ObjectiveTo identify the association of HLA-DQA1*0302 and DQB1*0303 alleles with vitiligo in Uygur nationality in Xinjiang Uygur Autonomous Region. MethodsPolymerase chain reaction with sequence-specific primers(PCR-SSP) was performed to analyze the distribution of HLA-DQA1*0302 and HLA-DQB1*0303 alleles among 300 patients with vitiligo and 300 normal human controls of Uygur nationality in Xinjiang region. ResultsA significant increase was observed in the frequency of HLA-DQA1*0302 and -DQB1*0303 alleles in patients with vitiligo compared with the controls(20.5％ vs. 13.83％, 30.17％ vs. 13.33％, both P ＜ 0.01 ). Increased frequency of HLA-DQA1*0302 and -DQB1*0303 alleles was also seen in patients with adult vitiligo (onset age ＞ 12 years) and those with childhood vitiligo (onset age ≤≤ 12 years) ascompared with the normal controls(both P ＜ 0.01). The frequency of DQB1*0303 allele was higher in both patients with and without family history of vitiligo than in the normal controls(both P ＜ 0.01), while that of DQA 1*0302 was higher in only patients without family history (P ＜ 0.01 ). No significant difference was observed in the frequency of HLA-DQA 1*0302 or HLA-DQB1*0303 between patients with adult vitiligo and those with childhood vitiligo or between patients with and without family history(all P ＞ 0.05). Conclusions HLADQA 1*0302 and DQB 1*0303 alleles may be associated with vitiligo in Uygur nationality in Xinjiang region,and there seems to be genetic heterogeneity between patients with adult and childhood vitiligo and between vitiligo patients with and without family history.

Objective Establishment and characterization of healthy donor's ??T cell clones.Methods ??T cells were cloned by limiting dilution after positive sorting with 60Co irradiated allogeneic PBMC as feeder cells.Flow cytometry analysis and molecular biology technique were then used to identify ??T cell clones.MTT assay was used to verify their proliferation after incubated with epitope peptides recognized by ??T cells.Results A ??T cell clone had been established.The subtype of this clone was V ?9 V ?2 without expression of CD4 and CD8.Further studies indicated that epitope peptide EP6 could not only specifically bind to ??T cell clone but also trigger the proliferation of ??T cell clone.Conclusion A healthy donor's ??T cell clone was successfully established,which laid a solid foundation for further study on ??T cells.

BACKGROUND:Proximal femoral nail anti-rotation has good biomechanical basis, and has obvious advantages for intertrochanteric fracture in aged patients, but there are some problems in the clinic, because of improper handling of material matching and operation details, which can impact therapeutic effects and functional recovery. OBJECTIVE:To analyze the efficacy and issues of proximal femoral nail anti-rotation in the treatment of intertrochanteric fracture in patients at more than 60 years old. METHODS:From July 2011 to July 2012, proximal femoral nail anti-rotation was used to treat 56 cases of intertrochanteric fractures. Clinical data bank was established to analyze intraopeative problems and postoperative complications. At 1, 3, 6, 9 and 12 months postoperatively, outpatient and telephone folow-up were carried out to evaluate therapeutic effects and functional recovery of hip joint. RESULTS AND CONCLUSION:Four patients died within 1 year. Seven patients lost within a year for other reasons. The remaining 45 patients were folowed with the time from 12 to 24 months, with an average time of 18.2 months. Harris score was (85.00±6.75) points. There were excelent in 26 cases, good in 15 cases, average in 3 cases and poor in 1 case, with an excelent and good rate of 91%. 18 cases were not satisfied with the position of fracture fragments. In 9 cases, proximal femur was not match with the proximal femoral nail anti-rotation. Seven cases were not satisfied because of the location and length of the spiral blade. Seven cases affected lateral cortex fracture. One case experienced postoperative pulmonary embolism. One case suffered from cardiovascular and cerebrovascular diseases. Nine cases suffered from local sweling. 13 cases experienced hip pain. Five cases affected the healing of fracture extended. Results showed that proximal femoral nail anti-rotation for intertrochanteric fracture in aged patients obtained good outcomes, but we should improve the separation of fracture fragments and reduce intraoperative and postoperative complications.

