Abstract:Objective:To investigate the relationship between oral squamous cell carcinoma (OSCC) and human herpesvirus 6 (HHV-6) infection. Methods:HHV-6 DNA in saliva from patients of OSCC and from health control people was amplified by nest PCR which was HHV-6 specific. Serum anti-HHV-6 IgG titer was tested by anti-HHV-6 IFA kit. HHV-6 glycoprotein B(gB) was detected by immunohistochemistry in OSCC tissues and peritumoral tissues,respectively. Results:The detectable rate of HHV-6 DNA in patients of OSCC is 65.3%,which was much higher than 40% of health control group. But there was no significant difference in clinicopathologic and background variables of OSCC between the PCR positive and PCR negative groups. The geometric mean titer of anti-HHV-6 IgG was significantly higher in the OSCC patients (1∶102.17) than in the health controls (1∶101.50). HHV-6 gB was detected in 6 out of 11 tissues of OSCC and in 3 out of 8 peritumoral tissues. The gB was condensated in tumour cells while was either negative or much lower expression in the intertumor issues. We isolated and cultured tumor cells from one tissue of OSCC which was positive for HHV-6 gB. And then the nude mouse was inoculated with those tumor cells. We could also detected HHV-6 gB in tumor tissues from the mouse. Conclusion:The results demonstrated the frequent presence of HHV-6 in OSCC,although which has no association with clinicopathologic and background variables of patients. An animal model which carry HHV-6 antigen were established and could be useful for understanding the role that the HHV-6 really play in the genesis and treatment of OSCC which is HHV-6 positive.

Abstract:Objective:To analyze the role of Rho GTPase activating protein (RhoGAP) domain of frequently deleted in liver(DLC-1) gene in suppression of proliferation and invasion in colon cancer HT29 cells. Methods:Recombinant pcDNA3.1 vector containing the DLC-1 gene and RhoGAP domain deleted ΔDLC1 subclone was constructed and transfected into HT29 cell line. The cell growth was measured by MTT and colony formation assay. The ability of cell migration of HT29 cells was examined by using Transwell migration assays. Furthermore,cell cycle was evaluated by flow cytometry. Results:After transfected successfully,pcDNA3.1-DLC1-HT29 cells showed a remarkable suppression of proliferation and invasion,as well as G1 sphase arrested and apoptosis induced,compared with wild or mock-vector HT29 cells. However,pcDNA3.1-ΔDLC1 subclone transfection had no effect on cell proliferation,invasion and cell cycle distribution. Conclusion:DLC-1 gene inhibits proliferation and invasion of colon cancer HT29 cells via its RhoGAP domain.

Abstract:Objective:To explore the effect of glutamine combined with ulinastatin on intestine damage and inflammatory mediators after severe burn injury. Methods:Total 174 Wistar rats were inflicted with full thickness burns in 30% total body surface area and then randomly divided into burned control (B) group,glutamine treatment (G) group,ulinastatin treatment (U) group and glutamine combined with ulinastatin treatment (G+U) group. Isonitrogenous and isocaloric nutrition supports were provided to all groups. Rats in G group were administered with glutamine at 30% total nitrogen. U group were treated with ulinastation 40 000 IU/kg via tail vein injection. G+U group were given both glutamine and ulinastation. Nitrogen balance,D-lactate,DAO,and the levels of plasma TNF-α and IL-6 were determined. Results:After burn injury,the nitrogen balance,D-lactate,DAO activity,and levels of TNF-α and IL-6 significantly increased. Compared with B group,nitrogen balance in G group was significantly higher,while plasma D-lactate,DAO activity and levels of TNF-α were significantly lower. Levels of TNF-α and IL-6 in U group were significantly lower than those in B group. In G+U group,all the indexes except for nitrogen balance were significantly lower than those in B group. Conclusion:Glutamine combined with ulinastatin treatment may relieve intestine damage after severe burn injury and reduce the level of plasma inflammatory cytokines.

Abstract:Objective:To investigate the effects of serum amyloid A(SAA) on the expression of E-Selectin in human umbilical vein endothelial cells(HUVECs) and the potential signaling pathway. Methods:HUVECs were incubated with recombinant human SAA (0-40 μg/ml). The transcriptional level of E-selectin was assayed by real-time PCR. The protein secretion was determined by ELISA assay. Results:SAA (20 μg/ml) markedly induced E-selectin expression of mRNA and protein,and promoted E-selectin secreting in HUVECs. Pretreatment with a specific NFκB inhibitor,Bay-11-7082,could block the SAA-dependent E-selectin induction. Furthermore,electrophoretic mobility shift assay (EMSA) results showed that SAA could directly induce the activation of NFκB. Conclusion:SAA induces E-selectin expression through the NFκB pathway in HUVECs.

