Objective To separate the SO-Rb 50cells antigen corresponding to the monoclonal antibody of anti-retinoblastoma. Methods The antigen corresponding to the monoclonal antibody of anti-retinoblastoma was separated elementarily by ion-exchange chromatography, and was identified by dot-blotting using the monoclonal antibody of anti-retinoblastoma. The target protein band of the antigen was separated in light of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results A special unmixed band of SO-Rb 50cells antigen was separated with the relative molecular weight of 83?103. Conclusion The antigen corresponding to the monoclonal antibody of anti-retinoblastoma could be separated from SO-Rb 50cells.

Objective To explore the effect of a novel colloidal gold immunochromatographic test strip for detection of group B streptococci (GBS).Methods A total of 202 cases of swab of vagina or neck of uterus were collected,and they were detec-ted by novel strip and control strip to evaluate their clinical applications.Results Sensitivity of novel strip was about 105 CFU/ml and the detection time was about 5 to 8 minutes,and it showed better sensitivity and shorter detection time com-pared with control strip.In the 202 cases of clinical samples,the detection results of 197 cases were in consistent with the control strip,however,the detection results of 5 cases were not in consistent.The positive coincidence rate and negative coin-cidence rate were 97.5% and 97.54% respectively,and the total coincidence rate and Kappa value were 97.52% and 0.948 respectively.The consistency test showed no significant difference between this strip and control strip.Conclusion This method was a effective technology for diagnosing of infection caused by GBS,and had high value in clinical application.

OBJECTIVE: To compare the effects of different factor combinations on diabetic nephropathy(DN)rats model and to determine the preferred plan for the establishment of DN rats model.METHODS:A total of 30 SD rats were randomly divided into 5 group(6 rats in each group):group A was assigned to receive placebo(normal control),group B to receive high-lipids and high-sucrose feeding,group C to receive high-lipids and high-sucrose feeding plus streptozotocin(STZ),group D to receive high-lipids and high-sucrose feeding plus doxorubicin hydrochloride and STZ,group E to receive high-lipids and high-sucrose feeding plus mononephroctomy and STZ 12 weeks later the model rats were put to death for the determination of serum parameters including blood lipid,glycosylated hemoglobin,renal function,aldose reductose(AR),glomerular filtration rate(GFR),SOD and MDA .RESULTS:The rats in group E showed significant increase in blood lipid,in particular the increase of AR,and which also showed significant differences in GFR and oxygen free radical indices as compared with normal group and the other model groups.CONCLUSIONS:The rats model that fed with high-lipids and high-sucrose feeding for 4wk prior to mononephroctomy followed by the postoperative intraperitoneal injection of low dose of STZ 2 weeks later proves to be the optimal one.

Objective To establish a single nucleotide polymorphisms genotyping (SNP) method for a convenient, accurate, and routine analysis of clinical samples. Methods Based on the design of oligonucleotide probe, the assay was performed through three steps:the conjunction of the detection probe, universal amplification, labeling and ELISA reaction. The genotype of each SNP was revealed by reading signals of each set of reaction tubes. This assay was applied to detect sixty-two plasma samples of lung cancer for circulating DNA for three SNPs of EGFR, c.2573T＞G(L858R), EGFR, c.2582T＞G＞T(G719C). Results were compared with those obtained by direct sequencing. Results The heterozygote mutation was identified for L858R by both methods, although no mutation was detected for L861Q and G719C. Six samples were identified as heterozygotes with the new method, and only two samples were unambiguously identified as heterozygotes by the direct sequencing. Two additional samples could not be identified as heterozygotes because the peak of mutant allele was very low compared with that of wild allele. Conclusion The developed method enabled accurate identification of SNP in a convenient manner, and which is adapted to routine analysis from heterogeneous samples unambiguously.

