RNA干扰抑制IL-23表达对感染后内脏高敏感小鼠肠黏膜固有层DC活化Th17细胞功能的影响

背景：前期研究发现感染后内脏高敏感小鼠肠黏膜固有层树突细胞（DC）诱导活化Th17细胞与肠道感染消退后肠黏膜免疫系统的持续激活有关。推测DC可能系通过分泌白细胞介素-23（IL-23）活化Th17细胞。目的：应用RNA干扰技术抑制DC分泌IL-23,探讨感染后内脏高敏感小鼠肠黏膜固有层DC活化Th17细胞的机制。方法：建立旋毛虫感染后内脏高敏感小鼠模型,以免疫磁珠分选肠黏膜固有层DC和脾脏CD4＋T细胞。构建、鉴定小鼠IL-23小发夹RNA（shRNA）干扰质粒,以脂质体法转染DC（A组）以抑制IL-23表达,同时设置转染空脂质体的DC（B组）和转染无关序列shRNA干扰质粒的DC（C组）作为对照。各组DC与CD4＋T细胞共培养120 h,以单独培养的CD4＋T细胞（D组）作为对照。以ELISA方法检测DC转染前后培养上清液中的IL-23水平,以及DC与CD4＋T细胞共培养上清液和CD4＋T细胞单独培养上清液中的IL-17水平。结果：A组DC培养上清液中的IL-23水平较转染前显著降低（P〈0.05）,B、C两组转染前后IL-23水平无明显变化。A、B、C组DC与CD4＋T细胞共培养上清液中的IL-17水平均较D组显著增高（P〈0.05）,其中A组显著低于B、C两组（P〈0.05）,B、C组间差异无统计学意义。结论：感染后内脏高敏感小鼠肠黏膜固有层DC可能通过分泌IL-23活化Th17细胞,参与维持肠道感染消退后肠黏膜免疫系统的持续激活。     

rhG-CSF未经动员与动员后供者外周血淋巴细胞与干细胞采集物的细胞构成及功能比较

目的比较重组人粒细胞集落刺激因子（rhG-CSF）动员后供者外周血干细胞（PBSC）采集物与未经动员供者外周血淋巴细胞采集物的细胞构成及功能。方法取异基因造血干细胞移植供者的rhG-CSF动员后PBSC采集物（A组）和未经动员的淋巴细胞采集物（B组）,以流式细胞术测定采集物组分、T细胞亚群、树突细胞（DC）及其亚群、CD14^＋细胞和CD19^＋细胞B7分子的表达、CD4^＋T细胞IL-4和IFNγ等细胞因子的分泌情况,四甲基偶氮唑盐法测定T淋巴细胞增殖能力。结果两组采集物的CD3^＋、CD4^＋、CD34^＋、CD14^＋细胞比例有明显差异,A组DC细胞及其亚群的比例明显高于B组,尤以DC2升高为著（P=0．000）,CD14^＋细胞上B7分子的表达A组明显低于B组,CD19^＋细胞上B7分子的表达无明显差异,分析两组CD4细胞内因子的分泌情况,A组的Ⅱ类细胞因子IL-4及IL-4／IFNγ均明显高于B组（P值分别为0．044,0．012）,经rhG—CSF动员后采集物的T淋巴细胞增殖能力明显下降。结论动员后的PBSC采集物较未经动员的淋巴细胞采集物富集了更多的CD34^＋、CD14^＋细胞,同时rhG—CSF动员后DC2比例的明显升高使得CD4细胞向Th2分化,PBSC含有更多的Ⅱ类细胞因子和其T细胞增殖能力的下降、共刺激分子的下调均提示PBSC较供者淋巴细胞输注更少地引起急性移植物抗宿主病的发生。     

溃疡性结肠炎患者肠黏膜及外周血淋巴细胞表型分析

目的　探讨淋巴细胞各亚群比例在溃疡性结肠炎（UC）发病中的变化及意义。方法 利用双色、三色荧光抗体流式细胞术及细胞内细胞因子检测法对19例UC肠黏膜及外周血淋巴细胞作表型分析。结果 ⑴UC患者外周血及肠黏膜T、B细胞百分比与对照组比较，均差异无显著性；⑵外周血CD4^ 、CD8^ T细胞百分比与对照组相比无改变，肠黏膜CD4^ T细胞百分比高于对照组，CD8^ 细胞百分比低于对照组；⑶外周血及肠黏膜Th1/Th2比值均低于对照组。结论 ⑴T细胞各亚群的变化在UC发病中起重要作用，对T细胞亚群的进一步研究有助于深入揭示其发病机制；⑵肠黏膜及外周血中Th1/Th2比值均明显下降，提示Th2细胞是UC病变中的主要辅助细胞群体，UC是一种Th2细胞占优势的炎症。     

