The progressive loss of renal function and accumulation of collagen leads to CKD. Human BM-MSCs are considered as an ideal therapeutic strategy for renal regeneration in the CKD. Polysaccharides extracted from Poria cocos, an edible medicinal mushroom, have been in use in the traditional Chinese herbal medicine as they exhibit antidiabetic, antioxidative, antitumor, and other pharmacological effects. Whether the polysaccharides of P. cocos could ameliorate the CKD via induction of BM-MSC differentiation remains to be explored. The data presented here show that the polysaccharides of P. cocos not only induced BM-MSC proliferation and differentiation, but also reduced the levels of proinflammatory cytokines and improved renal morphology.

目的 分析可溶性B7-H4在系统性红斑狼疮(systemic lupus erythematosus,SLE)患者以及疾病活动中的诊断价值.方法 收集60例SLE患者和健康体检者20例,SLEDAI评分评估SLE的疾病活动性,ELISA法检测SLE患者和健康体检者血清中可溶性B7-H4(soluble B7-H4,sB7-H4)的含量.结果 (1)SLE患者sB7-H4含量[(65.62±21.02)ng/mL]明显高于正常对照组[(55.4±11.2)ng/mL],差异有统计学意义(P<0.05);sB7-H4诊断SLE的敏感性为56％,特异性85％,ROC曲线下面积为0.65.(2)SLE患者疾病活动组sB7-H4含量[(70.99±19.85)ng/mL]明显高于稳定组[(59.06±20.89)ng/mL],差异有统计学意义(P<0.05);在SLE患者中sB7-H4诊断活动期SLE的敏感性为82％,特异性为56％,ROC曲线下面积为0.687.结论 sB7-H4有助于SLE的诊断,且对SLE疾病活动的预测有重要价值.

In the traditional Chinese medicine, polysaccharide of Hippophae rhamnoides (also known as sea buckthorn) has been widely used for the treatment of liver injuries or cardiovascular problems. The effect of the polysaccharide of H. rhamnoides on kidney injury in rats with severe acute pancreatitis induced by biliopancreatic duct injection with sodium taurocholate was examined. The rats with severe acute pancreatitis showed series of degenerative changes in pancreas and kidney including inflammatory cell infiltration, edema, and necrosis that were diminished following treatment with the polysaccharide of Hippophae rhamnoides. Furthermore, sodium taurocholate injection also aggravated renal function with enhanced levels of proinflammatory cytokines, blood urea nitrogen, serum creatinine, and serum levels of amylase and lipase that were ameliorated by the polysaccharide. There was an upregulation of cleaved caspase-3 and Bax, as well as downregulation of Bcl-2, in the rats model with SAP that diminished following polysaccharide treatment. Lastly, there was an activation of MAPK/NF-κB pathway in the rats exhibiting severe acute pancreatitis that was repressed by the polysaccharide of Hippophae rhamnoides.

Our previous study confirmed the defect of B7-H4 expression in peripheral blood and salivary glands of patients with primary Sjögren's syndrome (pSS). The aim of this study was to analyze the effect of the deficit expression of B7-H4 on CD4+ T cells.CD4+ T cells were purified by magnetic-activated cell sorting MACS. The proliferation and cytokine production of CD4+ T cells co-cultured with purified salivary gland epithelial cells (SGECs) from pSS or non-SS sicca syndrome were detected.By co-culturing the gland cells with CD4+ T cells, we found the proliferation of CD4+ T cells was significantly suppressed. The effect was weaker when SGECs from pSS patients were used compared to that from non-pSS sicca syndrome controls. Simultaneously, the productions of cytokines interleukin (IL)-5, IL-13, IL-17A, IL-6 in supernatant were reduced and also SGECs from pSS patients decreased them less than that from non-SS controls.The decrease of B7-H4 expression in salivary glands of SS patients contributes to the defect of negatively regulating the inflammation caused by CD4+ T cells, thereby providing new insights into the role of B7-H4 in the inflammatory process of salivary glands in SS.

