CD40 activation in human pancreatic islets and ductal cells

AIMS/HYPOTHESIS: CD40 expression on non-haematopoietic cells is linked to inflammation. We previously reported that CD40 is expressed on isolated human and non-human primate islets and its activation results in secretion of IL-8, macrophage inflammatory protein 1-beta (MIP-1beta) and monocyte chemoattractant protein-1 (MCP-1) through nuclear factor-kappaB and extracellularly regulated kinases 1/2 pathways. The objective of this study was to identify the pattern of gene expression, and to study viability and functionality affected by CD40-CD40 ligand (CD40L) interaction in human islets. Furthermore, we have studied the CD40-mediated cytokine/chemokine profile in pancreatic ductal cells, as they are always present in human islet transplant preparations and express CD40 constitutively. METHODS: CD40-CD40L gene expression modulation was studied by microarray on islet cells depleted of ductal cells. Selected genes were validated by quantitative RT-PCR. The cytokine profile in purified ductal cells was evaluated by Luminex technology, based on the use of fluorescent-coated beads, known as microspheres, and capable of multiplex detection of proteins from a single sample. Glucose-stimulated insulin secretion and islet viability were assessed by perifusion and 7-aminoactinomycin D membrane exclusion, respectively. RESULTS: Statistical analysis of microarrays identified 30 genes exhibiting at least a 2.5-fold increase across all replicate arrays. The majority of them were related to oxidative stress/inflammation. Prominently upregulated were chemokine C-X-C motif ligand 1 (CXCL1), CXCL2 and CXCL3 belonging to the CXC family of chemokines related to IL-8. CD40-mediated CXCL1 secretion was confirmed by ELISA. The viability or in vitro function was not affected by CD40 activation. In addition to previously reported IL-8, MIP-1beta and MCP-1, CD40 stimulation in ductal cells produced IL-1beta, IFN-gamma, TNF-alpha, granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. CONCLUSIONS/INTERPRETATION: CD40 activation in islets and ductal cells produces cytokines/chemokines with a broad-spectrum range of biological functions.     

Cytomegalovirus load at treatment initiation is predictive of time to resolution of viremia and duration of therapy in hematopoietic cell transplant recipients

BackgroundPreemptive antiviral therapy relies on viral load measurements and is the mainstay of cytomegalovirus (CMV) prevention in hematopoietic cell transplant (HCT) recipients. However, optimal CMV levels for the initiation of preemptive therapy have not been defined.ObjectivesThe objectives of our work were to evaluate the relationship between plasma CMV DNA levels at initiation of preemptive therapy with time to resolution of viremia and duration of treatment.Study designRetrospective analysis of HCT recipients undergoing serial CMV PCR testing between June 2011 and June 2014 was performed.Results221 HCT recipients underwent preemptive therapy for 305 episodes of CMV viremia. Median time to resolution was shorter when treatment was initiated at lower CMV levels (15 days at 135–440 international units (IU)/mL, 18 days at 441–1000 IU/mL, and 21 days at >1000 IU/mL, P < .001). Prolonged viremia lasting >30 days occurred less frequently when treatment was initiated at 135–440 IU/mL compared to 441–1000 IU/mL and >1000 IU/mL (1%, 15%, 24%, P < .001). Median treatment duration was also shorter in the lower viral load groups (28, 34, 37 days, P < .001).ConclusionInitiation of preemptive therapy at low CMV levels was associated with shorter episodes of viremia and courses of antiviral therapy. These data support the utility of initiating preemptive CMV therapy at viral loads as low as 135 IU/mL in HCT recipients.     

夏枯草有效组分结肠靶向片剂中迷迭香酸含量HPLC法测定

目的：建立以高效液相色谱法测定夏枯草有效组分结肠靶向片剂中迷迭香酸的含量测定方法,测定药物的回收率及含量等。方法 Diamonsil C18色谱柱(200 mm×4.6 mm,5μm),检测波长330 nm,流动相为甲醇-0.1%甲酸水溶液(45∶55),流速1.0 ml/min,进样量20μl。结果迷迭香酸甲醇溶液在2.99～119.6μg/ml浓度范围内线性关系良好,溶液72 h内稳定,精密度良好,加样回收率在95%～105%范围内,RSD均小于2%。结论建立夏枯草有效组分结肠靶向片剂的HPLC含量测定方法专属性强,符合含量测定要求。     

石榴皮、石榴子质量标准提高研究

目的：提高石榴皮、石榴子的质量标准。方法：采用显微鉴别法、薄层色谱法鉴别石榴子药材;按照《中国药典》2010年版（一部）附录方法,测定石榴子药材水分、总灰分、酸不溶性灰分、醇溶性浸出物含量;采用HPLC法测定石榴皮、石榴子的鞣花酸含量。结果：确定了石榴子药材的显微特征,建立了石榴子的薄层鉴别方法,建立了石榴皮、石榴子的鞣花酸含量测定方法。结论：本研究建立的定性、定量方法,可用于石榴皮、石榴子药材质量控制。     

