肾上腺髓质素对尾加压素Ⅱ刺激的血管平滑肌细胞增殖的影响

目的:观察肾上腺髓质素(ADM)对尾加压素Ⅱ(UⅡ)刺激的血管平滑肌细胞(VSMC)增殖的影响。方法:贴块法培养大鼠胸主动脉VSMC;[³H]-胸腺嘧啶([³H]-TdR)掺入测定反映VSMCDNA合成;[γ-³²P]-ATP标记的同位素法测定丝裂素活化蛋白激酶(MAPK)活性。结果:UⅡ(10^-8mol/L)显著促进VSMC-TdR掺入和激活MAPK比对照组分别高38%(P＜0.05)和260%(P＜0.01)。UⅡ加10^-10、10^-9、10^-8mol/LADM组VSMC-TdR掺入分别较UⅡ组低7%(P＞0.05)、32%(P＜0.05)和41%(P＜0.01)。MAPK活性分别低24%(P＞0.05)、32%(P＜0.05)和36%(P＜0.05)。结论:肾上腺髓质素抑制UⅡ诱导的VSMC增殖,可能与其抑制MAPK活性有关。     

亚低温缺血预处理对大鼠缺血再灌注肠保护作用的研究

目的:探讨亚低温缺血预处理对缺血再灌注肠的作用及机制。方法: 32只大鼠随机分为4组(每组8只),比较假手术对照组(sham)、缺血再灌注组(I/R)、缺血预处理组(IP)、亚低温预处理组(MHIP)小肠组织湿干重比、Ca²⁺-Mg²⁺-ATPase含量及血清乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平、总抗氧化(TAX)能力的变化,并观察各组肠组织超微机构及Bcl-2、Bax蛋白表达。结果: 缺血再灌注后I/R组小肠湿干重比值、LDH、MDA含量、Bcl-2和Bax蛋白表达吸光度(A)值明显高于sham组(P<0.01);Ca²⁺-Mg²⁺-ATPase、SOD活性及TAX能力明显低于sham组(P<0.01)。IP组小肠湿干重比值、LDH、MDA含量、Bax蛋白表达吸光度值明显低于I/R组(P<0.01);Ca²⁺-Mg²⁺-ATPase 、SOD活性及TAX能力、Bcl-2蛋白表达吸光度(A)值明显高于I/R组(P<0.01)。结论: 亚低温缺血预处理通过增加肠组织自身抗氧化能力、抑制脂质过氧化、上调 bcl-2 基因的蛋白表达与下调bax基因的蛋白表达以抑制肠组织细胞凋亡的发生等机制对抗肠缺血再灌注损伤。     

C-Jun 氨基端激酶与胰岛素抵抗

 C-Jun 氨基端激酶（c-Jun N-terminal kinase，JNK），又称应激活化蛋白激酶（stress-activated protein kinase，SAPK），是在哺乳动物体内发现的第 3 类促分裂原活化蛋白激酶（mitogen activated protein kinase，MAPK）的家族成员之一^[1]。JNK 广泛参与胚胎发育、细胞分化和凋亡、免疫反应以及胰岛素抵抗（insulin resistance，IR）等多种生理病理过程。JNK 蛋白可由 Jnk1、Jnk2、Jnk3 等 3 个基因编码，其中 Jnk1 基因和 Jnk2 基因在全身各组织中广泛表达，Jnk3 基因则选择性地在脑、心脏、睾丸组织中表达。Jnk 基因通过选择性剪接可表达 10 种不同形式的 JNK 蛋白，这些蛋白按其相对分子质量的不同可分为 46 000 亚型和 54 000 亚型^[2]。哺乳动物的单基因敲除实验表明，Jnk1 基因和 Jnk2 基因在多数情况下能够进行功能互补，同时又具有各自特定的生理功能，其中 Jnk1 基因在 IR 及胰腺 β 细胞功能衰竭的发生发展过程中发挥着重要的调节作用^[3]。进一步的研究还表明选择性地抑制 JNK 的活化将可能为糖尿病的治疗提供一个新途径^[4]。结合近年来相关文献，本文就 JNK 与 IR 关系的研究进展进行简要综述。。。。。。     