老年肺炎患者免疫状态初探

目的 观察老年社区获得性肺炎（CAP）患者免疫功能的改变.方法 以58例老年社区获得性肺炎患者为研究对象,以60例无肺炎体检老年人作为对照组,检测外周血淋巴细胞计数及各淋巴细胞亚群计数,放射免疫法测定外周血肿瘤坏死因子-α（TNF-α）、外周血白细胞介素-2（Interleukin-2,IL-2）等指标,观察老年肺炎患者机体免疫功能变化.结果 肺炎组老年患者与无肺炎组老年人比较,外周血淋巴细胞计数、T淋巴细胞计数（CD＋3）、CD4T淋巴细胞计数（CD＋3CD＋4）、CD＋4/CD＋8比值均明显低（P≤0.001）,NK细胞计数（CD＋16CD＋56）也偏低（P＜0.05）,而CD8T淋巴细胞细胞（CD＋3CD＋8）、B细胞计数（CD＋19）无明显差别（P＞0.05）.肺炎组老年患者与无肺炎组老年人比较外周血TNF-α明显升高（P〈0.001）,外周血IL-2明显偏低（P〈0.001）.结论 老年人免疫功能的减退改变与老年人CAP有着密切的关系.     

依那普利对急性冠脉综合征患者树突状细胞功能的影响

目的 探讨急性冠脉综合征（ACS）患者树突状细胞（DC）功能状态以及依那普利（Enapril）对DC功能的影响.方法 将42例ACS随机分为常规治疗组（21例）和常规治疗加依那普利治疗组（21例）,分别于治疗前及治疗后1个月取血分离外周血T淋巴细胞,用流式细胞仪检测各组DC细胞表面共刺激分子CD86表达（以CD86^＋CD11c^＋细胞比例表示）;混合淋巴细胞反应（MLR）检测DC细胞对正常献血者T淋巴细胞刺激作用;分析依那普利治疗后CD86^＋CD11c^＋细胞比例,MLR的变化及其与血清C-反应蛋白（CRP）的相关性.结果 与正常人比较,ACS患者DC细胞中CD86^＋CD11c^＋细胞比例明显增加（P＜0.001）;对T淋巴细胞增殖刺激作用明显增强（P＜0.001）;与治疗前比较依那普利治疗后CD86^＋CD11c^＋细胞比例和MLR明显下降（P＜0.01）;CD86^＋CD11c^＋细胞比例、MLR均与CRP呈正相关.结论 ACS患者DC细胞功能明显增强;依那普利可能通过抑制DC细胞功能,负向调节患者的特异性免疫而抑制斑块的炎症反应.     

反复呼吸道感染儿童T淋巴细胞亚群及免疫球蛋白检测的意义

目的研究反复呼吸道感染（RRI）患儿与外周血中T淋巴细胞亚群及免疫球蛋白水平的关系。方法检测35例临床确诊的反复呼吸道感染患儿外周血中总T淋巴细胞、B淋巴细胞、NK细胞、CD4＋、CD8＋、CD4＋/CD8＋比值及免疫球蛋白IgG、IgA、IgM水平,并与正常健康儿童对照组比较。结果 RRI儿童的总T淋巴细胞、CD4＋细胞百分率、CD4＋/CD8＋比值明显降低（P=0.014;P=0.000;P=0.000）,CD8＋细胞百分率明显升高（P=0.000）;血清IgG、IgA较健康对照组降低（P=0.000;P=0.000）。结论 RRI儿童的细胞免疫及体液免疫功能均存在紊乱,该结果对于临床免疫治疗有指导意义。     

