Objective To develop a rapid ,convenient ,sensitive and specific method for the detection of marine Vbrio vulnificus by using loop‐mediated isothermal amplification(LAMP) technique ,and evaluate the specificity and sensitivity of this method . Methods According to marine Vibrio vulnificus cytolysin(vvhA) gene sequence published by GenBank ,a set of LAMP primers were designed .LAMP and polymerase chain reaction(PCR) amplification were carried out in 47 strains of bacteria(including 20 strains of Vibrio) and specificities of LAMP and PCR were compared .Serial ten‐fold dilutions of an overnight Vibrio vulnificus M06 culture were prepared in sterile saline solution ,and compared the sensitivity of LAMP with that of PCR after extracting DNA tem‐plates .A recombinant plasmid containing fragment of vvhA gene was constructed and set as a standard positive control of LAMP assay .Results False‐positive results occurred in the conventional PCR ,while in the LAMP assay positive amplifications were only seen in strains of Vibrio vulnificus and no false‐positive or false‐negative results were generated among 47 strains of bacteria ,which indicated that primers had high specificity .Additionally ,the results of electrophoresis were consistent with those after adding the calcein .The detection limit of LAMP was 4 × 10 CFU each reaction for detecting vvhA gene in pure culture ,which was 10‐fold more sensitive than that of conventional PCR(4 × 102 CFU each reaction) .It indicated that LAMP assay had good sensitivity .Repeated the test twice ,the detection results of LAMP and PCR were both stable .Conclusion An LAMP detection method for marine Vibrio vulnificus has been developed ,which is highly specific ,sensitive ,convenient and suitable for rapid field detection and point‐of‐care testing .

Objective To investigate the expression and polymorphism of lipoprotein lipase (LPL) gene and their association with acute hyperlipidemic pancreatitis(HLP). Methods A total of 120 patients Were assigned to HLP group (n=20), acute pancreatitis (AP) group (n=50) and control group (n= 50). Serum levels of triglyceride (TG), cholesterol (Ch), free fatty acid (FFA), lipoprotein and apolipoprotein and serum LPL/HL activities were determined. The mRNA expressions of LPL/HL and LPL gene intron 8 polymorphisms were detected by RT-PCR and PCR-RFLP, respectively. Results The serum levels of TG, Ch, FFA and ApoE were significantly higher in HLP group than those in AP group and control group (P<0.05). The serum level of HDL was lower in HLP group than that in AP group and control group(P<0.05). The serum LPL/HL activities were significantly higher in HLP group than that in AP and control groups. The expression of LPL mRNA was up-regulated and intron 8 Hind Ⅲ H2 allele frequency was significantly inereased in the HLP group compared to control group(0.90/0.72, P<0.05). H1 allele frequency was significantly decreased in the HLP and AP groups compared to control group(0.10/0.28 and 0.14/0.28, respectively). Conclusions The high allele frequency of LPL gene intron 8 Hind H2 result in the increase of activities and expression of LPL mRNA, which exacerbate the development of HLP through changing TG metabolism such as FFA accumulation.

Objective To investigate the influence of Lipoprotein lipase (LPL)/Hepatic lipase (HL) on hyperlipidemie acute necrotizing pancreatitis (HLANP) in rats. Methods The rats were fed with hyperlipidemic feed for 4 weeks, then the rats were injected with 5% sodium taurocholate into pancreatic duct to induce HLANP model. Seventy-two rats were randomly assigned to HLANP and control groups, and then each group was subdivided into 6 subgroups (n = 6) at 0, 3, 6, 12, 24 and 48 h. Serum amylase, cholesterol, triglyeride (TG), free fatty acid (FFA) levels and serum LPL, HI. activities were determined. Under the light-microscopy and electron microscopy, the histopathologic and uhrastructure changes of pancreas were observed; the HL mRNA expressions were detected by RT-PCR; HL protein expressions HL were assessed by immunohistoehemical staining. Results The serum amylase levels reached peak values at 12 h after ANP induction in the two groups, the mean values were 7 176U/L and 6 366U/L, which were significantly higher than those of baseline values (P <0.05) ; serum levels of cholesterol in HLANP group at 0 ～ 12 h were higher than those of control group, however, only at 0 h the difference (1.19±0.49 vs 0. 32±0.14 mmol/L) was significant (P < 0.05) ; serum levels of FFA in HLANP group were not significantly different when compared with those of control group; serum levels of LPL and HL at 3 h were (17.5±7) U/L and (18.6±3.9) U/L, which were significantly higher than those of control group (8.9±3.4 U/L and 9.5±2. 1 U/L, P < 0.05). The pancreatic tissues necrosis levels were significantly increased in HLANP groups (3, 6, 24 and 48 h) than those of control group at corresponding time points (P < 0.05). lipid droplet deposition, rough endoplasmic reticulum distension, zymogen granule reduction, and chondriosome swelling in acinar cells of pancreatic tissues in HLANP group were found. The HI, mRNA expressions at 3 h and 6 h in HLANP group were 1.1±0.09 and 0.89±0.08, which were significantly higher than those in control group (0. 11 ± 0.01 and 0.15±0. 03, P <0.05). HL proteins were positively expressed in pancreatic tissues of two groups before ANP was induced, and HL proteins were strongly positively expressed after ANP induction. Conclusions Lipase (LPL/HL) expression increased in HLANP rats, and the content of serum protein increased, which resulted in lipids decomposition and increased the severity of ANP. LPL/HL may be one of the key lipids metabolic enzymes aggravating HLANP.