Objective To investigate the value of biliary tumor markers for differential diagnosis of the benign and malignant biliary tract diseases.Methods Tumor markers (CA19-9,CEA and CA242) examination and bacterial culture were performed in a total of 160 patients,who underwent therapeutic endoscopic retrograde cholangiopancreatography (ERCP) for biliary diseases.Resuts There were significant differences between malignant group and benign group in bile and serum in the level of CA19-9,CEA and CA242 (P ＜0.05) ; Cut-off value,according to ROC curve,was 239 ku/l in CA19-9,40 ng/ml in CEA and 60 ku/ml in CA242,respectively.There were significant differences between the bile marker and the serum marker in sensitivity,accuracy,negative predicative value of CEA (P ＜ 0.05).No significant differences was found in specificity between the serum group and the bile group.There were significant differences in bile CA19-9 level between cholangiocarcinoma,pancreatic cancer,duodenal papilla carcinoma with carcinoma metastasizing to bile duct,and hepatocellular carcinoma (P ＜ 0.05).Both in benign group and malignant group,there were significant differences in CA19-9 level between infectious bile and noninfectious bile (P ＜ 0.05).Conclusion The level of CA19-9,CEA and CA242 in bile can be applied to differentiate benign and malignant biliary diseases.The bile tumor markers do not have advantage over serum tumor markers in specificity for diagnosis.Bile bacterial infection can result in the elevation of bile CA19-9 while it does not have impact on differential diagnosis.

Objective To explore the change of Brain natriuretic peptide(BNP) and endothelin-1 (ET-1) levels in artial septal defect(ASD) patients and the relationship among BNP,ET-1 and pulmonary pressure.Methods 105 final diagnosed ASD patients were divide into non-pulmonary hypertension group (nPH group) and pulmonary hypertension group(PH group),and the PH group were divided into two subgroup:slight PH group,moderate and sever PH group.According to the altitude of habitation,105 ASD patients also were divided into 3 groups:＜ 2 500 m group,2 501-3 500 m group and ＞ 3 500 m group.Plasma BNP were measured by radioimmunity method and ET-1 were measured by ELISA.The data analysis used single factor analysis and Fisher least singnificant difference t test.Results Both the plasma BNP levels (152.34 ± 40.61) pg/ml and ET-1 level (137.69 ± 37.17) pg/ml of the ASD-PH group were significantly higher than those [BNP (126.70 ± 32.27) pg/ml,ET-1 (92.92 ± 32.3) pg/ml] of ASD-nPH group.There were strong difference in plasma BNP levels and ET-1 levels among different degree PH groups(F =6.782,P ＜ 0.05 ; F =8.475,P ＜ 0.05).Statistical difference were also shown in BNP(F =6.846,P ＜ 0.05) and ET-1 (F =9.327,P ＜ 0.05) levels by compared difference altitude groups.The BNP levels are positively correlated with mean pulmonary artery pressure (r =0.326,P ＜ 0.05),size of defect (r=0.301,P＜0.05) and the altitude of habitation (r =0.252,P＜0.05).Conclusion Plasma BNPand ET-1 levelsof ASD-PH group significantly higher than those of ASD-nPH group.By the increasing of the altitude and PH degree,the plasma BNP and ET-1 levels are increasing,which suggest that BNP and ET-1 play an important role on the proceeding and development of the PH and hypoxia promoted secretion of BNP and ET-1.

Objective The incidence of alcoholic fatty liver increases year by year in recent years .The aim of this study was to establish an animal model of AFL to investigate the pathogenesis of hepatic fibrosis . Methods This study involved 40 male Japa-nese rabbits aged (17.01 ±1.54) d and weighing 1.00-1.52 kg, which were equally randomized to an experimental group and a control group.The animals in the former group received lavage of 10 mL of 50%ethanol twice a day, with normal feedstuff and water, while those in the control group received normal feedstuff and water only .We performed ultrasonography for dynamic liver presentation before and at 12, 16, and 20 weeks after feeding, followed by pathological observation of the livers . Results After 12 weeks of eth-anol garage , fatty liver was observed in 18 of the rabbits and it deteriorated with the prolonged time of administration . The body weight was significantly decreased in the experimental rabbits as com-pared with the controls at 16 weeks ([2.48 ±0.30] vs [2.78 ± 0.15] kg, P<0.05) and 20 weeks ([2.61 ±0.44] vs [3.10 ± 0.13] kg, P<0.05).Ultrasound and pathological grading showed 1 mild, 3 moderate, and 13 severe cases of fatty liver in the experimen-tal group, but none in the control , and pathological examination re-
vealed similar results (1 mild, 4 moderate, and 12 severe cases of fatty liver) in the former group.At 20 weeks, alcoholic fatty liver was found mainly in the S3-S4 stage. Conclusion Alcoholic fatty liver models could be successfully established in rabbits by etha-nol garage and ultrasonography is useful for monitoring the development and progression of the condition .