参芪注射液对大鼠肠缺血再灌注肠壁免疫屏障的保护作用

目的探讨肠缺血再灌注损伤后参芪注射液（SQI）对肠黏膜免疫屏障的保护作用及机制。方法取60只Wistar大鼠制作肠缺血再灌注损伤模型后随机分为观察组及对照组各30只,术后均静滴生理盐水0.5ml/（100g.d）,观察组加用SQI0.2ml/（100g.d）。分别于造模后24、48、72h采用免疫组化双染色法测定两组小肠黏膜固有层内CD4＋、CD3＋、CD8＋淋巴细胞中p53基因表达。结果观察组48h和72hCD4＋、CD3＋淋巴细胞中p53基因表达低于对照组。结论SQI对肠损伤免疫屏障有保护作用,其机制可能为调控肠壁免疫细胞凋亡基因p53表达。     

胸腺基质淋巴细胞生成素对效应T细胞分化功能的影响

目的：探讨体外氧化型低密度脂蛋白（ ox-LDL）诱导人血管平滑肌细胞产生的胸腺基质淋巴细胞生成素（ TSLP）对效应T细胞分化功能的影响。方法分离并培养人血管平滑肌细胞、人树突状细胞（ DC）、初始CD4＋T细胞,随机分为4组,每组设5个样本。对照组、实验组人血管平滑肌细胞分别经PBS、ox-LDL处理后,取其上清液与DC、初始CD4＋T细胞共培养;TSLP中和抗体组、中和抗体对照组在实验组的基础上分别加入TSLP中和抗体、TSLP非特异性中和抗体与DC、初始CD4＋T细胞共培养。采用ELISA法检测各组细胞培养上清液中TSLP及Th17细胞因子IL-17、IL-22、TNF-α水平,流式细胞术检测Th17细胞构成比。结果与对照组比较,实验组培养上清液TSLP、IL-17、IL-22、TNF-α水平及Th17细胞构成比升高（ P均＜0．01）。与实验组、中和抗体对照组比较,TSLP中和抗体组TSLP、IL-17、IL-22、TNF-α水平及Th17细胞构成比降低（ P均＜0．01）。结论 Ox-LDL体外诱导人血管平滑肌细胞产生的TSLP可促进CD4＋T细胞向Th17细胞分化。     

抗CD137单克隆抗体对哮喘患者外周血CD4^＋CD25^＋T淋巴细胞死亡方式的影响

目的探讨抗CD啪单克隆抗体对哮喘患者外周血CD4^＋CD25^＋T淋巴细胞的影响。方法采用密度梯度离心法及尼龙棉柱法分离16例健康志愿者（对照组）及12例哮喘患者（哮喘组）外周血T淋巴细胞,磁性细胞分离器（MACS）分离得到CD4^＋CD25^＋T淋巴细胞,分别利用电镜及流式细胞仪观察、检测抗CD137单克隆抗体干预72h的细胞自噬率、凋亡率、胀亡率及FOXp3的表达。结果抗CD137单克隆抗体干预后两组外周血CD4^＋CD25^＋T淋巴细胞自噬率及凋亡率均增加,但哮喘组均低于对照组。结论抗CD137单克隆抗体可促进CD4^＋CD25^＋T淋巴细胞凋亡和自噬。     

不同阶段 HBV 相关 ACLF 患者外周血 DC 及T 淋巴细胞免疫功能特点研究

目的：比较不同阶段的 HBV 相关慢加急性（亚急性）肝衰竭（HBV-ACLF）患者外周血树突状细胞（DC）、T淋巴细胞（TC）相关细胞免疫功能,阐述 DC-TC 轴在 HBV-ACLF 发病过程中可能的细胞免疫学机制。方法HBV-ACLF患者30例,分为早期组15例与中晚期组15例,另设健康对照组8例,以外周血来源的 PBMC 体外分离诱导培养 DC 与TC,应用流式细胞计数检测 DC 细胞表型 HLA-DR、CD80、CD86、CD83、CD1α的表达率,及 TC 表面分子 CD3＋、CD4＋ T、CD8＋ T 淋巴细胞百分比,并检测 DC 上清液中 IFN-α、IL-4的分泌水平,比较不同阶段 HBV-ACLF 患者免疫细胞及炎性因子表达的差异。结果与健康人比较,HBV-CLF 患者 DC 表型 HLA-DR、CD1α、CD83、CD80、CD86表达率显著下降（t 值分别为5．3356、13．269、10．8742、13．3685和23．021,均 P ＜0．01）,DC 分泌因子 IFN-α显著升高（t 值为16．4569,P ＜0．01）;TC 表面分子 CD3＋、CD4＋ T、CD4＋／CD8＋细胞比值显著下降（t 值分别为7．4441、12．5557、11．0771,均 P ＜0．01）, CD8＋ T 细胞百分比显著上升（t＝4．4359,P ＜0．01）;HBV-ACLF 患者中晚期组 DC 表型 CD83、CD86表达率显著低于早期组（P 值分别为：0．0000,0．0057）,DC 分泌因子 IFN-α表达在早期组显著增多（P ＝0．0000）,IL-4表达在中晚期组显著增多（P ＝0．0000）,TC 表面分子中晚期组 CD4＋ T 细胞百分比、CD4＋／ CD8＋细 胞 比 值 显 著 下 降 （P 值分别为：0．0268、0．0002）,CD8＋ T 细胞百分比显著上升（P ＝0．0001）。结论不同阶段 HBV-ACLF 患者的 DC、TC 功能状态均表现为细胞免疫功能低下,中晚期患者的细胞免疫功能更为低下;HBV-ACLF 全病程存在促／抑炎性细胞因子功能紊乱,早期患者存在炎症因子过度释放,中晚期患者存在抗炎症细胞因子表达增强。     