以Pin1为靶点的肿瘤反义基因治疗研究进展

 肽基脯氨酰顺反式异构酶 1（peptidyl-prolyl cis/trans isomerase 1，Pin1）是进化上非常保守的肽基脯氨酰异构酶（peptidyl-prolyl isomerase，PPIase）家族的成员，属微小菌素蛋白^[1]，最初报道于 1996 年，在分离与 NIMA（never in mitosis gene A）相互作用的蛋白时得到^[2]。一些研究发现，Pin1 的作用靶点可以是参与细胞增殖、细胞凋亡和转录的蛋白，如细胞周期蛋白 E（cyclin E）、cyclin D1、β-连环蛋白（β-catenin）等，其中多数蛋白在肿瘤形成过程中会失调^[3-5]；并且 Pin1 在多种肿瘤中过表达（如宫颈癌、乳腺癌、前列腺癌等）^[6-14]，因此认为 Pin1 与肿瘤发生有关。有学者已经对 Pin1 在肿瘤治疗方面的应用进行了研究，包括靶向 Pin1 的小分子化合物和反义基因。本文主要就靶向 Pin1 的肿瘤反义基因治疗方面的研究进展做简要综述。 ......     

环胞霉素A抑制神经肽Y诱导大鼠心肌细胞肥大效应

目的:观察Ca²⁺/CaM依赖的钙调神经磷酸酶(CaN)抑制剂环胞素A(CsA)对神经肽Y诱导心肌细胞肥大效应的影响。方法:用神经肽Y(NPY)刺激Wistar乳鼠心肌细胞,并用环胞素A加以干预。应用氚-亮氨酸([³H]-Leu)掺入法测定心肌细胞蛋白质合成速率、RT-PCR法测心肌细胞c-junmRNA表达。结果:(1)心肌细胞氚-亮氨酸([³H]-Leu)掺入量测定:与对照组相比,NPY10nmol/L组氚-亮氨酸([³H]-Leu)掺入量有所增高,但与对照组比无显著差别,而NPY100nmol/L组心肌细胞氚[³H]-Leu掺入量较对照组明显增高(P＜0.05)。CsA组和对照组相比无显著差别。(2)心肌细胞内c-junmRNA表达:NPY组心肌细胞c-junmRNA的RT-PCR产物量明显高于对照组和CsA组(P＜0.01),对照组和CsA组间无显著差别。结论:NPY刺激心肌细胞蛋白质合成增加、心肌细胞原癌基因(肥大早期反应基因)c-junmRNA表达,提示NPY可诱导心肌细胞肥大;CaN抑制剂CsA可阻断NPY上述效应,说明Ca²⁺/CaM依赖的CaN信号途径在其中起重要作用。     

转基因动物育种研究的现状与趋势

 20 世纪 80 年代以来发展的转基因动物技术是指将已知的外源基因导入动物细胞并稳定整合到基因组中,使其得以表达的技术。转基因动物育种是指通过转基因的手段,从分子水平对动物进行改良,以期得到目标性状。1997 年,克隆羊 Dolly 的诞生^[1],开创了哺乳动物体细胞核移植技术的先河,随后,乳腺中表达人凝血因子 IX 的转基因克隆羊 Polly 培育成功^[2]。2005 年抗乳房炎转基因牛的诞生^[3],2006 年多不饱和脂肪酸转基因克隆猪的培育成功^[4],标志着转基因动物育种进入了新的发展历程。2009 年生产高比例的抗原特异性人源抗体的转染色体牛的出生^[5],是转基因动物生产药用蛋白的又一个里程碑。转基因动物技术自从诞生以来就在改良家畜生产性状、提高家畜抗病力以及生产非常规畜牧产品（如人药用蛋白和工业用酶）等方面显示了广阔的应用前景。随着基因工程技术的不断发展,转基因动物技术将会不断得到完善,从而在未来的动物育种中将发挥巨大的作用。......     

