巨细胞病毒感染对小鼠心肌组织TH1/TH2类细胞因子及其特异性转录因子T-bet/GATA-3表达的影响

目的动态观测MCMV感染对小鼠心肌组织TH1/TH2类细胞因子及其特异性转录因子T-bet／GATA-3表达的影响。方法建立小鼠巨细胞病毒感染心肌炎模型。观察小鼠血清删浓度变化，心肌组织病理损害和心肌组织细胞因子IFN-γ、IL-4表达水平以及脾组织中转录因子T-bet、GATA-3表达强度。结果心肌组织病理积分和血清cTnI浓度在MCMV感染后第3天增高，第7天达高峰，第14天仍维持较高水平；IFN-γ在MCMV感染第3天时表达迅速增高达到峰值，随后下降，第14天基本下落至正常水平；IL-4在MCMV感染第5天表达迅速增高达到峰值，随后下降，在第14天又升高；MCMV感染第3天转录因子T-bet蛋白表达达峰值，随后逐渐降低，到第14天时降低更为显著。转录因子GATA-3在MCMV感染第7天时表达明显增强，第14天时增强更为显著。结论MCMV感染通过上调GATA-3转录因子和叫表达，下调T-bet转录因子和IFN-γ表达，导致TH1/TH2平衡失调，表现为TH2细胞优势应答状态，使心肌损害加重。这可能是CMV感染致宿主特异性细胞免疫功能降低并造成组织损伤的机制之一。     

地塞米松对哮喘大鼠T-bet/GATA-3的影响

目的：探讨地塞米松对转录因子T-bet/GATA-3的影响。方法：将30只SD大鼠随机分为正常组、模型组与地塞米松组,每组10只。于第1、8天腹腔注射卵白蛋白,第15天雾化吸入卵白蛋白引喘,每日一次,共14次。地塞米松组于实验开始后第15天起,按0.5 mg/kg予以腹腔注射地塞米松,每日一次,共14次。流式细胞术测脾及外周血Th1、Th2含量,免疫组化法及免疫印迹法测T-bet、GATA-3蛋白表达,逆转录聚合酶链反应测T-bet、GATA-3mRNA的表达。结果：模型组与正常组比较,脾与外周血中Th1、Th2含量有非常显著性增多（P＜0.01）;肺组织中T-bet、GATA-3蛋白与mRNA的表达亦有非常显著性升高（P＜0.01）。而地塞米松组与模型组比较,T-bet/GATA-3有显著（P＜0.05）或非常显著（P＜0.01）性升高。结论：T-bet/GATA-3是影响Th1/Th2平衡的关键转录因子,地塞米松可提高T-bet/GATA-3比值,从而减轻哮喘的发作。     

白藜芦醇调节Th细胞失衡与T-bet、GATA-3途径作用机制的探讨

目的 探讨白藜芦醇对Jurkat T细胞T-bet/GATA-3表达的影响及其与调节Th1/Th2细胞分化作用机制之间的关系.方法 不同浓度的白藜芦醇刺激Jurkat T细胞48 h后,用ELISA法检测Th1/Th2细胞因子表达谱及用RT-PCR检测T-bet和GATA-3 mRNA表达的变化.结果 白藜芦醇抑制T-bet mRNA和GATA-3 mRNA的表达（F=6308,P〈0.05 F=3262,P〈0.05）.对Jurkat T细胞分泌细胞因子IFN-γ、IL-10的表达均起抑制作用,并具有浓度效应（F=2604,P〈0.05 F=2465,P〈0.05）.结论 白藜芦醇抑制Jurct T细胞由Th0向Th1细胞分化可能是通过降低转录因子T-bet表达水平进行调节,对Th2细胞的抑制作用则可能通过抑制转录因子GATA-3途径进行负调节,并具有浓度效应.     