Objective: To examine whether there are differences between dependent and independent individual’s shame and sad memories. Methods: Self -report measures were used. Results: ① There were significant interactions between emotion and cognitive style of the number of words recalled, number of times that others were mentioned. ②Field dependent individuals mention more emotional behaviors and presence of others. ③The subjects mentioned more emotion words and time in sad memories. ④In shame memories, there were less self-references than others-references, marginally less self in thinking than self in other descriptions. ⑤In sad memories, field dependent individuals reported less selfreferences than others-references, and less self in thinking than self in other descriptions. ⑥There were more emotional behaviors and self-thinking in sad memories than in shame memories. Conclusion: Field dependent and independent individuals demonstrate differences in emotional memory, and the difference is emotion specificity.

AIM:To investigate the effect of hepatic oval cells (HOCs) on the protein expression of TGF-?/Smad signaling pathway in the liver tissue of hepatic fibrosis rats.METHODS:The SD rat models of liver fibrosis were made by treating with carbon tetrachloride and combined factors.The HOCs was isolated from the model rats.HOCs suspension (0.5 mL at a density of 1 ? 1012cells/L) were transplanted via portal vein into the hepatic fibrosis rats at 8th week and observed continuously for 30 days.Meanwhile,WuLing capsules were used for positive control.The blood samples were collected through trail vein at 8th day,15th day,23th day and 30th day after transplantation of HOCs.The levels of aspartate aminotransferase (AST) and alamine aminotransferase (ALT) in serum were determined by enzyme method.The morphological changes of hepatic tissues were observed under microscope with HE and Musson staining.The protein levels of collagen type I (Col-Ⅰ),extracellular-signal regulated protein kinase (ERK),phosphory-lation extracellular regulatedprotein kinases (p-ERK),TGF-? receptor type Ⅰ (T?RⅠ),TGF-? receptor type Ⅱ (T?RⅡ),mothers against decapentaplegic homolog 2 /3 (Smad 2 /3) and mothers against decapentaplegic homolog (Smad 7) were assessed in liver tissues by Western blotting.RESULTS:In HOCs and WuLing capsules treated groups,the levels of ALT and AST decreased significantly at 15th day,23th day and 30th day after the transplantation of HOC.The damage degree of hepatic fiber hyperplasia of the liver histological structure reduced notably.The expression levels of Col-Ⅰ,ERK,p-ERK,T?RⅠ and T?RⅡ in liver tissues of hepatic fibrosis rats were down-regulated obviously while the expression of Smad 7 increased significantly.CONCLUSION:The implantation of HOCs prevents the progress of liver fibrosis in rats.The mechanism of action is to inhibit the protein expression of p-ERK,T?RⅠ,T?R Ⅱ for TGF-?/Smad signaling pathway of liver tissue.

Objective To investigate stem cell factor(SCF)expression in activated pancreatic stellate cell(PSC)and the effect of SCF on adhesion between PSC and mast cell(MC).Methods PSC of SD rat was separated and activated by pancreatic tissue culture;the expression of SCF mRNA and protein was detected bv RT-PCR and immunachemistry.SCF was blocked with different dose of anti-SCF antibody,the rate of adhesion between PSC and MC was determined by thiomn blue stain and β-hexasaminidase assay.PSC was stimulated by 10 μg/ml mitomycin-C and was co-cultured with MC,then the growth of MC was observed at 2,3,5 day,respectively.Results Activated PSC expressed SCF mRNA and protein,the number of adhered MC in the control,1mg/ml and 10 mg/ml antibody groups was 48±10,28±8,and 23±9,respectively;the difference between antibody and control group was statistically significant(P＜0.05).One day after PSC and MC was co-cultured without serum.there were MC adhesion;3 days later the rate of adhesion increased;5 days later the adhesion appeared bulk-like and lots of colonies were present,while PSC basically disappeared.Conclusions Activated PSC may synthesize,express SCF and induced adhesion and growth of MC.