Intestinal damage mediated by Kupffer cells in rats with endotoxemia

AIM：To determine the in vivo effects of phagocytic blockade of Kupffer cell(KC)on the release of proinflammatory cytokines in small intestinal lesion and on the integrity of intestinal tract by using gadolinium chloride(GdCl3)during early endotoxemia.METHODS:Wistar rats were divided into three groups:GropA,rats were injected with endotoxin(E.coliO111:B4,adose of 12mg·kg^-1)only;GroupB,rats were pretreated intravenously with 25mg of GdCl3per kg24hare given endotoxin;and Group C,sham operation only.All animals were sacrificed 4h after endotoxin injecton.In portion of the rats of three groups.bile duct was cannulated,which the bile was collected externally.Morphological changes of ileum were observed under light microscopy and electronic microscopy.The KC were isolated rfrom rats by collagenase perfusion and inKC,expression of TNF-αand IL-6mRNA were determined by RT-PCRanalysis.Plasma and bile TNF-αandIL-6Levels were determined by enzyme-linked immunosorbent assay(EISA)>RESULTS:In group A,there were neutrophil infiltration and superficial epitelial necrosis of the ilealvvilli,sloughing of mucosal epithelium.and disappear ance of somevilli.In groupB.the ileal mucosal damage was much reduced.which in groupC,no significant morphological changes were seen,GdCl3pretreatment decreased significantly the expression of TNF-αand IL-6mRNA ingroupB(4.32±0.47and4.05±0.43)when compared to groupA(9.46±1.21and9.04±1.09)(P<0.05).There was no significant expression of TNF-αand IL-6mRNA in groupC(1.03±0.14and10.4±0.13).In rats of groupA,the levels of TNF-αand IL-6in bile and plasma were 207±29ng·L^-1,1032±107ng·L^-1,213±33ng·L^-1,and 1185±127ng·L^-1,respectively.In groupB,they were113±18ng·L^-1,repectively.In groupC,they were 67±10ng·L^-1,72±13ng·L^-1,109±18ng·L^-1,and 118±22ng·L^-1respectively.There were significant difference between the three group(P<0.05).CONCLUSION:KC release cytokinesTNF-αand IL-6causing damage to the integrity of intestinal epithelium and play a crucial role in the initiation and progression of intestinal mucosal damage during early endotoxemia.     

