Objective To explore the effects of Shenmai injection combined with Xuebijing injection on the coagulation function of the early and inflammatory factor traumatic hemorrhagic shock.Method 86 cases of traumatic hemorrhagic shock patients select from December 2012 to December 2015 in our hospital were randomly divided into a control group(n=43)and experimental group(n=43).The control group was fluid resuscitation and Xuebijing injection treatment,the experimental group was increased Shenmai injection treatment in the control group basis.Compared with coagulation function and inflammatory factor levels before and after treatment in two groups of patients.Results 2 groups of patients was no difference in the index values before treatment,compared with before treatment,after treatment,the two groups of patients prothrombin time(PT),thrombin time(TT),activated partial thromboplastin time(APTT)and platelet count(PLT)were elevated,fibrinogen(FIB)was decreased,and the two groups were significantly different (P<0.05 or P<0.01),but in the control group was no difference in the index values after 0.5 h treatment;The inflammatory factor in two groups were significantly increased after treatment(P<0.01),the control group was significantly higher than the experimental group,a significant difference between the groups(P<0.01);The hemorheology indexes after treatment 7 d of 2 groups were significantly decreased compared with after treatment 1 d,and the experimental group was significantly lower than the control group(P<0.01).Conclusion The combined therapy of Xuebijing injection combined with Shenmai injection can effectively improve trauma patients with hemorrhagic shock early coagulation function,reduce the levels of inflammatory factors, improve hemorheology indexes,there are good clinical results.

Objective To study the effect of gastrin on the proliferation and angiogenesis of human umbilical vascular endothelial (HUVE) cell in vitro.Methods The immunocytochemistry assay,realtime-PCR,and Western blot were used to detect the gastrin receptor (CCK-BR) expression in HUVE cells.After HUVE cells were treated with 10 and 100 nmol/L gastrin for 72 h,MTT and soft agar colony formation assay were used to test the cell proliferation rate and colony formation rate,and the vascular-like structures were observed by three-dimensional culture of HUVE cells.The half-ring and ring vascular numbers were counted with five random visions.The vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1a (HIF-1a) mRNA and protein levels in HUVE cells were detected by realtime PCR and ELISA assay respectively.Results HUVE cells expressed CCK-BR.After the treatment with 10 and 100 nmol/L gastrin,the cell proliferation rate was increased by 48.48％ and 82.82 ％ compared to the control group,and colony formation rate was increased to 31.33％ and 45.67 ％ as compared with 15.33％ in the control group(P＜0.01).Relative expression quantities of VEGF and HIF-1a genes were 2.3 and 4.6 folds (VEGF) and 20.76 and 26.77 folds (HIF-1 a) than those in the control group.The concentration of VEGF protein in culture medium was 221 and 392 μg/mg protein higher than that in control group.The numbers of half-ring and ring vascular structures were (14.00 ± 3.00,39.33 ± 7.57 and 34.33 ± 4.50)/vision and (8.33 ± 2.51,41.33 ± 5.85 and 37.67 ± 3.51)/vision in control,10 and 100 nmol/L gastrin-treated groups,respectively (P＜0.01).Conclusion Gastrin up-regulates the expression of VEGF and HIF-1a genes in HUVE cells and promotes cell proliferation and vascular-like structure formation of HUVE cells in vitro by being combined to CCK-BR,which may be involved in the development and metastasis of gastric cancer.

Objective To study whether Helicobacter pylori CagA protein can control gastrin gene expression and the detailed mechanism. Methods First, pcDNA3. 1ZEO (-)/cagA7 was transfected into gastric cancer cell lines AGS and SGC-7901 cells. At the same time, culturing the Helicobacter pylori NCTC11637 and infecting AGS and SGC-7901 cells with it. Next, in the infected and transfecled AGS and SGC-7901 cells, respectively adding the JAK2 signaling pathway inhibitor AG490 and the ERK signaling pathway inhibitor U0126 to inhibit the two signaling pathway. Untreated gastric cancer cells and empty vector transfected cells as the control. Using real-time fluorescence quantitative PCR to detect the levels of gastrin mRNA in transfected and infected cells. Results After AGS and SGC-7901 cells were transfected with pcDNA3. lZE0(-)/cagA7 and infected with NCTC11637, the results showed that the expression of gastrin mRNA increased significantly (P ＜ 0. 05) in transfected and infected cells as compared with the control group, but after adding the inhibitor AG490 and U0126 respectively, the expression of gastrin mRNA decreased significantly(P＜0.05). Conclution These results suggest that CagA may up-regulate the expression of the gastrin gene, and CagA is one of the important proteins in regulating gastrin gene expression. The ERK/MAPK and JAK/STAT signaling pathways may be involved in controlling of gastrin gene expression by CagA.