AIM: To investigate the influence of compatibitity in Fengshi Maqian Tablet [ABD group(Semen strychni,Bombyx batryticatus,Herba ephedrae,Scorpio,Rhizoma atractylodis,Radix et Rhizoma glycyrrhizae) E group(Olibanum;Myrrha)]. METHODS: In the use of orthogonal design(5 factors and 2 levels).HPLC method were used to determine the main chemical componets such as strychnine,brucine and ephedrine.And anti-inflammation was adopted as pharmacological experiment. RESULTS: In the inhibitive experiment of granuloma,ABD group was superb to E group.In the tumescence experiment,otherwise. CONCLUSION: The minister,assistant and guide herbs in Fengshi Maqian Prescription had a significant effect on the toxicity of monarch medicines-Semen strychni.

Objective:To test the similarities and differences of shame experience between Chinese and American participants and explore the future research direction of shame in a cross-culture context.Methods:American college students were enrolled by convenient sampling to a semi-construct face-to-face interview about general and specific shame experience in their own culture.No new materials appeared after 8 students were interviewed.Then 8 Chinese college students were paired sampling to participate in the same interview.Finally all the in terview contents were coded according to the emotion,cognition and behavioral reaction of shame experience.Results:Both Chinese and American students reported shame experience with similar intensity in academic,personal relationship,body,group and transference shame situations.The expectations of self,peers,parents,teachers or supervisors could be sources of shame in both eastern and western cultures,and only Chinese students regarded expectation of a specific group as a source of shame.Chinese participants recalled more body reaction when feeling shame and had more cognition process related to the shame experience.Finally,besides the negative results such as avoidance that shame experience brought out,in both cultures,the most important influence of shame was reported to be the positive effects on improving or restricting improper behaviors.Conclusion:The participants from two cultures demonstrate a fairly coherence process of shame experience,and the results support the appraised-based model of self-conscious emotions.

Objective: To investigate effects of Wuling Pills and Capsules on immunological liver injury in mice. Methods: The rat liver fibrosis model was induced by carbon tetrachloride. The hydroxyproline (HYP) level in serum was measured by chemical method. The precollagen Ⅰ(PCⅠ) and Ⅲ(PCⅢ) levels in serum and hepatic homogenate were measured by radioimmunoassay. The expressions of tumor necrosis factor (TNF) and fibronectin (FN) in liver tissue were detected by histochemistry method. Moreover, the hepatic histomrphology was observed by HE staining. Results: Both of Wuling Pills and Wuling Capsules could greatly decrease PCⅠ, PCⅢ and HYP levels. The FN expression of liver tissue in Wuling Capsule group was obviously lower than that of the model group. The TNF and FN in liver tissues of rats in the Wuling Pill group and Capsule group showed strong positive. Proliferative kupffer cells and activated endothelial cells were also observed in those two groups. Conclusion: Wuling Capsules has the effect of anti liver fibrosis. Both of Wuling Pills and Capsules can stabilize hepatocyte membrane, and prevent liver cells from the effect of TNF and FN.

Objective:Express the recombinant human FGFR1 in order to screen the FGFs.Methods:A cDNA fragment encoding human fibroblast growth factor receptor 1(FGFR1) was isolated by RT PCR from human lung fibroblasts,and then cloned into pCR TM II plasmid and pFastBac1 donor plasmid.Through transposition,recombinant bacmid FGFR1 DNA was formed and was then transfected into Sf9 insect cells to produce recombinant Baculovirus.Sf9 insect cell were infected with recombinant Baculovirus.The harvested culture supernatant was subjected to Western Blot and ELISA analysis.Results:The size of FGFR1 cDNA fragment is 2 100 bp.The MW of expressed recombinant FGFR1 was 78 kD.ELISA showed that human recombinant FGFR1 was expressed on the membrane of insect cell Sf9.Conclusion:The recombinant human FGFR1 can be expressed on the membrane of insect cell Sf9 effectively.