objective To discuss the possible reasons responsible for the symptomrecurrence after operation for choledochocele, prevention and treatment. Methods By analyzing the symptom recurrent cases among 76cases of choledochocele who were treated in our hospital during the period from March 1966 to September 1997, we discuss the possible reasons responsible for the symptom recurrence after operation for choledochocele and its prevention and treatment. Result The possibility of symptom recurrence has no relationship with the age, sex and the type of the cyst. But it is closely related to the opportunity and method of operation (p

Objective To explore the advantages and characteristics of pathology experimental teaching based on WeChat public platform. Method Through the establishment ofhomogeneous pathol-ogy experiment platform—WeChat public platform, the students of clinical medicine major of Class 1 and 2 of Grade 2014 who participated in the pathological experiment course were divided into two groups. The experimental group (65 people) used the auxiliary teaching based on the WeChat public platform, and the control group (65 people) used the traditional teaching method. The students in the experimental group used the WeChat public platform in combination with pathology experimental teaching, pushing the experimental teaching by WeChat public platform, including the change of specimen and eye diseases under the micro-scope of typical pictures and videos, and at the same time pushing the typical disease image and the related text introduction, and auxiliary pathology experimental teaching. The effect of teaching was evaluated by student experimental examination and electronic questionnaire. The data were collated after the entry of SPSS 19.0, and the data comparison was performed by t test. Result The average score of the experimental group (14.80±0.24) was significantly higher than that of the traditional teaching group (13.79±0.33), and the difference was statistically significant (P=0.031). The experimental group students' evaluation on the learning of WeChat based public platform was higher than the control group's evaluation on traditional teaching in the aspects of learning interest, reducing learning pressure and feedback. Conclusion The application of WeChat public platform for the teaching of pathological experiments is feasible compared with the traditional teaching model and can improve the teaching effect effectively, and solve the problems of the pathology experiment teaching sample being insufficient or the teaching sample being not typical, which creates conditions for students to study independently and use specimens and pictures.

Objective To investigate the effect of continuous passive motion(CPM) and rehabilitation on flexion of the shoulder after intensive burn.Methods 36 patients with intensive burn on upper limb(including the skin around axilla fossa) were divided randomly into CPM group(n=19) and control group(n=17).The CPM group received both routine rehabilitation and continuous passive motion treatment,and the control group only underwent routine rehabilitation treatment.Their ranges of active or passive motion were assessed.Results There was a significant improvement both in the CPM group and the control group after treatment(P<0.01),but the CPM group improved more(P<0.05).Conclusion CPM can improve the efficacy of the routine rehabilitation on flexion of the shoulder after intensive burn.

Objective To explore the expression of negative costimulatory molecule B7-H1 of myeloid derived suppressor cells(MDSCs) and its roles in age-related immunosuppressive functions.Methods 36 C57BL/6 mice were divided into 2 groups:the elderly group(12-month-old,n=18) and the young group(4-8-week-old,n=18).The expressive level of negative costimulatory molecules of MDSCs in the two groups was measured by flow cytometry(FCM).Besides,MDSCs were sorted by using MDSC isolation kit and T cells were obtained by grinding lymph nodes.To explore the influence of B7-H1 on the MDSCs' immunosuppressive functions,T cell proliferation and intervention assay was conducted using the five following groups:T cells group,T cells + CFSE group [T cells with carboxyfluorescein diacetate succinimidyl ester(CFSE)],T cells +-CFSE cocultured with MDSCs from the young group,T cells + CFSE cocultured with MDSCs from the elderly group,and T cells + CFSE cocultured with MDSCs from elderly group and anti-B7-H1 blocking antibody.The effect of B7-H1 on T cell proliferation in MDSCs were observed.Results The expression level of B7-H1 of the MDSCs was significantly increased in the elderly group as compared with the young group(t=3.27,P＜0.05).T cell proliferation assay revealed that T cells were remarkably suppressed by MDSCs in elderly group as compared to the youth group(t=5.42,P＜0.05).The suppression of T cells by MDSCs was dramatically reversed by anti-B7-H1 blocking antibody in elderly group(t=8.28,P＜0.05).Conclusions B7-H1,a molecule expressed on the surface of MDSCs,plays a key role in the age-related immunosuppressive function of MDSCs.