AIM: To investigate the possible role of nuclear transcription factor kappa B (NF-?B), Bcl-2, Bax and caspase-3 in etodolac-induced apoptosis of liver tumor SMMC7721 cell line. METHODS: Cell apoptosis was determined by flow cytometry analysis with PI staining and DNA laddering. Expression of Bcl-2 and Bax protein was measured by Western blotting. Caspase-3 activity was evaluated by active caspase-3 apoptosis kit with flow cytometry. NF-?B activation was detected by ELISA-based TransAM~(TM) NF-?B p65/p50 kit. RESULTS: Etodolac, a selective COX-2 inhibitor, stimulated apoptosis in liver tumor SMMC7721 cell line significantly. Flow cytometry showed that the apoptotic rate was 16.3%?3.1%, 19.9%?3.6%, 22.9%?3.2%, 31.2%?3.3% with different concentrations of etodolac (0.25, 0.50, 1.0 or 2.0 mmol/L), while the apoptotic peak did not appear in the control group (0 mmol/L) (P

An in vitro detection method of the gastrointestinal absorption of Pilose Antler protein was established for mixed protein activity. Five bands of protein with molecular weight of 17.8-160 kD derived from the Pilose Antler were extracted and sufficiently labeled with FITC (FITC-PE). The stability and variation of FITC-PE in gastrointestinal circumstances were detected by native polyacrylamide gel electrophoresis and confocal laser scanning microscope. Results showed that the main component of FITC-PE kept invariant after being reacted with artificial gastric fluid and artificial intestinal fluid. The fluorescence signal was detected 20 min after administration in the valgus intestinal purse experiment, and three kinds of protein, with molecular weight of 45, 25, and 17.8 kD, were detected in the mixture of absorbent protein. The research laid the foundation for the further in vivo study of Pilose Antler protein. Meanwhile, it would be an in vitro screening method for the absorption, distribution and metabolism of mixed protein from traditional Chinese medicine.

Object To examine if Leonurus artemisia (Lour ) S Y Hu (Labiatae) contains ferulic acid Methods The samples were extracted by different solvents and the extracts examined by TLC and HPLC Results Ferulic acid was detectable in the extracts of L artemisia Conclusion L artemisia possibly contains free ferulic acid The use of 5% Na 2CO 3 as the extractant instead of methanol, as described in Chinese Pharmacopoeia seemed to be more simple, highly stable and with good reproducibility

Objective:To study the effects of ascorbic acid on BMP-2 mRNA expression of osteoblast cell sheets.Methods:Rat os-teoblasts were primaryly cultured and identified;osteoblast cell sheets were built by physical scraping method in vitro;the osteoblast cell sheets were cultured with 1 5,50 and 85 mg/L ascorbic acid for 1 and 2 weeks respectively,and the expression of BMP-2 mRNA of the cell sheets was detected by RT-qPCR.Results:The obtained cells were conformed to be osteoblasts.The osteoblast cell sheets could be rolled into tube in vitro.The expression of BMP-2 mRNA of osteoblast cell sheets in experiment group,whether in week one or week two was higher than that in control group,50 mg/L group showed the highest expression(first week P <0.05;second week P>0.05);the expression of any group in week two was higher than that in week one(P <0.05).Conclusion:Ascorbic acid may pro-mote the expression of BMP-2 mRNA in osteoblast cell sheets.

Objective To investigate the effects of blocking gastrin receptor on the proliferation,apoptosis and expression of key proteins in the related pathway in gastric cancer cell lines.Methods In the experimental group,the gastric cancer cell lines SGC-7901 and AGS cells were treated with 5 mmol/L proglumide,a kind of a gastrin receptor antagonist.And the normal cultured gastric cancer cells SGC-7901 and AGS were used in control group.The growth of each group was detected by MTT assay;the cell growth curve was drawn by flow cytometry;the cell cycle of each group was detected by flow cytometry.Annexin V-FITC/PI double staining was used to detect the cell growth of apoptosis.The relative mRNA expression of β-catenin,nuclear factor-P65,mammalian target of rapamycin and glycogen synthase kinase 3 beta in Wnt,NF-κB and PI3K-AKT-MTOR pathways were detected by RT-qPCR.The expression of β-catenin protein was detected by Western blotting.Results After treatment with proglumide,the growth of the cells in the experimental group was lower than that in the control group;and the proportion of S phase cells in the cell cycle was also lower than that in the control group,but the proportion of cells in G0/G1 phase was higher than that in the control group(P<0.05).The percentage of apoptotic cells was also increased after treatment with proglumide(P<0.05).Furthermore,proglumide treatment significantly reduced the expression of β-catenin at both mRNA and protein levels(P<0.05).Conclusion Blocking gastrin receptor can down-regulate the expression of β-catenin,inhibit the cell proliferation and promote the cell apoptosis in gastric cancer cells.

