氯沙坦和卡维地洛对缺血再灌注心肌细胞凋亡的影响

目的 :研究氯沙坦和卡维地洛对缺血再灌注心肌细胞凋亡的影响 ,比较氯沙坦和卡维地洛对缺血再灌注心肌损伤的保护作用。方法 :结扎Wistar大鼠左冠状动脉前降支 ,建立大鼠缺血再灌注动物模型 ,采用原位末端标记细胞凋亡法检测心肌细胞凋亡 ,并利用光学显微镜进行细胞计数。结果 :单纯缺血 再灌注组心肌细胞凋亡数较假手术组明显增多 (37.5 3± 9.2 2 /视野∶0 .18± 0 .0 9/视野 ,P <0 .0 5 ) ,氯沙坦和卡维地洛组心肌细胞凋亡数分别为 8.74± 3.5 1/视野和 7.6 3± 4 .0 5 /视野 ,较单纯缺血 再灌注组明显减少 (P <0 .0 5 ) ,氯沙坦和卡维地洛两组间无明显区别 (P >0 .0 5 )。结论 :氯沙坦和卡维地洛对缺血再灌注心肌细胞损伤具有相似的保护作用 ,可明显减少缺血再灌注心肌细胞凋亡     

丹参注射液对大鼠心肌缺血再灌注时心肌细胞凋亡的影响

目的 :研究丹参注射液对大鼠心肌缺血再灌注时心肌细胞凋亡的影响。方法 :用在体左冠状动脉前降支穿线结扎法制备心肌缺血再灌注模型。 6 0只SD大鼠分为 3组 :假手术组 (10只 ) ,缺血再灌注组 (2 5只 ) ,丹参注射液干预组 (2 5只 )。缺血 30min ,再灌注 12 0min。采用缺口末端标记法 (TUNEL)以及S P免疫组化法 ,检测心肌细胞凋亡指数 (AI)、Bcl 2、Bax基因蛋白表达。结果 :假手术组AI 2 .2± 0 .6 ,Bcl 2OD值 0 .112± 0 .0 18,BaxOD值 0 .10 2± 0 .0 31;缺血再灌注组AI 2 8.9± 3.6 ,Bcl 2OD值 0 .0 92± 0 .0 14 ,BaxOD值 0 .136± 0 .0 2 2 ;丹参注射液干预组AI 2 6 .2± 3.4 ,Bcl 2OD值 0 .10 4± 0 .0 2 1,BaxOD值 0 .12 2± 0 .0 12。与假手术组比较 ,缺血再灌注组AI增加 ,Bax阳性表达增强 ,Bcl 2阳性表达减弱 (P <0 .0 1) ;与缺血再灌注组比较丹参注射液干预组 ,AI、Bax阳性表达下降 (P <0 .0 1) ,Bcl 2阳性表达上调 (P <0 .0 5 )。结论 :丹参注射液通过抑制心肌细胞凋亡 ,下调Bax基因蛋白表达 ,上调Bcl 2基因蛋白表达 ,起到心肌缺血再灌注损伤的保护作用。     

人参皂甙Re对大鼠缺血再灌注心肌细胞凋亡及Fas基因表达的影响

目的 :观察人参皂甙Re(以下简称Re)对缺血再灌注心肌细胞凋亡及Fas基因表达的影响 ,探讨Re抑制心肌细胞凋亡的可能机制。方法 :结扎Wistar大鼠左冠状动脉前降支 ,建立大鼠缺血再灌注动物模型 ;采用透射电镜、缺口末端标记法检测心肌凋亡细胞 ,利用光学显微镜进行细胞计数 ;原位杂交及免疫组化分别检测Fas基因mRNA及蛋白的表达 ,并利用图像分析系统测量阳性表达区域平均光密度值 ,进行定量分析。结果 :①透射电镜发现缺血再灌注组缺血区出现心肌凋亡细胞 ,假手术组未发现心肌凋亡细胞 ;②缺血再灌注组心肌细胞凋亡数为 (134.4 5± 4 5 .6 1)个 /视野 ,Re治疗组细胞凋亡数 (90 .6 6± 19.2 2 )个 /视野 ,两组间差异有非常显著性意义 (P<0 .0 1) ;③原位杂交及免疫组化检测均发现Fas基因的表达缺血再灌注组较假手术组明显增加 (P <0 .0 1) ,Re治疗组较缺血再灌注组明显下降 (P <0 .0 5 )。结论 :心肌缺血再灌注诱导心肌细胞凋亡 ,Re治疗则可以显著减少缺血再灌注心肌细胞的凋亡。Re通过抑制Fas基因的表达而抑制心肌细胞凋亡     