OBJECTIVE:To examine amount of CD4+CXCR5+Tfh cells and B cells subsets in salivary gland and peripheral blood from patients with primary Sjogren's syndrome (pSS) and to analyze whether the frequency of CD4+CXCR5+Tfh cells is associated with pSS pathologic process.METHODS:The percentages of CD4+CXCR5+Tfh cells and B cell subsets were examined by flow cytometry. B-lymphocyte chemoattraetant (BLC; also called CXCL13), IL-21, IL-6 from the serum of pSS patients was assessed by polymerase chain reaction-enzyme-linked immunosorbent assay (ELISA).RESULTS:The percentages of CD4+CXCR5+Tfh cells in peripheral blood were increased in pSS patients, but decreased after treatment with glucocorticoid and/or immunosuppressive drugs. Abnormal B cell subsets appeared in salivary and peripheral blood of pSS patients. The frequency of salivary CD4+CXCR5+Tfh cells was positively correlated with CD19+CD27+ memory B cells and CD19+CD27high plasma cells. Also increase of salivary CD19+CD27high plasma cells was positively associated with serum ANA titer of pSS patients.CONCLUSIONS:CD4+CXCR5+Tfh cells are significantly increased in salivary and peripheral blood in pSS patients with aberrant CD19+CD27+ memory B cells and CD19+CD27high plasma cells, suggesting that CD4+CXCR5+Tfh cells may contribute to the pathogenesis of pSS by promoting the maturation of B cells.

目的 探讨类风湿关节炎(rheumatoid arthritis,RA)患者骨代谢指标、骨密度(bone mineraldensity,BMD)、骨矿物质含量(bone mineral conten,BMC)的变化及临床意义.方法 选择2011年3月至2012年7月安徽医科大学附属省立医院风湿免疫科RA患者(RA组)及健康者(对照组),采用双能X线骨密度仪检测各组受试者前臂BMD和BMC含量,检测2组骨钙素(bone gla protein,BGP)、Ⅰ型胶原交联羧基末端肽(C-terminal telopeptides of type Ⅰ collagen,CTX)、红细胞沉降率(erythrocyte sedimentation rate,ESR)、C反应蛋白(C reactive protein,CRP)、类风湿因子(rheumatoid factor,RF)和抗环瓜氨酸肽抗体(anti-cyclic citrullinated peptide antibody,抗-CCP).摄双手X线正位片,进行X线分期,计算患者疾病活动指数(disease activity score 28,DAS28).结果 RA组患者71例,对照组20例.RA组骨代谢指标CTX和BGP明显高于对照组,差异有统计学意义(均P ＜0.05).RA组前臂BMD、BMC,均低于对照组的BMD和BMC,差异有统计学意义(均P＜0.01).直线相关分析示RA组CTX与BMD和BMC均呈负相关(r=-0.301,P=0.014; r=-0.296,P=0.015);与病程、RF呈正相关(r=0.382,P=0.001;r=0.263,P=0.029);多元线性回归分析显示前臂BMD与RF、年龄呈负相关(r=-7.544、r=-3.254,P均＜0.01);前臂BMC与DAS28呈负相关(t=-4.440,P＜0.01);相关分析显示BMD、BMC与X线分期均呈负相关(r=-0.289、r=-0.284,P均＜0.01),CTX与X线分期成正相关(r=0.333,P=0.005).结论 RA患者骨代谢呈高转换型骨丢失,骨代谢指标及骨密度、骨矿盐含量可以预测RA患者出现骨破坏;RF可能为RA患者骨破坏的危险因素.

Background B7-H4 is a newly identified B7 family negative co-stimulate molecule, the expression and function of B7-H4, in the pathogenesis of lupus nephritis (LN) is still unclear. Objectives To investigate the expression of B7-H4 in patients with LN and the function of renal tubular epithelial cells (TECs)-associated B7-H4 in the regulation of T cell activation in vitro. Methods 90 LN patients, 3 acute kidney injury patients and 20 healthy donors were referred. Disease activity was assessed by SLEDAI scores. The expression of B7-H4 on kidney biopsies from patients with LN and acute kidney injury was measured using immunohistochemistry. In vitro, B7-H4 antigen on cultured HK-2 cells with or without stimulation by inflammatory factors was detected by flow cytometry. After co-cultured with HK-2 and purified CD4+T cells labeled with CFSE for 72 hours, T cell proliferation was detected by flow cytometry. The soluble B7-H4 in sera of LN patients and healthy donors were analyzed using ELISA. Results B7-H4 antigen expressed on tubular epithelium, the percentage of B7-H4 positive expression renal tubules from LN patients (45±16) % was lower than acute kidney injury patients (86±11) % (P<0.05). In vitro, the expression of B7-H4 on HK-2 cells was elevated upon stimulation by inflammatory factors, mixed lymphocyte reactions revealed that HK-2-related B7-H4 inhibits proliferation of co-cultured T cells. The soluble B7-H4 in serum of LN patients (61.45±29.38ng/ml) was not significantly lower than healthy controls (70.57±27.24ng/ml), but stronger association of serum soluble B7-H4 with serum creatinine levels was observed in LN patients (r=0.353, p=0.005). In addition, the mean concentration of soluble B7-H4 level in high activity group (74.40±31.95 ng/ml) was significantly higher than those in moderate group (51.89±21.79 ng/ml) (p=0.017) Conclusions B7-H4 molecules may play a role in the progress of LN, a clear understanding of its functional roles may further elucidate the pathogenesis of this disease. Disclosure of Interest None Declared

