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.

Adult ; Calcinosis ; Humans ; Male ; Pharyngeal Diseases

Objective To establish a new method for the diagnosis of active human cytomegalo- virus(HCMV)infection in renal transplant recipients and investigate its value in guiding antivial the- rapy.Methods The expression of HCMV phosphoprotein 65(HCMV pp65)antigen in peripheral blood leucocytes was detected by immunohistochemistry and catalyzed signal amplification(CSA). Results In 100 renal transplant recipients,44 were found to be positive for HCMV pp65 antigen.The mean number of antigen positive cells was(72?45)/2?10~5 WBC in 29 recipients suffering from symp- tomatic HCMV infection,while that of 15 asymptomatic patients was(46?25)/2?10~5 WBC(P

Analyzed in the paper is the current subsidy mechanism for public hospitals in Zhejiang province, with analysis of problems found. The authors recommended to build a mass fraction subsidy mechanism to cover up the insufficiency of government financial subsidy; to fully leverage the price compensation of medical services; to subsidize in view of the functional positioning of various medical institutions;and explore more channels for public hospitals′subsidies.

Objective To investigate the clinical and laboratory characteristics of SSc patients in China. Method The data of 119 consecutive SSc patients based on EUSTAR DATABASE in Peking Union Medical College Hospital from February 2009 to January 2010 were prospectively collected and analyzed. All patients fulfilled ACR classification criteria in 1980 for SSc. Thex2 test and t-test were used to analyze the data. Results (1) Demographic data. Sex ratio (F/M) was 109/10 and the age rang was (44±12) years. There were 65 diffuse cutaneous SSc (dcSSc) patients and 54 limited cutaneous SSc (1cSSc) patients. 112 patients (94.1%) had Raynaud's phenomenon (RP), and the age of RP occurrence was 36 years (13～76 years), among which it was the initial presentation in 91 patients (81.3%) and the disease duration from RP to other manifestation was 12 months.(2) Clinical manifestations. ① The gastrointestinal manifestations (70.6%), especially esophageal involvement (56.3%), articular involvement (54.6%), pulmonary interstitial fibrosis (PIF) (58.8%) were frequently observed, but renal crisis (2.5%), heart block (0) and reduced LVEF (0) were rarely detected. ② Twenty cases (28.6%) out of 70 PIF patients denied any respiratory symptom and were confirmed by HRCT screening. The disease duration from RP to PIF was 34 months(3～352months); 3 case of 24 pulmonary artery hypertension (PAH) patients had no clinical manifestations. The disease duration from RP to PAH was 25 months (4～343 months). Nineteen patients had PIF and PAH simultaneously. ③Peripheral artery disease: SSc patients had a lower ankle brachial index (ABI) level (0.91± 0.19 vs 1.09±0.08, P<0.00l). (3) Laboratory finding. All patients had positive ANA. The positive rate of anti-Scl-70 antibody and ACA was 56.0% and 14.7% respectively. There was no serum sample positive for anti-Scl-70 antibody and ACA. The positive rate of anti-RNA polymerase Ⅲ antibody was 13%. (4) Compared the clinical characteristics and laboratory findings between dcSSc and lcSSc patients, we found that there were significant differences between dcSSc and lcSSc patients in finger ulcer (40.0% vs 20.4%), ACA positive rates (7.7% vs 23.3% , P<0.05). Conclusion The system involvements besides skin in SSc is common, especially PIF and gastrointestinal involvement. According to our data, there are fewer cases with renal crisis and heart block. Because part of patients with PIF have almost no clinical manifestations, so early screening for PIF/PAH is important for early diagnosis and intervention.

Objective To evaluate the safety and efficacy of anti -CD20 monoclonal antibody (rituximab) in treating refractory connective tissue disease (CTD) patients. Methods Seven inpatients with refractory CTD from Peking Union Medical College Hospital (PUMCH) including four cases of SLE, one for primary SS,one for undifferentiated CTD and one for antiphospholipid syndrome (APS), who had been admitted in recent 3 years (Jan 2005～Feb 2008) were treated with rituximab in combination with corticosteroid and immunosuppressive agents. Responses to treatment as well as outcomes were investigated. Results Six patients were female and one was male with the age ranged from 21 to 58 yr. Disease duration ranged from 3 months to 20 years. Rituximab resulted in significant improvement in clinical manifestations such as lupus nephritis, autoimmune hemolytic anemia and thrombocytopenia. Four patients developed secondary infection including lung infections,herpes and septicemia during B cell depletion, and three patients died of severe lung infections and respiratory failure. Conclusion Our study has shown promising efficacy of rituximab therapy in refractory CTD patients, though it increases the risk of fatal infections, especially when in combination with immunosuppressive agents.

