5-LO were evaluated by real time PCR and Western blot respectively. Histological analysis was performed by microscopy observation. Fibrosis conditions in rat liver in each group were evaluated according to the semi-quantitative scoring system (SSS), Hyp in rat livers and the hepatic functional biochemistry were also detected. Results Along with the aggravation of liver fibrosis, the gene expression of 5-Lo,α-SMA,Collagen-1,MMP-2 and TIMP-1 increased gradually, and the level of ALT, TBA, Hyp and SSS score also increased gradually. With the administration of AA-861,the Mrna expression of 5-LO,α-SMA,Collagen-1,MMP-2,TIMP-1 and the protein expression of 5-LO decreased remarkably, and the reduction in TIMP-1 Mrna expression was more significant than that in MMP-2.Six weeks after AA-861 treatment, the level of ALT, TBA, Hyp and SSS score were also decreased significantly. Conclusion AA-861 ameliorates CCI4 induced rat liver fibrosis by inhibiting the 5-LO pathway and decreasing the expression of 5-LO,α-SMA,Collagen-1,MMP-2 and TIMP-1.

Objective: To investigate the inhibition of tanshinoneⅡA (TanⅡA) on the proliferation of lung cancer cells. Methods: Human lung cancer cell lines A549, SK-MES-1, H446 and H460 were cultured in vitro and treated with TanⅡA at concentrations of 0.3, 0.6, 1.3, 2.5, 5.0, 10.0 mg/mL, while untreated cells served as controls. Cell proliferation was measured by using CCK-8 assay, and apoptosis was measured using flow cytometry, while the expression of apoptosis-related proteins was determined using Western blotting. The apoptotic rate of lung cancer cells was compared between Tan ⅡA-treated cells and untreated cells. Results: Tan ⅡA inhibited lung cancer cell proliferation in a time-dependent and concentration-dependent manner, and the survival rates of A549, SK-MS-1, H446 and H460 cells reduced with the concentration of TanⅡA (t=4.503, 2.114, 2.103 and 3.567; all P<0.05) and the duration of TanⅡA treatment (t=5.189, 3.079, 3.023 and 3.845; all P<0.05). The 48 h half maximal inhibitory concentrations (IC50 values) of TanⅡA were (1.18±0.12), (0.78±0.08), (1.55±0.16) and (1.27±0.14) mg/mL against A549, SK-MES-1, H446 and H460 cells, respectively. Following 48 h treatment with Tan ⅡA at a concentration of 2.5 mg/mL, the apoptotic rates of A549, SK-MS-1, H446 and H460 cells were (34.97±3.78)%, (37.62±2.48)%, (18.27±2.98)% and (19.17±2.30)%, which were significantly significantly higher than those of untreated cells [(4.86±0.36) %, (3.21±0.48) %, (3.25±0.26)% and (2.66±0.19)%, all P<0.05]. Reduced Akt1 protein expression, elevated Bax/Bcl-2 expression ratio, and elevated caspase-9 and caspase-3 protein expression were detected in lung cancer cells treated with 2.5 mg/mL TanⅡA for 48 h relative to untreated cells Conclusion TanⅡA may inhibit lung cancer cell proliferation in a time- and concentration-dependent manner via the Bax/Bcl-2/caspase-9/caspase-3 pathway.

Objective To determine the probability of identification of differential expression of biliary proteins induced by cholangiocarcinoma using 2D-DIGE. Methods Bile was obtained from 12patients with obstructive jaundice (including 6 cases of cholangiocarcinoma and 6 of cholelithiasis).Each sample was labeled with three different CyDyes (y3,Cy5,Cy2) including one internal standard,pooled from all the samples, and separated with 2-D DIGE in triplicate experiments. MALDI-TOF-MS and bioinformatics were adopted to identify and elucidate the significance of differentially expressed proteins in bile induced by cholangiocarcinoma. Results 55 matched protein spots differences in abundance were detected with statistical variance of two groups(Average Volum Ratio ≥1.5, t-test, P＜0. 05). Among these proteins, 13 PMF were obtained by MALDI-TOF-MS analysis. Eight proteins were identified by searching a protein database. Conclusion The differentially displayed proteomes between the pathological bile obtained from benign and malignant obstructive jaundice indicates the potential application of 2D-DIGE to identify the biomarker of cholangiocarcinoma.