Abstract:Objective:To construct and indentify rat glutamine synthetase(GS) RNA interference expression vector and detect GS on cell adhesion of astrocytes. Methods:The rat GS specific siRNA sequence was designed by online software,and GS eukaryotic expression vector,GS-EGFP were co-transfected into Hela-G cells by LipofectamineTM 2000 regent. GFP expression level was used to identify the interference efficiency of GS siRNA. The effective siRNA sequence was inserted into eukaryotic expression vector pRNAT H1.l/Neo,and the vector was transfected into astrocytes. The expression of GS in astrocytes was detected by immunocytochemistry. After trasfected with GS siRNA vector,the effect of GS on cell adhersion was detected by actin specific immunocytochemistry. Results:GS siRNA inhibited the expression of GS-EGFP in Hela-G cells. The inserted sequence of GS siRNA was confirmed by DNA sequencing. The GS siRNA expression vector can specifically inhibit the expression of GS in astrocytes. GS RNA interference leads to a significant decrease of cell adhesion. Conclusion:The construction of rat GS siRNA eukaryotic expression vector was successful. The initial data showed GS level in astrocytes impacted on cell adhesion. The results of this study lay the foundation for further investigating the role of GS in reactive astrocytes after central nervous system injury.

Abstract:Objective:To investigate the possible association between the KLOTHO gene G-395A polymorphisms in promoter and acute coronary syndrome (ACS) in the Chinese Han population of Suwan region. Methods:This study was conducted with a case-control design including ACS group which is consisted of 326 patients with acute myocardial infarction (AMI) and unstable angina pectoris (UAP) and control group which is consisted of 219 subjects without coronary artery disease by coherence checks. The KLOTHO gene G-395A polymorphisms was determined by polymerase chain reaction(PCR) and gene microarray. Results:Compared with those in the control group,the frequencies of GG,GA,AA genotype and the A allele of the G-395A was not significantly different among ACS,AMI and UAP group,which were subgrouped according to gender and age (all P > 0.05). Conclusion:The G-395A polymorphisms in KLOTHO gene may not be the risk for ACS in the Chinese Han population of Suwan region.

Abstract:Objective:To the investigate the expression of TESTIN in human esophageal squamous cancer tissues,and discuss its value in the development of esophageal squamous cell cancer(ESCC) . Methods:The expression of TESTIN in 65 matched-pairs of ESCC tissues were detected by immunohistochemical staining,western blot,and FQ-RT-PCR methods. The correlations of TESTIN to clinicopathologic features of ESCC were analyzed. Results:The mRNA level of TESTIN was down-regulated in ESCC (P < 0.05).The positive rate of TESTIN protein was significantly lower in ESCC tissues than that in nomal tissues (36.9% vs 87.7 %,P < 0.01). The protein level of TESTIN was down-regulated in 45 (69.2%) of 65 cancers as compared with corresponding normal tissues. TESTIN expression was positively correlated with the differentiation of ESCC. Meanwhile,differences in gender,age,tumor size,TNM stage and lymph node metastasis were not significant. Conclusion:The decreased expression of TESTIN gene may play an importmant role in the development of esophageal squamous cancer. And thus TESTIN gene might be a novel candidate of tumor-suppressor. It may be an objective marker for prognostic factor and malignant level for ESCC.

Abstract:Objective:To evaluate the relationship between glutathione S-transferase class Theta-1(GSTT1) gene polymorphisms and oral Cancer. Methods:According to the theory and principles of systematic review,data from case-control studies regarding the association between GSTT1 gene polymorphisms and oral cancer were derived through electronic search of PubMed. To gain a more precise estimation of the relationship,a stratified meta-analysis with three subgroups was performed according to the races. Publication bias was also assessed by chi-square based Q test. Results:The overall effect between GSTT1 gene polymorphisms and oral cancer was OR=1.17 (95%CI:0.87~1.58). Furthermore,subgroup analysis were performed according to race. However,no significant effect was found[OR=0.99 (95%CI:0.71~1.36)in Asian;OR=1.07 (95%CI:0.69~1.64)in Caucasian;OR=1.89(95%CI:0.76~4.67)in Mixed race]. Conclusion:The association between GSTT1 gene polymorphisms and oral cancer was negative,and further large scale and multi-race epidemiological or biological functional studies were warranted to evaluate the results.