Therapeutic effect of Captopril on rheumatoid arthritis in rats

Objective:To investigate the therapeutic effect of the intervention treatment with different doses of Captopril on TNF-α contents in serum of rheumatoid arthritis(RA) rats,and to provide the theoretical proofs for clinical application of Captopril in treatments ol rheumatoid diseases.Methods:Fifty Wistar rats were randomly divided into 5 groups,namely.Group A,Group 13.Group C.Group D,Group E with 10 rats in each group.Injection of Freund's complete adjuvant was employed to establish adjuvant-induced arthritis model in rats.Group A was model group;after model establishment,rats were treated with 20 mL normal saline as placebo(ip.).Rats in Group B were treated with 8 mg/kg cyclophosphamide(ip.).Rats in Group C.D and E were intraperitoneally injected with 30 mg/kg.100 mg/kg and 300 mg/kg Captopril respectively.Rats in each group were subjected to continuous treatment for 3 weeks,and then sacrificed.Eyeballs of rats were excised and blood was collected.TNF- α content in serum were detected using ELISA:each group rats were compared for the hind legs arthrocele.Right ankle tissues of rats were collected to prepare section,and microscopic observation of pathological changes was performed.Results:TNF- α content in serum of Group A rats was significantly higher than that of rats in other 4 groups(P0.05).TNF- α content in serum of Group B rats was significantly lower compared with that of rats in Groups C.D and E.The highest TNF- α content in serum of rats treated with Captopril was found in Group C,followed by Groups D and E(P0.05).Right ankle arthrocele of rats in Groups B.C.D and E in early stage showed no statistical difference compared with that of Group A rats(P0.05).From Day 8,ankle arthrocele of rats in Groups B.C.D and E was obviously relieved compared with that of Group A rats:the anti-inflammatory effects were gradually enhanced with the extension of medication time.Treatments of Groups C.D and E showed significant activities against tardive aithrocele:the degree of ankle arthrocele in rats of these three groups was lower than that of Group A rats(P0.01).Histological observation showed that large amount of inflammatory cells and plasmocyte infiltration was found in ankle synovial tissues of Group A rats.Relief of hyperaemia and edema of right ankle synovial tissues as well as significant decrease in synoviocyte layer hyperplasia,intra—articular inflammatory cells infiltration and cartilage articularis damage degree etc.were observed in Groups B.C.D and E.Conclusions:Intervention treatment with Captopril can effectively reduce the TNF- α content in serum of rheumatoid arthritis rats and inhibit the generation of inflammatory factors,so as to achieve the therapeutic effect.     

携带肝细胞生长因子基因的重组腺病毒修复烟熏致大鼠肺气肿的作用

目的 通过构建大鼠肺气肿模型,观察携带肝细胞生长因子基因的重组腺病毒(Ad-HGF)对肺气肿病变的修复作用.方法 40只Wistar大鼠随机分为4组:肺气肿组(A组)、肺气肿+Ad-HGF 组(B组)、肺气肿+Ad-GFP组(c组)和正常对照组(D组);采用烟熏法建立大鼠肺气肿模型.A组给予0.5 ml生理盐水灌注治疗,B组给予1×10 9 pfu Ad-HGF,C组给予1×10 9 pfu Ad-GFP,D组正常饲养;干预后14 d取腹主动脉血,作动脉血气分析.取C组大鼠部分肺组织作常规冰冻切片,荧光显微镜下观察目的 基因的表达;取A,B,D组大鼠肺组织制作病理切片,观察并计算平均肺泡面积(MAA);采用脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记技术和增殖细胞核抗原免疫组织化学法对大鼠肺实质凋亡细胞、增殖细胞的阳性细胞率(AI、PI)进行检测.结果 与D组相比,A组、B组、C组大鼠的肺组织都存在不同程度的肺气肿病理改变,肺组织切片中Ad-HGF治疗组病理改变明显轻于模型组;各组大鼠动脉血气分析比较,差异无统计学意义(P>0.05);Ad-HGF治疗组MAA[-(4 227.63±156.01),μm2] 显著小于模型组[(6 346.35±148.60)μm2,P＜0.05] ,Ad-HGF治疗组AI[(18.95±2.27)%] 显著低于模型组[(23.41±3.55)%,P＜0.05] ,以上两组AI均高于正常对照组[(5.40±1.22)%,P＜0.05] ;Ad-HGF治疗组PI[(17.51±1.89)%] 显著高于模型组[(15.51±2.75)%,P＜0.05] ,以上两组PI均高于正常对照组[(5.44±1.80)%,P＜0.05] .结论 Ad-HGF对烟熏大鼠肺气肿有修复作用.     

Chemopreventive effect of a probiotic preparation on the development of preneoplastic and neoplastic colonic lesions: an experimental study