Bcl-2、Bax在NHE-1抑制导致的大鼠肺动脉平滑肌细胞凋亡中的作用

目的:探讨Bcl-2、Bax蛋白表达在Na⁺/H⁺交换器-1(NHE-1)抑制而诱导大鼠肺动脉平滑肌细胞(PASMC)凋亡中的作用。方法: 荧光指示剂(Fura-2/AM)测定法检测转染NHE-1特异性核酶基因的大鼠PASMC内Ca²⁺(i)变化;RT-PCR方法检测细胞内bcl-2和baxmRNA表达变化, 免疫组化法检测细胞内Bcl-2和Bax蛋白表达变化。 结果:转染NHE-1特异性核酶基因后, 大鼠PASMC内i显著升高, bcl-2mRNA及蛋白表达显著降低, baxmRNA和蛋白表达显著增加。结论: NHE-1抑制诱导的PASMC凋亡与i增加、bcl-2表达降低及bax表达增加有关。     

免疫酶技术在修饰的安卡拉痘苗病毒感染性滴度检测中的应用

 修饰的安卡拉痘苗（modified vaccinia Ankara，MVA）是MVA病毒在原代鸡胚成纤维细胞（CEF）中经过一系列的传代得到^[1]，在人体内可以表达 MVA 病毒蛋白及其携带的外源基因蛋白，但却不形成完整的病毒颗粒，为人体内复制缺陷性病毒^[2]，安全性好，即使在免疫功能缺陷病人和免疫抑制的恒河猴体内也未显示出明显的副作用^[3]。近年来，重组 MVA 病毒广泛地用于预防性疫苗和感染性疾病、癌症治疗的基础与临床研究^[4]，其中以 MVA 病毒作为载体的疫苗是最有希望用于预防人类艾滋病的活载体疫苗之一^[5]。由于 MVA 病毒本身的繁殖特点，其滴度的检测不适合采用常规的蚀斑法进行^[6]；而常规微量细胞病变（cytopathic effect，CPE）法相对耗时，对细胞培养要求高，往往受到主观因素的影响，因而不稳定且精确度降低^[7]，给检测工作带来不便。上世纪 90 年代初 Usuba 等^[8]建立了免疫酶技术测定流感病毒感染性滴度的方法，较常规方法省时、结果准确﹑重复性好。我们就免疫酶技术在 MVA 病毒感染性滴度检测中的适用性进行了探讨，旨在建立一种准确、快速的 MVA 病毒感染性滴度检测方法。......     

IGFBP7对人乳腺癌MCF-7细胞增殖的影响及其机制

目的: 探究胰岛素样生长因子结合蛋白7(IGFBP7)对人乳腺癌MCF-7细胞增殖的影响及其分子生物学机制。方法: 将质粒pCMV6-IGFBP7转染MCF-7细胞,构建稳定表达IGFBP7的MCF-7细胞系;采用Western blotting检测IGFBP7在MCF-7细胞稳定转染子的表达;采用软琼脂培养克隆形成实验检测IGFBP7对MCF-7细胞克隆形成能力的影响;采用流式细胞术检测IGFBP7对MCF-7细胞周期的影响;采用Western blotting检测IGFBP7对MCF-7细胞细胞外信号调节激酶1/2(ERK1/2)、p-ERK1/2、细胞周期素D1(cyclin D1)、细胞周期素依赖性激酶4(CDK4)、cyclin E、CDK2、p21^CIP1/WAF1、p27^KIP1、p53、视网膜母细胞瘤蛋白(Rb)和p-Rb蛋白含量的影响。结果: (1)只有稳定转染质粒pCMV6-IGFBP7的MCF-7细胞表达IGFBP7。(2)IGFBP7能够显著降低MCF-7细胞的克隆形成率(P<0.01),阻止细胞从G₁期进入S 期,使其停滞于G₁期(P<0.01)。(3)IGFBP7能够显著抑制ERK1/2的磷酸化(P<0.01)。(4)IGFBP7能够下调cyclin D1和cyclin E蛋白表达(P<0.01),上调p27^KIP1、p21^CIP1/WAF1和p53蛋白表达(P<0.01),抑制Rb的磷酸化(P<0.01)。(5)MEK1/2阻断剂PD98059可部分模拟IGFBP7的肿瘤抑制效应。结论: (1) IGFBP7可通过下调cyclin D1和cyclin E蛋白表达,上调p27^KIP1、p21^CIP1/WAF1和p53蛋白表达,以及抑制Rb磷酸化发挥抗肿瘤作用;(2) IGFBP7对cyclin D1和p27^KIP1的调节可能与其抑制ERK1/2信号通路有关。     