《中医体质分类和判定》标准质疑

2009年中华中医药学会颁布了《中医体质分类和判定》标准,并已进入推广应用过程。考察后发现,该标准的建立未能统一执行中医体质分类标准的原则和方法;体质分类特征交叉重叠,界限不清;单一体质内部各因素之间关联性较差。故建议不宜将中医体质学说拔高到中医体质学的高度;《中医体质分类和判定》标准尚不成熟,应慎重推广应用。     

Genetics in liver diseases

In recent years, few fields in medicine have witnessed discoveries as momentous as those pertaining to the liver. Dramatic advances have been made, particularly in the areas of molecular biology and genetics. A joint EASL/AASLD Monothematic Conference was held on June 23rd-24th, 2006, in Modena, Italy, to bring the latest breakthroughs in different fields of genetics to hepatologists. This article reports the highlights of the conference and summarizes the main conclusions and implications for clinical and experimental hepatology.     

狗皮膏(改进型)质量标准研究

目的:修订提高狗皮膏(改进型)的质量标准。方法:用薄层色谱法对狗皮膏(改进型)中当归进行定性鉴别;用气相色谱法对狗皮膏(改进型)中冰片、薄荷脑及水杨酸甲酯进行定性鉴别,并测定处方中冰片、薄荷脑及樟脑的含量;用高效液相色谱法测定处方中盐酸苯海拉明的含量。结果:当归的薄层色谱鉴别斑点清晰,重现性好,无干扰;龙脑在0.03~0.26μg,薄荷脑在0.07~0.71μg,樟脑在0.05~0.46μg,盐酸苯海拉明在0.56~5.59μg范围内线性关系良好,各成分平均回收率分别为99.74%(龙脑)、99.10%(薄荷脑)、99.08%(樟脑)、96.95%(盐酸苯海拉明)。结论:修订后的质量标准简便易行,可有效控制狗皮膏(改进型)的质量。     

山东省立医院2011-2013年门诊及住院患者乙型肝炎病毒感染状况分析

目的 分析山东省立医院门诊及住院患者乙肝五项定量检测结果,了解患者的乙肝感染状况,为乙肝的防治提供理论依据.方法 采用电化学发光免疫分析法对2011-2013年226 193例门诊及住院患者进行乙肝五项定量检测,分别按年份、性别、年龄和乙肝五项模式进行分组,应用SPSS 21.0统计软件进行统计分析.结果 共检出HBsAg阳性14 069例,HBsAg阳性率为6.22％(14 069/226 193).2011-2013年HBsAg阳性检出率分别为6.64％(4974/74 959)、6.18％ (4410/71 399)、5.87％ (4685/79 835),呈逐年下降趋势.男性HBsAg阳性检出率为7.43％(8855/119 181),明显高于女性的4.87％(5214/107012).0～19岁年龄组HBsAg阳性率最低,为1.28％ (364/28 355);30～39岁年龄组HBsAg阳性率最高,为8.21％ (2694/30 129).共检出12种乙型肝炎病毒感染模式,“乙肝小三阳”检出率最高,为60.63％ (8530/14 069);其次为“乙肝大三阳”,检出率为26.05％ (3665/14 069);再次为1、3、4、5阳,1、5阳,1、2、4、5阳,1、2、3、5阳,检出率分别为5.52％ (777/14 069)、2.84％ (400/14 069)、2.04％(287/14069)、1.26％(177/14069);还检出了6种罕见模式:1阳,1、2、3、4、5阳,1、3阳,1、2、5阳,1、2阳,1、2、3阳,检出率均低于1％,分别为0.80％(113/14069)、0.46％ (65/14 069)、0.23％(33/14069)、0.08％(11/14069)、0.07％ (10/14 069)、0.01％(1/14069).结论 山东地区乙肝感染具有性别差异、年龄差异,且呈逐年下降趋势,感染模式以小三阳和大三阳为主,多种感染模式并存.     

直接测序法和qPCR-HRM法检测NSCLC组织EGFR突变的比较研究

目的 比较直接测序法和实时聚合酶链反应-高分辨率融解曲线技术(qPCR-HRM)检测非小细胞肺癌(NSCLC)表皮生长因子受体(EGFR)基因突变的差异,探讨适用于临床检测EGFR基因突变的方法,为NSCLC患者的靶向治疗提供理论依据.方法 共收集NSCLC患者组织标本52例,分别采用qPCR-HRM法和直接测序法进行EGFR基因突变检测,采用x2检验对检测结果进行统计学比较.结果 52例NSCLC患者组织标本中,直接测序法检测的EGFR基因突变率为30.8％(16/52),qPCR-HRM法检测的EGFR基因突变率为32.7％ (17/52),两种方法检测结果相比较,差异无统计学意义(x2=0.04,P ＞0.05).结论 与直接测序法相比较,qPCR-HRM法是一种操作简便、费用低、速度快、灵敏度高的检测方法,对于临床筛选适合EGFR-TKI治疗的NSCLC患者亚群,预测EGFR-TKI疗效都具有重要意义,值得临床进一步推广应用.