前列腺素E1对梗阻性黄疸大鼠肝脏缺血再灌注损伤时白介素-1β表达的影响

目的:探讨前列腺素(PG)E1对梗阻性黄疸肝脏缺血再灌注损伤的保护作用以及对IL-1β表达的影响.方法:将♂Wistar大鼠随机分为PGE1处理组(PG组,n=18)和生理盐水对照组(NS组,n=18).参照Yoshidome法结扎并切断胆总管建立梗阻性黄疸模型,1 wk后Pringle法阻断肝门15 min,再灌注后建立胆道再通.于缺血前15 min至再灌注60 min,PG组经门静脉持续泵入PGE1 0.5 μg/(kg·min),NS组给予等量生理盐水.于再灌注1、6和24 h 3个时点取材,检测血清ALT,AST,总胆红素(total bilirubin,TBIL),直接胆红素(direct bilirubin,DBIL)水平,测定肝组织GSH,MDA含量.ELISA法测定肝组织IL-1β表达,并观察肝脏病理组织学改变.结果:再灌注各时点PG组血清ALT水平(1 h:1939±1427 nkat/L vs 5596±2975 nkat/L;6 h:3409±1708 nkat/L vs 9279±4404 nkat/L;24 h:1434±274 nkat/L vs 2264±630 nkat/L)和AST(1 h:21746±12083 nkat/L vs 37552±12382 nkat/L;6 h:55039±35471 nkat/L vs 98811±11126 nkat/L;24 h:9394±1662nkat/L vs 27664±15856 nkat/L)、肝组织MDA含量(1 h:0.89±0.18 μmol/g vs 1.21±0.24 μmol/g;6 h:1.08±0.23 μmol/g vs 1.45±0.13 μmol/g;24 h:1.03±0.08 μmol/g vs 1.45±0.26 μmol/g)以及肝组织IL-1β表达水平(1 h:304.1±67.9 ng/L vs 362.8±137.1 ng/L;6 h:376.8±74.6 ng/L vs 618.8±217.8 ng/L;24 h:273.0±69.0 ng/L vs 373.0±71.7 ng/L)均显著低于NS组(P＜0.05),而肝组织GSH含量显著高于NS组(1 h:945.1±121.2 mg/g vs 720.0±80.1 mg/g;6 h:753.5±118.6 mg/g vs 553.6±140.0 mg/g;24 h:768.0±135.9 mg/g vs 596.3±36.4 mg/g,P＜0.05),但两者胆红素水平无明显差异(P＞0.05).PG组肝脏病理组织学损伤程度也较NS组明显减轻.结论:PGE1下调肝组织IL-1β表达,对梗阻性黄疸肝脏缺血再灌注损伤具有保护作用.     

番石榴叶提取物对三硝基苯磺酸诱导大鼠结肠炎结肠组织的保护作用

目的:研究番石榴叶提取物对大鼠结肠炎组织损伤的保护作用及其机制.方法:用健康Wistar大鼠建立大鼠结肠炎模型,灌胃用药2 wk后,评价大鼠结肠黏膜损伤指数(CMDI),检测结肠组织髓过氧化物酶(MPO)及超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量和细胞肿瘤因子(TNF)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)水平.结果:不同剂量番石榴叶提取物(200,500,800 mg/kg)灌胃均能不同程度降低模型大鼠CMDI(2.62±0.47,2.06±0.54,1.83±0.43 vs 3.15±0.39,P＜0.05,P＜0.01,P＜0.01),MPO活性(2240.78±479.26,2078.42±377.91,1748.85±236.71 nkat/g vs 2695.37±624.79 nkat/g P＜0.05,P＜0.01,P＜0.01),减少MDA含量(2.16±0.41,1.97±0.35,1.96±0.30,P＜0.05,P＜0.01,P＜0.01),提高SOD活性(3618.06±538.44,3756.92±577.45,4197.51±375.74 nkat/g vs 2663.37±603.79,P＜0.05,P＜0.01,P＜0.01),降低TNF(2.65±0.40,2.51±0.47,2.21±0.41 μg/L vs 3.30±0.63 μg/L,P＜0.05,P＜0.01,P＜0.01)、IL-8(1.20±0.33,1.05±0.24,0.92±0.13 μg/L vs 1.53±0.38 μg/L,P＜0.05,P＜0.01,P＜0.01)、IL-10(39.36±9.65,50.26±11.32.59.68±13.65 ng/L vs 30.74±12.91 ng/L,P＜0.05,P＜0.01,P＜0.01)水平,且与用药剂量呈一定量效关系.结论:番石榴叶提取物通过拮抗氧化、免疫调节、损伤修复作用缓解结肠炎大鼠炎症反应,减轻结肠损伤.     

