Objective To understand the residual risk of transfusion blood donation in the native region and to conduct the sim-ple comparison of Roche diagnostic company′s first generation and second generation nucleic acid combined detection reagents . Methods The menstrual blood specimens qualified by the routine test in this center (serology test and enzyme immunodetection as-say results were negative) from May to July 2013 were selected and performed the 6-specimen mixed sample test by adopting the Roche Diagnostics company′s first-generation and second-generation nucleic acid combined detection reagents in the United Roche COBAS s 201 operating system ,if the test had the reactivity ,the nucleic acid identification tests and other complementary serologi-cal testing were performed .Results 9 417 specimens were detected ,8 cases of nucleic acid positive were screened out with the posi-tive rate of 0 .85 ‰ ,through the detection of nucleic acid identification reagents ,4 specimens were confirmed as positive for HBV nucleic acid ,1 case was positive for HCV nucleic acids and the rest 3 cases of uncertain HBV-positive .In addition ,with the first generation reagent (reference reagents) as the relative standard ,the second generation reagents (assessment reagent) had the over-all positive coincidence rate of 66 .67% ,the negative coincidence rate of 99 .98% and the total coincidence rate of 99 .96% .In addi-tion ,no HIV nucleic acid positive specimens was found by the nucleic acid test .Conclusion The nucleic acid detection technique can effectively shorten window period,further improve the safety of blood transfusions ,at the same time the overall conformance situ-ation of the Roche′s first-generation nucleic acid combined detection reagents and the second generation combined detection reagents is good ,but there are still lesser differences in the detection of HBV .

OBJECTIVE To explore and evaluate the efficacy of preventing the central venous catheter-related infection using catheter-sealing separately with gentamicin and heparin.METHODS One hundred and thirty six hemodialysis patients with temporary indwelling central venous catheters were enrolled in this study,and randomly assigned into 3 groups: Group A(catheter-sealing separately with gentamicin and heparin,n=46),Group B(catheter-sealing with gentamicin mixted heparin,n=45) and Group C(catheter-sealing with heparin,n=45).Complications such as infection were monitored.RESULTS Rate of catheter-related infection and intravenous catheter infection were without significant difference between Groups A and B,but were obviously lower than that in Group C(6.5% and 4.4% vs 22.2%,P

Objective To explore the diagnostic significance of C reactive protein(CRP)in diagnosing haemorrhagic fever with renal syndrome(HFRS).Methods 96 cases of patients with HFRS of different stages were enrolled in this study,and serum speci-men were collected.30 cases of patients with fever of unknown origin(fever of unknown origin group)and and 30 healthy individu-als(healthy control)were selected as control.Serum levels of CRP,alanine aminotransferase(ALT),aspartate amino-transferase (AST),creatine phosphokinase(CK),lactate dehydrogenase(LDH)and creatinine(Cr)were detected.Results Serum levels of CRP in patients with HFRS of different stages were lower than that in the fever of unknown origin group,had statistically significant differences(P <0.05).The variation trend of CRP in each stage of HFRS was consistent with the trend of AST,CK,LDH and Cr. Conclusion CRP has clinical significance in differentiating HFRS from fever of unknown origin.

Objective:To obtain some effective objective markers used to predict the early liver metastasis of colorectal tumor,the relationship of liver metastasis of colorectal tumor with associate detection three markers such as CK20mRNA、CD44V6 and PCNA was studied. Methods:The expression of CK20mRNA in portal venous blood from 30 colorectal cancer patients was detected by fluorescent quarto RT-PCR,and the results of CD44V6 and PCNA in colorectal cancer tissue were determined by means of immunohistochemistry, and then compared with control groups through statistics analysis. Results:The rate of positive expression of CK20mRNA in colorectal cancer patients' portal venous blood was obviously superior to the level of benign pathological changes controls(P

Objective To construct a naive human Fab fragment phage display library,from which the anti-gp96 antibodies may be panned by the gp96 purified from the tissue of urothelial carcinoma in the urinary bladder and provide a basis to new therapy for the malignant tumors.Methods Peripheral blood lymphocytes were isolated from 800 ml of blood,which was obtained from four healthy blood donors.The heavy chain Fd and light chain cDNA synthesized from the total RNA of lympbocytes were amplified by PCR with variable regions 5' and 3' primers of heavy and light chain, and the amplification products were ligated into the phagemid vector pComb3, then the ligated sample was transformed into competent E.coli XL1-Blue by electroperation.The transformed cells were infected with VCSM13 helper phage to yield recombinant phage antibody Fabs.The phagemids abstracted from amplified E.coli were cut with endonucleases such as Sac Ⅰ,Xba Ⅰ,Xho Ⅰ and Spe Ⅰ, and both the phage antibody Fabs and phage-raids abstracted from amplified E.coil were amplified by PCR to monitor the insertion of the genes of light chain or heavy chain Fd fragment.The gp96 purified from the urothelial carcinoma tissue of the bladder by affinity chromatog-raphy on eoncanavalin-A sephnrose and DEAE-sephnrose ion exchange chromatography were utilized as antigens to process three rounds of panning to the original Fab antibody library.Results The quantity of total RNA and cDNA were qualified.By combination of light chain and heavy chain genes, an antibody library containing 6.6×106 clones was obtained, and both the cutting of enzymes and PCR showed that there were the genes of light chain or heavy chain Fd fragment in the phagemids.The gp96 protein was obtained from urothelial carcinoma tissue in the urinary bladder.After having been panned by gp96, the original antibody library gained enrichment by 68 times.Conclusion Utilizing the technology of phage surface display, specific antibody can be gained from the human naive Fab phage display library,which can be used for immunological therapy for tumors.