Objective To study the expressions and clinical significances of activating transcription factor 3 (ATF3) and Runt-related transcription factor 2 (Runx2) in breast carcinoma tissues.Methods The expressions of ATF3 and Runx2 were detected in 105 cases of primary breast invasive ductal carcinoma (IDC) and their matched para-tumor breast tissues by immunohistochemistry.The relationships between the two transcription factors and the clinical pathological features of IDC patients were analyzed.Results The positive expression rates of ATF3 and Runx2 in IDC group were 80.95％ (85/105) and 69.52％ (73/105) respectively,much higher than those in para-tumor breast tissues 11.43 ％ (12/105) and 8.57％ (9/105),with statistically significant differences (x2 =102.097,P =0.000;x2 =11.595,P =0.001).ATF3 and Runx2 expressions showed significant relationships with histological grade (x2 =14.623,P =0.001;x2 =24.891,P =0.000),lymph node metastasis (x2 =7.059,P =0.008;x2 =6.358,P =0.012) and pTNM stage of IDC (x2 =5.807,P =0.016;x2 =4.902,P =0.027),while both were not correlated with patients' age (x2 =0.274,P =0.601;x2 =1.554,P =0.213) and tumor size (x2 =2.476,P =0.290;x2 =5.261,P =0.072).There was a significant positive relationship between ATF3 and Runx2 in breast carcinoma (C =0.498,P =0.000).Conclusion The over-expressions of ATF3 and Runx2 may participate in the tumorigenesis,invasion,metastasis and clinical stage of breast carcinoma,which suggests that they may be key factors to evaluate malignant degree and prognosis of breast carcinoma.

Objective To investigate the effect of Rho kinase inhibitor,fasudil,on pulmonary fibrosis induced by paraquat in rats in order to elucidate the underlying mechanisms.Methods A total of 72 SpragueDawley male rats of specific pathogen free (SPF) were randomly (random number) divided into four groups:the normal control group (NS group,n =18),fasudil control group (FS control group,n =18),paraquat poisoning group (PQ poisoning group,n =18) and fasudil intervention group (FS intervention group,n =18).On days 7,14,28 after paraquat exposure,six rats were respectively selected from each group.These rats were anesthetized and sacrificed immediately,and their lung tissues were collected.The hydroxyproline (HYP) in the lung tissue was detected by using alkaline hydrolysis.The expressions of type Ⅰ,Ⅲ collagen protein,connective tissue growth factor (CTGF) and ROCK1 mRNA in Rho/ROCK signaling pathway were assayed by using the real-time quantitative PCR (RT-PCR),and the levels of type Ⅰ,Ⅲ collagen protein,connective tissue growth factor (CTGF) and Rho / ROCK signaling pathway ROCK1 protein were measured by using Western blotting.The pathological changes of lung tissue were observed under light microscope.Results There were no significant differences in the observed biomarkers between FS control group and NS group (P ＞ 0.05).While in PQ poisoning group and FS intervention group on days 7,14,28 (all P ＜ 0.05),the amount of HYP increased obviously (P ＜ 0.05),the expressions of type Ⅰ,Ⅲ collagen protein,CTGF,ROCK1 mRNA and protein levels were increased significantly (P ＜ 0.05).Compared with the PQ poisoning group,the amount of HYP decreased significantly,and the expressions of type Ⅰ,Ⅲ collagen protein,CTGF,ROCK1 mRNA and protein levels were decreased significantly in FS intervention group on days 7,14,28 (all P ＜ 0.05).The pathological changes of lung tissue revealed that the degree of pulmonary fibrosis in the PQ poisoning group were most serious on 28 d after paraquat exposure,and the degree of pulmonary fibrosis were lessened in FS intervention group on days 7,14,28.Conclusions ROCK inhibitor,fasudil,has obvious therapeutic effects on paraquat-induced lung fibrosis,by regulating Rho / ROCK signaling pathway with downregulated expression of CTGF,and decrease in the levels of type Ⅰ,Ⅲ collagen protein,thus reducing protein deposition.