OBJECTIVE To investigate the effect of tranilast on cardiac fibroblasts proliferation and phenotype transformation incubated with a high concentration of glucose and the possible mechanism. METHODS The cardiac fibroblasts were divided into seven groups in accordance with different nutrient solutions:normal control group(5.5 mmol · L-1 glucose),hypertonic group(5.5 mmol · L-1 glucose+mannitol 25 mmol · L- 1),high glucose group (25 mmol · L- 1 glucose),tranilast intervention groups (25 mmol·L-1 glucose+tranilast 50,100 and 200μmol·L-1),and activin receptor-like kinase 7(ALK7) inhibitor group(25 mmol · L-1 glucose+10μmol · L-1 SB431542). The cell proliferation was detected by MTT method. The transformation of cardiac fibroblasts was determined by immunfluorescence staining. The expression of fibroblast specific protein 1 (FSP-1),α-smooth muscle actin (α-SMA),transforming growth factor-β1(TGF-β1)and ALK7 was assessed by Western blotting. RESULTS Compared with nor?mal control group,A492 nm of cells in high glucose group was significantly increased(P<0.01),while the expression of α-SMA,TGF-β1 and ALK7 protein in high glucose group was significantly increased(P<0.01),but FSP-1 protein was significantly decreased(P<0.01). There was no difference between normal control group and hypertonic group. Compared with high glucose group,A492 nm of cells in tranilast or SB431542 intervention groups were decreased(P<0.05),and the expression of α-SMA,TGF-β1 and ALK7 protein was significantly decreased(P<0.05),but the expression of FSP-1 protein was increased (P<0.05)in tranilast or SB431542 intervention groups. CONCLUSION Tranilast can inhibit the proliferation and phenotype transformation of cardiac fibroblasts induced by high glucose,which may be related to down-regulation of the expression of ALK7.

BACKGROUND: In according to ISO-10993-4 and GB/T 16886.4, the in vitro hemo-compatibility evaluation on biomaterieisincludes thrombosis, coagulation factors, platelets and platelet functions, hematology and complement system. However, in thecase of China, the in vitro hemo-compatibility evaluations were performed only thrombosis, coagulation factors and plateletattachment, the investigation on evaluation of platelet and complement activations is less reported.OBJECTIVE: To evaluate the effect of polyethylene, polyvinyl chloride and polymethylvinylsiloxane tubes on platelet activation,and establish a useful method to evaluate the effect of tubular materials on platelet activation.METHODS: Tubes of polyethylene, poiyvinyl chlorid and silastic were established by 3.7 mm inner diameter, 3.5 mm externaldiameter, and 35 cm length, respectively. 1 mL blood was injected into the tube of polyethylene, polyvinyl chlorid and silasUc,respectively. The tubes were connected using a two-way tube, shaken at 140 r/min by 30° sloping for 3.5 hours at 37 ℃.Radiolmmunoassay was employed to detect α-granules protein level of platelet poor plasma, while flow cytometry was used todetect the percentage of positive plateiet of o-granules protein and that of activated gp Ⅱb/Ⅲa composite.RESULTS AND CONCLUSION: Radiolmmunoassay showed that o-granules protein level of plateiet poor plasma in thepolyethylene and polyvinyl chlorid tubes was significantly greater than that in the silastic tube (P < 0.05). There were no significantdifferences in o-granules protein between polyethylene and polyvinyl chlorid (P > 0.05). Flow cytometry indicated that percentageof positive platelet of o-granules protein in the polyethylene and polyvinyl chlorid tubes was significantly greater than that in thesilastic tube (P < 0.05); the percentage in the polyethylene tube was significantly greater than that in the polyvinyl chlorid tube (P <0.05). There was no significant differences in the percentage of positive plateiet of activated gp IIb/IIIa composite between thethree materials (P > 0.05). A useful blood-material contact model was established, and it was considered that o-granules protein isan available parameter for evaluating platelet activation. The percentage of positive platelet of o-granules protein determined byflow cytometry was a more sensitive parameter for evaluating platelet activation.

Objective To develop a method to determine Pb and Cd in Qingkailing Injection by graphite furnace atomic absorption spectrometry, and search their source. Methods The samples, digested via microwave, were determined the contents of Pb and Cd by graphite furnace atomic absorption spectrometry in raw material, intermediate and finished product of Qingkailing Injection. Results The standard curve of Pb was Y=0.007 3X+0.011 6, and Cd was Y=0.056 7X+0.060 4. The regression equation of Pb and Cd was 97.0% and 95.6%, respectively. Content determination of Pb and Cd in Qingkailing Injection revealed that Pb and Cd in finished product came from raw materials. Conclusion The method is rapid, high sensitive and accurate, and can be applied to the inspection of Pb and Cd in Qingkailing Injection.