Objectives To investigate etiology, clinical characteristics and outcome in children with epilepsia partialis continua (EPC). Methods Sixty-three pediatric patients with EPC were retrospectively analysed. The patients aged (5.53±3.65) years old, with brain CT scans or MRIs after diagnosis, basic laboratory tests, cerebrospinal lfuid analysis and electroencephalog-raphy. The average follow-up time was (22.19±21.19) months (6-72 months). Results The median duration of EPC was 11 days (1-180 days). The causes of EPC were inlfammatory and immune-mediation (36 cases, 57.14%, Rasmussen’s encephalitis included), metabolic disorders (8 cases, 12.70%), brain structure abnormalities (5 cases, 7.94%), vascular malformation (5 cases, 7.94%), dual causes (3 cases, 4.76%), post brain surgery (2 cases, 3.17%) and cryptogenic pathogenesis (4 cases, 6.35%). Neurological dysfunc-tions were observed in 44 cases (69.84%). Age, routine cerebrospinal lfuid abnormalities, the presence of inlfammation and im-mune mediated, EPC long duration, involving the right upper extremity were the risk factors of poor prognosis. Conclusions The most common causes of childhood EPC are inlfammation and immune-mediated central nervous system diseases. Patients with early age of onset, a great tendency of longer duration of EPC and cerebrospinal lfuid abnormalities, involving the right upper ex-tremity have a poor prognosis.

Objective To evaluated the effect of hyperbaric oxygenation therapy(HBO)on the RhoA ex- pression and nerve function after transient focal cerebral ischemia in a rat model of middle cerebral artery occlusion. Methods One hundred and twenty-six healthy Sprague-Dawley rats were used and randomly divided into a sham op- eration group(shame group,n=42),a treatment group(n=42),and a control group(n=42).The animal model of middle cerebral artery occlusion(MCAO)was established by using the Zea-Longa method with the animals in the treatment and the control groups,and sham operation was performed with those in the sham group.HBO was applied to the animals in the treatment group.The RhoA protein expression was observed by using immunohistochemistry technique,and the neurological function was evaluated by Bederson's scale at different time points after MCAO.Re- sults(1)Weakly positive expression of RhoA could be located in bilateral cortex and the basal ganglia in the sham group.The expression of RhoA in the treatment group and control group was increased as early as 6 hours after MCAO when compared with that of the sham group,and peaked at 48 h after MCAO and decreased after then,but was still higher than that of the shame group at 7th day to 14th day after MCAO.It was also found that the expression of RhoA of the treatment group was significantly lower than that of the control group(P

Blood stasis theory (BST) is widely used in the department of Chinese medical dermatology. Skin lesion we often see and modern medical examination results can be used as evidence for diagnosing BST and indications for using it. Better efficacy could also be obtained by using BST in treating wind evil or heat evil induced skin disease, and itching, hemorrhagic and stubborn dermatoses as well.
Humans ; Medicine, Chinese Traditional ; Pruritus ; Skin Diseases ; therapy