转录因子T-bet/GATA-3比率评价哮喘患者Th1/Th2失衡及CpG ODN干预机制的研究

目的: 探讨哮喘患者PBMCs中转录因子T-bet/GATA-3比率与Th1/Th2细胞失衡的关系及体外CpG干预后T-bet/GATA3比率的变化及其对Th1/Th2细胞平衡的调节作用。方法: 用RT-PCR法测定30例发作期哮喘患者（哮喘组）及20例慢性阻塞性肺病患者（COPD组）及20例正常人（对照组） PBMCs中T-bet mRNA、GATA-3 mRNA及TLR9 mRNA的表达强度，用ELISA法测定血浆IL-4、IL-5、IL-13和IFNγ的表达水平，以观察哮喘患者PBMCs中转录因子T-bet/GATA-3比率与Th1/Th2细胞失衡的关系。将20例发作期哮喘患者的PBMCs分为2部分，一份加入CpG ODN和PHA（CpG组），一份仅加入PHA（对照组），培养48 h后分别收集培养液和细胞，采用RT-PCR法测定细胞中T-bet mRNA、GATA-3 mRNA及TLR9 mRNA的表达强度，ELISA法测定培养液中IL-4、IL-5、IL-13和IFN-γ的表达水平。结果： 哮喘患者PBMCs中T-bet/GATA-3的比率显著低于COPD患者和正常人，IL-4、IL-5、IL-13的水平显著高于COPD患者和正常人，与T-bet/GATA-3的比率呈负相关，而IFN-γ的水平显著降低，与T-bet/GATA-3的比率呈正相关。哮喘组TLR9的表达强度显著弱于COPD患者和正常人。CpG干预组细胞培养液中IL-4、IL-5和IL-13的水平分别显著低于对照组，IFN-γ的水平显著高于对照组。CpG干预组细胞中T-bet mRNA和TLR9 mRNA的表达强度显著强于对照组，GATA-3 mRNA的表达强度显著低于对照组；T-bet/GATA-3的比率显著高于对照组。结论: 哮喘患者T-bet/GATA-3的比率降低，可以作为评价Th1/Th2细胞失衡的精确指标。CpG ODN可以上调T-bet的表达，下调GATA-3的表达，从而上调T-bet/GATA-3的比率，逆转Th1/Th2细胞失衡，是一种很有前景的哮喘治疗手段。     

母-胎免疫调节机制在妊娠高血压综合征病理过程中的作用

目的：分析妊娠高血压综合征（PIH）患者外周血中免疫细胞因子水平变化及滋养层细胞中免疫相关转录因子的基因、蛋白表达情况,探讨母 胎免疫调节机制在妊娠高血压综合征病理过程中的作用。方法：纳入PIH患者50例作为观察组,选择同期分娩的正常妊娠妇女40例作为对照组。ELISA法检测两组孕妇外周血中IFN-γ、IL-4、IL-17及TGF β水平;实时荧光定量PCR（RT-qPCR）法和蛋白质印迹法（Western blot）分别测定两组孕妇胎盘滋养层细胞中转录因子T-bet、GATA-3、RORC、Foxp3的mRNA及蛋白表达情况。结果：①观察组外周血中IFN-γ和IL 17水平明显高于对照组,TGF β水平明显低于对照组（P＜0.05）;两组间IL 4水平无明显差异（P＞0.05）;观察组IFN-γ/IL-4、IL-17/TGF β比值均明显大于对照组（P＜0.05）。②与对照组比,观察组T bet、RORC的mRNA及蛋白表达水平均显著升高（P＜0.05）,而GATA 3、Foxp3的mRNA及蛋白表达水平均显著降低（P＜0.05）,T-bet/GATA-3及RORC/Foxp3比值均显著增大（P＜0.05）。结论：PIH具有明显的Th1/Th2及Th17/Treg比例失衡现象,其病理机制可能与Th细胞介导的母 胎免疫耐受机制功能紊乱有关。     

以T-bet/GATA-3的比率作为评价哮喘患者Th1/Th2失衡指标的探讨

目的：探讨哮喘患者PBMCs中转录因子T-beL／GATA-3的比率与Th1／Th2细胞失衡的关系。方法：采用RT-PCR法测定30例发作期哮喘患者(哮喘组)及20例慢性阻塞性肺病患者(COPD组)及20例正常人(对照组)PBMCs中T-bet和GATA-3的活性，ELISA法测定血浆IL-4、IL-5、IL-13和IFN-γ的表达水平。结果：哮喘组PBMCs中T-bet／GATA-3的比率较COPD组和对照组明显降低，Th2细胞因子IL-4、IL-5、IL-13的水平增高，与T-bet／GATA-3的比率成负相关，而Th1细胞因子IFN-γ的水平降低，与T-bet／GATA-3的比率成正相关。结论：哮喘患者T-bet／GATA-3的比率降低，可以作为评价Th1／Th2细胞失衡的精确指标。     