砄bjective:To investigate the effects of salivary statherin on adherence of two kinds of main cariogenic bacteria. Methods: Human whole salivary statherin was separated and purified by high performance hydrophobic interaction chromatography (HPHIC) and was further identified by SDS PAGE electrophoresis and amino acid analysis.Then the adherence of Streptococcus mutans ( S.mutans serotype c,7H) and Sreptococcus sanguis (S.sanguis ,ATCC 10557)to hydroxyapatite (HA), which was covered with purified statherin or whole saliva (positive control) or PBS buffer (negative control) as experimental pellicles respectively,was studied by bacteria counting.Results:①More S.sanguis adhered to the experimental pellicles than S.mutans ( P

TcdA and B toxins secreted by Clostridium difficile( CD) are two important causes of diseases in organisms. The expression of tcdA and tcdB genes is regulated by a few factors located in the pathogenicity locus ( PaLoc) .Studies have indicated that the tcdC gene is likely to act as a negative regulator of toxin gene expression.So far, it has been debatable whether tcdC gene is regarded as a negative regulator.The mechanism of tcdC gene in pathogenesis remains unclear.In this paper, the structure and function of the tcdC gene are summarized, which will help study the mechanism of tcdC gene and obtain optimal drug targets.

Objective To explore the clinical significance of T lymphocyte subsets and NK cells' detection in peripheral blood of patients with non-Hodgkin' s lymphoma (NHL).Methods 62 newly diagnosed NHL and 30 healthy adults were enrolled for analyzing.T lymphocyte subsets and NK cells were deteced by flow cytometry and compared between different groups according to age,sex and pathology.Multiple linear regression was used to analyze the correlation among the levels of T lymphocyte subsets and NK cells and several clinical factors,such as clinical stage,IPI and B symptoms.Results The proportions of CD3,CD4 and NK cells in peripheral blood of NHL group [(64.13±19.83) ％,(27.10±8.20) ％,(13.51± 10.59) ％] were significantly lower than those of normal control [(65.78±10.69) ％,(31.95±12.74) ％,(18.76± 7.36) ％] (P ＜ 0.05),while the proportions of CD8 and Treg cells in peripheral blood of NHL [(32.15±13.83) ％,(10.44±3.00) ％] were significantly higher than those of normal control [(29.25±12.35) ％,(7.51±4.36) ％] (P ＜ 0.05).In NHL,the proportions of CD3 and NKT cells [(67.06±19.24) ％,(4.91±3.69) ％] in ≤60 years old group were significantly higher than those in ＞ 60 years old group [(59.18±20.33) ％,(4.89±3.05) ％] (P ＜ 0.05).The proportions of Treg cells in T-NHL was significantly higher than that in B-NHL [(8.17±6.41) ％ vs (7.11±2.53) ％] (P ＜ 0.05).The proportions of CD3,CD4,NK and NKT cells in peripheral blood of NHL were significantly correlated with clinical stage (P ＜ 0.05),that of CD8 cells were significantly correlated with IPI (P =0.000),that of Treg cells were significantly correlated with IPI and B symptoms (P ＜ 0.05).Conclusion The cellular immune is suppressed in NHL patients.The proportions of T lymphocyte subsets and NK cells in peripheral blood of patients with NHL are different between different groups according to age,sex and pathology.The correlations among that and clinical factors need to be further explored.

