Objective To evaluate the practical value of Serology, PCR-SSP and PCR-SSOP for HLA-B typing. Methods A total of 30 samples, the blood of patients and donors waiting kidney transplantation, were used in the study. HLA-B typing was performed by one-step monoclonal antibody typing, micro PCR-SSP typing and PCR-SSOP reverse hybridisation. Results All samples were successfully typed by three methods. The difference between serological and PCR-SSP typing was 13%. The difference between PCR-SSOP and PCR-SSP typing was 3%. Conclusion Serology is high discrepancy rate and low-resolution, but cheap, simple and rapid. PCR-SSP and PCR-SSOP typing are high specific and accuracy. SSP is suitable for several samples, and SSOP is for lots of samples simultaneously although it needed long time.

Objective To investigate the significance of enzyme-linked immunosorbent assay (ELISA) in cross-match test of donor-recipient before organ transplantation.Methods HLA glycoproteins were prepared by solubilizing the lymphocytes of donor with a non-ionic detergent, and bond to the monoclonal antibody specific for HLA class Ⅰ or Ⅱ immobilized in the ELISA tray. ELISA with addition of recipients' serum and complement-dependent cytotoxicity (CDC) were compared.Results Cross-match test was performed by two methods for lymphocytes of 40 donors and sera of 72 recipients. All samples were successfully tested. The results of one pair of donor-recipient cross-match test by two methods were different.Conclusion ELISA for cross-match test is simple, convenient and time-saving, but more sensitive and specific than CDC.

Objective To evaluate the effect of Danshen Chuanxiongqin injection combined with Xuesaitong capsule on blood viscosity in patients with pulmonary heart disease.Methods 80 patients with pulmonary heart disease who were admitted to our hospital between January 2013 and November 2015 were studied,the patients were randomly divided into the combined group and the control group according to the order of admission,40 cases in each group.Control groups were treated with Xuesaitong capsule,and the combined group were treated with Danshen Chuanxiongqin injection on the basis of the control group.The clinical efficacy,blood viscosity(whole blood viscosity,plasma viscosity,whole blood viscosity ratio),blood indexes(blood platelet aggregation rate,erythrocyte aggregation index,erythrocyte deformation index),cardiac function and safety were observed.Results There were no significant differences in blood rheological indexes,LVEF,MAP and HR between the two groups before treatment.After treatment,the whole blood viscosity,plasma viscosity,whole blood viscosity ratio,platelet aggregation rate,erythrocyte aggregation index,erythrocyte deformation index,MAP and HR in the two groups were significantly lower than those before treatment,while LVEF was significantly higher,and the changes in the combined group were more significant than those in the control group(P<0.05).After treatment,the clinical total effective rate of treatment in the combined group(88.89％)was significantly higher than that in the control group(62.50％).Conclusion Danshen Chuanxiongqin injection combined with Xuesaitong capsule is effective in the treatment of pulmonary heart disease.It can significantly reduce blood viscosity,blood related indexes and improve cardiac function.

BACKGROUND:There are a lot of reports about the association between estrogen receptor α polymorphism and osteoarthritis susceptibility, but the results are stil some controversial. OBJECTIVE:To investigate the relationship between the estrogen receptor α gene PvuII, XbaI site polymorphisms and genetic susceptibility of osteoarthritis. METHODS: A computer-based search of PubMed, web of science, Wanfang, CNKI, Weipu and China Biology Medicine Disc was performed for the published case-control studies addressing the association between estrogen receptor α gene PvuII, XbaI site polymorphism and osteoarthritis susceptibility. Odds radio (OR) and 95% confidence interval were used to analyze the correlation between estrogen receptor α gene PvuII, XbaI site polymorphism and osteoarthritis. Fixed or random effect models were selected for pooledOR calculation. Publication bias was assessed. Al statistical analysis was constructed with Revman5.1 software. RESULTS AND CONCLUSION:Nine case-control studies including 3 228 cases of osteoarthritis and 6 327 healthy controls were included. Overal, the pooledOR values of PvuII loci aleles and genotypes (Cvs. T; CTvs. TT; CCvs. TT; CT+CCvs. TT; CCvs. CT+TT) were less than 1; the pooled OR values of Asian which grouped by region were greater than 1 (except CTvs. TT); the pooledOR values of Europe and the Americas were less than 1. The pooledOR values of XbaI loci aleles and genotypes (Gvs.A; GAvs. AA; GGvs. AA; GA+GGvs. AA; GGvs. GA+AA) were less than 1; the pooledOR values of Asian which grouped by region were less than 1; the pooledOR value of Europe and the Americas were less than 1 (except GGvs.GA+AA). Estrogen receptor α gene PvuII, XbaI site polymorphism is not associated with osteoarthritis susceptibility. However, the susceptibility of PvuII loci in the Asian is a little higher compared with that of the Europeans and American population. On the contrary, the susceptibility of XbaI loci in the recessive genetic model of Europeans and American population is a little higher compared with that of the Asian, suggesting a possible role of ethnic differences in genetic backgrounds.

