Ex Vivo Costimulatory Blockade to Generate Regulatory T Cells From Patients Awaiting Kidney Transplantation

Short‐term outcomes of kidney transplantation have improved dramatically, but chronic rejection and regimen‐related toxicity continue to compromise overall patient outcomes. Development of regulatory T cells (Tregs) as a means to decrease alloresponsiveness and limit the need for pharmacologic immunosuppression is an active area of preclinical and clinical investigation. Nevertheless, the immunomodulatory effects of end‐stage renal disease on the efficacy of various strategies to generate and expand recipient Tregs for kidney transplantation are incompletely characterized. In this study, we show that Tregs can be successfully generated from either freshly isolated or previously cryopreserved uremic recipient (responder) and healthy donor (stimulator) peripheral blood mononuclear cells using the strategy of ex vivo costimulatory blockade with belatacept during mixed lymphocyte culture. Moreover, these Tregs maintain a CD3⁺CD4⁺CD25⁺CD127^lo surface phenotype, high levels of intracellular FOXP3 and significant demethylation of the FOXP3 Treg‐specific demethylation region on allorestimulation with donor stimulator cells. These data support evaluation of this simple, brief Treg production strategy in clinical trials of mismatched kidney transplantation.     

Mechanisms of Tolerance Induced by Donor-Specific Transfusion and ICOS-B7h Blockade in a Model of CD4+ T-Cell-Mediated Allograft Rejection

The inducible co-stimulatory molecule (ICOS) has been shown to play a critical role in T-cell activation and differentiation, and the regulation of alloimmune responses in vivo. Using an MHC class II mismatched model of CD4(+) T-cell-mediated rejection, we found that treatment of mice with DST and ICOS-B7h blockade induced long-term skin allograft survival and donor-specific transplantation tolerance. ICOS blockade, either during antigen priming or during the effector phase, previously shown to alter the outcome of the immune response, had a similar effect on graft survival. DST and anti-B7h mAb reduced the frequency of IFN-gamma-producing allospecific cells but did not produce deviation to a T(H)2 phenotype. In an adoptive transfer model using ABM TCR transgenic mice directly reactive to I-A(bm12), DST and anti-B7h mAb reduced the number of allospecific CD4(+) T cells and increased CD4(+) T-cell apoptosis. These data demonstrate that DST and anti-B7h mAb induces transplantation tolerance to MHC class II mismatched skin grafts by a reduction of the alloreactive clone size that is, at least in part, dependent on apoptosis of host alloantigen-specific CD4(+) T cells.     

亚麻子水提液对DPP-4及α-葡萄糖苷酶活性抑制实验研究

目的通过观察亚麻子水提液对二肽基肽酶-4（DPP-4）、α-葡萄糖苷酶抑制作用,为该药防治糖尿病提供实验依据。方法①以DPP-4酶、缓冲液、底物建立DPP-4抑制剂的体外筛选体系,对亚麻子水提液进行抑制实验,采用发色底物法测定吸光度（OD）,计算DPP-4抑制率及IC50值;②以蔗糖为底物建立α-葡萄糖苷酶活性抑制模型,采用葡萄糖氧化酶法测定亚麻子水提液对α-葡萄糖苷酶的抑制作用,计算其抑制率和IC50值。结果①亚麻子具有轻度DPP-4抑制作用,其IC50值738.20mg/L;②亚麻子具有α-葡萄糖苷酶抑制作用,其IC50值为365.9mg/mL。结论亚麻子水提液可一定程度地抑制DPP-4及α-葡萄糖苷酶活性。     

TLIF术式在布氏杆菌性脊柱炎病人中的临床疗效及安全性分析

目的 对比分析单纯后路内固定+一期经腰椎间孔病椎间病灶清除(TLIF)与经典的前后联合手术在布氏杆菌性脊柱炎患者中的临床疗效及安全性。 方法 对我院2015年1月至2017年12月收治的93例布病性脊柱炎患者的临床资料进行分析。按手术方式分为观察组(45例)和对照组(48例)。对两组患者的基础数据、临床指标、术前术后各项指标水平以及术后并发症、植骨治愈情况。 结果 观察组与对照组基础数据比较,差异无统计学意义(P>0.05)。观察组患者的手术时间、住院天数、术中出血量及术后下床时间均明显低于对照组(P<0.01)。两组患者术后3个月的ODI、VAS、CRP、ESR及Cobb角均明显低于术前(P<0.05);术后3个月,观察组患者的ODI、VAS、CRP、ESR及Cobb角均明显低于对照组(P<0.05)。观察组术后并发症发生率(4.4%)明显低于对照组(25.0%)(Χ²=7.674,P<0.01)。 结论 TLIF治疗布氏杆菌性脊柱炎患者的临床疗效突出,安全性较好,更有利于患者术后身体的恢复。     