促凋亡基因 puma 的研究进展

 p53 上调节的细胞凋亡调控因子（p53 up-regulated modulator of apoptosis，PUMA）是 2001 年由 Yu 等^[1]和 Nakano 等^[2] 2 个独立研究小组同时发现的，是 Bcl-2 蛋白家族的促凋亡成员之一，具有强大的促凋亡作用，可被内、外源性 p53 快速诱导活化。PUMA 基因定位于 19q，cDNA 全长 1.9 kb，转录本由 4 个外显子（1a、2、3、4）组成，编码 1 个由 193 个氨基酸组成的蛋白，该蛋白定位于线粒体膜上。Bcl-2 蛋白家族中促凋亡蛋白的共同点是都含有 1 个由 9 个氨基酸（LRRMADDLN）组成的 BH3 保守结构域。其中 PUMA 与 Bik、Bad、Bid、Bim、Hrk/DP5 及线虫 Eg-1 等促凋亡蛋白，都只存在 1 个 BH3（Bcl-2 homology3）结构域，统称为 BH3 仅有蛋白。从秀丽隐杆线虫到小鼠的程序性细胞死亡的遗传学研究显示，BH3 仅有蛋白是程序性细胞死亡的重要启动因子，BH3 仅有蛋白通过其 BH3 结构域与 Bcl-2 蛋白家族中的抑凋亡蛋白表面的沟槽相互作用从而启动凋亡，提示 BH3 结构域对 BH3 仅有蛋白与 Bcl-2 等抑凋亡蛋白结合并启动凋亡起着至关重要的作用^[3]。PUMA 的 BH3 结构域位于其氨基酸序列的第 141 ~ 149 位，缺少此结构域的 puma 突变体也将丧失诱导凋亡的功能^[4]。近年来，PUMA 的诱导凋亡作用与肿瘤的相关性研究成为热点，本文仅就 PUMA 的促凋亡作用在肿瘤研究中的应用进行综述。......     

Intelectin contributes to allergen-induced IL-25, IL-33, and TSLP expression and type 2 response in asthma and atopic dermatitis

The epithelial and epidermal innate cytokines IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) have pivotal roles in the initiation of allergic inflammation in asthma and atopic dermatitis (AD). However, the mechanism by which the expression of these innate cytokines is regulated remains unclear. Intelectin (ITLN) is expressed in airway epithelial cells and promotes allergic airway inflammation. We hypothesized that ITLN is required for allergen-induced IL-25, IL-33, and TSLP expression. In two asthma models, Itln knockdown reduced allergen-induced increases in Il-25, Il-33, and Tslp and development of type 2 response, eosinophilic inflammation, mucus overproduction, and airway hyperresponsiveness. Itln knockdown also inhibited house dust mite (HDM)-induced early upregulation of Il-25, Il-33, and Tslp in a model solely inducing airway sensitization. Using human airway epithelial cells, we demonstrated that HDM-induced increases in ITLN led to phosphorylation of epidermal growth factor receptor and extracellular-signal regulated kinase, which were required for induction of IL-25, IL-33, and TSLP expression. In two AD models, Itln knockdown suppressed expression of Il-33, Tslp, and Th2 cytokines and eosinophilic inflammation. In humans, ITLN1 expression was significantly increased in asthmatic airways and in lesional skin of AD. We conclude that ITLN contributes to allergen-induced Il-25, Il-33, and Tslp expression in asthma and AD.     

Intrathymic lymph nodes in humans

An unusual lymph node exists in the centre of the human thymus. This lymph node, which we call an intrathymic lymph node (ITLN), possesses some interesting morphological characteristics. In ontogeny, this node seems to appear at the latter half of fetal period. The function of the ITLN is still unknown, but it is assumed that it may play a different role in the immune system than other peripheral lymph nodes by its characteristics.     