BACKGROUND/AIMS: A number of studies have suggested a key role played by certain resident gut bacteria in the development of large bowel cancer. The aim of the present study was to test the effect of a novel symbiotic preparation, which has been recently shown to beneficially modify gut ecosystem and systemic immunity, on either preneoplastic and neoplastic changes in a colon carcinogenesis model. METHODOLOGY: Sprague-Dawley rats were fed a standard diet for 1 week and then were randomly assigned to three groups. The control diet was given to groups A and B, whereas in group C, the same diet plus 2 mL of a probiotic mixture was given throughout the experiment. Thirty rats (groups B, C) each received a weekly subcutaneous injection of azoxymethane at a dose of 15 mg/kg of body weight for 10 weeks. Group A served as a control group and received a subcutaneous injection of saline for 10 weeks. Forty-five rats were sacrificed at 3-week observation and 60 rats at 20-week observation for assessing metaphase index together with aberrant crypt foci and intestinal immune system markers from one hand and tumor occurrence from the other, respectively. RESULTS: Group A showed a significantly increased metaphase index either in aberrant crypt foci or in "normal appearing" crypts when compared to group A (p < 0.01). Group B rats caused a significant decrease at both sites (p < 0.05). The numbers of lymphocytes derived from the mesenteric lymph nodes in group B rats were significantly decreased (p < 0.01) as compared to either control and to group C. The percentage of CD8 lymphocytes in group C was significantly higher than that in group B. Group C showed a significantly reduced ratio of aberrant crypt foci/colon and of aberrant crypt per colon and per each single focus (p < 0.05). A total of 18 (90%) group B and 10 (50%) group C rats had colon tumors, this difference was significant. The mean number of colon tumors per rat was 2.2 and 1.0 in group B and C, respectively. CONCLUSIONS: Effective probiotics treatment, through mechanisms still to be fully elucidated (decreased fecal pH, specific reduction of carcinogenetic bacterial enzymes, modulation of gut-associated and systemic immune system etc.) has the potential to exert significant antimutagenic properties against colon cancer.     

谷氨酰胺对铜绿假单胞菌所致肺部感染大鼠的防护作用

目的探讨谷氨酰胺在全胃肠外营养（TPN）中对铜绿假单胞菌所致肺部感染大鼠的保护作用。方法SD大鼠40只,随机分为4组,A组大鼠向气管内注入生理盐水0．3ml。B组大鼠向气管内注入0．3ml的铜绿假单胞菌悬浮液,并使细菌直接进入肺内。C组大鼠静脉泵标准的不含N（2）L-丙氨酰-谷氨酰胺（商品名为力肽）的TPN溶液160ml／kg,5d;D组大鼠静脉泵含力肽的TPN溶液160ml／kg,5d;C、D组大鼠第6天向气管内注入0．3ml的铜绿假单胞菌悬浮液,并使细菌直接进入肺内。观察大鼠生命活动。注菌48h后取外周血计数白细胞;行支气管肺泡灌洗,取灌洗液,计数其中的白细胞,测其中的TNFα、IL-1、IL-10水平及总蛋白含量。取肺、肝、回肠小块组织,光镜下观察病理改变。结果（1）实验开始时4组大鼠体重无明显差别,第8天时B、C、D组大鼠体重较A组轻;C组死亡的鼠数多于D、B组。（2）C组大鼠外周血WBC计数、血清中TNFα、IL-1高于D组,而IL-10低于D组。C组大鼠支气管肺泡灌洗液中WBC计数、总蛋白含量、TNFα高于D组,而IL-10低于D组。（3）光镜下见C组大鼠肺、肝、回肠组织病理改变明显重于D、B组,A组基本正常。结论谷氨酰胺可保护TPN大鼠的胃肠道功能,提高肺部抗感染能力,并减轻由严重感染所致其他重要器官的损害。     