Abstract:Objective:To provide basis for the early diagnosis and prevention of coronary atherosclerotic heart disease (CHD) by calculating the ratio of pulse pressure (PP)/ankle-brachial index(ABI). Methods:Total 120 cases of hypertension patients were randomly selected. They were divided into two groups according to coronary angiography(CAG)and clinical manifestation,group A:CHD group,group B:non-CHD group. The average of 24-hour systolic blood pressure(SBP),diastolic blood pressure(DBP),PP,PP/ABI and some other risk factors of CHD between the two groups were compared;the predictive value of CHD was judged by Logistics regression analysis with the above factors,and the ratio of PP/ABI was analyzed;the cutoff value,specificity,sensitivity,positive predictive value and negative predictive value of in diagnosing CHD were predicted with the method of receiver operator characteristic(ROC) analysis. Results:There were no significant differences in SBP,DBP,PP and ABI between the two groups,but there existed great differences in total cholesterol,low density lipoprotein,diabetes and so on. PP and ABI correlate with PP/ABI. The ratio of PP/ABI differed between the two groups,which had statistical significance(P=0.001). Correlation and regression analysis indicated that there was independent predictive value of the PP/ABI ratio in the diagnosis of CHD. ROC analysis indicated that when the cut point equalled to or above 55.9,the sensitivity of predicting hypertensive patients with CHD is 65.9%,the specificity is 32.9%,the positive predictive value is 53.7% and the negative predictive value is 77.3%. Conclusion:In the diagnosis of hypertension patients with CHD,the ratio of PP/ABI is more sensitive than ABI or PP. The ratio of PP/ABI has clinical value in the early identification of hypertension patients with CHD.

Abstract:Objective:To compare the eradication rates of H.pylori among 7-day levofloxacin- based triple therapy,10-day levofloxacin-based sequential regimen and conventional sequential therapy. Methods:The total 64 patients with H.pylori were randomly divided into three groups. Eighteen patients in group A were treated with proton pump inhibitor(PPI),amoxincillin and levofloxacin for 7 days. Tweenty-five patients in group B were treated with PPI and amoxicillin for 5 days,and then with PPI,clarithromycin and tinidazole for another 5 days. Tweenty-one patients in group C were treated with PPI and amoxicillin for 5 days;and then with PPI,levofloxacin and tinidazole for another 5 days. Results:The eradication rate of 7-day levofloxacin-based triple therapy was 83.33%;the eradication rate of conventional sequential therapy was 84%;and that of 10-day levofloxacin-based sequential regimen was 76.19%. Conclusion:There were no significant difference in eradication rate of H.pylori among 7-day levofloxacin-based triple therapy,10-day levofloxacin-based sequential regimen and conventional sequential therapy.

Abstract:Objective:To investigate some surgical problems about the prevention and treatment of small-for-size syndrome after living donor liver transplantation(LDLT). Methods:LDLT was performed in 59 cases from February 2002 to October 2008. The clinical data were analyzed retrospectively. Four cases whose graft-recipient weight ratio (GRWR)were≤0.8%,were evaluated as small-for-size graft. Angioplasty and vascular graft provide unobstructed outflow,microsurgery,vascular graft and artery plasty were applied in hepatic artery reconstruction. Results:One case was diagnosed as small-for-size syndrome only and recovered four weeks later with suitable drug treatment. Recipients were followed up for 17 to 90 months with no death. Conclusion:On the premise of donor’s safety,much liver graft procurement,a large outflow, high-quality reconstruction of hepatic artery and portal vein are the key points to prevent the incidence of small-for-size syndrome.

