4-1BB co-stimulation enhances human CD8(+) T cell priming by augmenting the proliferation and survival of effector CD8(+) T cells

Interactions between 4-1BB and its ligand, 4-1BBL, enhance CD8(+) T cell-mediated antiviral and antitumor immunity in vivo. However, mechanisms regulating the priming of CD8(+) T cell responses by 4-1BB remain unclear, particularly in humans. The 4-1BB receptor was undetectable on naive or resting human CD8(+) T cells and induced in vitro by TCR triggering. Naive cord blood cells were therefore primed in vitro against peptides or cellular antigens and then co-stimulated with 4-1BBL or agonistic antibodies. Co-stimulation enhanced effector function such as IFN-gamma production and cytotoxicity by augmenting numbers of antigen-specific and effector CD8(+) T cells. OKT3 responses also showed reduced cell death and revealed that the proliferation of CD8(+) T cells required two independently regulated events. One, the induction of IL-2 production, could be directly triggered by 4-1BB engagement on CD8(+) T cells in the absence of accessory cells. The other, expression of CD25, was induced with variable efficacy by accessory cells. Thus, suboptimal accessory cells and 4-1BB co-stimulation combined their effects to enhance IL-2 production and proliferation. Reduced apoptosis observed after co-stimulation in the presence of accessory cells correlated with increased levels of Bcl-X(L) in CD8(+) T cells, while Bcl-2 expression remained unchanged. Altogether, 4-1BB enhanced expansion, survival and effector functions of newly primed CD8(+) T cells, acting in part directly on these cells. As 4-1BB triggering could be protracted from the TCR signal, 4-1BB agonists may function through these mechanisms to enhance or rescue suboptimal immune responses.     

4-1BB在人外周血γδT细胞活化中的协同刺激作用

采用结核杆菌(Mtb)低分子多肽刺激人外周血单个核细胞,流式细胞术(FCM)检测不同活化时相γδT细胞膜表面4-1BB分子的表达;用阻断型4-1BB配体(4-1BBL)单抗阻断4-1BB/4-1BBL信号,FCM检测γδT细胞的增殖比率和细胞内产生IFN-γ的情况,同时与阻断CD28/B7-1信号相比较。结果显示,静止的γδT细胞膜表面不表达4-1BB分子,Mtb抗原刺激后6 h,4-1BB即有明显表达(29.71%),48 h达到高峰(49.79%);与未阻断组相比,阻断4-1BB/4-1BBL信号,γδT细胞的增殖效应和细胞内IFN-γ的产生均明显下降(P<0.01),与CD28/B7-1信号阻断组相比,差异无显著性(P>0.05)。提示4-1BB/4-1BBL信号同CD28/B7-1信号一样,可为γδT细胞活化提供协同刺激作用。     

Genetic dissection of lymphopenia from autoimmunity by introgression of mutated Ian5 gene onto the F344 rat

Peripheral T cell lymphopenia (lyp) in the BioBreeding (BB) rat is linked to a frameshift mutation in Ian5, a member of the Immune Associated Nucleotide (Ian) gene family on rat chromosome 4. This lymphopenia leads to type 1 (insulin-dependent) diabetes mellitus (T1DM) at rates up to 100% when combined with the BB rat MHC RT1 u/u genotype. In order, to better study the lymphopenia phenotype without possible confounding effects of diabetes or other autoimmune disease, we generated congenic F344.lyp rats by introgression of lyp on diabetes-resistant MHC RT1 lv1/lv1 F344 rats. Analysis of thymic CD4 and CD8 T lymphocytes revealed no difference in the percentage of CD4(-)CD8(+)and CD4(+)CD8(-)subsets in lyp/lyp compared to +/+ F344 rats. The same subsets was however dramatically reduced in blood (P=0.005), spleen (P=0.019) and mesenteric lymph nodes (MLN) (P<0.0001). Compared to F344 +/+ rats double positive CD4(+)CD8(+)T cells were increased only in lyp/lyp spleen (P=0.034) while double negative CD4(-)CD8(-)were increased in thymus (P=0.033), spleen (P=0.012), MLN (P<0.0001), and peripheral blood (P<0.0001). There were no signs of inflammatory lesions in organs and tissues in F344.lyp/lyp rats examined at 120 days of age or older. We thus conclude that the lymphopenia phenotype was reconstituted by introgression of lyp on to F344 rats without subsequent development of organ-specific autoimmunity. The congenic F344.lyp rat should prove useful to dissect the mechanisms by which the Ian5 frameshift mutation affects T cell selection, differentiation and maturation without organ-specific autoimmunity.     