Effect of WeiJia on carbon tetrachloride induced chronic liver injury

AIM:To study the effect of WeiJia on chronic liver injuryusing carbon tetrachloride(CCl_4)induced liver injuryanimal model.METHODS:Wistar rats weighing 180-220g were ran-domly divided into three groups:normal control group(Group A),CCl_4 induced liver injury control group(GroupB)and CCl_4 induction with WeiJia treatment group(GroupC).Each group consisted of 14 rats.Liver damage andfibrosis was induced by subcutaneous injection with 40?l_4 in olive oil at 3 mL/kg body weight twice a week foreight weeks for Groups B and C rats whereas olive oilwas used for Group A rats.Starting from the third week,Group C rats also received daily intraperitoneal injectionof WeiJia at a dose of 1.25 μg/kg body weight.Animalswere sacrificed at the fifth week(4 male,3 female),andeighth week(4 male,3 female)respectively.Degree offibrosis were measured and serological markers for liverfibrosis and function including hyaluronic acid(HA),typeIV collagen(CIV),γ-glutamyl transferase(γ-GT),alanineaminotransferase(ALT)and aspartate aminotransferase(AST)were determined.Alpha smooth muscle actin (α-SMA)and proliferating cell nuclear antigen(PCNA)immunohistochemistry were also performed.RESULTS:CCl_4 induction led to the damage of liver anddevelopment of fibrosis in Group B and Group C ratswhen compared to Group A rats.The treatment of WeiJiain Group C rats could reduce the fibrosis condition sig-nificantly compared to Group B rats.The effect could beobserved after three weeks of treatment and was moreobvious after eight weeks of treatment.Serum HA,CIV,ALT,AST and γ-GT levels after eight weeks of treatmentfor Group C rats were 58±22 μg/L(P<0.01),57±21 μg/L(P<0.01),47±10 U/L(P<0.01),139±13 U/L(P<0.05)and 52±21 U/L(P>0.05)respectively,similar to normalcontrol group(Group A),but significantly different fromCCl_4 induced liver injury control group(Group B).An in-crease in PCNA and decrease in α-SMA expression levelwas also observed.CONCLUSION:WeiJia could improve liver function andreduce liver fibrosis which might be through the inhibi-tion of stellate cell activity.     

腺苷A_1受体激动剂R-苯异丙基腺苷介导的大鼠心肌延迟预适应

目的　研究腺苷A1受体激动剂R 苯异丙基腺苷 (R PIA)能否介导大鼠心脏产生延迟预适应。方法　取雄性Wistar大鼠 48只 ,随机分为 3组。A组 :生理盐水 0 .2ml/只 ;B组 :R PIA 0 .0 3mg/kg ;C组 :R PIA 0 .0 3mg/kg +DPCPX 0 .2mg/kg。大鼠经皮下注射给药后 2 4h开胸 ,结扎左冠状动脉前降支 30min、再灌注 12 0min ,摘取心脏 ,用于梗死范围测定 (TTC染色法 )、原位心肌细胞凋亡 (TUNEL法 )检测。结果　大鼠体重及左室重量组间差别无显著性 (P >0 .0 5 )。R PIA组心肌梗死范围显著低于生理盐水组及R PIA +DPCPX组 (P均 <0 .0 1) ,而生理盐水组与R PIA +DPCPX组比较差异无显著性意义 (P >0 .0 5 )。生理盐水组、R PIA组及R PIA +DPCPX组心肌凋亡细胞计数 (个 /高倍视野 )组间差异无显著性意义 (P >0 .0 5 )。结论　R PIA可使大鼠在体心脏产生延迟预适应 (显著降低心肌梗死范围 ) ;R PIA对 2 4h后缺血再灌注损伤所致的心肌细胞凋亡可能无影响。     

支气管哮喘大鼠CD4+CD25+T细胞的变化及地塞米松干预作用的研究

目的 研究支气管哮喘(简称哮喘)大鼠模型支气管肺泡灌洗液(BALF)、血液、脾脏CD4+CD25+T细胞的变化,及地塞米松对CD4+CD25+T细胞的影响.方法 50只SD大鼠随机分为5组,空白对照(A)组,哮喘(B)组,地塞米松1(C)组、地塞米松2(D)组,地塞米松3(E)组.A组第l天给予腹腔注射生理盐水l ml,第15～21天每天给予生理盐水雾化.B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原l ml(卵蛋白1 mg+灭活百日咳杆菌9×106个+氢氧化铝干粉100 mg)混悬液,第15～21天给予1%的卵蛋白雾化30 min,C、D、E组于雾化后分别给予腹腔注射地塞米松0.2 mg/kg、1 mg/kg、2 mg/kg.采用流式细胞仪检测的方法 ,观察大鼠体内BALF、外周血、脾脏CD4+CD25+T细胞的变化及使用不同剂量地塞米松后对其的影响.结果 B组BALF、外周血、脾脏CD4+CD25+T细胞表达占CD4+T细胞的百分比分别是(42.21±5.62)%、(12.69±2.70)%、(11.15±1.05)%,A组结果 分别是(18.76±5.85)%、(6.21±1.73)%、(7.85±2.13)%.B组与A组比较,差异均具有统计学意义(P＜0.01,P＜0.01,P＜0.05);C组、D组、E组BALF中CD4+CD25+T细胞占CD4+T细胞的百分比表达分别是(10.49±4.03)%、(13.28±5.12)%、(7.51±5.39)%,显著低于A组和B组,(P＜0.05,P＜0.01);外周血中,C组(6.03±1.43)%、D组(4.88±0.95)%与A组(6.21±1.73)%比较,差异无统计学意义,E组(3.49±0.62)%与C组、A组比较,差异有统计学意义(P＜0.05).脾脏中,C组(7.25±1.82)%、D组(8.63±3.18)%与A组(7.85±2.13)%比较,差异无统计学意义,E组(3.38±1.37)%与C组、D组、A组比较,差异有统计学意义(P＜0.05).结论 CD4+CD25+T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一.地塞米松可以抑制CD4+CD25+T细胞的表达.BALF内CD4+CD25+T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4+CD25+T细胞变化可了解肺部情况.     

