B cells as a target of immune modulation

B cells have recently been identified as an integral component of the immune system; they play a part in autoimmunity through antigen presentation, antibody secretion, and complement activation. Animal models of multiple sclerosis (MS) suggest that myelin destruction is partly mediated through B cell activation (and plasmablasts). MS patients with evidence of B cell involvement, as compared to those without, tend to have a worse prognosis. Finally, the significant decrease in new gadolinium-enhancing lesions, new T2 lesions, and relapses in MS patients treated with rituximab (a monoclonal antibody against CD20 on B cells) leads us to the conclusion that B cells play an important role in MS and that immune modulation of these cells may ameliorate the disease. This article will explore the role of B cells in MS and the rationale for the development of B cell–targeted therapeutics. MS is an immune-mediated disease that affects over 2 million people worldwide and is the number one cause of disability in young patients. Most therapeutic targets have focused on T cells; however, recently, the focus has shifted to the role of B cells in the pathogenesis of MS and the potential of B cells as a therapeutic target.     

两性霉素B微乳的制备及体外经皮渗透研究

 目的制备水包油型两性霉素B微乳,并对其进行体外经皮渗透性能考察。方法用相转变温度法制备两性霉素B微乳,用光散射粒度仪测定微乳的粒径,电子透射电镜观察微乳的形态。用药物渗透扩散仪进行两性霉素B透皮吸收研究,高效液相色谱法测定两性霉素B含量。结果用相转变温度法制备了Tween80和异丙醇重量比为1∶1,IPM和混合表面活性剂之比为1∶9,水含量64.6%,载药量为0.247%的两性霉素B微乳。所制微乳符合微乳特征,调节pH值有助于两性霉素B由分散介质向微乳表面转移。水性月桂氮酮对两性霉素B的促渗效果最好,pH值为5.12,月桂氮酮浓度为1%时,两性霉素B微乳经离体兔皮的JS为4.627μg·cm^-2·h^-1,经离体大鼠皮肤的JS为4.484μg·cm^-2·h^-1。结论开发微乳透皮给药制剂有望提高两性霉素B的疗效,降低其毒副作用。     

Novel azapeptide inhibitors of cathepsins B and K. Structural background to increased specificity for cathepsin B

Abstract: We have designed and synthesized a new series of azapeptides which act as potential inhibitors of cathepsin B and/or cathepsin K. Their structures are based upon the inhibitory sites of natural cysteine protease inhibitors, cystatins. For the synthesized azapeptides, the equilibrium constants for dissociation of inhibitor–enzyme complex, K_i, were determined. Comparison of these values indicated that all of the azainhibitors act much stronger toward cathepsin B. Z‐Arg‐Leu‐His‐Agly‐Ile‐Val‐OMe ( 7 ) proved to be approximately 500 times more potent for cathepsin B than for cathepsin K. To be able to explain the obtained experimental values we used the molecular dynamics procedures to analyze the interactions between cathepsin B and compound 7 . We also determined the structure of the most potent and selective cathepsin B azainhibitor by means of NMR studies and theoretical calculations. In this report, we describe SAR studies of azapeptide inhibitors indicating the influence of the conformational flexibility of the examined compounds on inhibition of cathepsins B and K.     

Immune reactivity towards insulin, its amyloid and protein S100B in blood sera of Parkinson's disease patients

Peripheral immune responses can be sensitive indicators of disease pathology. We evaluated the autoimmune reactions to endocrine (insulin) and astrocytical (S100B) biomarkers in the blood sera of 26 Parkinson's disease (PD) patients compared with controls by using ELISA. We found a statistically significant increase of the autoimmune responses to both antigens in PD patients compared with controls with a mean increase of 70% and 50% in the autoimmune reactions towards insulin and S100B, respectively. Heterogeneity of the immune responses observed in patients may reflect the modulating effect of multiple variables associated with neurodegeneration and also changes in the basic mechanisms of individual autoimmune reactivity. We did not detect any pronounced immune reactions towards insulin amyloid fibrils and oligomers in PD patients, indicating that an amyloid-specific conformational epitope is not involved in immune recognition of this amyloid type, while sequential epitope of native insulin is hidden within the amyloid structures. Immune reactions towards S100B and insulin may reflect the neurodegenerative brain damaging processes and impaired insulin homeostasis occurring in PD.     

