Objective To expand the NKT cells in vitro and to determine the cytokine releasing of NKT cells during proliferation.Methods Several kind of methods were established to expand TCRV ?24 +/TCRV ?11 + NKT cells from PBMC or purified T lymphocyte.The levels of IL 4,IFN ?and TNF ? of NKT cells were determined by flow cytometry.Results After 19 days' expansion,the largest number of NKT cells increased (23.0?16.7)?10 3 times and the largest ratio of NKT cells to total T lymphocyte was (25.5?7.2)%.The ratio of TCRV ?11 + NKT cells that secreted IL 4,IFN ?and TNF ? was higher than the TCRV?11 T lymphocytes.Conclusion TCRV ?24 +/TCRV ?11 + NKT cells can be expanded in vitro with ? Galcer presented by CD 1d molecule.The NKT cells can also be expanded from PBMC because the monocyte lineage cells within PBMC can express CD 1d molecule and present specific glycolipids like ? Galcer to them.

Objective To explore the protective effect of vitamin E on the rabbit bladder after partial outlet obstruction artificially setup. Methods A total of 28 New Zealand white male rabbits were divided into 4 groups (group A in 6,group B in 6,group C in 8 and group D in 8).Group A,B and C were fed a regular diet,and group D were placed on a diet enriched with 600 mg vitamin E.After 4 weeks partial outlet obstruction was created in groups C and D,while group B underwent sham operation. After 4 weeks of obstruction each rabbit was sedated and cystometry was repeated.After cystometry the bladder was weighed.The gene expression of sarcoplasmic endoplasmic reticulum,calcium,magnesium,adenosine triphosphatase (SERCA2) in bladder was detected by using RT-PCR assay,while the protein level of SERCA2 was measured by Western blot analysis. Results All parameters measured were approximately identical in nomal rabbits(Group A) and shum operation rabbits(group B).Thus,these 2 groups were combined as the control group(Group A and B).Partial outlet obstruction resulted in bladder weight increased significantly in obstructed groups given vehicle group C(13.07?1.71)g and those vitamin E group D(11.80?2.01)g,4-fold higher than in the control group (2.81?0.30)g(P

BACKGROUND:Many internal fixations,such as dynamic hip screw,Gamma screw,proximal femoral nail,angle steel plate,as well as locking proximac femoral plate,are utilized in treating intertrochanteric fractures,especially the dynamic hip screw However,the failure rate is gradually increased.OBJECTIVE:To explore the application and clinical efficacy of total hip replacement for the treatment of elderly intertrochanteric fracture fixation after failure of dynamic hip screw.METHODS:A total of four cases with intertrochanteric fractures were treated by total hip replacement after failure of dynamic hip screw fixation was selected.According to Evans typing,one case were type Ⅱ,two cases were type Ⅲ A,and one case was typeⅢB.Internal fixation displacement could be found at half to 1 year after dynamic hip screw fixation.Because of bone disunion,coxa adducta and pain,the patients could not walk.Sequentially,total hip replacement was performed with 45°abduct angle and10°-15°anteversion angle The clinical efficacy was evaluated by Harris scoring criteria.RESULTS AND CONCLUSION:All the cases were operated smoothly,with 1.5-2 hour operation duration and 400-600 mL blood loss.No case appeared allergic response to bone cement.By 3-12 months follow-up,4 successful operative cases do not appear prosthesis loosening,subsidence and are satisfied with well hip function.The average Harris score were 81 points.The results revealed that application of total hip replacement after the failure of DHS in elderly intertrochanteric fracture fixation,which shortened the time patients stay in bed to reduce complications and improved the hip joint function.

