Objective To establish a kind of simple,rapid,accurate and reliable method to analyze the concentration of alcohol in blood by headspace gas chromatography (HS-GC) with dual-column and dual-detector.Methods The samples were pre-treated by headspace sampler,which was the basis on the extraction principle of the gas extracting volatile substances.Next,these samples were analyzed by HS-GC that the tertiary butyl alcohol was acted as the internal standard substance.The HS-GC was equipped with two chromatographic column (the DB-ALC2 chromatographic column of 001 channel;the DB-ALC1 chromatographic column of 002 channel).At the same time,the HS-GC was also equipped with two hydrogen flame ionization detector (FID1 detector;FID2 detector).The retention time of the peak was finally performed as qualitative parameter and the standard curves method of internal standard were acted as quantitative basis.Results The liner range of the method was 0.2-2.0 mg/mL.The linear regression equation of 001 channel was Y=1.057 7X+0.048 2 and the correlation coefficient was R2=0.999 05.Besides,the linear regression equation of 002 channel was Y=1.039 5X+0.046 5 and the correlation coefficient was R2=0.999 25.In short,the average recovery rate of the method was 99.70%.Relative standard deviation(RSD) was less than 4% between the analysis results of 001 channel and 002 channel for the determination of the plan sample.Conclusion The method shown satisfactorily that it could not only be applied to determine the alcohol of blood of forensic toxicological analysis,but also be applied to determine the plan sample of ability test and verify of laboratory ability accreditation.

OBJECTIVE:To study the influence of oscillator on the cleanliness of class 100 clean bench in PIVAS. METH-ODS:Using sedimentated bacteria and the number of dust particle as index,in common drug configuration room,antibiotics con-figuration room and risk drugs configuration room including biological safety cabinet and horizontal laminar flow,the cleanliness of class 100 clean bench were monitored when oscillator was set at clean bench and different positions in work and non-working state. RESULTS & CONCLUSIONS:In working and non-working state of oscillator,there was no difference in sedimentated bacteria and the number of dust particle which was in line with the requirements of 2010 edition of GMP,i.e. the application and location of oscillator didn't influence the cleanliness of class 100 clean bench. From a view of safety,it is suggested to place the oscillator in the left(or right)posterior wall of clean table when biological safety cabinets is used to dispense antibiotic and risk drugs.

To investigate the relationship of the variation of virulence and the external capsid proteins of the pandemic duck hepatitis A virus type 1(DHAV-1) isolates,the virulence,cross neutralization assays and the complete sequence of the virion protein 1(VP1) gene of nine virulent DHAV-1 strains,which were isolated from infected ducklings with clinical symptoms in Shandong province of China in 2007-2008,were tested.The fifth generation duck embryo allantoic liquids of the 9 isolates were tested on 12-day-old duck embryos and on 7-day-old ducklings for the median embryonal lethal doses(ELD50s) and the median lethal doses(LD50s),respectively.The results showed that the ELD5s of embryonic duck eggs of the 9 DHAV-1 isolates were between 1.9 × 106/mL to 1.44 × 107/mL,while the LD50s were 2.39 × 105/mL to 6.15 × 106/mL.Cross-neutralization tests revealed that the 9 DHAV-1 isolates were completely neutralized by the standard serum and the hyperimmune sera against the 9 DHAV-1 isolates,respectively.Compared with other virulent,moderate virulent,attenuated vaccine and mild strains,the VP1 genes of the 9 strains shared 89.8％-99.7％ similarity at the nucleotide level and 92.4％-99.6％ at amino acid level with other DHAV-1 strains.There were three hypervariable regions at the C-terminus(as 158-160,180-193 and 205-219) and other variable points in VPI protein,but which didn't cause virulence of DHAV-1 change.

