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 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.

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 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.

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.

Objective To discuss the clinical application and significance of non-enhanced computed tomography axis rotating movie imaging technique in PCNL for complex renal calculi. Methods Thirty-one cases unilateral and 2 cases bilateral multiple and staghorn renal calculi with mild or mediurn hydronephrosis patients were performed bilateral kidneys non-enhanced CT scanning,three dimensional reconstruction and the axis rotating movie composition were carried on by computer software,PCNL accesses were designed and the residual stone were predicted referred to the access-calyces angle measured in axis rotating movie image,PCNL were performed after while.Comparing between preoperation accesses design and residual stone prediction with in-operation practice were carried out.Results The first PCNL access was constructed via posterior middle upper minor calyces in 22 renal units and via posterior middle lower minor calyces in 13 renal units,which was consistent with pre-operation design according to CT axis rotating movie image.The second PCNL accesses were constructed via lower calyx posterior upper minor calyces in 9 renal units and via lower calyx posterior lower minor calyces in 5 renal units,nephrolithotomy were performed in the same operation,clinical stone clearance rate was 80%(28/35),other 7 cases with residual stone were consistent with pre-operation prediction,No blood transfusion was necessary and no severe complication happened in all 33 cases.Conclusions Non-enhanced CT axis rotating movie imaging provided the detail three dimensional shape and spatial structure of complex renal calculi intuitively) that was benefit for designing appropriate PCNL accesses for complex renal calculi patients, guiding for searching stone fragments in operation, predicting residual stone, and ensuring operation safety.

Objective To evaluate the feasibility of European Organization for Research and Treatment of Cancer (EORTC) risk tables in non-muscle invasive bladder cancer in Chinese patients.Methods A retrospective analysis was performed on the data from 185 patients with non-muscle invaaive urothelial bladder cancer from January 2003 to February 2009. Among the 185 patients, 128 patients were stage Ta compared with 57 patients who were stage T1. There were 87, 53 and 45 patients with grade G1, G2 and G3 respectively. Transurethral resection of the bladder tumor was performed on all the patients and all the patients received routine post-operative intravesical instillation. A telephone interview follow-up was conducted on all the patients, and the average follow-up period was 36 months. EORTC risk tables were used to calculate risk scores for recurrence and progression for each patient. The recurrence and progression rates of different risk groups were recorded and compared with the estimated rates by EORTC risk table. Statistical analysis was used for comparison. ResultsTotal 1-year recurrence rate and progression rate for these patients were 25.9% and 3.8% respectively. According to calculated values of the patients, the 1-year recurrence rates of Group 0, Group 1-4, Group 5-9, Group 10-17 were 10.4%(5/48), 21. 5%(14/65), 35. 2% (19/54), 55.6%(10/18), respectively. The 1-year progression rates of Group 0, Group 2-6, Group 7-13, Group 14-23 were 0% (0/43), 1.5% (1/67), 6. 7% (4/60), 13. 3% (2/15). There was no significant difference between the real rates and estimated rates of the EORTC risk tables (P＞0. 05). However,the 1-year recurrence and progression rates between the low risk group, the medium risk group and the high risk group showed significant differences respectively (P ＜ 0. 05 ). Conclusions The EORTC risk tables are feasible to evaluate the recurrence and progression risk of non-muscle invasive bladder cancer in the present cohort. Nevertheless, the long term value and feasibility need more research to confirm.

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.

OBJECTIVETo explore the role of histone H3 acetylation in cleft palate induced by 2, 3, 7, 8-Tetrachlorodibenzo-p-dioxin (TCDD) in C57BL/6J mice, and its mechanism.

METHODSOn gestation day 10 (GD10), 36 pregnant mice were randomly divided into two groups as the treated group(n = 18) and the control group( n = 18). The mice in the treated group received intragastric administration with TCDD 28 μg/kg, while the mice in the control group received equivalent corn oil. The pregnant mice were sacrificed on GD13. 5, GD14. 5 and GD15. 5, collecting fetal palates to determine the activities of histone acetyltransferases (HATs) by Colorimetric and the expression level of acetylated histone H3 (Acetylated histone H3, Ac-H3) by Western-blot.

RESULTSThe activity of HATs was 0.409 7 ± 0.0147, 0.522 3 ± 0.017 1 and 0.643 5 ± 0.013 9 in control group on GD13.5, GD14.5 and GD15.5; 0.865 0 ± 0.0129, 0.719 1 ± 0.017 8 and 0.551 2 ± 0.016 8 in TCDD group. The activity of HATs in TCDD group was higher than that in control group on GD13. 5, GD14. 5, showing significantly difference between the two groups (t = - 56. 932, t = - 19. 516, P < 0.01); however, the activity of HATs in TCDD group was significantly lower than that in control group on GD15. 5 (t = 10. 382, P < 0.01). The expression level of Ac-H3 was 0.745 0 ± 0.113 5, 1.055 9 ± 0.249 4 and 1.795 5 ± 0.081 9 in control group on GD13. 5, GD14. 5 and GD15. 5; while 1.4490 ± 0. 1460, 1. 641 8 ± 0.099 7 and 1. 512 1 ± 0. 150 2 in TCDD group. The expression of Ac-H3 in TCDD group was higher than that in control group on GD13. 5, GD14. 5, showing significantly difference( t = -6. 593, -3. 779, P <0. 01, P <0.05) ; However, the expression of Ac-H3 in TCDD group was statistically lower than that in control group (t = 2. 870, P <0. 05).

CONCLUSIONThe acetylation of histone H3 was involved in the cleft palate of C57BL/6J mice induced by TCDD, which may be one of the mechanisms in TCDD-induced cleft palate.

Acetylation ; drug effects ; Acetyltransferases ; metabolism ; Animals ; Cleft Palate ; chemically induced ; metabolism ; Dioxins ; Female ; Fetus ; Histones ; metabolism ; Humans ; Mice ; Mice, Inbred C57BL ; Polychlorinated Dibenzodioxins ; Pregnancy ; Random Allocation ; Teratogens