Objective To establish two PCR assays to rapidly and simultaneously detect U.parvum and U.urealyticum.Methods Two PCR assays were established through designing two sets of specific primer targeting urease gene B of U.parvum and U.urealyticum,respectively.The standard strains of U.parvum and U.urealyticum and the clinical samples were detected by these PCR assays and PCR products of the standard strains and two clinical specimens were directly sequenced and carried out specific and sensitive assays.Forty-eight clinical specimens were tested for culture and the PCR assays and tow methods were compared.Results The standard strains of U.parvum,U.urealyticum and the clinical specimens were successfully and differentially detected by the PCR assays and the sequencing results were found to have a complete similarity to the GenBank sequences.The 14 strains of other bacterial genera including 5 strains that produced urease were not amplified by these PCR assays.Each assay had a detection limit of 10 copies/?l of plasmid DNA.The positive rate in 48 clinical specimens by PCR(54.2%)was higher than that by culture(39.6%)(P

Objective To investigate the similarities and differences of 3 methods for detecting hepatitis B virus S 1(PreS1) ,and select the appropriate method for detecting clinical samples .Methods The PreS1 of the serum samples from chronic hepatitis B pa-tients were tested with enzyme immunoassay analyzer ,time-resolved method and the manual method .To compare the repetition rate ,select PreS1 antigen strongly positive serum and weak positive serum were detected 15 times by three methods ;To compare the explicit rate ,the reaction temperature was raised or lowered by 3 ℃ .Results The positive rate of three methods was 93 .53% , 92 .81% ,92 .81% .Automated ELISA reproducibility CV strong positive CV3 .62% ,weakly positive CV was 13 .42% ,CV of time-resolved method for the 2 kinds of samples were 5 .10% ,7 .92% ,manual methods CV 11 .10% 29 .88% ;changing the reaction incu-bation temperature 3 ℃ ,automatic detection ELISA all specimens S/CO value decreased ,increasing the chance of a false negative . The manual methods and time-resolved detection method for all specimens S/CO values increased ,increasing the chance of a false positive .Conclusion The detection rate and repeatability of automated ELISA were better .The time-resolved method followed and the manual methods were poor .

Objective To study the diagnostic value of multiplex protein microarray and ELISA for human serum pathogens.Methods Totally 230 national standard samples were tested by multiplex protein microarray and ELISA respectively for the pathogens of HBV,HIV,HCV and treponema pallidum(TP).Results The matched pair test for numeration data showed ?2=0.5,P=0.125,indicating that no statistical significance between the two methods.Conclusion Multiplex protein microarray shows high specificity and sensitivity to the four pathogens so that it is worth extensive application.

Objective To establish an immunoassay based on microfluidic chip for detection of TSH and total T4.Methods The direct-write laser micromachining technology was used to fabricate microfluidic chip,luminol-H2O2 as luminator.This method and Roche E2010 electrochemiluminescence immunoassay were used to examine TSH by double-antibody sandwich method and total T4 by competitive principle in 50 serum samples collected at our outpatient clinic.Results The detection was completed within 25 min.Ten microliter sample and reagent was required.The detection range of TSH was 0-50 ?IU/ml and its average intraassay and interassay precision was 4.7% and 4.6% respectively and its average recovery rate was 95.3%.The detection range of T4 was 9.375-300 ng/ml,and its average intraassay and interassay precision was 4.9% and 4.7% respectively and its average recovery rate was 91.27%.Conclusion Rapid detection of TSH and T4 by microfluidic chip-based immunoassays is convenient,stable and accurate and of less reagent and time consuming.

Objective　To investigate the significance of TGF-β1, TGFRl and TGFR2 in the pathogenesis and prognosis in patients with myelofibrosis. Methods　The expression of TGF-β1 and its receptors (TGFR1 and TGFR2 ) in bone marrow tissues and the level of TGF-β1 in the blood of 23 patients with myelofibrosis were detected by SABC immunocytochemistry and ELISA repectively. Results　Expression of TGF-β1 and TGFR 1 was significantly higher in primary and secondary myelofibrosis patients than that of the control. No significant difference of TGFR2 expression was found between the groups of myelofibrosis and the control (P>0.05). The level of TGF-β1 in the blood of the patients with myelofibrosis was significantly higher than that of the control (P<0.01) and more obvious in secondary cases while TGF-β1 decreased nearly to the normal level when patients were in clinical remission. Conclusion　TGF-β1 and it's receptors may be involved in the pathogenesis of myelofibrosis and might be of importance for the prognosis of the patients with myelofibrosis.