Objective To investigate the expression and sensitivity of 6 muhidmg resistance gene products(MDR) : GST-π、LRP、MRP、Topo Ⅱ、P-gp、TS in gastric cancer during neoadjuvant chemotherapy. Methods Expression of the 6 muhidrug resistance gene was detected by immunohistoehemistry in 35 cases of stomach cancer tissues before and after neoadjuvant chemotherapy. The relationship between muhidrug resistance gene and neoadjuvant chemotherapy was analyzed. All patients were given FOLFOX4, and the effects were evaluated according to World Health Organization criteria. Results The overall response rates to chemotherapy was 46%, expression of the 6 mtdtidrug resistance gene in 35 cases of stomach cancer tissues were as follows:GST-π was 40% ,LRP was 69% ,MRP was 34% ,Topo Ⅱ was 37% ,P-gp was 86%, TS was 40%, expression of the 6 muhidrug resistance gene did not change during neoadjuvant chemotherapy; GST-π、LRP、MRP、Topo Ⅱ、P-gp were not correlated with chemotherapy sensitivity(P > 0.05), while TS was significantly correlated with chemotherapy sensitivity (P = 0.0048). Conclusion FOLFOX4 does not effect a change in the expression of the 6 muhidrug resistance gene: GST-π、LRP、MRP、Topo Ⅱ、P-gp and TS, while TS is significantly correlated with gastric cancer chemotherapy sensitivity.

Objective To study the changes condition and clinical significance of serum cystatin C(Cys‐C) ,β2 microglobulin(β2‐MG) and retinol binding protein (RBP) detection in different stages of hemorrhagic fever with renal syndrome(HFRS) .Methods The levels of serum Cys‐C ,β2‐MG and RBP were detected in 22 patients with HFRS and 30 cases of healthy physical examination and the detection results were statistically analyzed by the SPSS13 .0 software .Results The serum Cys C and β2‐MG levels in every stage of HFRS were increased compared with the healthy control group(P< 0 .05) ,which in the oliguria stage was most significant (P< 0 .01) .RBP had no significant differences in the fever stage and shock stage compare with the healthy control group (P >0 .05) ,while RBP was significantly increased in the oliguria stage ,polyuria stage and convalescence stage(P< 0 .01) ,but no statisti‐cally significant differences among the three stage(P> 0 .05) .Conclusion β2‐MG is more sensitive than Cys‐C in the early stage of HFRS ;RBP has the clinical guidance significance in the progression of HFRS .

Objective Discuss the interference of injection cefotiam on vanadate oxidation method and dry chemical method assay total bilirubin .Methods Collected 60 examples ,include total bilirubin concentration 20 examples less than 20 μmol/L ,20 examples between 150-220 μmol/L and 20 examples between 350-410 μmol/L ,add an equal volume of various concentrations of cefotiam in each case ,formulated into cefotiam final concentrations of 300 ,150 ,75 mg/L of serum samples as the test group ,add an equal volume of water in each serum samples as the control group ,determine all the samples total bilirubin concentration respectively by vanadate oxidation method and dry chemical method ,compared the interference of cefotiam on determined total bilirubin by two method ,analyze the data by SPSS13 .0 .Results Determined total bilirubin by dry chemical method ,the test group higher than the control group ,the difference was statistically significant(P<0 .05) ,at the same total bilirubin levels ,with cefotiam concentrations decreased ,increased rate of total bilirubin concentration were decreased in the experimental group .Determined total bilirubin by vanadate oxidation method ,when the total bilirubin concentration between 150 -220 μmol/L ,the test group was higher than the control group ,the difference was statistically significant(P<0 .05) .Conclusion Interference of injection cefotiam on determined to‐tal bilirubin by dry chemical method is strong ,and with the drug concentration increased ,effect is more obvious ,but determination of total bilirubin by vanadate oxidation method has almost no effect .

AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and “bystander effect” were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The “bystander effect” was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G0-G1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of “bystander effect” was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.

AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and "bystander effect" were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The "bystander effect" was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G 0-G 1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of "bystander effect" was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.