Objective To identify interleukin 17 (IL-17) and B cell activating factor (BAFF) that could influence B cell biology by detecting the expression of BAFF in the serum and labial salivary glands from primary Sj(o)gren's syndrome (pSS) patients and to test the apoptosis rates of B cells cultured with Th17 cells which were transfected with IL-17-siRNA,BAFF-siRNA.Methods A total of 40 patients with pSS who were referred to the Department of Rheumatology and Immunology at Anhui Provincial Hospital from June 2011 to June 2012 were enrolled into this study.The expression of BAFF on salivary gland and serum from pSS patients and healthy controls were detected by ELISA and immunohistochemical examination (22 patients with pSS).Flow cytometry was used to detect B cell's apoptosis,BAFF and IL-17 interfered with amplified Th17 cells,and co-cultured with B cells.Immunoblot was used to detect supernatant antibody in 5 patients with pSS.Independent samples t test was used for statistical analysis.Results In all pSS specimens,infiltrating inflammatory cells expressed BAFF,so did some ductual cells,but acinar cells did not express these markers.There was no expression of BAFF in the controls.BAFF-positive cell numbers in the labial salivary glands of pSS patients with focal infiltrating lymphocytes were more than that with non-focal infiltrating lymphocytes (888±372 vs 164±161,t=5.94,P＜0.05),and the percentage of BAFF-positive lymphocytes over the total infiltrating lymphocytes in the salivary glands of pSS patients with focal infiltrating lymphocytes [(0.18 ±0.08) ％] was higher than those with non-focal infiltrating lymphocytes [(0.09 ±0.07) ％] (t =3.03,P＜0.05).The level of soluble BAFF in patients with pSS [(6.0±2.8) ng/ml] was significantly higher than the controls [(3.8±1.7) ng/ml,t=3.26,P＜0.05].BAFF or IL-17 transfected group,B cell apoptosis rate [(24± 5)％,(23±5)％] were significantly higher than the non-transfected group [(7±4)％],t=4.6,4.4; P＜0.05].And there was no significant difference when compared with cultured B cells (P＞0.05).Compared with the controls,no antibody could be detected in the supernatants.Conclusion BAFF may be involved in the process of local inflammatory damage of the pSS,it may have a synergistic effect with IL-17 on abnormal B cell function.

Objective To investigate the changes and clinical significance between bone mineral density(BMD) and bone mineral conten(BMC) in patients with rheumatoid arthritis(RA).Methods The BMD and BMC of patients of RA and normal control group were measured by means of dual energy X-ray absorptiometry in the forearm.The correlations of indexes were analyzed by linear correlation analysis and multiple linear regression analysis.Results The BMD and the BMC of patients with RA were significant deceased compared to normal control group(P0.05).The BMD in forearm was negatively correlated with DAS28 score,ESR and RF(r=-0.357,-0.390,-0.255,P0.05),respectively.There was a significant negative correlation between BMC in forearm and DAS28 score,ESR(r=-0.344,-0.401,P0.05).And multiple linear regression analysis showed that the BMD in forearm with RF and age were negatively correlated;The BMC in forearm was negatively correlated with the DAS score.The patients of RA with X-ray for the period I have 15 cases,which 6 patients showed osteoporosis,bone mass decreased by 2 cases,bone loss and osteoporosis in patients with a total of 8 cases.Conclusions The declines of BMD and BMC of patients with RA are more obvious;BMD and BMC loss may be closely related with disease activity;The BMD may be reflected the loss of bone mass in patients with RA more reflected earlier than X-ray.