BALB/c小鼠感染弓形虫后Th1/Th2免疫漂移的实验研究

目的：动态分析BALB/c小鼠感染弓形虫后Th1/Th2免疫失衡及免疫漂移特点,并探讨转录因子T-bet和GA-TA-3在此过程中的改变及其意义。方法：90只BALB/c小鼠随机分为正常对照组30只,弓形虫感染组60只。于感染后奇数天每天处死感染组小鼠2只,对照组小鼠1只,采用ELISA法动态检测各组小鼠血清中IFN-γ和IL-4的水平,同时应用荧光定量PCR方法检测小鼠脾细胞中T-bet和GATA-3 mRNA的表达情况。结果：感染组小鼠中,血清IFN-γ于感染后第4天开始显著升高,第5~7天维持在高峰值,从第8天开始下降,第9天降至正常水平;IL-4于感染后第8天开始显著升高,第9天升至峰值,从第14天开始下降,第15天降至正常水平;脾细胞T-bet mRNA的表达在感染后第3天升高,第5天达高峰后于第9天降至正常水平;脾细胞GATA-3 mRNA的表达在感染后第7天升高,第11天达高峰,于第13天降至正常水平。正常对照组小鼠在实验期内IFN-γI、L-4水平没有明显变化,维持在正常的较低水平。结论：BALB/c小鼠感染弓形虫后诱导的免疫应答在感染急性期（第1~8天）以Th1应答为主,第9至13天,宿主免疫应答以Th2细胞应答为主,之后Th1/Th2应答基本恢复平衡。Th1应答向Th2应答的漂移与T-bet和GATA-3 mRNA的表达相关并受其调控,Th1/Th2型免疫应答的发生时相和效应强度可能影响弓形虫感染的最终结局。     

慢性阻塞性肺疾病的炎症反应与Foxp3、T-bet、GATA3表达失衡有关

目的观察慢性阻塞性肺疾病(COPD)大鼠调节性T细胞(Treg)、叉头状转录因子3(Foxp3)、T-bet、GATA连接蛋白3(GATA3)的变化。方法将30只大鼠随机分为对照组、模型组,每组15只。模型组大鼠采用烟熏加脂多糖(LPS)气管滴入方法建立COPD模型。模型复制成功第28天,采用ELISA检测大鼠血清、支气管肺泡灌洗液(BALF)白介素4(IL-4)、γ干扰素(IFN-γ)水平,流式细胞术检测外周血Treg表达,分别采用逆转录PCR及Western blot法检测肺组织Foxp3、T-bet、GATA3mRNA和蛋白表达。结果模型组大鼠肺泡腔及肺间质内大量炎性细胞浸润,肺组织结构破坏。与对照组比较,模型组大鼠血清IFN-γ表达升高(P0.01),IL-4、CD4+CD25+Treg、CD4+CD25+Foxp3+Treg降低(P0.05);模型组Foxp3、GATA-3mRNA、蛋白表达降低,T-bet mRNA和蛋白升高(P0.01)。相关性分析显示,T-bet、GATA-3、Foxp3基因和蛋白表达与IL-4、IFN-γ分泌相关(P0.05)。结论 COPD炎症反应与T-bet、GATA-3、Foxp3表达失衡有一定关系。     

哮喘患儿DC源性IL-12对T细胞GATA-3和T-bet的影响

目的 了解哮喘中树突状细胞(Dc)分泌的细胞因子IL-12与转录因子T-bet和GATA-3的相互关系.方法 用rhGM-CSF和rhIL-4诱导培养外周血来源的DC并予鉴定,ELISA法检测其分泌的细胞因子IL-12以及与自身T细胞反应后,RT-PCR检测T-bet和GATA-3 mRNA含量,流式细胞术检测T细胞分泌的胞内细胞因子IL-4和IFN-γ水平.结果 DC诱导培养第8天,哮喘组Dc分泌的IL-12水平低于正常对照组,差异有显著性意义(P<0.01);混合培养第7天,哮喘组IFN-γ和T-bet mR-NA水平均低于正常对照组(P<0.05),而IL-4、GATA-3水平和比值IL-4/IFN-γ、GATA-3/T-bet均高于正常对照组,有显著性差异(P<0.01);IL-12与T-bet呈显著正相关,与GATA-3和GATA-3/T-bet呈显著负相关.结论 转录因子T-bet、GATA-3调控ThO细胞分化受DC分泌的细胞因子IL-12的影响.IL-12如何影响T-bet、GATA-3的表达有待进一步研究.     