Abstract:Objective:To compare the effect of different endometrial preparation methods on the clinical outcome of frozen-thawed embryo transfer (FET). Methods:Total 1 372 FET cycles from January 2007 to July 2009 were retrospectively analyzed and divided into two groups:natural cycle group(n=1 014)and ovulation induction cycle group(n=358). The latter was further divided into three Sub groups:HMG cycle group(n=127),clomiphene citrate + HMG cycle group(n=77) and letrozole+HMG cycle group(n=154). The patient age,duration of infertility,basal FSH,LH and E2 levels,number of embryo transferred,embryo quality and endometrial thickness,the rates of embryo implantation,biochemical pregnancy,clinical pregnancy,early abortion,late abortion and live births were compared between these different groups. Results:FET cycle outcomes were similar between the nature cycle group and the ovulation induction cycle group on early abortion rates(9.3% vs 9.8%,P > 0.05),late abortion rates (1.8% vs 1.6%,P > 0.05) and ectopic pregnancy rates(1.6% vs 0.5%,P > 0.05). There were significant differences of the rates of embryo implantation (26.7% vs 31.5%,P < 0.05),biochemical pregnancy (50.2% vs 57.5%,P < 0.05),clinical pregnancy (43.5% vs 51.1%,P < 0.05) and live births (38.0% vs 44.9%, P < 0.05) between the two groups. The clinical pregnancy rates of three ovulation induction cycle groups were 54.5%,51.9%,42.8%,respectively in letrozole+HMG cycle group,the HMG group and the clomiphene citrate+HMG group. There were no significant differences among them. Conclusion:Endometrial preparation of natural cycle or proper ovulation induction cycle are very suitable for FET.

Abstract:Objective:To detect serum levels of placenta growth factor (PLGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) during different periods in normal pregnancy and patients with preeclampsia and to evaluate their role in the pathogenesis of preeclampsia. Methods:Women enrolled prospectively in following groups:healthy pregnant control(n = 200) and preeclampsia patients,who had mild preeclampsia(n = 31) or severe preeclampsia(n = 26) . The serum PLGF and sFlt-1 levels were measured by enzyme linked immunoassay(ELISA). Results:Maternal serum PLGF levels during all periods were significantly lower in the preeclamptic women than in normal controls(P < 0.01). The levels of maternal plasma sFlt-1 were significantly higher in the preeclamptic women than in normal controls during the latter three periods of pregnancy(P < 0.01). The serum sFlt-1/PLGF ratio was significantly higher in the preeclamptic women than in normal controls(P < 0.01). Conclusion:Alterations in the levels of sFlt-1 and free PLGF in maternal serum were associated with the occurrence and development of preeclampsia.

Abstract:Objective:To evaluate the clinical value of rapid culture of Mycoplasma pneumoniae (MP) in diagnosis of MP for children and explore the potential influence factors related to the MP rapid culture. Methods:The rapid MP culture method was used for throat swabs of acute respiratory infection,and passive agglutination assay was used to detect serum MP antibody(Ab). Meanwhile the multiple related factors which can influence the MP infection were analyzed. Results:Female children patients’ positive rate of MP cultivation was 46.26%,higher than male children’s (42.16%). The positive rate of rapid MP cultivation for children with 3~5 years old were 54.05%,highest among all groups with different years old(P < 0.001). The results of rapid MP cultivation was different among 4 seasons. The positive rates of rapid MP cultivation in winter and spring were 48.05% and 49.03%,respectively,obviously higher than those in summer and autumn (26.26% and 36.97%,respectively)(P < 0.001). Moreover,the positive rate of rapid culture for MP was in line with the results of serum MP-Ab detection (P=0.556). No evidence was found between the common clinical symptoms such as fever,cough,expectoration and infection of MP. The increasing percentage of blood neutrophile granulocyte was similar to the respiratory infection resulting from the other pathogenic microorganisms,suggesting this can not be viewed as an independent index for diagnosis of MP inflection. Conclusion:MP inflection is common in winter and spring. Children and old people is easily infected. Children with 3~5 years old is susceptible to MP infection. The patients with MP infection is lack of special clinical symptoms. The rapid MP cultivation method plays a significant role in early rapid diagnosis of MP infection.

Abstract:Objective:To observe the effect of valdecoxib on PGE2 secretion of human gingival fibroblasts(HGFs) which were treated with lipopolysaccharides(LPS). Methods:The quantification of cell proliferation and cell viability was analyzed by ELISA assay to determine the influences of LPS and valdecoxib on the secretion of PGE2 in HGFs cultured in vitro. Results:LPS can stimulate the HGFs secreting PGE2 in a dose-dependent manner;and valdecoxib can significantly inhibit the effect of LPS on the secretion of PGE2,which is also in a dose-dependent manner. Conclusion:Valdecoxib can inhibit PGE2 secretion of HGFs stimulated with LPS.