OBJECTIVETo detect the expression of B7-H4 in salivary gland and sera in patients with primary Sjogren's syndrome (pSS).METHODSA total of 40 pSS patients were referred to our department from June 2009 to January 2011. Immunohistochemistry and flow cytometry were used to detect the expression of B7-H4 in salivary gland from pSS patients and disease controls. Enzyme-linked immunosorbent assay (ELISA) was used to detect soluble B7-H4 of serum from pSS patients and healthy donors.RESULTSImmunohistological staining revealed that B7-H4 antigen was restricted to tubular epithelium. The B7-H4 positive expression of tubules in salivary gland biopsies from pSS patients (18 ± 14)% were lower than that of controls (85 ± 13)% (P < 0.05). Flow cytometry revealed that the B7-H4 expression of cell suspensions from salivary gland from pSS patients (42 ± 21)% were lower than that of controls (48 ± 22)% (P < 0.01). And the serum level of soluble B7-H4 detected in pSS patients (49 ± 31)µg/L significantly decreased than that in healthy donors (71 ± 27) µg/L (P < 0.05) and positively correlated with saliva and tear flow rates (P < 0.01) respectively.CONCLUSIONThe expression of B7-H4 molecule may play some roles in the progression of pSS. And a further understanding of its mechanism helps to elucidate the pathogenesis of pSS.

Background Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by immunological intolerance, hyperactivity of various immunocyte and production of a variety of autoantibodies. Although etiology of the disease remains unclear, there is an increasing evidence that the presence and accumulation of dendritic cells in target tissues may play a role in the pathogenesis of SLE. Objectives To explore the changes and significance of plasmacytoid dendritic cells (PDC), myeloid dendritic cells (MDC) and cytokines in peripheral blood in patients with SLE. Methods The propotions of PDC and MDC in peripheral blood from 38 SLE and 15 normal individuals were measured by flowcytometer. The concentrations of IFN-α, TNF-α and IL- 6 were detected using ELISA. Results 1. The levels of PDC and MDC in peripheral blood of SLE patients decreased significantly comparing with normal controls (P<0.01). 2. It was found that the percentage of PDC in SLE patients with stable disease was significantly higher than patient with active disease (P<0.05). No significant difference was found in MDC between patients with and without disease activity. 3. SLE patients with proteinuria had lower percentage of PDC comparing with patients without proteinuria (P<0.05). In addition, anti-dsDNA negative patients had higher percentage of PDC than anti-dsDNA positive patients (P<0.05). 4. The percentage of PDC was correlated significantly with SLEDAI, C3 and ESR (with r value of -0.363, 0.451, -0.453 and p value of 0.031, 0.001, 0.008 respectively). 5. Further study found SLE patients had higher level of IFN-α, IL-6 and TNF-α than controls (P<0.05). No correlation was found between the percentage of PDC, MDC and level of IFN-α, IL-6 and TNF-α. Conclusions The percentages of PDC and MDC in peripheral blood are decreased in patients with SLE. The reduction of PDC is correlated with disease activity and kidney damage of SLE. Disclosure of Interest None Declared

Objective To investigate the expression of B7-H4 on renal tissue and serum in the patients with LN.Methods A total of 90 LN patients,3 acute kidney injury patients,20 SLE patients without nephritis and 20 healthy donors were referred.Disease activity was assessed by SLEDAI scores.The expression of B7-H4 on kidney biopsies from patients with LN and acute kidney injury was measured using immunohistochemistry.The sB7-H4 of serum in LN and healthy donors was analysed using ELISA,and the correlation between sB7-H4 of serum in LN with clinical data was analysed too.Results Immunohistological staining revealed that B7-H4 antigen expressed on tubular epithelium,the percentage of B7-H4 positive expression renal tubules from LN patients(45±16)% was lower than that from acute kidney injury patients(86±11)%(P﹤0.05),and was negatively correlated with the index of tubulointerstitial damage(r=-0.476,P=0.02).The sB7-H4 in serum of LN patients(58.20 ng/ml,median) was not significantly lower than that of healthy controls(70.57±27.24 ng/ml),but was strongerly associated with SLEDAI with LN(r=0.353,P=0.006).Besides,the mean concentration of sB7-H4 level in high activity group(74.40±31.95 ng/ml) was significantly higher than that in moderate group(51.89±21.79 ng/ml)(p=0.017).Conclusion The expression of B7-H4 molecules probably has effects during the progress of LN,and a clear understanding of its functional roles may further elucidate the pathogenesis of this disease.