Objective To discuss prevention and treatment of the late complications of deteniaceacal-colon continent urinary reservoir. Methods 105 patients who underwent detenia ceacal-colon con-tinent urinary reservoir were followed up for 9 -82 months(mean 40.5 months).Clinical data of these wereretrospectively analyzed. Results Late complications occurred in 25.7%(27/105) of the cases and 9were treated with operations,including difficulty with catheterization in 4 cases(3.8%),3 treated with dila-tion and 1 with efferent limb reconstruction with ileal,urinary pouch stones in 5(4.8%),4 treated with li-thotomy,urinary pouch perfotation in 1 (0.95%),treated with repair,urinary pouch superdistention in 1(0.95%),treated with reduction, single or double lateral hydronephrosis in 8(7.6%),caused by strictureof single or double lateral lower ureters in 4 (2 treated with excision lesion and anastomosis ) and by refluxin 1,1 case suffering from azotemia preoperatively worsened to uraemia treated with regular haemodialysis,hyperchloremia in 7 (6.7%),only 1 needed drugs treatment,repeated urinary infection in 11,amomg them,10 were complicated by the other complications,all treated with sensitive antibiotic. Conclusions Most ofpatients underwent detenia ceacal-colon continent urinary reservoir had good curative effects. Late complica-tions afflicting these patients mostly had immediate or mediate relations with symptomatic urinary infection.Prevention and treatment of symptomatic urinary infection can reduce the occurrence of late complications ofthis operative method,regular and adequant pouch washing and catheterization is efficient to prevention oflate complications of this operative method.

Objective To explore operation method,skills,and curative effect of orthophofia microscope discectomy(OMD)for the treatment of lumber disc herniation.Method Retrospective analysis of the 64 patients(74 herniated discs)of lumbar disc herniation heated by OMD,which included 38 segments posterior-lateral extrusion,32 segments central type herniated,and 4 segments extra-lateral extrusion.Resuits All patients were followed-up 6 to 8 months,average 7 months,54 cases were good and 4 cases were fair and no case was bad according to the Nakai scale.The choiceness rate was 93.75%,and no serious complications.Conclusions Orthophoria microscope discectomy has a better third dimension,good operation field of vision,less invasion and best clinical curative effect.Orthophoria microscope discectomy may be the best way of discectomy for the treatment of lumber disc hemiation.

Objective To construct targeted ultrasound contrast agent carried goat anti-mouse IgG antibody (UCA-IgG) and evaluate the effectiveness of its targeted adhesion using parallel plate flow chamber. Methods The ultrasound contrast agent targeted to mouse IgG was designed by conjugating monoclonal antibodies against mouse lgG to the lipid monolayer shell of the agent using biotin-streptavidin. The binding of IgG antibodies to the ultrasound contrast agent were identified by fluorescence in vitro. The attachment and detachment of UCA-IgG to mouse IgG immobilized on a culture dish were assessed in a parallel-plate flow chamber. While the plate lacked mouse IgG,or blocked with large number of goat anti-mouse IgG were served as two control groups. Results UCA-IgG issued a bright green fluorescence, while the contral lipid ultrasound contrast agent didn't show fluorescence. The number of UCA-IgG bound to mouse IgG of experimental group was greater than two control groups,increased with increasing coverslips surface antibody concentrations (P<0. 05),and there was significant positive correlation between the number of UCA-IgG bound to mouse IgG and time of combination (P<0.05). The adhesion rate of experimental group increased with shear stress before 0. 5×10-5 N/cm2 (P<0.05) and then decreased (P<0. 05). There was limited adherence of control groups to the UCA-IgG. The stess of half-maximal detachment was increased with increasing coverslips surface antibody concentrations (P<0.05). Conclusions UCA-IgG could adhere to mouse IgG in the physical conditions. It may provide strong supports for studying other targeted ultrasound contrast agent preliminary and fatherly in vitro.

