To investigate the changes in the activated T-lymphocyte CD3/HLA-DR and CD3/CD(16+56) populations in peripheral blood of the patients with allogeneic hand transplantation, lymphocytes from peripheral blood of the patients at different time points were immunologically labeled with dual color fluoresecent monoclonal antibodies CD3/CD(16+56) and CD3/HLA-DR, mono-color fluoresecent monoclonal antibody CD25. CD25, CD3/CD(16+56), and CD3/HLA-DR were determined with flow cytometry (FCM). The levels of activated T-lymphocyte (CD25 +,CD3 +/HLA-DR + ), silent T-lymphocyte [CD3 +/CD(16+56) -,CD3 +/HLA-DA - ] decreased significantly during the first week after transplantation and then increased gradually to the pre-operafive level. Nature killer cells [CD3 -/CD(16+56) +] increased significantly at the first day after transplantation, then decreased sharply and maintained a lower level. The results suggest that immunosuppressive agents have significantly effects on lymphocyte subsets in allogenaic hand transplanted patients, and dynamic determination of HLA-DR, CD3 /CD(16+56) could be valuable in immunomonitoring after allogeneichand transplantation.

Objective To discuss hemodynamic and electrolyte changes associated with irrigation fluid absorption during percutaneous nephrolithotripsy(PCNL). Methods Eithty nine upper urinary tract lithiasis patients underwent PCNL assisted with pressure irrigation. Sixty five cases were with renal calculi and 24 cases were with ureteral calculi. There were 62 males and 27 females. Nor mal saline was used as irrigation fluid. Heart rate(HR),central venous pressure(CVP),cardiac out put(CO),stroke volume(SV),systemic vascular resistance(SVR),thoracic fluid content(TFC) wererecorded before operation and every 30 min during irrigation. Serum Na+,K+,CI ,Ph,BE weredetected before and after irrigation. One way ANOVA,linear correlation and paired t test were usedas statistic analysis. Results The mean irrigation time was 105 min. Mean irrigation fluid volumewas 18 391 ml and mean irrigation velocity was 174.46 ml/min. HR,CO,SV,SVR and blood Na+ ,K+,C1 did not change significantly during and after irrigation. CVP and TFC significantly increasedduring irrigation. The increasing of CVP and TFC were correlated with irrigation time, volume andvelocity. CVP and TFC increased rapidly in 5 patients with calyx laceration and recovered after diuret ic injection. No serious complication was detected. Conclusions Irrigation fluid absorption is observed during PCNL with pressure irrigation. Generally, no significant changes in hemodynamic andelectrolyte balance are found in patients with normal cardiac and renal function.

BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.

Objective To study dynamic changes of T cell subsets and CD3＋HLA-DR＋T cells in peripheral blood of composite tissue transplantation-hand allograft recipients. Methods The levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 in peripheral blood of hand allograft recipients in different periods were examined by using flow cytometry.The recipients before transplantation were as the control groups.Results The first day after transplantation,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 all began to descend.The 3th to 5th day after transplantation,the levels were the lowest.The 8th day,the levels of CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD3＋HLA-DR＋T cells and ratio of CD4/CD8 were eventually rised and go to stable 15th day after transplantation.But the levels of CD3＋、CD3＋CD4＋and value of CD4/CD8 were still lower than those of the contols,and the levels of CD3＋CD8＋、CD3＋HLA-DR＋T cells were higher distinctly.Conclusion Dynamic changes of T cell subsets and active T cells in peripheral blood of composite tissue transplantation-hand allograft recipients were accordant with that of renal allograft recipients with stable period and with stable clinical state of the hand transplantation recipients.

Tetraspanins are a class of cell surface glycoproteins that are expressed very broadly, which can form a complex tetraspanins net by combining with many kinds of cell surface molecules such as integrins, signal proteins, growth factors and so on.In recent years, more and more studies suggest that tetraspanins are closely related to the invasion and metastasis of malignancies and show a certain clinical value, which can be used as new diagnosis and prognosis indicators of malignancy.