Objective To analyze the effects of individualized lifestyle intervention on compliance and metabolic status of patients with type 2 diabetes mellitus (T2DM).Methods Two hundred T2DM patients were selected and randomly divided into experimental and control groups of 100 patients respectively.The experimental group was given individualized lifestyle intervention for 6 months in addition to conventional oral medications.The intervention was to prescribe diet control and exercise therapy according to the patients' individual conditions.The control group was given conventional treatment and verbal lifestyle intervention for 6 months.Comparison was made in patients compliance and various metabolic markers between the two groups.Results The percentage of conduction of diet control and exercise therapy in experimental group was significantly higher than control group ( Diet control:80 vs.52,x2=7.08,P=0.029;Exercise therapy:78 vs.44,x2=11.207,P=0.004).After intervention,the fasting plasma glucose (FPG),2-hour postprandial blood glucose (2hPG),glycated hemoglobin ( HbA1c),body mass index ( BMI),triglyceride ( TG),total cholesterol (TC),low-density lipoprotein ( LDL-C ),and insulin resistance index ( HOMA-IR ) in experimental group decreased significantly,and high-density lipoprotein ( HDL-C ) increased significantly [FPG:( 8.45 ± 1.46 ) mmol/L vs.(6.66 ± 0.67) mmol/L,P=0.000;2hPG:( 12.76 ± 2.25 ) mmol/L vs.(8.22 ± 1.79) mmol/L,P=0.000;HbA1c:(7.68 ± 1.06 ) ％ vs.( 6.48 ± 0.69 ) ％,P=0.000;BMI:( 25.90 ± 1.72 ) kg/m2 vs.( 22.81 ±1.41 ) kg/m2,P=0.016;TG:(2.57 ±0.68) mmol/Lvs.( 1.88 ±0.35) mmol/L,P=0.006;TC:(5.72 ±0.13) mmol/L vs.(5.14 ± 1.38) mmol/L,P=0.043;LDL-C:(3.28 ±0.10)mmol/L vs.(2.81 ±0.57)mmol/L,P=0.009;HOMA-IR:7.58 ± 0.19 vs.4.58 ± 1.98,P=0.000;HDL-C:( 1.29 ± 0.04) mmol/L vs.( 1.62 ± 0.27 ) mmol/L,P=0.003].The levels of FPG,2hPG,HbA1c,BMI,TG,HOMA-IR also decreased in control group after intervention compared with before intervention [FPG:( 8.67 ± 2.71 ) mmol/L vs.( 7.26 ± 1.21 ) mmol/L,P=0.001;2hPG:( 12.82 ± 2.15 ) mmol/L vs.( 10.85 ± 1.98 ) mmol/L,P=0.000,HbA1c:( 7.75 ± 1.08 ) ％ vs.( 7.01 ± 0.87 ) ％,P=0.002;BMI:( 25.82 ± 1.74 ) kg/m2 vs.( 24.23 ± 1.36 ) kg/m2,P=0.024;TG:(2.47 ±0.75) mmol/L vs.(2.13 ± 0.43 ) mmol/L,P=0.018;HoMA-IR:7.88 ± 0.20 vs.6.15 ± 2.01,P=0.042].No significant difference was found on the values of TC,HDL-C and LDL-C before and after intervention in control group (P ＞ 0.05).The effect of intervention of experimental group was more obvious when compared with control group ( FPG:P=0.036;2hPG:P=0.000;HbA1c:P=0.045;BMI:P=0.037;TG:P=0.022;HoMA-IR:P=0.000).Conclusion Individualized lifestyle intervention can improve the compliance of T2DM patients,and was in favor of control metabolic status of T2DM patients to delay the occurrence and development of complications.

Objective To investigate the role of the Wnt/LRPS/3-catenin signaling pathway in the pathogenesis of postmenopausal osteoporosis.Methods Fifty female Wistar rats aged 6-month-old,were randomly divided into control group ( NS,n =24 ) and ovariectomized group ( NOVX,n =26),NOVX underwent bilateral ovariectomy.At 0,4 and 8 weeks,all of rats were measured blood estrogen ( E2 ) and bone mineral density (BMD),4 and 8 weeks,low density lipoprotein receptor-related protein 5 (LRP5),3-catenin and Runx2 mRNA in bone were measured respectively by reverse transcription (RT)-PCR.Results In 4 and 8 weeks,compared with NS which had (117±29) and (114+15) pmol/L in E2 level,(0.098 ± 0.016 ) and (0.095 ± 0.028 ) g/cm2 in BMD,NOVX had significantly decreased to ( 92±15 ) and(95±22) pmol/L in E2 level ( P＜0.05 ),( 0.076 ± 0.016) and ( 0.052 ± 0.013 ) g/cm2 in BMD values ( P ＜ 0.01 ).And bone tissue LRP5,β-catenin and Runx2 mRNA expression was 1.02 ± 0.06,1.04 ±0.05,1.07 ±0.21 in NS,NOVX was significantly reduced to 0.97 ± 0.04,0.58 ± 0.05,0.86 ±0.03 (P＜0.05).Conclusion Wnt/LRP5/β-catenin signaling pathway may be important in the pathogenesis of postmenopausal osteoporosis.