Background Retinal neovascularization disease is a common cause of blinding.Retinal neovascularization is related to enhancing proliferation of vascular endothelial cells.So how to inhibit proliferation of vascular endothelial cells is a hot burning issue.p21 is known to be involved in the regulation of cell cycle and therefore inhibit the cell proliferation.However,the relationship of p21 and the proliferation of vascular endothelial cells in retinal neovascularization disease is for further study.Objective The aim of this experiment was to study the proliferation of rhesus retinal vascular endothelial cells(RF/6A) and expression of p21 in RF/6A cells under the hypoxia condition,and discuss their association.Methods The RF/6A cells were cultured and passaged in vitro,then they were randomly divided into normoxia culture group(5％ CO2 +95％ O2) and hypoxia for 1 hour,3,6,12 hours group(1％ 02+5％ CO2 +94％ N2).Flow cytometer(FCM) was used to check the distribution of RF/6A cell cycle in the normoxia culture group and hypoxia for 1 hour,3,6,12 hours groups.MTT assay was used to detect and compare the cell proliferation(A570)among the various groups.The expression of p21 in the cells was analyzed by Western blot.Results FCM showed that the cells proportion of G0/G1 stage was reduced initially and then increased afterward in hypoxia for 1 hour and 3,6,12 hours groups,showing a significant difference among 5 groups (F =20.083,P =0.000),and the cells proportion of S stage and G2/M stage were increased firstly and then declined in different hypoxia groups with statistical significances (F =7.861,P =0.001 ; F =10.305,P =0.003).Compared with normoxia culture group,cells proportion of G0/G1 stage was declined and that of S stage and G2/M stage were raised after hypoxia culture,showing statistically signifcant differences(P＜0.05).MTT showed that cell multiplication capacity(A570 value)strengthened firstly and then weakened in hypoxia groups with time prolongation,showing a significant difference among all the groups(F=7.768,P=0.001),and A570 value in hypoxia for 3 hours and 6 hours groups (0.315± 0.062,0.365 ± 0.064) was significantly higher than that of the normoxia group (0.205 ± 0.063),respectively(P＜0.05).Western blot showed that the expression of p21 in the cells down-regulated at the beginning and then up-regulated with the increase of hypoxia time,and there was statistical significance (F =16.738,P=0.000).The p21 relative levels in different hypoxia groups were reduced in comparison with the normoxia group,showing statistical signifcances(P＜0.05).Conclusions Short-term hypoxia could reduce the expression of the p21 in RF/6A and induce cell proliferation initially,then p21 increases and cell proliferation is inhibited with the prolongation of hypoxia time.

Objective To explore a highly sensitive and highly specific method to detect the serum MG7 antigen (Ag) level for early gastric cancer diagnosis.Methods The serum MG7-Ag level was detected by enzyme-linked immunosorbent assay (ELISA) method in 116 preoperative gastric cancer patients,63 postoperative gastric cancer patients,41 patients with precancerous lesion,37 patients with precancerous diseases,50 healthy individuals and 281 patients with other cancers.Meanwhile,the expression of MG7-Ag was also examined with immunohistochemistry in patients with gastric cancer or precancerous lesion.Chi-square test was used for comparing positive rates of the two detection methods.Results The positive rate of MG7-Ag determined by ELISA was 83.6％(97/116) of preoperative gastric cancer patients,45.2％(28/62) of lung cancer patients,45.5％(20/44) of rectal cancer patients,17.6％ (12/68) of colonic cancer patients,14.2％ (6/42) of breast cancer patients,47.6％ (30/63) of postoperative gastric cancer patients,19.5 ％ (8/4 1) of patients with precancerous lesions,5.4 ％ (2/37) of patients with precancerous diseases and 0 of healthy individuals.The sensitivity of ELISA (83.6 ％) was similar with that of immunohistochemistry (94.0％)(P＞0.05).However,the false positive rate of ELISA (12.8 ％) was lower than that of immunohistochemistry (51.3 ％) (x2 =26.491,P＜0.01).There was statistically significant difference in MG7 Ag expression in gastric cancer with different clinical stages (x2=15.564,P＜0.01).Conclusion This ELISA method might be a non-invasive screening method for population with high risk of gastric cancer.

Objective To evaluate the value of sonourethrography in the diagnosis of dysuria after bipolar transurethral plasmakinetic prostatectomy. Methods Sixty male patients with dysuria after bipolar transurethral plasmakinetic prostatectomy underwent sonourethrography and re-operation.The clinical data of these patients were reviewed. Results The sonourethrographic findings were similar with the operative findings in 57 cases.In the 60 cases,there were 11 cases with bladder neck closure,10 cases with bladder neck stricture,30 cases with urethral stricture (16 located at membranous urethra,12 located at pars cavernosa urethra and 2 in external orifice of urethra),5 cases with prostate remnant,1 case with calculi in prostatic urethra,2 cases with dysfunction of detrusor of bladder and 1 case with flap of internal urethral orifice,Conclusions Sonourethrography could be a reliable diagnostic method for dysuria after bipolar transurethral plasmakinetic prostatectomy.It may be helpful for clinical treatment.