ObjectiveTo investigate the effects and problems of problem-based learning ( PBL ) teaching methods that are adopted in the laboratory medicine practice teaching. MethodsOne hundred and four undergraduate students of 5-year system of laboratory medicine were selected to use the PBL teaching methods during the laboratory medicine practice. ResultsIt showed that the PBL teaching methods obtained good probation effect. But teachers and students are required to make greater efforts in PBL teaching mode than in traditional mode, and the teaching methods was also needed to be consummated.ConclusionsUsing the problem-based teaching methods the comprehensive ability of the student is enhanced. Therefore it deserves to be generalized during laboratory medicine practice teaching.

Objective To construct an immunosensor for detecting CD4+ T lymphocytes without labeling .Methods The staphy-lococcus protein A(SPA) method was adopted to conduct the oriented immobilization of CD4 monoclonal antibodies on the gold in-terdigitated microelectrode surface for capturing CD4+ T lymphocytes .Then cyclic voltammetry(CV) method was used to conduct the representation of modification situation on the gold interdigitated microelectrode surface .Finally the electrochemical impedance spectroscopy(EIS) was used to detect the impedance of CD4+ T lymphocytes captured by the immunosensor .The standard curve was drawn by the impedance values change obtained by the equivalent electric circuit fitting .Results The linear range of this im-munosensor for detecting CD4+ T lymphocytes was (5 × 103 -5 .0 × 106 )/mL ,with lower detection limit of 5 .0 × 102/mL .Conclu-sion The constructed immunosensor has accurate and reliable detection results uhidn is simple to operate accurate ,convenient and cheap ,which might be expected to be used in the real-time detection system ,and offers help for realizing rapid ,accurate and inex-pensive CD4+ T lymphocyte count .

Objective To investigate the effect and the mechanism of the GITR-antibody(glucocorticoid-induced tumor necrosis factor receptor-ligand antibody) on the mouse leukemia model induced by L615.Methods The mouse leukemia models induced by L615 cells were divided into 4 groups: negative controls(peritoneal injection of normal saline,0.2 ml/d),GITR group(GITR,100,infused through caudal vein 2 d before leukemic lymphocytes inoculation,again at dose of 50 ?g/each mouse after inoculation),Cyclophosphamide group(200 mg?kg~(-1)?d~(-1),intraperitoneal injection from the 3~(rd) day after inoculation for 3 d),GITR+ Cyclophosphamide group(100 mg?kg~(-1)?d~(-1) Cyclophosphamide instead).The survival time,leukocyte counting in the peripheal blood,liver and spleen index were calculated and the pathological examination of liver,spleen were performed.Results GITR-ligand could prolong the survival time of mouse leukemia model,lead the necrosis and apoptosis of leukemic cells in bone marrow,decrease the liver and spleen index,decrease and relieve the leukocyte increase of peripheal blood and the irregular swelling of liver and spleen.Conclusion Through immunoregulation,GITR-antibody can inhibit the L615 leukemic cells effectively,therefore inhibit the progress of leukemia to some extent.

Objective To explore how the antibody of glucocorticoid-induced tumor necrosis factor receptor (GITR) exerts inhibitory effect on the L615 leukemia cells by strengthening the activation of the NK cells. Methods The 24 established L615 leukemia mice were equally and randomly divided into 4 experimental groups according to different drugs given intraperitoneally, groupⅠ (normal saline), Ⅱ (GITR), Ⅲ (cyclophosphamide), and Ⅳ (GITR +cyclophosphamide).Then the NK cells were extracted from the spleen of mice as effective cells, and L615 leukemia cells served as the target cells. The changes of the NK cells’killing activation was observed in vivo. The mRNA levels of 3 proteins tightly related to the NK cells’activation Perforin, IFN-? and Fas mRNAs were detected with RT-PCR. Results The GITR-antibody enhanced the killing activity of the NK cells obviously, with the expressions of the 3 proteins increasing obviously. Conclusion By regulation of the Treg cells, the GITR-antibody can inhibit the L615 leukemia cells through enhancing the NK cells' killing activity.

Objective: To study the effects of pharmaco-serum from rabbits administrated intragastrically with Fuzheng Jiedu decoction(FJD) on injured human bronchial epithelial(16HBE) cells exposed to nickel sulfate(NiSO4).Methods: Cultured 16HBE cells were treated with both NiSO4 and different doses of FJD pharmaco-serum in vitro.The detections of cell activities and viabilities were carried out.Meanwhile,activities of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD) as well as maleic dialdehyde(MDA) contents were measured.Results: FJD pharmaco-serum could decrease mortalities and increase viabilities of 16HBE cells exposed to NiSO4.In cells co-treated with NiSO4 and FJD pharmaco-serum,the content of MDA was decreased,while GSH-Px and SOD activities were increased at the same time.High dose of FJD pharmaco-serum had the most dramatic effect.Conclusion: This study suggested that FJD could antagonize NiSO4-induced toxicity,which may be involved in its antioxidant function.