中药肠炎清治疗小鼠葡聚糖硫酸钠所致结肠炎的机制

目的:探讨肠炎清的疗效及其机制.方法:以葡聚糖硫酸钠(DSS)饮水法复制小鼠实验性结肠炎40只,随机平均分为4组:肠炎清中药组、柳氮磺胺吡啶(SASP)西药组、肠炎清和SASP中西药结合组和模型组.观察肠炎清(灌胃剂量为0.2mL/(20g?d)、1次/d、疗程7d)对疾病活动指数(DAI)和肠组织髓过氧化物酶(MPO)活性及TNF-α、IL-1β和IL-6mRNA表达的影响.结果:与模型组相比,肠炎清可降低DAI(1.413±0.835vs2.167±0.911,P<0.05)和MPO活性(72.4±0.590nkat/gvs117.0±0.902nkat/g,P<0.05),并降低肠组织TNF-α(0.841±0.190vs1.320±0.282,P<0.05)、IL-1β(0.641±0.095vs0.920±0.082,P<0.05)和IL-6mRNA(1.241±0.247vs1.620±0.312,P<0.05)的表达,中西医结合组以上指标下降更为明显(DAI:0.608±0.449;MPO:27.3±0.211;TNF-α:0.339±0.081;IL-1β:0.239±0.073;IL-6:0.639±0.141)(P<0.01).肠炎清与柳氮磺胺吡啶(SASP)的作用相当(P>0.05).结论:肠炎清可治疗DSS结肠炎,其降低肠组织TNF-α、IL-1β和IL-6mRNA的表达可能是其疗效机制之一.     

脂质体介导核因子-kappaB诱捕物寡聚脱氧核苷酸对重症急性胰腺炎大鼠胰腺炎症因子mRNA表达和胰腺损伤的影响

目的探讨脂质体(liposome)介导核因子-κB(nuclear factor κB,NF-κB)诱捕物(decoy)寡聚脱氧核苷酸(ODN)对重症急性胰腺炎大鼠胰腺NF-κB活性及受其调控炎症因子基因mRNA表达和胰腺损伤的影响.方法除假手术组(n=10)外,其余SD大鼠以牛磺胆酸钠(STC)诱导建立重症急性胰腺炎模型后,分别于建模后1 h静脉注射裸ODN(n=10)、脂质体/decoy ODN复合物(n=10)、脂质体/scrambled ODN复合物(n=10)和生理盐水(n=10),注射4 h应用电泳迁移率变动分析(EMSA)NF-κB的活性,利用逆转录-聚合酶链反应(RT-PCR)法检测胰腺组织ICAM-1,IL-1α,IL-2,TNF-α,VCAM-1 mRNA表达,同时检测血淀粉酶、胰腺组织湿/干重比率和胰腺组织髓过氧化物酶(MPO).结果EMSA显示,脂质体/decoy ODN复合物组NF-κB活性明显低于生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组,均有显著性差异(P＜0.05).RT-PCR结果显示,脂质体/decoy ODN复合物组ICAM-1,IL-1α,IL-2,TNF-α和VCAM-1 mRNA表达低于生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组,均有显著性差异(ICAM-10.75±0.13 vs 1.39±0.15,1.37±0.16,1.32±0.17,P＜0.05;IL-1α0.64±0.09 vs 1.34±0.20,1.30±0.14,1.25±0.20,P＜0.05;IL-20.23±0.08vs 0.74±0.13,0.71±0.12,0.69±0.14,P＜0.05;TNF-α0.41±0.13 vs 1.30±0.17,1.26±0.17,1.23±0.20,P＜0.05;VCAM-10.21±0.06 vs 0.68±0.13,0.69±0.15,0.63±0.13,P＜0.05).与生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组相比,脂质体/decoy ODN复合物组淀粉酶、胰腺组织湿/干重比率和胰腺组织MPO活性显著性降低(淀粉酶50931.85±22432.15 nkat/L vs 188024.26±38659.56,188412.68±37988.26,183119.95±33636.23nkat/L,all P＜0.05;湿/干重比率5.76±0.20 vs 6.77±0.18,6.72±0.18,6.35±0.12,P＜0.05;MPO活性46.68±3.00 nkat/g vs 99.02±2.50,98.19±2.83,98.52±2.50 nkat/g,P＜0.05).结论NF-κB decoy ODN可特异性抑制胰腺NF-κB活性及其调控的炎症因子ICAM-1,IL-1α,IL-2,TNF-α和VCAM-1 mRNA的表达,减轻胰腺损害.     