蛋白酶抑制剂对全氟异丁烯吸入致 ALI 大鼠氧化应激的影响

目的：研究探讨蛋白酶抑制剂(抑肽酶)对全氟异丁烯(PFIB)吸入致 ALI 大鼠炎症因子的影响。方法健康清洁级成年雄性 Sprague-Dawley 大鼠40只,根据体质量随机分成5组：生理盐水对照组(简称 A 组)、ALI 组(简称 B 组)、抑肽酶治疗组(5 mg/kg、15 mg/kg 和30 mg/kg,简称 C、D、E 组,),每组8只。将 B、C、D、E 组大鼠经头部暴露于自制 PFIB 动态吸入染毒系统装置染毒5 min。染毒30 min 后,各组大鼠均经腹腔注射生理盐水或配置好的不同浓度抑肽酶溶液。24 h后结束实验,处死大鼠,获取 BALF、肺组织样本,检测肺湿重与肺干重比(W/D),BALF 总蛋白含量,肺组织匀浆中超氧化物歧化酶(SOD)、丙二醛(MDA)的含量,光镜下肺组织病理学和免疫组织化学检查,并将数据进行统计学分析。结果 B 组、C 组、D 组、E 组的肺湿与干重比、BALF总蛋白含量和 MDA 含量升高,与 A 组比较差异有统计学意义(P <0.05),B 组、C 组、D 组、E 组的 SOD 含量下降,与 A 组比较差异有统计学意义(P <0.05)。E 组肺湿与干重比、BALF 总蛋白及MDA 含量均低于 B 组、C 组、D 组,差异有统计学意义(P <0.05);E 组 SOD 含量高于 B 组、C组、D 组,差异有统计学意义(P <0.05)。肺组织病理学检查显示,C 组、D 组、E 组较 B 组的肺组织水肿情况减轻、炎性细胞浸润减少,肺损伤程度降低,以高剂量最为显著。免疫组织化学检查示, C 组、D 组、E 组的核因子κB 的表达较 B 组为低。结论蛋白酶抑制剂抑肽酶能通过抗氧化应激及抑制炎性反应,来减轻 PFIB 吸入导致的 ALI。     

Prenylamine inhibition of isoproterenol induced myocardial lesions. Histochemical and ultrastructural findings.

Eighty Wistar rats were divided into 4 groups. All of them received 10 mg/kg i.p. isoproterenol (ISP). The animals were sacrificed at 5 min. (groups A and B) and 24 hours (groups C and D). Groups A and C served as control of group B and D. Group B also received 9 mg/kg prenylamine (P), 30 min before ISP and group D 9 mg/kg P, 1 hour before and 1 hour after ISP. Transversal slices of each heart underwent the following procedures: hematoxilineosin, Barbeito-Lopez trichromic, "ischemia stain", acid phosphatase stain and standard electron microscopy. Group A and C showed positivity for ischemic techniques; necrotic and ischemic zones represented, in average 25.1 +/- 1.4% of the myocardial volume. Group B and D (ISP + prenylamine) showed no significant lesions. Group D (ISP + prenylamine, sacrificed at 24 hours) presented complete absence of "infarct-like" lesions in 17 out of the 20 studied animals. No evident changes in acid-phosphatase enzymes were detected. Animals treated with ISP alone presented "contraction bands" at electron microscopy as well as myofibrillar fragmentation. Those receiving also P showed only light alterations, in the electron microscopy specimens. A primary action of ISP on the calcium pump might explain the infarct-like lesions found in our study. The inhibition of these above mentioned lesions by prenylamine, a drug which acts by slowing down the ca transport, also supports this interpretation.