Background and purpose: Three-incision esophagectomy for patients with esophageal cancer has been increasingly used, but the incidence of either postoperative anastomotic leak or stricture is higher than that in intrathoracic gastroesophageal anastomosis. The purpose of this study was to investigate the clinical effects of the side-to-side stapled cervical gastroesophageal anastomosis in preventing anastomotic leak and stricture after three-incision esophagectomy. Methods: One hundred and twenty-seven patients undergoing three-incision esophagectomy for esophageal cancer with gastric replacement were reviewed. A side-to-side stapled cervical gastroesophageal anastomosis was performed in 71 cases and manually sewn anastomosis in 56 cases. The incidence of postoperative anastomotic leak and stricture was compared between these two groups and the results were statistically analyzed using SPSS11.0 soft.Results: Anastomotic leakage was noted in seven patients (9.9%) in the stapler group and fourteen patients (25.0%) in the manually sewn group (P=0.04). After the operation two patients (2.8%) in the stapler group and nine patients (16.1%) in the manually sewn group developed a benign esophageal stricture (P=0.02).The incidence of either postoperative anastomotic leak or stricture in cases of the stapler group was significantly lower than that in the manually sewn group (P<0.05). Conclusion: Construction of the cervical esophagogastric anastomosis with a side-to-side stapled technique provides a larger luminal diameter which significantly reduces the incidence of postoperative anastomotic strictures. The surgery also greatly reduces the incidence of anastomotic leaks and strictures, so it could be used as an alternative strategy for cervical esophagogastric anastomosis after esophagectomy for esophageal cancer

Objective:To study the relationship between CYP4F2 gene polymorphism and coronary heart disease (CHD) in Mongolia patients ,and investigate clinical characteristics of these patients . Methods:All subjects received questionnaire . Gene amplification and genotyping were performed in 234 Mongolian CHD patients (CHD group) and 221 non-CHD pa‐tients (normal control group) using high temperature ligase detection reaction technique .The relationship between Mongo‐lian CHD and CYP4F2 gene polymorphisms of two single nucleotide polymorphism (SNP) sites (rs1558139 ,rs2108622) was analyzed .Results:Compared with normal control group ,there were significant rise in percentages of male (41.18% vs . 67.95% ) ,smoking history (32.13% vs .41.88% ) ,body mass index [BMI ,(21.66 ± 4.53 ) kg/m2 vs .(25.34 ± 5.37 ) kg/m2 ] and triglyceride level [ (1.66 ± 0.90) mmol/L vs .(1.92 ± 1.38) mmol/L] ,and significant reduction in level of high density lipoprotein cholesterol [ (1.18 ± 0.28) mmol/L vs .(1.07 ± 0.29) mmol/L] in CHD group , P<0.05 or <0.01.There were no significant difference in genotype and allele frequencies of rs 1558139 and rs2108622 between two groups . Conclusion:Clinical characteristics of Mongolian CHD patients include high male percentage ,smoking history ,high body mass index and high triglyceride level .CYP4F2 gene polymorphisms of rs1558139 and rs2108622 are not related to coronary heart disease in Mongolian patients .

Objective:To investigate the effect of Rab7 on cytokine induced by TLR7 (Toll like receptor-7) R848 activated in Raw264.7,and discusses the influence of Rab7 on MAPK signal transduction.Methods: TLR7 downstream cytokines such as TNF-α,IL-6,IFN-α,IFN-β and IP-10 activated by R848 were detected through Q-PCR in Rab7 silenced mouse macrophages,and then analysis of phosphorylation of MAPK determined with Western blot showed the effect of Rab7 on signal transduction of MAPK.Results: Rab7 inhibit production of cytokine activated by TLR7,and also,Rab7 had an inhibitory effect on MAPK signal pathway.Conclusion: The experimental results further illustrate that the Rab7 is the TLR7 signal transduction pathway negative regulatory factor,and to participate in MAPK signaling pathway.