Objective To study the effects of hSSTR2 gene in vitro transfection on differential proteins expression in pancreatic cancer cell line Panc-1 and search new sensitive therapeutic targets of pancreatic cancer. Methods The full length hSSTR2 cDNA was introduced into pancreatic cancer cell line Panc-1 by adenovirus vector ( Ad. CMV. hSSTR2. GFP) mediated transfection. The differential expressed proteins between the hSSTR2 transfection group, vector control and mock control were isolated and screened by 2D-DIGE analysis. Protein identification was performed by peptide mass finger printing with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF). Results The hSSTR2 gene was transfected into Panc-1 pancreatic cancer cells in vitro successfully, and fluorescence difference protein expression patterns were established between hSSTR2 negative and positive expression of Panc-1 cell. Analysis by DeCyder v6.5 software showed a total of 18 protein spots ( > 1.3-fold) and these protein spots were identified by mass spectrometry as 13 proteins. Proteins with lower abundance levels included GMP synthase, stress induced phosphoprotein 1, glutamate dehydrogenase 1, Septin-11, vimentin, Isocitrate dehydrogenase [NAD] subunit alpha, Import inner membrane translocase subunit TIM50. Proteins with high abundance levels included Elongation factor 1-alpha-1, Isoform M2 of Pyruvate kinase isozymes M1/M2, Enoyl-CoA hydratase,tripartite motif-containing 28 protein, Mitofilin, HSP105. Conclusions The proteins expression changed after hSSTR2 gene in vitro transfection in Panc-1 cells, and the function of difference proteins involved the process of metabolism of sugar, fat and nucleic acid, and the regulation of cell growth. The present study paved the way for searching new sensitive therapeutic targets of pancreatic cancer.

Objective To investigate the role of TLR4/NF-kB signaling pathway in inhibited invasion ability of pancreatic cancer cells caused by triptolide (TP).Methods PANC1 cells were divided into parental cells group,TP group,lipopolysaccharide (LPS) group and TP + LPS group.50 ng/ml of TP was added in culture medium in TP group,and 1 μg/ml of LPS was added in culture medium in LPS group,while 50 ng/ml of TP was pretreated for 2 h and 1 μg/ml of LPS was added in culture medium in TP + LPS group.All the ceils were cultured for 24 h.The TLR4 and matrix metalloproteinase-9 (MMP-9) mRNA and protein expression were evaluated by real-time PCR and Western blot.The NF-kB activity was determined by dual-luciferase reporter assay system.The invasion ability of pancreatic cancer cells was evaluated by transwell invasion chamberassay.Results The TLR4 mRNA expressions in parental cells group,TP group,LPS group and TP + LPS group were 0.41 ± 0.06,0.46 ± 0.10,0.20 ± 0.04,0.25 ± 0.06 ; the TLR4 protein expressions were 0.55 ±0.06,0.55 ±0.06,0.18 ±0.04,0.13 ±0.00; the activities of NF-kB were 13.0 ±3.0,31.6 ±4.3,7.3 ±1.5 and 10.8 ± 2.1,and the numbers of invasion cell were (56.8 ± 8.6),(104.5 ± 12.8),(32.0 ± 5.7) and (46.8 ± 7.0) ; the MMP-9 mRNA expressions were 0.36 ± 0.05,0.58 ± 0.07,0.18 ± 0.03,0.30 ± 0.004 ;the MMP-9 protein expressions were 0.31 ± 0.04,0.53 ± 0.08,0.11 ± 0.02,0.15 ± 0.00.In LPS group,TLR4 mRNA and protein expressions were not statistic significant when compared with those in parental cells group,but the activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions were statistically increased when compared with those in parental cells group (t =8.654,7.593,6.655,4.982,P ＜0.01).TLR4 mRNA and protein expressions,activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions in TP group were significantly lower than those in parental cells group (t =-7.609,-9.948,-4.176,-5.915,-8.179,-9.948,P＜ 0.01).TLR4 mRNA and protein expressions,activities of NF-kB,the numbers of invasion cell,MMP 9 mRNA and protein expressions in TP +LPS group were significantly lower than those in LPS group (t =-4.437,-14.805,-10.506,-9.700,-9.055,-8.932,P＜ 0.01).Conclusions TP can inhibit pancreatic cancer cell invasion,and the mechanism is related to the inhibition of TLR4/NF-kB signaling pathway and down-regulation of MMP-9 expression.