急性肝损伤大鼠肝脏血红素加氧酶-1/一氧化碳系统的变化及意义

目的:研究急性肝损伤时大鼠肝脏HO-1/CO系统的变化规律,探讨HO-1和内源性CO的作用机制及其病理生理意义.方法:采用D-氨基半乳糖(GalN)和脂多糖(LPS)联合腹腔注射制备大鼠急性肝损伤模型,动态测定各时间点(3,6,12,24,36 h)大鼠肝脏HO-1活性和蛋白表达情况,测定肝脏CO浓度以及血清ALT,AST水平和肝组织SOD活性、MDA含量变化.结果:Ga1N和LPS联合注射成功诱导了大鼠急性肝损伤,表现为染毒24 h时大鼠血清ALT,AST水平以及肝组织MDA浓度显著升高(10872.5±708.5 nkat/L,9246.8±814.7 nkat/L,5.06±1.21 μmol/g vs 1043.5±247.4 nkat/L,1278.6±273.8 nkat/L,2.03±0.59 μmol/g,均P＜0.01),SOD活性明显下降(813.7±168.3nkat/mg vs 1248.2±84.9 nkat/mg,P＜0.01),HE染色显示肝细胞出现严重损伤.染毒3-24 h大鼠肝脏HO-1活性明显增强,6-24 h呈现一定的时间依赖方式(4.02±0.74,5.97±1.51,6.13±1.18 μmol/g vs 2.86±0.41 μmol/g);Westernblot测定结果亦显示,HO-1蛋白表达显著增强,24 h时明显高于正常对照组(1.87±0.39 vs0.37±0.09,P＜0.01).正常大鼠肝脏的CO浓度极低,染毒后以时间依赖方式开始升高,并显著高于正常对照组(0.373±0.112,0.474±0.152,0.513±0.193 μmol/g vs 0.172±0.041 μmol/g,P＜0.01),这与HO-1表达情况相一致.结论:大鼠急性肝损伤时出现HO-1活性增加和蛋白表达持续上调以及CO浓度迅速增高,提示HO-1/CO系统参与急性肝损伤的病理生理过程,其表达增加可能对机体有重要调节作用.     

脂质体介导核因子-κB诱捕物寡聚脱氧核苷酸对重症急性胰腺炎大鼠胰腺炎症因子mRNA表达和胰腺损伤的影响

目的:探讨脂质体(liposome)介导核因子-κB(nuclear factor κB,NF-κB)诱捕物(decoy)寡聚脱氧核苷酸(ODN)对重症急性胰腺炎大鼠胰腺NF-κB活性及受其调控炎症因子基因mRNA表达和胰腺损伤的影响.方法:除假手术组(n=10)外,其余SD大鼠以牛磺胆酸钠(STC)诱导建立重症急性胰腺炎模型后,分别于建模后1 h静脉注射裸ODN(n=10)、脂质体/decoy ODN复合物(n=10)、脂质体/scrambled ODN复合物(n=10)和生理盐水(n=10),注射4 h应用电泳迁移率变动分析(EMSA)NF-κB的活性,利用逆转录-聚合酶链反应(RT-PCR)法检测胰腺组织ICAM-1,IL-1α,IL-2,TNF-α,VCAM-1 mRNA表达,同时检测血淀粉酶、胰腺组织湿/干重比率和胰腺组织髓过氧化物酶(MPO).结果:EMSA显示,脂质体/decoy ODN复合物组NF-κB活性明显低于生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组,均有显著性差异(P＜0.05).RT-PCR结果显示,脂质体/decoy ODN复合物组ICAM-1,IL-1α,IL-2,TNF-α和VCAM-1 mRNA表达低于生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组,均有显著性差异(ICAM-1:0.75±0.13 vs 1.39±0.15,1.37±0.16,1.32±0.17,P＜0.05;IL-1α:0.64±0.09 vs 1.34±0.20,1.30±0.14,1.25±0.20,P＜0.05;IL-2:0.23±0.08vs 0.74±0.13,0.71±0.12,0.69±0.14,P＜0.05;TNF-α:0.41±0.13 vs 1.30±0.17,1.26±0.17,1.23±0.20,P＜0.05;VCAM-1:0.21±0.06 vs 0.68±0.13,0.69±0.15,0.63±0.13,P＜0.05).与生理盐水组、脂质体/scrambled ODN复合物组和裸ODN组相比,脂质体/decoy ODN复合物组淀粉酶、胰腺组织湿/干重比率和胰腺组织MPO活性显著性降低(淀粉酶:50931.85±22432.15 nkat/L vs 188024.26±38659.56,188412.68±37988.26,183119.95±33636.23nkat/L,all P＜0.05;湿/干重比率:5.76±0.20 vs 6.77±0.18,6.72±0.18,6.35±0.12,P＜0.05;MPO活性:46.68±3.00 nkat/g vs 99.02±2.50,98.19±2.83,98.52±2.50 nkat/g,P＜0.05).结论:NF-κB decoy ODN可特异性抑制胰腺NF-κB活性及其调控的炎症因子ICAM-1,IL-1α,IL-2,TNF-α和VCAM-1 mRNA的表达,减轻胰腺损害.     