Objective To investigate the therapeutic effect of Saccharomyces boulardii on antibiotics associated diarrhea (AAD)in infant pneumonia.Methods 302 hospitalized infant patients (1 month-3 years) with pneumonia but without gastroenteric disease were selected , and antibiotics were administrated intravenously at least 5 continuous days for each patient .Patients were all giv-en antibiotics and randomly divided into three groups:control group with no Saccharomyces boulardii administration ( group A, 60 ca-ses);Saccharomyces boulardii were applied as addition on the occurrence of diarrhea (group B, 92 cases), and Saccharomyces boular-dii and antibiotics were co-administrated (group C, 150 cases).Incidences of AAD in all groups were carefully examined and differ-ences of therapeutic effect between groups were compared and analyzed .Results The incidence of AAD in Group C was significantly lower than that in other groups .In term of diarrhea severity, no significant difference was observed in all groups (P>0.05).Howev-er, duration of diarrhea showed significant different between groups (P<0.001):group A with the longest duration and group C with the shortest .The results indicated that the total efficiency of anti-diarrhea in group C was significantly higher than that in control group (P<0.05).However, group B presented no significant difference compared with neither control group nor group C in term of therapeu -tic effect on diarrhea .Conclusion Administration of Saccharomyces boulardii Sachets could shorten the duration of diarrhea on the oc-currence of AAD in infant pneumonia .According to our results , Saccharomyces boulardii was effective in both preventing the develop-ment of AAD and shortening duration of diarrhea , and therefore improved therapeutic effect on ADD .

Objective:To explore the inhibitory effects of seabuckthorn polysaccharide on hepatic oxidative stress in a mice model of acute liver injury induced by intraperitoneal injection of LPS and D -GalN and detect the expression on hepatic BCL-2/Bax and PPAR-γ.Methods: C57BL/6 male mice were randomly divided into six groups:control group ( CTRL), model group ( L/G), dexamethasone positive control group ( DXM ) , low ( SPL ) , medium ( SPM ) and high dose group ( SPH ) of seabuckthorn polysaccharide.Mice in the SPL,SPM and SPH group were gavaged with 50,100 and 200 mg/kg seabuckthorn polysaccharide for 14 days respectively.Acute liver injury model were established by intraperitoneal injection of LPS (10 μg/kg) and D-GalN (700 mg/kg) .Serum and liver samples were collected 4 h after model establishment .Serum levels of ALT and AST and the content of MDA were de-tected.Hepatic expression of SOD 2 BCL-2 and Bax was determined by Western blot and the expression of PPAR-γwas detected by im-munohistochemistry .Results:ALT and AST levels significantly increased in the model group and decreased dose-dependently after pre-treatment with seabuckthorn polysaccharide .The level of MDA in the model group increased significantly as compared with the control group and decreased in seabuckthorn polysaccharide groups ,while the level of SOD 2 decreased in the model group and recovered in sea-buckthorn polysaccharide groups .The expression of Bax decreased after pretreatment with seabuckthorn polysaccharide .There was no obvious effect on BCL-2 expression after sea buckthorn polysaccharide supplementation .The expression of PPAR-γreduced in the sea-buckthorn polysaccharide group as compared with the model group .Conclusion:Seabuckthorn polysaccharide protects against LPS /D-GalN-induced liver injury.The effect is associated with an upregulation of SOD 2 and downregulation of Bax .

Objective:Using the macrophage cell lines RAW264.7 stably expressing Rab5a and its dominant negative mutant Rab5aN133I as models to analyze the effect and the mechanism of Rab 5a,Rab5aN133I on CpG-induced production of pro-inflammatory cytokines and type Ⅰ IFN.Methods: The eukaryotic expression vectors of Rab5a and Rab5aN133I were transfected into RAW264.7 cells by liposome,and screened with G418.The G418-resistant colonies were obtained and amplified.The transformed cell lines were i-dentified by RT-PCR,Real time-PCR and Western blot.The production of cytokines were measured after transformed cell lines of Rab5a and Rab5aN133I was stimulation with CpG for 8 h.Results: Rab5a expression in transfected cells was significantly higher than the control cell group (P<0.05).Overexpression of Rab5a significantly promoted the production of TNF -α,IL1-β(P<0.01) and IFN-β( P<0.05) in CpG stimulated RAW264.7.The production of cytokines was restored in Rab 5aN133I transfected cell line.Conclusion:Rab 5a promotes CpG-induced pro-inflammatory cytokines and typeⅠIFN in macrophages,it may be act as a positive regulator in TLR9 signaling pathway.

Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.