Objective To discuss the feasibility and safety of retroperitoneal laparoscopic nephrectomy for treatment of kidney tuberculosis. Methods From March 2005 to February 2009, 28 patients with kidney tuberculosis underwent retroperitoneal laparoscopic nephrectomy. The patients′ data were reviewed and analyzed. Results There were 18 men and tencwomen with an average age of 36 (26-51) in the cohort. Sixteen patients had lesions on the left kidney and 12 on right kidney. All patients had a normal renal function on the contra lateral side. The severely impaired renal function of the lesion side was confirmed before operation. Anti-tuberculosis chemotherapy was administered to patients for two weeks to six months in advance of the surgery. No active lesion of tuberculosis was found and ESR level was normal before operation. All the operations were successfully performed without switching to open surgery. The average operative time was 170 (121-258) minutes, blood loss was 110 (70-250) ml and average postoperative hospital stay was 5.7 (5-14) days. Peritoneum injury was seen in three patients and incision infection in two patients. No severe complications were observed. Anti-tuberculosis chemotherapy was continued for three months. Twenty-four patients were followed-up, and the average follow-up time was 12.5 (6-20) months. All patients recovered without any lesion remaining. Conclusions Retroperitoneal laparoscopic nephrectomy could be a safe and reliable method for the treatment of non-functioning kidney due to tuberculosis.

BACKGROUND:In recent years, tissue engineering has made great progress, and skin tissue engineering is especialy noteworthy. Artificial dermis (PELNAC) is relatively used widely, but there is a lack of relevant reports on wound repair in children. OBJECTIVE:To investigate the clinical efficacy of Pelnac? METHODS:In a retrospective study, 22 patients with the wound of severe trauma were treated with Pelnac as skin graft dressings on treatment of the wounds of severe trauma in children. ? graft, negative-pressure wound therapy and split-thickness skin graft as experimental group (Pelnac? group), and another 19 patients treated with granulation formation dressing and split-thickness skin graft as control group. We colected data including the graft livability, the required re-operative times and the epithelization time after the skin graft. During the folow-up, the skin color and texture of survival skin, subcutaneous fulness, scar hyperplasia and the joint function were also evaluated. RESULTS AND CONCLUSION: In the Pelnac ? group, the graft livability was up to 90% within 10-14 days after grafting. The secondary split-thickness skin graft was required in two cases in the Pelnac? group and in eight cases in the control group. There was a significant difference in the graft livability (P < 0.05). The average epithelization time after the skin graft was (13.86±3.09) days in the Pelnac? group, which was significant shorter than the control group, (19.10±4.62) days, after the first time operation (P< 0.05). During the 10 months folow-up, the survival skin color and skin elasticity in the Pelnac? group was significantly better than that in the control group (P< 0.05). Better subcutaneous fulness and milder scar hyperplasy in the injured sites were obtained in the Pelnac? group compared with the control gorup. Five cases had certain joint function limitation in the Pelnac? group, compared to 10 cases in the control group, with a significant difference (P < 0.05). Artificial dermis Pelnac? has a stronger anti-infectious ability and higher graft livability. Pelnac? graft combined with negative-pressure wound therapy, granulation culture and split-thickness skin graft can shorten the epithelization time, improve wound healing and aleviate harm to the joint function after the skin graft.

ObjectiveTo assess the diagnosis and treatment of staphylococcal scalded skin syndrome (SSSS)in children.MethodsA total of 177 children whh SSSS were eligible for this study.Clinical data were retrospectively reviewed to assess the patients' age and geographical areas,predisposing factors,prodrome,clinical features,results of laboratory examination(including bacterial isolation and drug susceptibility test),treatment and prognosis.ResultsThe median age at onset was 23 days past 15 months.Infants aged 1 to 3 years were the most commonly affected population.The male to female ratio was 8:9.SSSS occurred in summer or autumn in 71.2％ of the patients.Most patients came from countryside.Of these patients,148 (83.6％) had no definite inducement,13 (7.3％) had fever or cold history before the occurrence of SSSS.Skin lesions often began in the face(71.6％).Almost all cases presented with erythema and blisters.Abnormal myocardial enzymes were observed in 104(80.6％) of 129 patients,and Staphylococcus aureus was isolated from 38.7％ of 142 patients.Susceptibility testing of 21 Staphylococcus aureus isolates from patients in the past 3 years showed that 95.2％ and 95.0％ were sensitive to cephalosporins and amoxicillin/clavulanic acid respectively.Double-antibiotic therapy was effective in 72.9％ of the patients.Intravenous gamma globulin was given to 36.2％ of the patients in the early stage.Myocardial-protecting drugs were also administrated early in patients with abnormal myocardial enzymes.All the patients healed without scarring.ConclusionsFor infants with acute facial erythema and exfoliative skin lesions,SSSS should be suspected and timely antibiotic treatment should be given.Cephalosporins and semi-synthetic penicillins are effective and the first choice for the treatment of SSSS.Meanwhile,early addition of gamma globulin and myocardial-protecting drugs may be beneficial to disease recovery.

