Acupuncture Points ; Acupuncture Therapy ; Chronic Disease ; therapy ; Diarrhea ; therapy ; Female ; Humans ; Middle Aged

Objective： To investigate the effect of oxLDL and VitE on the expression of CD40 and CD40 ligand(CD40L) in cultured human monoc ytes. Methods： The expression of CD40 and CD40L on monocytes su rface were measured by indirect immunorescence technique in combination with flo w cytometry. Results： Low concentration of oxLDL（≤200 μg/L） significantly increased the expression of CD40 and CD40L in a dose and time dep endent manner. High concentration (>200 μg/L）of oxLDL markedly reduced the exp ression of CD40 and CD40L. When VitE was added, it significantly reduced the ex pression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose-depe ndent manner. Conclusion：It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the for mation of atherosclerosis. Antioxidan VitE can partially inhibit the high expres sion of CD40 and CD40L on monocytes surface induced by oxLDL.

Objective: To investigate the effects of hyaluroni dase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothel ial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was dete cted by MTT assay. The expression of CD44 and DNA content of the cells were meas ured by flow cytometry (FCM). Results: HAase (50 μg/ml) stimula ted cell proliferation ［(50.10±1.23）% vs control, P<0.01］, incre ased S phase cell rate and induced the expression of CD44, but HA (100 μg/ml) i nhibited cell proliferation and the expression of CD44. Conclusion: HAase may degrade antiangiogenic HA of extracellular matrix, which may stim ulate proliferation of endothelial cells and enhance the curative effect of grow th factors to myocardial ischemia.

In order to improve the teaching quality of experimental teaching of laboratory diagnosis and to cultivate the clinical ideation of medical students,we have been trying a series of reform in this course and obtained good results.Here,our experience in the selection of teaching contents,the improvement of teaching methods,and the setting of experimental assessment have been summarized and investigated.

Objective To explore the clinical effect of autogenous bone,allograft bone and BMP synthetic bone in the treatment of lum-bar spondylolisthesis,and provide more basis to choose transplant material.Methods A total of 96 patients with lumbar spondylolisthesis were chosen as research subjects,who were cured in our hospital from January 2014 to January 2015.They were divided into group A(who were treated with autogenous bone),group B(who were treated with allograft bone)and group C(who were treated with BMP synthetic bone), according to prospective study method.The indicators of the operation,postoperative adverse reactions,change of intervertebral disc height and bone graft fusion rate of three groups were compared.Results Difference of the operation indexes of three groups had no statistical sig-nificance(P >0.05).The incidences of adverse reactions in group A and group C had no statistically significant difference(P >0.05),but both less than that in group B,with statistically significant difference(P <0.05).The intervertebral disc height after 6,9,12 and 18 months in group A and group C had no statistically significant difference(P >0.05),but both more than that in group B,with statistically significant difference(P <0.05).Bone graft fusion rate of group C was faster than that of group A and goup B,and the graft fusion rate in 18 month in group A and C had no statistically significant difference(P >0.05).And the fusion rates of group A and C in each period were significantly higher than that of group B,with statistically significant difference(P <0.05).Total effective rate of neurologic improvement in group A and group C had no statistically significant difference(P >0.05),but both better than that of group B,with statistically significant difference (P <0.05).Conclusion BMP synthetic bone used in lumbar spondylolisthesis has the same clinical effect as autologous bone.But BMP synthetic bone has faster bone graft fusion rate than autologous bone.And it is beneficial to patients’recovery.

AIM: To investigate whether human umbilical vein endothelial cells and human atherosclerotic plaque lesions can coexpress CD40 and CD40 ligand . METHODS: The expression of CD40 and its ligand CD40L on human endothelia cells were measured by fluorescence microscope , flow cytometry(FCM), reverse transcription PCR(RT-PCR) and Western blotting, respectively. Both CD40 and CD40L expression in atheroma plaques were determined by immunohistochemistry. RESULTS: Cultured human endothelial cells constitutively coexpressed CD40 and CD40L in mRNA and protein levels. Stimulation with interleukin-1?, interleukin-6, tumor necrosis factor-? and interferon-? increased expression of CD40 and CD40L on endothelial cells . Human atherosclerotic lesions( n =6) showed coexpression of immunoreactive CD40 and CD40L. However, no expression of CD40 and CD40L in nonatherosclerotic human arteries. CD40L mainly expressed on the shoulder and base of the plaque. But CD40 was widespreadly expressed in plaque. CONCLUSION: Our results demonstrated that CD40 and CD40L coexpressed on human umbilical vein endothelial cells and in human atherosclerotic plaque lesions. These findings suggest a previously unsuspected role for CD40-CD40L interactions in atherosclerosis.