Objective To know the Tanshinone Ⅱ A induce human small cell lung cancer cells (H446 cells) apoptosis,and discuss the possible molecular mechanism.Methods Determined by MTT method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells proliferation;Determined by Hoechst 33258 method of different concentrations Tanshinone Ⅱ A influence on human small cell lung cancer H446 cells apoptosis;Determined by qPCR method of different concentrations of Tanshinone Ⅱ A impact on human small cell lung cancer H446 cells apoptosis gene;Determined by Westen blot method of different concentrations of Tanshinone lⅡ A effect on lung cancer H446 cells apoptosis.Results Different drug concentrations of Tanshinone Ⅱ A (0.3,0.6,1.25,2.5,5 and 10μg/ml) inhibit human small cell lung cancer H446 cells proliferation and depend by the concentration and time;With the increase of the concentration of the drug in the formation of human small cell lung cancer H446 cells apoptosis corpuscle number increase,significant difference was found in high concentration group (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promote apoptosis gene (Bax,caspase-9,caspase-3) expression,and inhibit apoptosis inhibiting genes (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05);High concentration group of Tanshinone Ⅱ A promoting apoptosis protein (Bax,caspase-9,caspase-3)expression,and inhibit apoptosis inhibiting protein (Akt,the Bcl-2) expression and there were significant differences (P ＜ 0.05).Conclusion Tanshinone Ⅱ A may be through Akt-Bax/Bcl-2-caspase-9-caspase-3 signaling pathways induced human small cell lung cancer H446 cells apoptosis.

Objective To compare the performance of four commercial anti-dsDNA antibody assays,i.e,BioPlex 2200 (BioPlex),Farr radioimmunoassay (Farr),MESACUP DNA-Ⅱ TEST ds [MBL-enzyme linked immunosorbent assay (ELISA)] and Anti-dsDNA-NcX ELISA (IgG) (EURO-ELISA),Antoantibodies Profile Assay Kit (HOB-Chemiluminescent Immunoassay) in disease activity assessment of systemic lupus erythematosus (SLE).Methods SLE patients (n=119) as well as healthy controls (n=200) and disease controls (n=100) were recruited and their serum anti-dsDNA antibodies were detected by BioPlex,Farr,MBL-ELISA,EURO-ELISA,and a standard Crithidia luciliae indirect immunofluorescence test (CLIFT).The consistency between above four methods to CLIFT was analyzed.The correlation of anti-dsDNA antibody level of these four methods to SLE disease activity was assessed.All data analyses were performed with Statistical product and service solutions (SPSS) 16.0 (SPSS.Inc) and GraphPad Prism 4.0.3 (GraphPad).Unless otherwise specified,all data in this study were expressed as mean±standard deviation.Cut-off values of the anti-dsDNA quantification methods were set by the manufacturers.Chi square and kappa coefficients were adopted to assess the agreement determination and correlation analysis between anti-dsDNA level and SLE disease activity (SLEDAI).Receiver-operator characteristic (ROC) curve analysis was used to compare the specificity and sensitivity of the anti-dsDNA assays.Student's t test was adopted for the comparison of anti-dsDNA levels by different methods between SLE and SLE+LN groups.A p value small than 0.05 was considered statistically significant.Results Using cut-off values set by the manufacturers,BioPlex demonstrated the highest overall agreement with CLIFT,while MBL-ELISA and EURO-ELISA showed the highest positive agreement with CLIFT.Disease activity correlation analysis showed that SLEDAI score correlated poorly with anti-dsDNA level in Farr assay,but strongly with the other three assays.Bioplex had a better performance in terms of SLE activity index corelation (r=0.297 6,P=0.001 2).Moreover,anti-dsDNA level differed in SLE patients with renal lupus nephritis in BioPlex assay (P=0.026 8),but not in the other assays.In ROC curve analysis,BioPlex showed the largest area under the curve (AUC) over other assays.Conclusion Bio Plex assay has better sensitivity and specificity than Farr,MBL-ELISA and EURO-ELISA and correlates well with SLE disease activity.

Sugarcane molasses containing large amounts of sucrose is an economical substrate for succinic acid production. However, Escherichia coli AFP111 cannot metabolize sucrose although it is a promising candidate for succinic acid production. To achieve sucrose utilizing ability, we cloned and expressed cscBKA genes encoding sucrose permease, fructokinase and invertase of non-PTS sucrose-utilization system from E. coli W in E. coli AFP111 to generate a recombinant strain AFP111/pMD19T-cscBKA. After 72 h of anaerobic fermentation of the recombinant in serum bottles, 20 g/L sucrose was consumed and 12 g/L succinic acid was produced. During dual-phase fermentation comprised of initial aerobic growth phase followed by anaerobic fermentation phase, the concentration of succinic acid from sucrose and sugarcane molasses was 34 g/L and 30 g/L, respectively, at 30 h of anaerobic phase in a 3 L fermentor. The results show that the introduction of non-PTS sucrose-utilization system has sucrose-metabolizing capability for cell growth and succinic acid production, and can use cheap sugarcane molasses to produce succinic acid.
Bioreactors ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; genetics ; Fermentation ; Membrane Transport Proteins ; genetics ; Metabolic Engineering ; Molasses ; Saccharum ; chemistry ; Succinic Acid ; chemistry ; Sucrose ; chemistry