Objective To explore the role of glycogen synthase kinase (GSK)-3β/β-catenin signaling pathway on human immunodeficiency virus 1 (HIV-1) negative factor (Nef) protein promoting of human herpesvirus-8 (HHV-8) viral interleukin-6 (vIL-6)-induced angiogenesis.Methods GSK-3β mutant plasmid GSK-3β-S9A,dominant negative (DN) form GSK-3β-DN and the control vector pcDNA3.1+ were transfected into endothelial cells which stably expressed HHV-8 vIL-6 or HIV-1 Nef,or co-expressed vIL-6 and Nef protein.Microtubule formation assay was performed to explore microtubule formation ability.A chick embryo chorioallantoic membrane (CAM) model was used to detect angiogenesis.The expression of GSK-3β/β-catenin signaling pathway-related kinases in transfected cells and CAM tissue were further detected by Western blot.The measurement data were compared by t test.Results The activity of GSK-3β was decreased and the ability of HIV-1 Nef protein was enhanced by transfection with GSK-3β-DN in promoting vIL-6 induced microtubule formation (3.42 vs 2.51,t =3.67,P＜0.01) and angiogenesis (6.25 vs 3.97,t=4.06,P＜0.01).In contrast,the activity of GSK-3β was significantly increased and these functions of HIV-1 Nef protein mentioned above were inhibited by transfection with GSK-3β-S9A (0.62 vs2.51,t=8.48,P＜0.01; 0.39 vs 3.97,t=8.59,P＜0.01).The results of Western blot showed that with the elevated level of,β-catenin (in cells:3.53 vs 2.07,t=6.60,P＜0.05; in tissues:2.76 vs 1.74,t=17.40,P＜0.01) and vascular endothelial growth factor (VEGF,in cells:2.68 vs 1.87,t=4.28,P＜0.01; in tissues:2.20 vs 1.39,t=7.08,P＜0.01) were increased in the GSK-3β-DN transfected cells or tissues,while the opposite results were achieved in the GSK-3β-S9A-transfected cells (GSK-3β phosphorylation:0.50 vs 1.47,t=7.33,P＜0.01; β-catenin:1.05 vs 2.62,t=29.50,P＜0.01; VEGF:0.74 vs 2.16,t=20.95,P＜0.01) or tissues (GSK-3β phosphorylation:0.35 vs 1.97,t=10.72,P＜0.01; β-catenin:0.79 vs 1.77,t=5.72,P＜0.01; VEGF:0.43 vs 1.65,t=11.89,P＜ 0.01).Conclusion GSK-3β/β-catenin signaling pathway is involved in vIL-6-induced angiogenesis promoted by HIV-1 Nef protein,which would be valuable for the therapy of Kaposi's sarcoma,an acquired immunodeficiency syndrome,as a potential molecular target.

Objective To describe our experience in laparoscopic radical prostatectomy (LRP)for incidental prostate cancer after TURP. Methods From April 2007 to July 2010, 5 patients with incidental prostate cancer after TURP were treated with a mean age of 73 years. The patients underwent LRP (2.8± 1.1) months after TURP. Results The five cases of LRP were performed successfully, with 1 case of transperitoneal approach and 4 cases of extraperitoneal approach. Mean operation time was (227.6±38.4) min, mean blood loss was (130±152.5) ml, and the mean follow-up was (16.1 ± 15.9) months. All five patients survived, and their urinary function was good without any incontinence. Conclusions Previous TURP represents a technical challenge when performing LRP, but highly skilled surgeons trained to perform LRPs can handle it.