The quantitative identification and enrichment of viable regulatory T cells (Treg) requires reliable surface markers that are selectively expressed on Treg. Foxp3 is the accepted marker of nTreg, but it cannot be used to isolate cells for functional studies. In this study, we compared four staining profiles of Treg, including CD4+CD25high T cells, CD4+CD39+ T cells, CD4+CD73+ T cells, and CD4+CD25+CD127low/− T cells. We found that CD4+CD25+CD127low/− T cells expressed the highest level of Foxp3 and had the strongest correlation with CD4+CD25+Foxp3+ T cells, the accepted identifying characteristics for “real” nTreg cells. Moreover, functional data showed that CD4+CD25+CD127low/− T cells could effectively suppress the proliferation of CD4+CD25− T cells, suggesting that compared with the other three populations, CD4+CD25+CD127low/− T cells best fit the definition of naturally occurring regulatory T cells in human peripheral blood. Finally, we showed that CD4+CD25+CD127low/− can be used to quantitate Treg cells in individuals with systemic lupus erythematosus supporting the use of CD4+CD25+CD127low/− to identify human Treg cells.

T细胞激活需要两种信号途径:一种是通过T细胞表面受体(TCR)-人类主要组织相容性复合体(MHC)-肽复合物特异性抗原信号,第二种是通过抗原提呈细胞(APC)表面分子提供的共刺激信号.后者需要共刺激分子的参与,B7家族就是这一类分子,包括B7-H1、B7-H2、B7-H3、B7-DC和B7-H4.B7类分子主要与T细胞表面相应受体结合,在抗原特异性免疫应答过程中发挥正向或负向调控作用[1]。

目的研究系统性红斑狼疮（SLE）患者外周血CD4＋CXCR5＋T细胞占CD4＋T细胞百分率以及糖皮质激素对其的影响，探讨其在SLE发病机制中的作用。方法采用流式细胞术检测45例活动期、20例缓解期SLE患者及20名健康对照外周血中CD4＋CXCR5＋T细胞占CD4＋T细胞的百分率，比较其在各组中的差异及糖皮质激素治疗对其的影响，同时检测各组中CD19＋B细胞上CXCR5的表达。2组间比较用独立样本t检验，3组间比较采用多变量方差分析，与临床指标之间的相关性分析采用非参数的sDearman相关分析，治疗前后的差异用重复测量的方差分析。结果①SLE患者外周血CD4＋CXCR5＋T细胞占CD4＋T细胞的比例高于健康对照组[（16±7）％与（12±3）％，P〈0．01]，其中活动组[（18±7）％]高于健康对照组（P〈0．05），而缓解组[（11±4）％]和健康对照组之间差异无统计学意义（JD〉0．05）；狼疮肾炎组高于非狼疮肾炎组，但差异无统计学意义[（18±7）％与（14±7）％，P=0．05]。②CD4＋CXCR5＋T细胞百分率与SLE疾病活动指数（SLEDAI）、抗核抗体滴度和红细胞沉降率（ESR）呈正相关，与补体c3呈负相关（P均〈0．05），与C反应蛋白、病程、免疫球蛋白无相关性（P〉0．05）。抗双链DNA抗体升高组与正常组之间、抗Sm抗体、抗SSA／SSB抗体阴性组和阳性组之间差异无统计学意义（P均〉0．05）。③活动期SLE患者CD19＋B细胞卜CXCR5的表达比例低于健康对照组[（85±11）％与（94±3）％，P〈0．05]。④10例初发、未接受治疗的活动期患者在接受地塞米松（20mg／d）治疗后第1、3、7天外周血中CD4＋CXCR5＋T细胞百分率均低于治疗前（P均〈0．05）。治疗前后CD19＋CXCR5＋B细胞的百分率无变化（P均〉0．05）。结论外周血CD4＋CXCR5＋滤泡辅助性T细胞样细胞的异常可能参与SLE的发病。

机体是免疫活化与免疫抑制平衡的整体,CD4+调节性T细胞(Tr)是主要负责负性调节的细胞,但是目前关于CD4+Tr细胞特征性的标记仍众说纷纭.转录因子Foxp3是目前公认的Tr细胞的标记,CD4+CD25+Foxp3+T细胞是经典的Tr细胞标记,然而Foxp3定位在细胞内,透膜标记后细胞丧失功能及活性,所以用此群细胞进行功能研究受到限制[1].后来不同的报道表明CD4+CD39+T细胞、CD4+CD73+T细胞以及CD4+CD25+CD127low/-T细胞也高表达Foxp3并具有免疫抑制功能[2-4],但对于各群细胞的特异性并未做比较研究.本研究通过检测正常人外周血中CD4+CD25highT细胞、CD4+CD39+T细胞、CD4+CD73+T细胞、CD4+CD25+CD127low/-T细胞占CD4+T细胞的百分率、Foxp3蛋白的表达及抑制功能试验,寻找更好的Tr细胞表面的特异性标志。