Objective To investigate the dynamic change and the clinical curative effect evaluation of plasma (1-3)-beta glucan D (BG) in the patients with pulmonary disease complicating fungal infection .Methods The MB-80 miroorganism dynamic rapid de-tection system and fungi BG detection kits were adopted to detect plasma BG content before and after treatment in 87 cases of pul-monary disease complicating fungal infection and the controls .The sputum culture in the patients was performed before and after treatment .Results Plasma BG levels before antifungal therapy ,at 1 ,2 weeks after treatment in 87 patients were (162 .81 ± 70 .03) , (15 .89 ± 30 .88) and (4 .58 ± 7 .87)pg/mL ,which in the control group was (5 .62 ± 1 .83)pg/mL ,plasma BG level had statistical differences between before treatment and at 1 ,2 weeks after treatment in the patients with the control group (P<0 .05);Plasma BG levels between at 1 week after treatment with at 2 weeks after treatment and the control group had statistically significant differ-ences (P<0 .05) .Among 87 patients ,66 cases were positive sputum culture at 1 week after antifungal drug treatment and 9 cases were positive sputum culture at 2 weeks after treatment .Conclusion Continuously monitoring the patient′s plasma BG level com-bined with the sputum fungal culture results ,clinical symptoms and lung shadow in X-ray has certain clinical value to judge the anti-fungal effect .

Objective To investigate the value of multi-slice spiral CT(MSCT) in diagnosis of laryngocarcinoma in early stage.Methods MSCT data of 28 patients with laryngocarcinoma confirmed pathologically were analyzed retrospectively.Images quality was evaluated and the results obtained with various windows and CT virtual laryngoscopy (CTVL) were compared.Results 20 cases(69%) could be displayed with conventional soft tissue windows,24(81%) could be demonstrated with lung windows and 27(93%) could be demonstrated with CTVL.Conclusion MSCT can effectively demonstrate laryngocarcinoma,and can be applied routinely in examination of laryngocarcinoma.

Objective:To establish the technique for real-time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-PCR)by DNA melting curve analysis for detecting the CDR3 shewing of TCR alpha gene repertoire in human peripheral blood.Methods:Total RNA of peripheral blood mononuclear cell(PBMC)from 4 healthy donors and 2 patients with lymphomatous leukemia were transcripted reversely into cDNA.The cDNA of 32 TRAV gene family CDR3 was amplified by FQ-PCR.Analysis of the monoclonal/oligoclonal/polyclonal CDR3 spectratyping with DNA melting curve.Results:The FQ-PCR products of 32 TRAV family CDR3 were showedas a blur land at the predicted of products size in healthy donors and parts of TRAV family CDR3 products disappeared in patients on 1.5% agarose gel by Gold-View staining.The 32 TRAV family CDR3 were showed with different frequencies by relative fluorescence quantitative in healthy donors and the patients.The CDR3 spetratyping for 32 TRAV families was showed as polyclonal peak(Gaussian distribution)in healthy donors but showed as different monoclonal/oligoclonal/polyclonal peak in the patients with lymphomatous leukemia with DNA melting curve analysis(we called "melting curve spectratyping of CDR3")Conclusion:The study suggests that the technique of "FQ-PCR with DNA melting curve analysis be convenience and celerity for detecting the CDR3 skewing of TCR alpha gene repertoire in human peripheral blood.