新生儿缺氧缺血性脑病患儿脐血肌酸激酶脑型同工酶和神经元特异性烯醇化酶变化及临床意义

目的观察新生儿缺氧缺血性脑病（HIE）时脐血肌酸激酶脑型同工酶（CK-BB）及神经元特异性烯醇化酶（NSE）水平变化,早期诊断新生儿缺氧缺血性脑损伤,为临床早期诊断新生儿缺氧缺血性脑损伤及协助HIE分度,估计预后提供一定的依据。方法20例足月儿,出生后1分钟Apgar〉9分的新生儿为对照组,32例诊断为新生儿缺氧缺血性脑病的新生儿为病例组,生后立即收集脐血,检测CK-BB及NSE值。结果新生儿HIE组脐血CK-BB及NSE水平明显高于对照组,差异显著（P〈0.01）,且和HIE程度呈正相关。结论新生儿缺氧缺血性脑病（HIE）组脐血CK-BB及NSE水平高于对照组,差异显著。CK-BB,NSE水平变化可作为反映早期脑损伤的生化指标。协助HIE分度,估计预后,进行早期干预,具有一定的临床价值。     

任务驱动+翻转课堂教学法在程序设计类课程中的应用

以BB网络平台为载体,为学生提供一个自由、开发的学习环境,将任务驱动式教学法和“翻转课堂”教学理念相结合,对中医药院校的程序设计类课程的教学方法、教学流程、教学模式等进行改革,培养学生分析问题、解决问题的能力,以及自主学习和终生学习的技能。经过课程组两轮的实验,教学效果良好,对其他课程的教学模式和教学结构的研究和改革具有借鉴意义。     

蛋白酶抑制剂联合CRRT对脓毒症患者FABP3、GPBB水平的影响

目的 探讨蛋白酶抑制剂联合连续肾脏代替治疗(CRRT)对脓毒症患者心型脂肪酸结合蛋白(FABP3)、糖原磷酸化酶同工酶BB(GPBB)水平的影响。方法 选取2019年2月至2020年6月我院收治的脓毒症患者134例,按照随机数字表法分为CRRT组与联合治疗组,每组各67例。其中CRRT组患者进行CRRT治疗,联合治疗组患者进行CRRT联合蛋白酶抑制剂进行治疗。使用流式细胞仪对CD4+、CD8+、CD4+/CD8+指标水平进行测定与比较,采用酶联免疫法检测白介素-8(IL-8)、肿瘤细胞因子-α(TNF-α)、白介素-6(IL-6)及脂肪酸结合蛋白3(FABP3)、糖原磷酸化酶同工酶BB(GPBB)水平,并比较,免疫组化法检测D-乳酸、二胺氧化酶(DAO)水平,并对两组患者治疗疗效进行评价。结果 治疗后,与CRRT组相比,联合治疗组CD4+指标水平与CD4+/CD8+均较高,CD8+指标水平较低(P0.05);与CRRT组相比,联合治疗组IL-8、TNF-α、IL-6水平均较低(P0.05);联合治疗组D-乳酸水平及DAO水平均低于CRRT组(P0.05);与CRRT组相比,联合治疗组FABP3水平较高、GPBB水平较低(P0.05);与CRRT组相比,联合治疗组治疗总有效率较高(P0.05)。结论 使用蛋白酶抑制剂联合CRRT对脓毒症患者进行治疗,能够显著改善患者免疫功能,抑制炎症反应,同时能够降低肠道黏膜损害程度,使得FABP3水平升高、GPBB水平降低,从而减轻脓毒症患者心肌损伤程度。     

Physico-chemical characterization of a novel group of dopamine D(3)/D(2) receptor ligands, potential atypical antipsychotic agents

The fundamental physico-chemical properties such as ionization and lipophilicity of twelve alkyl-aryl-piperidine and aryl-piperazine derivatives have been determined. Compounds are members of a recently identified, new class of potent dopamine D(3)/D(2) receptor ligands as potential atypical antipsychotic agents and were used in the development of a promising drug candidate (RGH-188) being present currently in clinical phase II investigations. The ionization constant (pK(a)) and the partition coefficient in octanol/water (logP(oct)) and cyclohexane/water systems (logP(ch)) were measured by validated analytical methods. Based on the highly precise physico-chemical data the structure-property relationships (SPR) were studied. The effect of the polar and apolar heteroatoms as well as polar and apolar surface areas on the partition in the two solvent systems was investigated by linear regression and multivariate linear regression analyses. Brain/blood concentration ratios (BB values) as a function of time were determined by HPLC analyses on plasma and brain homogenates of Wistar rats. A linear relationship has been found between DeltalogP values (logP(oct)-logP(ch)) and experimental logBB values, verifying that physico-chemical data can predict pharmacokinetic behaviour.     