米非司酮治疗后VEGF-165在异位和在位子宫内膜中的表达

目的探讨米非司酮对VEGF-165在子宫腺肌症中表达的影响。方法43例子宫腺肌症患者分为对照组(n=21)和米非司酮治疗组(n=22),采用放射免疫法测定对照组(月经第3天)和米非司酮组(术前)血清中FSH、LH、E2、PRL、P及T的水平。采用免疫组化法测定两组患者在在位和异位子宫内膜中的VEGF-165蛋白水平。结果米非司酮组较对照组血清FSH、LH、E2、P明显降低(P<0.05),而血清PRL和T的水平两组间无统计学差异(P>0.05)。VEGF-165在子宫内膜腺上皮细胞内的表达,异位内膜均明显高于在位内膜(P<0.05);而其在间质细胞内的表达,异位内膜与在位内膜无统计学差异(P>0.05)。米非司酮组异位和在位内膜腺上皮细胞、间质细胞中VEGF-165的表达水平均较对照组明显降低(P<0.05)。VEGF-165在对照组异位内膜腺上皮细胞中的表达,增殖期高于分泌期(P<0.05)。结论米非司酮治疗后,VEGF-165在异位和在位内膜中的表达明显下降,可能是通过抑制VEGF蛋白合成,降低血管通透性,抑制新生血管生成,从而有效地控制子宫内膜异位症的发生发展。     

慢性乙肝患者肝组织纤维化程度与HBV-DNA复制水平的相关性研究

目的:探讨HBV-DNA复制水平与肝纤维化之间的相关性。方法:对210例慢性乙型肝炎患者进行HBV-DNA和肝纤维化血清学标志透明质酸(HA)、层粘蛋白(LN)、III型前胶原(PCIII)、IV型胶原(IV-C)进行定量检测。应用SPSS10.0统计软件对结果数据进行分析处理。结果:随慢性乙肝临床类型的加重,肝纤维化血清学标志逐渐升高(P<0.01),而肝纤维化血清学标志与HBV复制水平呈正相关(P<0.05);结论:HBV复制水平与肝纤维化之间呈正相关。     

米非司酮对不同月经周期异位和在位子宫内膜雌激素受体α和孕激素受体表达的影响

目的 探讨米非司酮对雌激素受体α(ERα)和孕激素受体(PR)在不同月经周期的子宫腺肌症中表达的影响.方法 47例行全子宫切除术的子宫腺肌症患者分为米非司酮组(n=24)和未用药组(n=23);无腺肌症的正常子宫内膜作为对照组(n=15).运用免疫组化的方法测定异位和在位子宫内膜及正常子宫内膜在不同月经周期的腺上皮及间质细胞中ERα和PR水平.结果 未用药组ERα和PR在异位子宫内膜腺上皮及间质细胞中的表达均低于在位内膜及对照组正常内膜(P<0.05).对照组及未用药组ERα和PR在在位内膜腺上皮细胞中的表达,增生期高于分泌期(P<0.05),而在未用药组异位内膜,差异无统计学意义(P>0.05).ERα和PR在异位和在位子宫内膜中的表达,米非司酮组低于未用药组(P<0.05).结论 ERα和PR在异位子宫内膜中的表达与在位子宫内膜不同;异位内膜丧失了正常内膜的周期性变化规律;米非司酮通过下调其性激素受体(ERα和PR)含量治疗子宫腺肌症.     

Beta 2-integrin and L-selectin are obligatory receptors in neutrophil aggregation [see comments]

We have recently found that antibodies to L-selectin, the homing receptor on neutrophils, are as effective as those to beta 2-integrin at blocking formyl peptide-stimulated aggregation. Therefore, we investigated the requirements for expression of L-selectin and beta 2- integrin on adjacent cells during aggregation. Fluorescence flow cytometry allowed characterization of aggregates on the basis of size and color, as well as antibody binding to these two adhesive molecules. Formyl peptide-stimulated aggregate formation was measured for individual populations fluorescently labeled red (LDS-751) or green (CD44-FITC), and interpopulation red-green cell conjugates. Blocking either the beta 2-integrin or L-selectin adhesive epitope with monoclonal antibody on individual cell populations resulted in an approximately 50% reduction in two-color aggregation as compared with that in unblocked samples. Shedding the L-selectin on a cell population by preincubation with complexes of lipopolysaccharide and its plasma membrane binding protein also decreased aggregation to a control population by approximately 50%. We examined the aggregation of neutrophils from patients genetically deficient in beta 2-integrin and clinically leukocyte adhesion deficient (LAD). LAD adhesion to normal neutrophils was dependent on the expression of L-selectin on LAD cells and beta 2-integrin on normal cells. Thus, the minimum requirement for adhesion between two mixed populations of neutrophils was that one population expressed the beta 2-integrin and the other expressed the L- selectin adhesive epitope.     

地格达-4味汤中獐牙菜苦苷在急性肝损伤模型大鼠体内药代动力学研究

目的:建立急性肝损伤模型大鼠血清中獐牙菜苦苷的HPLC血药浓度测定方法,并对其药代动力学进行研究.方法:SD大鼠腹腔注射D-半乳糖胺(D-GIaN) (400 mg·kg-1),制备急性肝损伤模型,按高、中、低(0.39,0.77,1.54 g·kg-1)剂量灌胃给予地格达-4味汤,收集不同时间点含药血清,HPLC测定血药浓度,采用Winnonlin 5.1药动学软件拟合房室模型,计算其药动学参数.结果:獐牙菜苦苷在0.000 6 ～0.019 2 mg ·L-1(r =0.998 9)线性关系良好,平均回收率均在80％以上,方法学考察其他项均符合实验要求.地格达-4味汤高、中、低剂量组吸收半衰期基本一致,药时曲线下面积(AUC)与给药剂量之间呈现良好的线性关系,属于线性动力学过程.结论:獐牙菜苦苷在急性肝损伤模型大鼠体内的药时曲线符合二室模型特征;所建立的方法准确,灵敏度高,专属性好,可作为獐牙菜苦苷体内血清药物浓度分析方法.