Objective:Preformed anti-human leukocyte antigen(HLA)specific antibodies are a major risk for antibody-mediated rejection.The aim of this study is to detect and analyze anti-HLA specific antibodies in sera of renal transplant candidates for evaluating the status of presensitization.Methods:A total of 3 500 patients awaiting renal transplantation in our hospital from 1998 to 2007 were included in the study.Panel reactive antibody(PRA)in sera of 694 candidates was detected by complement-dependent cytotoxicity(CDC)before March 2000,then the sera of other 2 806 recipients were screened by enzyme-linked immune absorbent assay(ELISA)alternatively.The polymorphism,specificity and relevant factors of anti-HLA antibodies were analyzed.Results:Only IgG type of antibodies specific to HLA classⅠcould be detected by CDC,whereas both anti-HLA classⅠand classⅡ of IgG class could be found out by ELISA.Lower PRA positive rate by CDC(8%)was shown when compared to that by ELISA(17%).Furthermore,multiple types of specific antibodies against HLA-A(20),B(37),CW(8),DR(14)and DQ(7)were determined by ELISA.Some types of the antibodies presented higher frequencies,such as anti-HLA-A2,24,68,23 and 32;B27,56,57 and 7;DR7,4,9,13,17 and 12;CW1,2,6,4 and 8.These high frequenies of anti-HLA antibodies were somewhat different from the distribution of HLA antigens in South China population.There were significantly different positive rates of anti-HLA antibodies between the male and the female,as the male were sensitized mainly through blood infusion and the female were sensitized by either blood infusion or pregnancy.Conclusion:Specific antibodies against HLA can be detected out by ELISA accurately,whereby to find high freguencies of the antibodies and to avoid donor-recipient mismatching at HLA-loci.Detection of preformed anti-HLA and reducion of HLA-mismatched blood infusion to reduce production of anti-HLA antibodies may be the valuable pathway to improve graft survival.

Apoptosis is particularly important for the body to inhibit or eliminate Mycobacterium tuberculosis ( Mtb) infection. A series of mechanisms to regulate macrophage apoptosis are triggered after macrophages are infected by Mtb. In Mtb-infected macrophages, there are differentially expressed microRNAs (miRNAs), which can directly bind to the binding sites in the 3′-untranslated region (3′ UTR) of apoptotic genes to regulate the expression of apoptotic genes and the apoptosis of macrophages through the mitochondrial or death receptor pathway. This paper reviewed the miRNAs associated with the apoptosis of Mtb-infected macrophages and the major mechanisms.

Objective To investigate the effect of different duration skeletal muscle denervation on acetylcholine receptor activity in rats.Methods Fourteen Balb/c mice weighing 18-22 g were used in this study.The denervation model was established by excising sciatic nerve.Two rats were chosen before(T0 ) and at days 1,4,7,14,21 and 28 after excising sciatic nerve (T1～6),and flexor digitorum brevis of the hindfoot was acutely isolated Skeletal muscle cells were isolated ( five cells in each rat),the acetylcholine currents were recorded using whole-cell patch-clamo technique.Extracellular fluid containing 30 μmol/L acetylcholine was first applied to skeletal muscle cells for 10 s,acetylcholine currents (11)were recorded,then the ceils were washed out using extracellularfluid.Skeletal muscle cells were balanced using extracellular fluid containing 0,0.1,1,10,30,100,1000,3000,or 10 000 nmol/L atracurium for 3 min respectively,then perfused using extracellular fluid containing 30 μmol/L acetylcholine and differents concentrations of atracurium mentioned above for 10 s respectively,and acetylcholine currents were recorded,then the cells were washed out,and 30 μmol/L acetylcholine was perfused again and currents(I2 ) were recorded.The mean value of I1 and I2 was taken as control current,and inhibitory percentage of control current was calculated,and the inhibition concentrations for the half-maximal response (IC50) of atracurium were determined by nonlinear regression analysis.Results Compared with T0,IC50 significantly increased at T1～6 ( P ＜ 0.05).IC50 was increased gradually at T1～3 ( P ＜ 0.05).Compared with T3,IC50 was decreased at T4～6 ( P ＜ 0.05).IC50 was decreased gradually at T4～6 ( P ＜ 0.05 ).Conclusion Skeletal muscle denervation can inhibite acetylcholine receptor activity,which is relate to the denervation time.