罗格列酮对大鼠溃疡性结肠炎肠黏膜NF-κB,ICAM-1表达的影响

目的:探讨罗格列酮对大鼠溃疡性结肠炎肠黏膜核转录因子-κB(NF-κB),细胞间黏附分子-1(ICAM-1)表达的影响.方法:应用三硝基苯磺酸(TNB)/乙醇灌肠制备大鼠溃疡性结肠炎模型.实验设正常对照组,模型对照组,阳性药物组(柳氮磺吡啶SASP组,100mg/kg),罗格列酮组(2,4,8mg/kg),每天灌胃给药1次,给药时间从造模后第2天开始至实验结束共8d,观察大鼠疾病活动指数(DAI)和结肠黏膜损伤指数(CMDI),生化法检测大鼠结肠组织髓过氧化酶(MPO)活性,免疫组化法检测大鼠肠黏膜NF-κBp65和ICAM-1蛋白的表达.结果:与正常组相比,模型组DAI、CMDI评分及结肠组织MPO活性明显升高(2.11±1.29vs0.11±0.17,2.67±0.82vs0.33±0.52,1.26±0.36U/gvs0.27±0.07U/g,P<0.01),结肠黏膜NF-κBp65及ICAM-1表达明显增强(0.7081±0.0671vs0.2293±0.0474;0.4846±0.0366vs0.1783±0.0201,P<0.01).罗格列酮中、高剂量组DAI,CMDI评分及MPO活性较模型组有明显下降(DAI:1.11±0.50,0.61±0.25vs2.11±1.29,P<0.05;CMDI:1.67±0.52,1.17±0.75vs2.67±0.82,P<0.05;MPO:0.82±0.13,0.51±0.10U/gvs1.26±0.36U/g,P<0.01),NF-κB及ICAM-1表达也有不同程度降低(NF-κB:0.4544±0.0379,0.2577±0.0131vs0.7081±0.0671,P<0.01;ICAM-1:0.3854±0.0277,0.2830±0.0234vs0.4846±0.0366,P<0.01).大鼠结肠黏膜NF-κB的表达与ICAM-1表达呈正相关(r=0.927,P<0.01),ICAM-1的表达与结肠组织MPO活性也呈正相关(r=0.580,P<0.01).结论:罗格列酮对大鼠溃疡性结肠炎有保护作用,其作用机制可能与抑制NF-κB活化,减少黏附分子ICAM-1产生以及降低中性粒细胞浸润有关.     

青黛颗粒对溃疡性结肠炎大鼠结肠黏膜MUC2和iNOS基因表达的影响

目的:观察青黛颗粒对TNBS诱导的溃疡性结肠炎(UC)大鼠结肠黏膜黏蛋白(MUC2)及诱导型一氧化氮合成酶(iNOS)基因表达的影响,探讨其治疗UC的可能作用机制.方法:将54只SD实验大鼠随机分为正常对照组、模型对照组、阳性药物治疗组(SASP,500 mg/kg)、青黛颗粒600、900、1 200 mg/kg治疗组.造模后第3天开始灌胃给药,共给药10d.实验第14天,处死大鼠,剖取其病变结肠组织,比较各组大鼠的DAI积分和CMDI评分,用逆转录聚合酶链反应(RT-PCR)法检测MUC2及iNOS基因的表达.结果:较模型组青黛颗粒900、1200 mg/kg治疗组能显著降低实验大鼠DAI积分和CMDI评分,上调结肠组织中MUC2的基因表达(2.06±0.70 vs 1.24±0.47;2.34±0.86 vs 1.24±0.47.均P<0.01),且青黛颗粒1 200 mg/kg治疗组能下调iNOS的基因表达(0.35±0.12vs 0.62±0.31.P<0.05).结论:青黛颗粒可能通过上调结肠黏膜MUC2的基因表达并下调iNOS的基因表达而起到抗TNBS诱导的大鼠溃疡性结肠炎的作用.     