复方丹参注射液干扰素治疗乙肝后肝纤维化

目的探讨复方丹参注射液联合干扰素治疗慢性乙肝患者的疗效。方法110例慢性乙肝患者，按随机方法分成①对照组30例，应用普通保肝药物治疗，疗程6个月；②丹参组30例，应用复方丹参注射液（每ml含丹参、降香各1g）30ml加入10％葡萄糖溶液300ml中静脉注射1个月；③IFN组30例，应用IFN—α 3MU，隔日一次肌内注射，3个月；④联合组20例，应用复方丹参注射液30ml加10％葡萄糖溶液300ml静脉注射1个月，IFN-α 3MU，隔日一次肌内注射，3个月。丹参组，IFN组和联合组保肝药物治疗同对照组。四组病例在性别、年龄、病程，治疗前肝功能等方面均无统计学差异。治疗前检测肝功能，肝炎病毒标志，血清HA、IV—C、PCI—Ⅱ，部分病例进行肝穿病理检查。治疗开始后每月检测肝功能，3个月（治疗后）和6个月（随访时）时检测血清HA、IV—C、PCⅢ及乙肝病毒标志，治疗后1年行肝穿病理检查。结果治疗前四组患者血清HA、PCⅢ、IV—C水平无统计学差异；治疗后丹参组、IFN组、联合组血清HA、FCⅢ、IV—C水平较治疗前及对照组有不同程度的降低。结论复方丹参注射液联合IFN治疗可使血清HA、PCⅢ、IV—C有明显下降，肝组织病理改变明显改善，为目前有效的慢性乙肝治疗措施。     

Bile-pancreatic juice exclusion promotes Akt/NF-kB activation and chemokine production in ligation-induced acute pancreatitis

Using a unique surgical model (the donor rat model), we showed previously that duodenal replacement of bile-pancreatic juice, obtained fresh from a donor rat, ameliorates ligation-induced acute pancreatitis. We hypothesize that bile-pancreatic juice exclusion from gut exacerbates Akt/nuclear factor-kB (NF-kB) pathway activation and induces chemokine production in ligation-induced acute pancreatitis. We compared rats with bile-pancreatic duct ligation to those with duodenal bile-pancreatic juice replacement fresh from a donor rat beginning immediately before duct ligation. Sham control rats had ducts dissected but not ligated. Rats were killed 1 or 3 hours after operation (n=7/group). Akt activation (immunoblotting, immune-complex kinase assay, and ELISA), inhibitory protein I-kB (I-kB) activation (immunoblotting), and production of chemokines MCP-1 and RANTES (ELISA) were measured in pancreatic homogenates. NF-kB was quantitated in nuclear fractions using electrophoretic mobility shift assay. Duct ligation produced significant increases in pancreatic Akt, IkB, and NF-kB activation and production of MCP-1 and RANTES. Activation of the Akt/NF-kB pathway and increased MCP-1 and RANTES production in response to duct ligation were significantly reduced by bile-pancreatic juice replacement (ANOVA, P<0.05). Bile-pancreatic juice exclusion stimulates Akt/NF-kB pathway activation and increases chemokine production in ligation-induced acute pancreatitis. Presented at the annual meeting of The Society for Surgery of the Alimentary Tract, Chicago, Illinois, May 16, 2005 (poster).     

丹参酚酸B盐对内皮素诱导的人肝星状细胞收缩的抑制作用及机制研究

目的 以培养的人肝星状细胞（HSC）为研究对象，观察内皮素-1（ET-1）对HSC的收缩作用及丹参酚酸B盐（SA-B）对ET-1的抑制作用，并探讨SA B抑制HSC收缩的作用环节和可能作用机制。 方法 采用酶消化、Nycodenz密度梯度离心的方法分离人HSC，以胶原凝胶收缩法观察ET-1对于传代HSC的收缩作用，以及低、中、高3种剂量的SA B对其不同的干预作用；将ET-1、SA-B直接添加于传代HSC的无血清培养液中，以激光共聚焦显微镜技术观测HSC内钙离子浓度的变化。 结果 胶原凝胶收缩实验显示，ET-1可诱导HSC收缩（P<0.01）；3种剂量的SA-B可明显抑制ET-1诱导的HSC收缩（均P<0.01）；加入ET-1后，HSC形态学改变明显，细胞数量减少，但SA B对这些改变也有抑制。激光共聚焦显微镜下观察，ET-1可刺激细胞内钙离子短暂迅速增加，加入SA-B后，该作用被明显抑制。 结论 SA-B能显著抑制ET-1诱导的HSC收缩，其抑制收缩的机制与降低HSC内的钙离子浓度有关。SA-B的抑制HSC收缩作用可能是其抗肝纤维化及门脉高压的机制之一。     

慢性乙型肝炎肝郁脾虚证的logistic回归及判别分析

目的归纳慢性乙型肝炎肝郁脾虚证的辨证要点。方法对慢性乙型肝炎肝郁脾虚证、肝郁证、脾虚证的症状及舌、脉进行logistic回归及判别分析。结果肝郁脾虚证logistic回归提取的症状为腹胀、倦怠乏力、腹痛、食欲不振、紧脉、便溏、胁肋疼痛、胁肋不适；判别方程包含的症状则为腹痛、腹胀、倦怠乏力、便溏、黄苔。结论肝郁脾虚证的辨证要点包括胁肋疼痛或不适、腹胀、腹痛、乏力、食欲不振、便溏。利用logistic回归和判别分析归纳本证诊断要点是可行的。