Objective To construct the murine allogeneic acute GVHD model.Methods C57BL/6 (H-2b) mice were used as the donors and Balb/c (H-2d) mice as the recipients in allogeneic bone marrow transplantation (BMT).Groups were set as total body radiation (TBI) control group (n =4),GVHD group (n =10),simple BM transplantation group (n =10) and normal control group (n =4).For TBI control group,mice were subjected to TBI but did not receive BMT after radiation.For GVHD group,5 days before TBI,gentamycin (320 mg/L) and erythromycin (250 mg/L) were added into the drinking water,and on the day of transplantation,mice received one total dose of 8.0 Gy 60Coγ TBI,and within 5 h,2 × 106 C57BL/6 BM cells and 1 × 107 C57BL/6 spleen cells were transfused per mouse via the tail vein.For simple BMT group,the pretreatment was the same as GVHD group,and mice received only 2 × 106 C57BL/6 BM cells per mouse via the tail vein.The mental status,activity,posture,fur,weight,and stool were observed after transplantation.Survival time of each mouse was recorded,survival rate was calculated,and survival curve was drawn.Pathological examination was done for the liver,skin,small intestine and BM on the brink of death.Results The median survival time (MST) in TBI control group,GVHD group and BMT group was (9.0 ± 0.7),(32.0 ± 3.2) and ( 17.5 ± 1.6) days respectively,and there was significant difference between every two groups (P ＜ 0.01 ).Pathological examination in TBI control group showedhematopoiesis exhaustion.GVHD group showed acute GVHD symptoms 10-13 days after allo-BMT,and the pathological changes of the skin,liver and small intestine corresponded to those of Ⅰ to Ⅱ degree of GVHD.Simple BMT group also showed acute GVHD symptoms 10-13 days after alloBMT,but their GVHD manifestation and histological changes were less serious and only 0 to Ⅰ degree of GVHD could be seen.ConclusionStable acute GVHD model can be constructed by transfusion of allogeneic BM cells and spleen cells into Balb/c mice after lethal TBI.

Objective To observe the function of immature CD8α+ dentritic cells (DCs) in vitro. Methods The bone marrow and spleen of C57BL/6(H-2b) and Balb/c (H-2d) mice were got to prepare immature CD8α+ DCs and spleen lymphocytes,and treated by mytomycin. MTT test was used.MLR group, MLR plus variable density syngeneic CD8α+ DC group, MLR plus variable density allogeneic CD8α+ DC group,MLR plus variable density CD8α+ DC supernatant group,CD8α+ DC plus syngeneic T cell group and negative control group were established. MLR group was set up by responder cell ratio of 0.2,0.5,0.8,1.0,to build the MLR plus syngeneic and allogeneic CD8α+ DC experimental groups. Culture supernatant from different density (1 × 105/ml - 5 × 106/ml) of CD8α+DCs was added into MLR to build CD8α+ DC supernatant group. CD8α+ DCs were co-cultured with syngeneic T cells to build CD8α+ DCs plus syngeneic T cells group. 2 × 105/well responder cells served as the negative control group. ELISA was used to detect the concentrations of IFN-γ and IL-10 in the DCs could both suppress MLR (P＜0. 05), and the difference was not statistically significant (P＞0. 05). When CD8α+ DCs were increased, the suppressive effect was enhanced. When CD8α+ DC/responder cell ratio ＞0. 2, the inhibitory effect could be observed, and this effect reached the peak when the ratio was 1.0. The CD8α+ DCs had weak ability to stimulate syngeneic lymphocyte proliferation in vitro, and certain stimulating effect could be seen only when CD8α+ DC/responder cell ratio ＞2 (P＜0. 05). Its culture supernatant also showed suppressive effect (P＜0. 05), and the supernatant with a cell density of 5 × 105/ml showed the maximum effect. IL-10 concentration in the concentration was 1.0 ± 1.2 pg/ml. Conclusion The in vitro function of immature CD8α+ DCs was immunosuppression/tolerance,and they could secret high level of IL-10. The CD8α+ DCs and their culture supernatant could suppress MLR in vitro.

Objective To analyze the correlation between miR-31 and ESCC in expression of miR-31 in the plasma of ESCC in Xinjiang Kazak and Han nationality patients. Methods The plasma samples were collected respectively from patients with ESCC in 20 cases and healthy subjects in 20 cases. The relatively expression of miR-31 was detected by real-time Q-PCR. Results The expression of miR-31 with ESCC were higher than those of the normal control group, which related to the degree of tumor differentiation in Kazak ESCC patients (P < 0.01); the levels of miR-31 relative expression in Kazak were higher than that of Han (P = 0.008, P = 0.027). Conclusion miR-31 may be involved in the occurrence of ESCC in Han and Kazak nationality. miR-31 might be another risk factor in high incidence of ESCC in Kazak than Han nationality.

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).