Objective: To investigate the effects of hyaluronidase (HAase) and hyaluronan (HA) on proliferation of vascular endothelial cells and its mechanism. Methods: The cultured bovine aortic endothelial cells (BAEC) were treated with HAase or HA. Cell proliferation rate was detected by MTT assay. The expression of CD44 and DNA content of the cells were measured by flow cytometry (FCM). Results: HAase (50 ?g/ml) stimulated cell proliferation [(50.10? 1.23)% vs control, P

200 ?g/L)of oxLDL markedly reduced the expression of CD40 and CD40L. When VitE was added, it significantly reduced the expression of CD40 and CD40L on monocytes surface induced by oxLDL in a dose dependent manner. Conclusion:It is an important mechanism that the high expression of CD40 and CD40L induced by oxLDL may be contributed to the formation of atherosclerosis. Antioxidan VitE can partially inhibit the high expression of CD40 and CD40L on monocytes surface induced by oxLDL. [

Objective To explore the effect of IFN-γ, IL-10 and IL-4 on B cell activating factor (BAFF) expression in human HL-60 cells, a kind of myeloid tumor cell lines, and its possible regulation mechanism. Methods Cultured human HL-60 cells were treated with IFN-γ, IL-10 and IL-4 for 1-3 days. The expression of membrane-bound BAFF on HL-60 cells was examined by flow cytometry, the amount of soluble BAFF was detected by ELISA assay, and the level of BAFF mRNA was tested by real-time PCR method. A functional 1021 bp fragment of the 5'-tlanking region of the human BAFF gene (-1349 to -329 bp) was cloned and investigated with serial 5'-deletion. The 5'-deleted promoters were recombinated with chloramphenicol acetyltransferase (CAT) as reporter gene. These five recombinant plasmids were transiently transfected to HL-60 cells with liposomal transfectian method. Promoters activities were determined by CAT reporter gene assay(CAT-ELISA) in those transfected cells treated with different cytokines. Results The results showed that the expression of membrane-bound BAFF, soluble BAFF and BAFF mRNA in human HL-60 cells were significantly elevated (P ＜ 0. 05) after incubated with IFN-γ and IL-10. In addition, IFN-γ and IL-10 showed significantly (P ＜ 0. 05) increased effects on promoter activity in human BAFF gane. And the cytokines-responsive sequences were located between -929 and -719 bp of the BAFF promoter region. Conclusion The enhancement of IFN-γ and IL-10 on BAFF expression and synthesis were regnla-ted by promoter activation. Our in vitro studies also raise the possibility to investigate the mechanisms regula-ting BAFF expression in other tumor cells of myeloid origin under pathological circumstances.

Objective To investigate the population and role of IL-10+ CD19+ regulatory B cell (Breg) in patients with chronic hepatitis B.Methods Patients with acute hepatitis B (AHB) (n =28),chronic hepatitis B (CHB) (n =31) and normal subjects (n =25) were collected from Changshu No.2 People's Hospital between 2011 June and 2012 October.Peripheral blood mononuclear cells (PBMC) were isolated and stimulated with CpG ODN 2006 and PMA.Flow cytometry was used to analyze the population of IL-10-CD19 + Breg,CD4 + CD25high Treg,and ELISA was used to analyze the concentration of IL-10 in culture supernatant.Results The population of Breg in Peripheral blood of the CHB group [1.28％ (1.05％-2.20％)] was higher than that in the AHB group [0.87％(0.55％-1.22％)] and the HCs group [0.89％ (0.51％-1.37％)] (P =0.001,0.006),and the difference between the AHB group and the HCs group was not statistically significant (P=0.669).Breg in the CHB group [14.30％ (10.70％-16.70％)] was higher than that in the AHB group [10.30％ (7.05％-13.30％)] and the HCs group [10.40％(6.85％-12.60％)] (P =0.003,0.001),treg in the CHB group [5.80％ (4.20％-9.10％)] was also higher than that in the AHB group [4.05％ (2.53％-5.40％)] and the HCs group [4.50％ (2.55％-5.50％)] (P ＜0.001,P =0.005),and there was no significantly difference between the AHB group and the HCs group (Breg:P =0.796 ; Treg:P =0.227).Spearman correlation analysis showed that Breg was positively correlated with Treg in the CHB group (r =0.50,P =0.004),however there was no significantly correlation in the AHB group and the HCs group (r =-0.15,P =0.462; r =0.09,P =0.669).The concentration of IL-10 in the CHB group was higher than that in the AHB group and the HCs group (P ＜ 0.001),and the difference between the AHB group and the HCs group was not statistically significant (P=0.341).Spearman correlation analysis showed that IL-10 were positively correlated with the population of Breg in the CHB group (r =0.409,P =0.022).Conclusion The poluations of regulatory B cell and regulatory T cell increased in patients with chronic hepatitis B,and Breg cell might play the immune regulation role through secreting IL-10 in chronic HBV infection.