Phase I metabolism of the highly potent synthetic cannabinoid MDMB‐CHMICA and detection in human urine samples

Among the recently emerged synthetic cannabinoids, MDMB‐CHMICA (methyl N ‐{[1‐(cyclohexylmethyl)‐1H ‐indol‐3‐yl]carbonyl}‐3‐methylvalinate) shows an extraordinarily high prevalence in intoxication cases, necessitating analytical methods capable of detecting drug uptake. In this study, the in vivo phase I metabolism of MDMB‐CHMICA was investigated using liquid chromatography‐electrospray ionization‐tandem mass spectrometry (LC‐ESI‐MS/MS) and liquid chromatography‐electrospray ionization‐quadrupole time‐of‐flight‐mass spectrometry (LC‐ESI‐Q ToF‐MS) techniques. The main metabolites are formed by hydrolysis of the methyl ester and oxidation of the cyclohexyl methyl side chain. One monohydroxylated metabolite, the ester hydrolysis product and two further hydroxylated metabolites of the ester hydrolysis product are suggested as suitable targets for a selective and sensitive detection in urine. All detected in vivo metabolites could be verified in vitro using a human liver microsome assay. Two of the postulated main metabolites were successfully included in a comprehensive LC‐ESI‐MS/MS screening method for synthetic cannabinoid metabolites. The screening of 5717 authentic urine samples resulted in 818 cases of confirmed MDMB‐CHMICA consumption (14%). Since the most common route of administration is smoking, smoke condensates were analyzed to identify relevant thermal degradation products. Pyrolytic cleavage of the methyl ester and amide bond led to degradation products which were also formed metabolically. This is particularly important in hair analysis, where detection of metabolites is commonly considered a proof of consumption. In addition, intrinsic activity of MDMB‐CHMICA at the CB₁ receptor was determined applying a cAMP accumulation assay and showed that the compound is a potent full agonist. Based on the collected data, an enhanced interpretation of analytical findings in urine and hair is facilitated. Copyright © 2016 John Wiley & Sons, Ltd.     

Enhanced SRSF5 Protein Expression Reinforces Lamin A mRNA Production in HeLa Cells and Fibroblasts of Progeria Patients

The Hutchinson Gilford Progeria Syndrome (HGPS) is a rare genetic disease leading to accelerated aging. Three mutations of the LMNA gene leading to HGPS were identified. The more frequent ones, c.1824C>T and c.1822G>A, enhance the use of the intron 11 progerin 5′splice site (5′SS) instead of the LMNA 5′SS, leading to the production of the truncated dominant negative progerin. The less frequent c.1868C>G mutation creates a novel 5′SS (LAΔ35 5′SS), inducing the production of another truncated LMNA protein (LAΔ35). Our data show that the progerin 5′SS is used at low yield in the absence of HGPS mutation, whereas utilization of the LAΔ35 5′SS is dependent upon the presence of the c.1868C>G mutation. In the perspective to correct HGPS splicing defects, we investigated whether SR proteins can modify the relative yields of utilization of intron 11 5′SSs. By in cellulo and in vitro assays, we identified SRSF5 as a direct key regulator increasing the utilization of the LMNA 5′SS in the presence of the HGPS mutations. Enhanced SRSF5 expression in dermal fibroblasts of HGPS patients as well as PDGF‐BB stimulation of these cells decreased the utilization of the progerin 5′SS, and improves nuclear morphology, opening new therapeutic perspectives for premature aging.     

浅谈Blackboard教学平台对教师角色转变和能力发展的影响

实施信息化教育既是国内外教育改革与发展的趋势与潮流,也是高等教育教学实践中面临的重要课题。目前推广使用的Blackboard教学平台是网络时代教育发展的要求,是教学现代化的鲜明体现。同时,以Blackboard教学平台为代表的信息化教育对教师教学能力提出了新的要求和挑战,高校教师必须具备相应的教学能力才能有效地实施信息化教育。探讨了Blackboard教学平台推广使用的背景下,教师教学角色的转变和教学能力的新发展。