BALB/c小鼠感染弓形虫后Thl/Th2免疫漂移的实验研究

目的:动态分析BALB/c小鼠感染弓形虫后Thl/Th2免疫失衡及免疫漂移特点,并探讨转录因子T-bet和GA.TA-3在此过程中的改变及其意义.方法:90只BALB/c小鼠随机分为正常对照组30只,弓形虫感染组60只.于感染后奇数天每天处死感染组小鼠2只,对照组小鼠1只,采用ELISA法动态检测各组小鼠血清中IFN-γ和IL-4的水平,同时应用荧光定量PCR方法检测小鼠脾细胞中T-bet和GATA-3 mRNA的表达情况.结果:感染组小鼠中,血清IFN-γ于感染后第4天开始显著升高,第5~7天维持在高峰值,从第8天开始下降,第9天降至正常水平;IL-4于感染后第8天开始显著升高,第9天升至峰值,从第14天开始下降,第15天降至正常水平;脾细胞T-bet mRNA的表达在感染后第3天升高,第5天达高峰后于第9天降至正常水平;脾细胞GATA-3 mRNA的表达在感染后第7天升高,第11天达高峰,于第13天降至正常水平.正常对照组小鼠在实验期内IFN-γ、IL-4水平没有明显变化,维持在正常的较低水平.结论:BALB/c小鼠感染弓形虫后诱导的免疫应答在感染急性期(第1-8天)以Th1应答为主,第9至13天,宿主免疫应答以Th2细胞应答为主,之后Thl/Th2应答基本恢复平衡.Thl应答向Th2应答的漂移与T-bet和GATA-3 mRNA的表达相关并受其调控,Thl/Th2型免疫应答的发生时相和效应强度可能影响弓形虫感染的最终结局.     

Polymerization of cyclopentene, 3-methylcyclopentene,and 3-methylcyclohexene

The cycloolefins cyclopentene, 3-methylcyclopentene, and 3-methylcyclohexene have been polymerized. The polymerization rates were very low. Depending on the particular monomer, ZIEGLER and/or cationic catalysts were effective. Cyclopentene polymerizes both by double bond opening (producing enchained cyclopentane rings) and by ring opening to give cis and trans polypentenamers. The resulting polymers are partially crystalline and have birefringence melting points between 120 and 150°C. The 3-methylcyclopentene and 3-methylcyclohexene monomers polymerize (at least partly) by 1,3-addition following a 3,2-hydride shift. The 3-methylcyclohexene polymers are also partly crystalline and have birefringence melting points between 200 and 250°C. Little ring-opening polymerization of the methylcycloolefins occurs. Polymerization mechanisms are discussed.     

Topography of 3H-DHA, 3H-QNB, 3H-Dopamine, and 3H-DAGO Binding Sites Distribution in the Central Part of the Sinoatrial Node in Rat Heart

The topography of distribution of ³H-dihydroalprenolol, ³H-quinucledinyl benzilate, ³H-dopamine, and ³H-DAGO binding sites in the central part of the sinoatrial node in rat heart was studied by autoradiography after electrophysiological identification of the dominant pacemaker region location. Receptor asymmetry between the lateral and median regions of the central part of the sinoatrial node was shown. The dominant pacemaker region lay in the lateral area of the sinoatrial node; the number of binding sites for all four ligands was minimum in it. The number of binding sites gradually increased in the cranial and caudal directions from the dominant pacemaker region along the sinoatrial node artery (more smoothly in the caudal direction). The relative densities of bindings sites for ³H-dihydroalprenolol and ³H-dopamine were higher in the lateral region compared to the perinodal working myocardium, while the densities for ³H-quinucledinyl benzilate and ³H-DAGO were virtually the same. The distribution of binding sites along the artery in the median region of the sinoatrial node was even for ³H-quinucledinyl benzilate and ³H-DAGO. For ³H-DAGO these parameters were close to those in the perinodal atrial myocardium, for ³H-quinucledinyl benzilate somewhat lower. Curves presenting the distribution of binding site densities for ³H-dihydroalprenolol and ³H-dopamine in the median region of the sinoatrial node were similar, with a pronounced peak in the region contralateral to the dominant pacemaker region, and significantly higher binding parameters compared to those for the perinodal atrial myocardium. The difference consisted in higher density of ³H-dopamine binding sites in the median region of the sinoatrial node in comparison with the lateral region. Binding activity was maximum in the wall of the sinoatrial node artery. The distribution of binding sites for ligands to the main autonomic nervous system neurotransmitters in the rat heart sinoatrial node is heterogeneous. __________ Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 140, No. 10, pp. 472–477, October, 2005     