表没食子儿茶素没食子酸酯对乙酸诱导的大鼠结肠炎抗炎作用机制的研究

背景：表没食子儿茶素没食子酸酯（EGCG）具有抗氧化作用,能抑制炎症信号通路,如抑制可诱导促炎因子表达的核因子（NF）-κB。目的：探讨EGCG对乙酸诱导的大鼠结肠炎的抗炎作用机制。方法：60只Sprague-Dawley（SD）大鼠随机分为正常对照组（n=10）、模型安慰剂组（n=20）、EGCG治疗组（n=15）和柳氮磺吡啶（SASP）治疗组（n=15）。正常对照组常规饲养;以8%乙酸制备结肠炎模型后,模型安慰剂组、EGCG治疗组和SASP治疗组每日分别予0.9%NaCl溶液2ml、EGCG50mg/kg、SASP0.25g/kg灌胃治疗7d,第8d处死大鼠。酶联免疫吸附测定（ELISA）检测大鼠血清肿瘤坏死因子（TNF）-α和干扰素（IFN）-γ含量,免疫组化SP法测定结肠组织NF-κBp65表达。结果：与模型安慰剂组和SASP治疗组相比,EGCG治疗组大鼠血清TNF-α和IFN-γ以及结肠组织NF-κBp65水平显著降低（P〈0.01）。结论：EGCG可通过调节免疫因子表达而达到抗炎的治疗作用,使结肠炎症反应减轻,其疗效优于传统药物SASP。     

活血化瘀注射液Ⅰ号预处理与缺血预处理改善肝缺血再灌注损伤

目的:研究活血化瘀注射液Ⅰ号(HHI-Ⅰ)预处理与缺血预处理(ischemic preconditioning,IP)对大鼠肝脏缺血再灌注(ischemia and reperfusion,I/R)损伤的改善作用,并比较两者的作用效果.方法:健康♂SD大鼠80只,随机分为假手术对照组(Sham组)、缺血再灌注组(I/R组)、缺血预处理组(IP组)、HHI-Ⅰ预处理组(HHI-Ⅰ组),每组20只.建立大鼠部分肝缺血模型,各组在I/R后1,3,6,24 h分别取材,测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、乳酸脱氢酶(LDH)的水平;取左肝测组织丙二醛(MDA)、超氧化物歧化酶(SOD)的含量.I/R后1 h RTPCR检测肝组织中肿瘤坏死因子α(TNF-α)和细胞间黏附分子-1(ICAM-1)mRNA的表达,I/R后3 h行组织学观察.结果:I/R组、IP组、HHI-Ⅰ组的ALT,AST,LDH活性、MDA值和TNF-α,ICAM-1 mRNA表达水平均明显高于Sham组.IP组、HHI-Ⅰ组低于I/R组.HHI-Ⅰ组所有时间点ALT,AST,LDH明显低于IP组(ALT:2378.8±303.4 nkat/Lvs 2840.6±248.4 nkat/L;AST:2887.2±270.1nkat/L vs 4567.6±275.1 nkat/L;LDH:10550.4±710.1 nkat/L vs 12164.1±735.1 nkat/L;P均＜0.05).HHI-I组MDA值明显低于IP组(17.35±1.39 nmol/g vs 21.66±1.84 nmol/g,P＜0.05).HHI-Ⅰ组TNF-α,ICAM-1 mRNA表达水平低于IP组(TNF-α:0.54±0.06 vs 0.78±0.08;ICAM-1:0.43±0.03 vs 0.69±0.11,P均＜0.01).各组SOD值均低于Sham组(P＜0.05),IP组、HHI-Ⅰ组均高于I/R组(P＜0.05),HHI-Ⅰ组SOD(1,3,6 h)明显高于IP组(136.00±12.50 nmol/g vs 124.70±9.32 nmol/g,P＜0.05).Sham组光镜下肝小叶结构正常;I/R组肝小叶结构紊乱,肝细胞水肿变性;IP组肝细胞水肿明显,部分肝细胞变性;HHI-Ⅰ组肝小叶结构基本正常,肝细胞无明显水肿.结论:HHI-Ⅰ预处理与IP均可改善I/R对肝脏造成的损伤,前者的效果优于后者.HHI-Ⅰ的保护机制可能在于改善肝脏微循环,减轻组织缺氧状态,并通过抑制TNF-α和ICAM-1等细胞因子和细胞黏附分子的转录表达,减少肝组织中中性粒细胞的浸润.     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.