A comparative review of intelectins

Intelectin (ITLN) is a new type of glycan-binding lectin. It has been demonstrated to agglutinate bacteria probably due to its carbohydrate-binding capacity, suggesting its role in an innate immune response. It is involved not only in many physiological processes but also in some human diseases such as asthma, heart disease, inflammatory bowel disease, chronic obstructive pulmonary disease and cancer. Up to now, intelectin orthologs have been identified in placozoans, urochordatas, cephalochordates and several vertebrates, such as cyclostomata, fish, amphibians and mammals. Although the sequences of intelectins in different species are conserved, their expression patterns, quaternary structures and functions differ considerably among and within species. We summarize the evolution of the intelectin gene family, the tissue distribution, structure and functions of intelectins. We conclude that intelectin plays a role in innate immune response and there are still potential functions of intelectin awaiting discovery.     

Arborization of the inferior laryngeal nerve and internal nerve on the posterior surface of the larynx

The morphological patterns of the inferior laryngeal nerve and internal laryngeal nerve display complex arborizations. This paper attempts to identify and clarify these patterns. Dissections were performed on 105 adult Japanese cadavers, and observations were made on 201 sides. Results showed that the communications between the inferior laryngeal nerve (ILN) and internal laryngeal nerve (ITLN) could be classified into two types and three subtypes. Also, the ITLN displayed three characteristic patterns at the arytenoid cartilage. These communications produce complex arborizations of the ILN as it enters the larynx. This may explain the variety of potential clinical symptoms observed after thyroid surgery or neck dissections. © 1995 WiIey-Liss, Inc.     

IgA antibodies in the bile of rats. III. The role of intrathoracic lymph nodes and the migration pattern of their blast cells.

Eight weeks after rats had had their mesenteric lymph nodes (MLN) removed surgically, they were found to be still able to generate substantial titres of biliary IgA-antibodies after antigens were injected into their Peyer's patches. This suggested that systemically significant IgA production could be induced in extra-abdominal lymphoid tissue. It was found that the intrathoracic lymph nodes (ITLN) were an important source of IgA production. These nodes could be stimulated to produce biliary antibody by introducing antigen either into the peritoneal cavity or directly into the thorax. Cells forming IgA were identified in the ITLNs by haemolytic plaque assays and immunoperoxidase techniques. In spite of this, immunoblasts obtained from the ITLNs, and labelled with 125IUdR did not localize in the gut after i.v. injection to anywhere near the extent that immunoblasts from the MLN did. Instead they seemed to have a predilection for localizing in the lungs.     

AP-4、EZH2基因表达量与子宫内膜癌病灶中细胞凋亡、 上皮间质转化的相关性研究

目的 研究AP-4、EZH2基因表达量与子宫内膜癌病灶中细胞凋亡、上皮间质转化的相关性。方法 回顾分析2017年1月~2018年1月在我院进行切除术治疗的35例子宫内膜癌患者临床资料,收集子宫内膜癌病灶和癌旁病灶适量,提取RNA后分别测定AP-4、EZH2基因表达量,进一步测定病灶中细胞凋亡、上皮间质转化基因的表达量。结果 子宫内膜癌病灶内AP-4、EZH2基因表达量高于癌旁病灶,差异有统计学意义（P＜0.05）;子宫内膜癌病灶内SRPX2、RLIP76、ZEB1、SALL4、TET1、RANKL、N-cadherin的mRNA表达量高于癌旁病灶,Fat-1、ITLN-1、Caspase-3、α-catenin的mRNA表达量低于癌旁病灶;AP-4高表达病灶内SRPX2、RLIP76的mRNA表达量高于低表达病灶,Fat-1、ITLN-1、Caspase-3的mRNA表达量低于AP-4低表达病灶;EZH2高表达病灶内ZEB1、SALL4、TET1、RANKL、N-cadherin的mRNA表达量高于EZH2低表达病灶,α-catenin的mRNA表达量低于EZH2低表达病灶。结论 宫内膜癌病灶内AP-4、EZH2基因高表达量,可以抑制癌细胞凋亡、促进癌细胞上皮间质转化,值得临床予以重视和关注。     

Common variants of SLAMF1 and ITLN1 on 1q21 are associated with type 2 diabetes in Indian population