Objective To study the alternations of regulatory T cells in early HIV infected patients and its association with disease progression.Methods Fifty-one untreated HIV infected patients were enrolled and divided into 3 groups according to their infection time and CD+4 T cell levels(30 early HIV infected patients,15 typical progressors,6 AIDS patients).Twenty normal controls were enrolled.There were no significant differences between the age and sex among four groups.Blood was drawn by venipuncture from each subject in EDTA tubes and the levels of CD+4 CD+25 Foxp3 + regulatory T cells were detected by FACSAria flow-cytometry.Spearman correlation was used to detect association between CD+4 CD+25 Foxp3 + regulatory T cells and the absolute CD+4 T cells,viral load and activation of T cells.Results The levels of CD+4 CD+25Foxp3+ regulatory T cells showed the tendency of increasing tendency from normal control to early HIV infected patients,asymptomatic HIV infected patients and AIDS patients.Early HIV infected patients was significantly lower than that in AIDS group [3.79(2.11 - 5.43) % vs 8.09(4.90 - 8.90) %,Z = - 2.29,P = 0.022].The levels of CD+4 CD+25 Foxp3 + Treg cells were associated with viral set point(r = 0.479,P =0.038) and inversely associated with CD+4 T cells(r = -0.455,P =0.011) and closely associated with HLA-DR expression on CD+3 T cells(r = 0.533,P = 0.002).Conclusions The ratio of CD+4 CD+25 Foxp3 +regulatory T cells of early HIV infected patients was significantly increased and associated with viral set point and CD+4 T cell counts,which indicate that alternation of regulatory T cell may be an important factor contributing to the disease progression in early HIV infection.

Objective To explore the immune property and possible mechanism of the CD71+ CD235a+ nucleated erythroid cells from peripheral blood in elderly,as compared with those of healthy young.Methods Peripheral blood obtained by venipuncture from healthy young(n=59,mean age=28 years) and elderly(n=78,mean age=68 years)donors were measured by flow cytometry to evaluate the frequency of the CD71+ CD235a+ cells in peripheral blood mononuclear cells (PBMC).In vitro assays,CD71+ CD235a+ cells were sorted by flow cytometry and T cells were sorted using CD4+T cell isolation kit,then the T cell proliferation assays were conducted by following groups:T cell + CFSE group;T cell +CFSE co-cultured with CD71+ CD235a+ cells from the young donors;T cell + CFSE co-cultured with CD71+ CD235a+ cells from the older donors.Real-time PCR were used to identify the expression of the immune cytokine secreted by CD71+ CD235a+ cells.Results (1)the results showed a significant increase in the percentage of CD71+ CD235a+ cells in the elderly compared with the young [(9.93± 2.95)％ vs (1.96 ± 1.16)％,t =3.37,P ＜ 0.001];(2)the CD71+ CD235a+ cells from the older donors could suppress the proliferation of the T cells (t =2.91,P＜ 0.05)while from the young group we could not observe this phenomenon(t =0.387,P＞0.05).(3)The results of Real-time PCR revealed that,compared with CD71+ CD235a+ cells from the young,CD71+ CD235a+ cells from the elderly expressed higher levels of Arg-2 (t =9.04,P＜0.01),IL-1β,IL-6 and TGF-β(t =4.51,5.46,6.92,all P＜0.05).Conclusions There are higher frequencies of CD71+ CD235a+ cells in aged samples than the young.And the CD71+CD235a cells from the elderly could secrete more Arg-2,IL-1β,IL-6 and TGF-β to suppress the T cell proliferation.

Objective To study characteristics and immune mechanisms of CD11b+ GR-1-myeloid-derived suppressor cells (CD11b+ GR 1+ MDSCs) in elderly mice,as compared with those of healthy young mice.Methods Totally 20 healthy C57BL/6 young mice (aged 1-2 months) and 20 elderly mice (aged over 18 months) were randomly chosen and splenetic CD11b+ GR-1+ MDSCs were sorted with the MDSCs Isolation Kit.In vitro assays,the effects of young and elderly CD1 1b+ GR 1+ MDSCs on the proliferation of T cells were determined by Brdu Elisa.Transwell co-culture and real-timePCR were used to identify the mechanisms of different immune suppressive functions of CD11b+GR 1+ MDSCs sorted from young mice and elderly mice.Results Compared with young MDSCs,elderly MDSCs could evidently suppress the proliferation of T cells (t=8.67,P＜0.001),and this function could be reversed by trans-well co-culture (t=6.93,P＜0.001).The results of realtime PCR revealed that,compared with young MDSCs,elderly MDSCs expressed higher levels of arginase-1 (ARG-1),inducible nitric oxide synthase (iNOS),reactive oxygen species (ROS),interleukin 10 (IL-10),IL13 and transforming growth factor (TGF)-β (t=9.04,4.86,7.04,6.92,4.51,5.46,respectively,P＜0.05 or P＜0.01).Conclusions CD11b+GR-1+MDSCs sorted from healthy elderly mice can evidently suppress the proliferation of T cells through cell-cell contact and secretion of suppressive medium.