Objective To estimate the characteristics of gastroesophageal reflux (GER) and its clinical association with pulmonary arterial hypertension (PAH) in systemic sclerosis (SSc) patients.Methods Two hundred and five patients with SSc,who fulfilled the American College of Rheumatology criteria were consecutively recruited.GER was recognized in patients with symptom of heartburn or regurgitation.Demographic,clinical,and laboratory data were analyzed.A six minute walk test,pulmonary function test and modified Rodnan's skin score (mRSS) were also calculated for GER and non-GER groups.x2 test,Fisher's exact test and t-test were used for statistical analysis.Logistic regression test was used for the analysis of risk factors.Results There were 90 patients with GER among 205 patients,the prevalence of GER was 43.9％.The presence of PAH (23.3％ vs 9.6％),Raynaud's phenomenon (98.9％ vs 92.2％ ) and fingertip ulcers (56.7％vs 51.3％) were significantly higher in patients with GER than those without GER.There was no difference in autoantibody profile between GER patients and non-GER patients (P＞0.05).The New York Heart Association (NYHA) functional class of SSc-related GER was worse than patients without GER (P=0.015).Pulmonary function test showed that diffuse capacity (DLCO)％,forced vital capacity (FVC)％,and forced expiratory volume (FEV1)％ were lower and the FVC％/DLCO％ ratio was higher in patients with GER than non-GER (P＜0.05).GER was an independent risk factor of PAH in SSc patients (P=0.047,OR=3.41 ).Conclusion GER frequently occurs in SSc patients,SSc patients presenting with GER should be screened for PAH.Targeting the underlying vascular dysfunction might prevent not only PAH,but also GER in SSc patients.

Objective To detect the expression of scleroderma-related autoantibodies, such as anti-Scl-70, anli-centromere antibody ( ACA)and anti-RNA polymerase Ⅲ ( ARA) , and their relationship with clinical features in Chinese systemic sclerosis (SSc) patients. Methods One hundred and thirty-five Chinese SSc patients from the clinical database of the Scleroderma Trials and Research Group proposed by European League Against Rheumatism's Scheroderma Trial and Research Group( EUSTAR) were consecutively enrolled. The expression of ARA, anti-Scl-70 and ACA were detected through linear immunoblotting, double immunodiffusion and indirect irnmunofluorescence, respectively. The relevance between the existing of autoantibodies and clinical manifestations was analyzed statistically. Results Among the 135 Chinese SSc patients, the prevalence of anti-Scl-70, ACA, ARA were 49. 6% , 13.3 % and 8.9% respectively. Patients with anti-Scl-70 antibody had significantly shorter disease course [(71 ±59) month vs (90 ± 103) month, P = 0.041] , higher proportion of interstitial lung disease ( P = 0. 031) but lower of pulmonary arterial hypertension (P =0.042). Modified Rodnan's skin score (P=0.008) and prevalence of facial and cervical cutaneous sclerosis (P = 0. 002) , distal (to elbow/knee ) cutaneous sclerosis ( P = 0. 004 ) and digital pitting scarring/disappear of digital pad were all significantly higher in anti-Scl-70 positive group. Patients with AC A had longer disease course ( P = 0. 036) , lower IgM level ( P = 0. 045) and were less prevalent of interstitial lung disease ( P =0. 045). Patients with ARA had higher serum creatinine and urea nitrogen level ( P ＜ 0.001) although otherwise features had unremarkable differences. Conclusion Scleroderma-related autoantibodies have relevance with different clinical manifestation and detection of these autoantibodies may be helpful to the diagnosis of SSc, organ involvement evaluation and predicting outcomes. The clinical relevances of autoantibodies in Chinese SSc patients may differ from other areas or races.

Objective To figure out the effect of resveratrol (Res) in skin and lung pathology of systemic sclerosis (SSc) animal model and find a new target of anti-fibrosis therapy in SSc.Methods First, we establish ed SSc animal model by daily subcutaneous injection of bleomycin (BLM) for 4 weeks in BALB/c mice.Then we fed the mice with Res.We observed the pathological changes in skin and lung and the expression of the deacetylase SIRT1.We observed the following parametrs.The pathologicalchanges in injected skin and lung which shown by hematoxylin-eosin (HE) staining, the expression of α-smooth muscle actin (α-SMA) in injected skin and lung which measured by immunohistochemistry, the expression of SIRT1 and pro-collagen Ⅲ mRNA which assessed by Real Time polymerase chain reaction (PCR).For the homosce dasticity data.We used one-way analysis of variance (ANOVA) and LSD-t test to compare between the groups.Results Daily subcutaneous injection of BLM for 4 weeks in Balb/c mice could successfully establish a mouse model of SSc.The thickening of skin and alveolar septum, the infiltration of inflammatory cells in lung, and even fibrosis insome area of lung could be observed.The number of α-SMA positive cells and the expression of pro-collagen Ⅲ mRNA were increased (P＜0.05).Meanwhile, the expression of SIRT1 mRNA was decreased [the number of α-SMA positive cell: skin 26.4±5.9 vs 4.4±2.2, lung 14.6±4.6 vs 2.4±1.1, cells per view, P＜0.01;the expression of pro-collagen Ⅲ mRNA: 1.06±0.24 vs 0.45±0.14, relative to glyceraldehyde phosphate dehydrogenase (GAPDH), P＜0.05].Meanwhile, the expression of SIRT1 mRNA was decreased (1.01±0.51 vs 5.03±1.59, relative to GAPDH, P＜0.05).Treated with Res, the pathological changes in skin and lung were alleviated and the number of α-SMA positive cells in lung and skin was decreased [skin 26.4±5.9 vs 10.0±3.5 (high dosage group), 26.4±5.9 vs 13.4±4.4 (medium dosage group);lung 14.6±4.6 vs 8.8±3.5 (high dosage group), cells per view, P＜0.05].There was were no significant differences in the expression of SIRT1 and pro-collagen Ⅲ mRNA in lung after treated by Res.Conclusion Daily subcutaneous injection of BLM for 4 weeks could successfully establish a mouse model of SSc.Res could be a new medicine for the treatment of SSc.