Though multiple studies link chromosomal regions 1q21-q23 and 20q13 with type 2 diabetes, fine mapping of these regions is yet to confirm gene(s) explaining the linkages. These candidate regions remain unexplored in Indians, which is a high-risk population for type 2 diabetes. Hypothesizing regulatory regions to have a more important role in complex disorders, we examined association of 207 common variants in proximal promoter and untranslated regions of genes on 1q21-23 and 20q13 with type 2 diabetes in 2115 North Indians. Further, top signals were replicated in an independent group of 2085 North Indians. Variants-rs11265455-SLAMF1 (odds ratios (OR)=1.32, P=1.1 × 10(-3)), rs1062827-F11R (OR=1.36, P=1.7 × 10(-3)) and rs12565932-F11R (OR=1.35, P=1.8 × 10(-3)) were top signals for association with type 2 diabetes whereas rs1333062-ITLN1 (OR=1.28, P=3.4 × 10(-3)) showed strongest association in body mass index-stratified analysis. Replication of these four variants confirmed associations of rs11265455-SLAMF1 (OR=1.27, P=9.1 × 10(-3)) and rs1333062-ITLN1 (OR=1.25, P=1.1 × 10(-3)) with type 2 diabetes. Meta-analysis further corroborated the association of rs11265455-SLAMF1 (OR random effect=1.29, P random effect=3.9 × 10(-5)) and rs1333062-ITLN1 (OR random effect=1.19, P random effect=1.8 × 10(-4)). In conclusion, the study demonstrates that variants of SLAMF1 and ITLN1, both implicated in inflammation, are associated with type 2 diabetes in Indians.     

Novel genes in Human Asthma Based on a Mouse Model of Allergic Airway Inflammation and Human Investigations

Purpose

Based on a previous gene expression study in a mouse model of asthma, we selected 60 candidate genes and investigated their possible roles in human asthma.

Methods

In these candidate genes, 90 SNPs were genotyped using MassARRAY technology from 311 asthmatic children and 360 healthy controls of the Hungarian (Caucasian) population. Moreover, gene expression levels were measured by RT PCR in the induced sputum of 13 asthmatics and 10 control individuals. t-tests, chi-square tests, and logistic regression were carried out in order to assess associations of SNP frequency and expression level with asthma. Permutation tests were performed to account for multiple hypothesis testing.

Results

The frequency of 4 SNPs in 2 genes differed significantly between asthmatic and control subjects: SNPs rs2240572, rs2240571, rs3735222 in gene SCIN, and rs32588 in gene PPARGC1B. Carriers of the minor alleles had reduced risk of asthma with an odds ratio of 0.64 (0.51-0.80; P=7×10^-5) in SCIN and 0.56 (0.42-0.76; P=1.2×10^-4) in PPARGC1B. The expression levels of SCIN, PPARGC1B and ITLN1 genes were significantly lower in the sputum of asthmatics.

Conclusions

Three potentially novel asthma-associated genes were identified based on mouse experiments and human studies.     

A method for the quantitative analysis and standardization of interleukin-1 bioactivity

A method is presented for the reproducible quantitation of the biological activity of interleukin 1 (IL-1). This method provides diagnostic tools which give insights into the qualitative aspects of the binding of IL-1 and of the resulting activation of the responder thymocytes; for example, whether the lymphokine and/or the responder population is heterogeneous, or whether a threshold level exists. It establishes under what circumstances the assumptions on which it is based are reasonably adhered to and, consequently, quantitative estimation in the manner it prescribes is justified. It also gives a simple way to calculate both the maximal response attainable for each preparation in an assay and the dilution of a particular preparation that would produce a half-maximal response, the accepted unit of activity of IL-1. This empirical technique provides an improved means of comparing the activities of various preparations of IL-1 in bioassays using various stocks of responder cells and reagents. It should also be applicable to the evaluation of the biological activity of lymphokines in general.     

TBX1基因与染色体22q11.2微缺失综合征

染色体22q11·2微缺失综合征(22q11·2DS)是人类最常见的染色体微缺失综合征。TBX1基因作为一个T-BOX家族转录因子,其单拷贝缺失可能是22q11·2DS的主要原因之一,它对该综合征临床表征的出现可能有重要作用。TBX1在胚胎生长发育是胚胎咽部分节、咽弓和动脉弓形成、心脏流出道生长与排列及分隔等过程所必需的基因。TBX1受一系列调控基因调节,其本身也调控一系列基因。