Objective To investigate the level of subpopulations of myeloid-derived suppressor cells (MDSCs) in elderly tumor-bearing mice versus elderly tumor-free mice,and to study the difference in immune suppressive functions between different subpopulations and their mechanisms.Methods A total of 20 healthy C57BL/6 elderly mice(aged 18-20 months) were randomly chosen to establish Lewis lung cancer models.The amount of monocytic-MDSCs (MO-MDSCs) and polymorphonuclear granulocytic-MDSCs(PMN-MDSCs) in tumor-free and tumor-bearing elderly mice was evaluated by using flow cytometry.MO-MDSCs and PMN-MDSCs were separated with Magnetic-Activated Cell Sorting (MACS) MicroBeads and their morphological characteristics were observed after May-Grunwald-Giemsa staining.The effects of MO-MDSCs and PMN-MDSCs on the proliferation of T cells were determined by Brdu-enzyme-linked immunosorbent assay(ELISA).And the immune suppressive mediators secreted by the subpopulations were detected by Real-time polymerase chain reaction(PCR).Results Compared to the tumor-free group,the proportion of MO-MDSCs in the spleen of tumorbearing group were increased [(12.44± 1.20) ％ vs.(38.42±3.66) ％,t=5.67,P＜0.001],while PMN-MDSCs were not [(10.34±0.68) ％ vs.(12.18±1.27) ％,t=2.21,P=0.09].The result of Brdu-ELISA showed that MO-MDSCs could suppress the proliferation of T cells [(0.30 ± 0.18) vs.(3.38±0.96),t=8.33,P＜0.001],while PMN MDSCs could not [(2.69±0.45)vs.(3.38±0.96),t =1.72,P=0.11].The result of PCR showed that as compared with PMN-MDSCs,Mo-MDSCs had the increased expression levels of arginase-1 (ARG-1),inducible nitric oxide synthase (iNOS),interleukin-10 (IL-10),interferon-γ(IFN-γ) (t =4.31,8.89,1.70,3.13,respectively,P ＜ 0.01 or 0.05),while the expression levels of interleukin-13 (IL-13),transforming growth factor-β(TGF-β) had no differences (t=4.94 and 2.75,P =0.39 and0.47).Conclusions MO-MDSCs are significantly increased in elderly Lewis lung cancer mice models.MO-MDSCs could mediate lung tumor evasion by suppressing the proliferation of T cells through highly expressing ARG-1,iNOS,IL-10 and IFN-γ.

Objective: To use autophagy inhibitors combined with radiation to treat the oral squamous cell carcinoma CAL-27 and KB cells,and to explore the influence of autophagy in the oral cancer radiation sensitivity and its mechanisms. Methods:The human oral squamous cell carcinoma CAL-27 and KB cells were divided into control group,CQ group,3-MA group,IR group,CQ+IR group,and 3-MA+IR group. The survival rate was detected by MTT method and the autophagy of CAL-27 cells was observed by immunofluorescence method and laser scanning confocal microscope.The expression levels of LC3 and beclin-1 were detected by Western blotting method. The apoptotic rate was determined by flow cytometry with Annexin Ⅴ/PI doulde staining. Results:Compared with IR group,the survival rates in 3-MA + IR and CQ+ IR groups were signifcantly decreased (P < 0.05 ).The autophagy levels of cells in IR group were significantly higher than those in control group, CQ group, 3-MA group,CQ+IR group,and 3-MA+IR group (P <0.05).The expression levels of LC3 and beclin-1 proteins in IR group were significantly higher than those in control group,CQ+ IR group,and 3-MA+ IR group (P < 0.05). The apoptotic rates in IR,3-MA+ IR,and CQ+ IR groups were markedly higher than those in control group. Compared with IR group,the apoptotic rates in CQ+IR and 3-MA+IR groups were significantly increased (P <0.05).Conclusion:Radiotherapy can induce the increase of autophagy level of oral squamous cell carcinoma cells. Inhibiors of autophagy can increase the radio-sensitivity of oral squamous cell carcinoma cells by inhibiting the proliferation and inducing the apoptosis.

Objective:To detect the expression of BTLA on Treg cells of HIV-infected patients and investigate the role of BTLA in HIV infection.Methods: Forty-four HIV-1-infected patients (twenty-four early HIV infection,fourteen chronic HIV-infected patients with CD4+ T counts> 200 cells/μl,AIDS patients with CD4+T counts<200 cells/μl) and nine healthy people served as normal controls were selected to detect the expression of BTLA on Treg cells by flow cytometry.The correlations between BTLA expression on Treg cells and disease progression or immune activation were studied.Results: There was a higher percentage of BTLA on Treg cells in chronic HIV patients and AIDS patients than that in early HIV infected patients(P<0.05,P<0.01),and the expression of BTLA on Treg cells in AIDS patients was higher than that in normal controls(P<0.05).The expression of BTLA on Treg cells was negatively correlated with CD4+T lymphocyte counts and positively correlated with viral load (P<0.001,P<0.01).The percentage of BTLA on Treg cells was positively correlated with CD4+CD38+T lymphocytes and CD4+HLA-DR+T lymphocytes(P<0.001,P<0.001).Conclusion: Increased BTLA expression on HIV-infected Treg cells is associated with disease progression,suggesting that it may accelerate disease progression by enhancing Treg cells inhibitory function and may provide intervention information for HIV infection in the future.