Objective To explore the curative effect of fibular flap with limbs composite soft-tissue.Methods From February,2013 to February,2016,13 cases with body severe trauma patients were treated,which including 5 cases of upper limbs and 8 cases of lower limbs,and all existed bone defect,soft tissue defect and trunk vessel defect.Three cases with limbs distal non blood supply were emergency treated with debridment and flow-through fibular flap transplantation renovation,peroneal artery repairing defective blood vesscls to rcstorc limbs distal blood supply,fibular flap repairing bone defect,skin flap repairing soft tissue defect.The limb blood supply for other 10 cases were in good condition,but one case with main artery defect did the second phase of fibular flap transplantation and repaired defective blood vessels,bone and skin soft tissue synchronously according to wound condition.According to the postoperative observation for flap survival and appearance,X-ray films to observe fracture healing after 6 weeks,three months and 6 months of operation as well as evaluating limb function recovery,then analyzed the results.Results Flaps survived successfully for 11 cases,and flaps for the other 2 cases were partial necrosis.One Case was edge flap necrosis,heal scabby after dressing,and the other case was necrosis for 1/3 of the area,but the deep fascia survival,and the skin graft healing after dressing.One case with forearm rolling was in vascular crisis after operation,but tbe crisis was relieved after detection,and fingers blood supply was recovered.All the patients were followed up for 6 to 36 months(mean,14 months).All flaps were survived,fractures healed well and limbs distal blood supply was good.Bone healing time was 8 to 24 weeks,and patients with lower limbs injury could bear load after 3 to 8 months.Lower limbs restored walking function.Upper limbs and hands restored rotation function.Transplant flapshad good elasticity and satisfactory appearance.Conclusion Using fibular flap to repair defective blood vessels,bone and soft tissue synchronously,not only can rescue the limbs on the verge of amputation,but also can repair defective composite tissue and get a good prognosis.It is an effective method for open injuries severely treatment in clinic.

Objective To investigate the characteristics of BK virus (BKV) infection in renal transplant recipients. Methods A total of 243 renal recipients from our clinic within 48 months after transplantation were enrolled as the trial group and 82 healthy people as the control group. Urine and peripheral blood samples of these two groups were harvested for urinary sediment BKV cytology by Decoy cell counting and BKV DNA by real-time PCR. Results The positive rates of urinary Decoy cell, BKV viruria and viremia were 35.4%, 36.6% and 16.9% in trial group, and 4.9%, 20.7% and 2.9% in control group, respectively. In trial group, the medians of urinary Decoy cell, urinary BKV and peripheral blood BKV were 6/10 HPF, 1.00×104 copy/ml and 6.87×103 copy/ml respectively, while in control group, they were 2/10 HPF, 1.10×104 copy/ml and 2.24×1(3 copy/ml. Compared with the healthy people, the positive rates and the levels of BKV DNA in urine and peripheral blood of recipients were significantly higher. The amount of urinary Decoy cells was positively correlated to urinary BKV load (r=0.636, P＜0.01). Conclusions BKV replication is easier to happen in renal recipients as compared to healthy people. Counting of urinary Decoy cells is convenient, useful and sensitive to evaluate BK viruria and viremia in renaltransplant recipients. BKV DNA detection in urine and peripheral blood can be used to screen the evidence of BK reaction in order to prevent irreversible graft damage by BKV.[ Key words ] Kidney transplantation; BK virus; Kidney diseases; Decoy cells

Biomarkers, Tumor ; metabolism ; Bronchoalveolar Lavage ; methods ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; pathology