In vitro biocompatibility and degradation of terpolyester 3HB-co-4HB-co-3HHx, consisting of 3-hydroxybutyrate, 4-hydroxybutyrate and 3-hydroxyhexanoate

A terpolyester consisting of 3-hydroxybutyrate (3HB), 4-hydroxybutyrate (4HB) and 3-hydroxyhexanoate (3HHx), abbreviated as P(3HB-co-4HB-co-3HHx), was studied for possible application as an implant biomaterial. L929 mouse fibroblasts, MC3T3-E1 murine osteoblasts and a human cell line of immortalized human keratinocyte (HaCat cells) were used to study the biocompatibility of P(3HB-co-4HB-co-3HHx). Cell morphology and cell activity were studied using scanning electron microscopy (SEM) and the MTT assay, respectively. All three cell types showed higher activities when grown on films of P(3HB-co-4HB-co-3HHx) compared with their growth on poly(lactic acid) (PLA), co-polyester PHBHHx films and on polylysine-coated plates (blank), respectively. The three cell types grown on the terpolyester also demonstrated a well-spread cell shape and large number of pseudopods due to strong cell-cell and cell-material interactions. It was clearly observed that P(3HB-co-4HB-co-3HHx) had a much faster degradation rate than PHBHHx after 15 weeks of incubation in phosphate-buffered saline under dynamic conditions. The results proved that the terpolyester had favorable biocompatibility and biodegradability compared with the well-studied polyesters PLA and PHBHHx.     

In vitro biocompatibility and degradation of terpolyester 3HB-co-4HB-co-3HHx,consisting of 3-hydroxybutyrate, 4-hydroxybutyrate and 3-hydroxyhexanoate

A terpolyester consisting of 3-hydroxybutyrate (3HB), 4-hydroxybutyrate (4HB) and 3-hydroxyhexanoate (3HHx), abbreviated as P(3HB-co-4HB-co-3HHx), was studied for possible application as an implant biomaterial. L929 mouse fibroblasts, MC3T3-E1 murine osteoblasts and a human cell line of immortalized human keratinocyte (HaCat cells) were used to study the biocompatibility of P(3HB-co-4HB-co-3HHx). Cell morphology and cell activity were studied using scanning electron microscopy (SEM) and the MTT assay, respectively. All three cell types showed higher activities when grown on films of P(3HB-co-4HB-co-3HHx) compared with their growth on poly(lactic acid) (PLA), co-polyester PHBHHx films and on polylysine-coated plates (blank), respectively. The three cell types grown on the terpolyester also demonstrated a well-spread cell shape and large number of pseudopods due to strong cell–cell and cell–material interactions. It was clearly observed that P(3HB-co-4HB-co-3HHx) had a much faster degradation rate than PHBHHx after 15 weeks of incubation in phosphate-buffered saline under dynamic conditions. The results proved that the terpolyester had favorable biocompatibility and biodegradability compared with the well-studied polyesters PLA and PHBHHx.     

急性阑尾炎围手术期血清FT3、FT4、T3、T4、rT3测定的临床意义

目的:探讨急性阑尾炎围手术期甲状腺激素水平的变化规律及其临床意义.方法:选择82例急性阑尾炎患者,分别于手术前、手术后1、3、5、7、14d取空腹静脉血,利用放射免疫分析检测FT3、FT4、T3、T4、rT3及TSH水平,同时分析甲状腺激素水平与临床病理分型、并发症的关系.结果:本文患者手术前均存在不同程度的低FT3、FT4、T3、T4血症和高rT3血症,且以坏疽性和穿孔性阑尾炎最明显.这种现象均于术后(5～7)d恢复正常,但当出现感染性并发症时血清FT3、FT4、T3、T4持续低下,而临床治愈后则恢复正常水平.结论:急性阑尾炎患者甲状腺内分泌功能均受到不同程度的抑制,围手术期动态观察FT3、FT4、T3、T4、rT3血液激素水平,有利于判断病情轻重、临床病理分型和感染性并发症的存在,有利于临床治疗方案的制定.