血管紧张素Ⅱ及血管紧张素Ⅱ1型受体反义寡核苷酸对心肌细胞bax、bcl-2基因表达

目的　探讨血管紧张素 (AngⅡ )及血管紧张素Ⅱ 1型受体 (AT1R)反义寡核苷酸 (AS ODN)对心肌细胞凋亡调节蛋白bax、bcl 2的影响。方法　将培养的乳鼠心肌细胞分为正常组、AngⅡ组、AT1R AS ODN组 :AngⅡ组用 10 - 6 mol/LAngⅡ刺激 2 4小时 ;AT1R AS ODN组在转染反义寡核苷酸后也用 10 - 6 mol/LAngⅡ刺激 2 4小时 ;正常组不予任何刺激。刺激 2 4小时后用免疫细胞化学方法观察AngⅡ及AT1R AS ODN对心肌细胞凋亡调节蛋白bcl 2、bax表达的影响。结果　与正常组相比 ,AngⅡ组心肌细胞bcl 2蛋白光密度值下降显著 (0 0 72 5± 0 0 0 65vs 0 0 975± 0 0 0 83 ,P <0 0 5) ,bax蛋白光密度值增加 (0 0 941± 0 0 0 42vs 0 0 82 3± 0 0 0 2 4,P <0 0 5) ;与AngⅡ组相比 ,AT1R AS ODN组bcl 2蛋白光密度值增加 (0 0 90 0± 0 0 0 16vs 0 0 72 5± 0 0 0 65,P <0 0 5) ,bax蛋白光密度值降低 (0 0 863± 0 0 0 19vs 0 0 941±0 0 0 42 ,P <0 0 5)。结论　AngⅡ可以诱导心肌细胞发生凋亡 ,而AT1R AS ODN可以逆转AngⅡ诱导的心肌细胞凋亡。     

缺血-再灌注诱导家兔房室结细胞凋亡的研究

为探讨缺血 再灌注对在体家兔房室结细胞凋亡的作用 ,以及凋亡相关调控基因蛋白 (Fas、bcl 2、bax)的表达 ,以家兔为实验对象 ,麻醉后开胸暴露心脏 ,结扎右冠状动脉 ,建立房室结缺血 再灌注模型。于预定时间处死动物 ,快速取出房室交界区组织 (3mm× 2mm× 1mm) ,10 %福尔马林磷酸盐缓冲液固定。采用TUNEL法检测房室结细胞凋亡 ;免疫组化法检测Fas、bcl 2、bax基因蛋白表达。结果 :①对照组及单纯缺血 10 ,30min组均未检测到细胞凋亡 ,缺血 6 0min组可见少量散在的凋亡细胞 ,缺血 12 0min房室结可见大量的细胞凋亡 ;②缺血 再灌注组 :不同缺血时间组均出现细胞凋亡 ,凋亡细胞数随缺血时间的延长而增加 (P <0 .0 5 ) ;③随缺血时间的延长 ,Fas、bax的表达明显增加 (P <0 .0 1) ,而bcl 2的表达则无明显增加 (P >0 .0 5 ) ;④缺血 再灌注组较相应时间单纯缺血组细胞凋亡数、Fas、bax表达均明显增加 (P <0 .0 1)。结论 :单纯缺血及缺血 再灌注均可诱导房室结细胞发生凋亡 ,这可能与该条件下Fas、bax基因蛋白的过量表达有关。     

卡维地洛和美托洛尔对大鼠缺血再灌注损伤心肌细胞凋亡的影响

目的观察卡维地洛和美托洛尔对大鼠缺血再灌注损伤心肌细胞凋亡及caspase-3、p53的影响。方法取SD大鼠32只,随机分为假手术组、I/R组、卡维地洛组、美托洛尔组,每组8只。通过结扎和放松大鼠左冠状动脉前降支(LAD)造成心肌缺血再灌注损伤。采用末端标记原位细胞凋亡法检测心肌凋亡细胞,采用免疫组化法测caspase-3、p53蛋白表达。结果 I/R组caspase-3、p53表达和心肌细胞凋亡指数较假手术组明显升高(P<0.05),卡维地洛组和美托洛尔组各项指标较I/R组明显降低(P<0.05),卡维地洛组各指标较美托洛尔组明显降低(P<0.05)。结论 p53和caspase-3在缺血诱导的心肌细胞凋亡中发挥重要作用。卡维地洛和美托洛尔均可抑制心肌细胞凋亡,对缺血再灌注损伤心肌细胞有保护作用,且卡维地洛作用优于美托洛尔。     

黄芪注射液对肠缺血再灌注肝损伤大鼠bcl-2、Bax蛋白表达影响的实验研究

目的　探讨大鼠肠缺血再灌注肝损伤后肝组织bcl 2、Bax蛋白的表达及黄芪注射液对其表达的影响。方法　通过夹闭肠系膜上动脉建立肠缺血再灌注肝损伤模型 ,采用免疫组织化学方法与医学图像分析检测假手术组、模型组及黄芪注射液组大鼠肝组织bcl 2、Bax蛋白表达的阳性表达指数。结果　①与假手术组比较 ,模型组肝组织bcl 2、Bax蛋白的阳性表达指数升高 (P <0 0 1)。②与模型组比较 ,黄芪注射液组肝组织bcl 2蛋白的阳性表达指数升高 (P <0 0 1) ,Bax蛋白的阳性表达指数降低 (P <0 0 1)。结论　大鼠肠缺血再灌注肝损伤后肝组织bcl 2、Bax的表达明显升高 ;黄芪注射可增强bcl 2的表达、抑制Bax的表达。这可能是黄芪防护大鼠肠缺血再灌注肝损伤的机理之一。     

缺血后处理对心肌缺血再灌注损伤过程中内质网应激的影响研究

目的 探讨缺血后处理(IPostC)对心肌缺血再灌注损伤(MIRI)中细胞凋亡的影响,以及特异性内质网应激(ERS)损伤相关蛋白Grp78(葡萄糖调节蛋白78)和Caspase 12(半胱氨酸蛋白酶12)的蛋白表达水平的变化和意义。方法 Wistar 大鼠24只随机分为假手术组、I/R组(缺血再灌注组)、IPostC组(缺血后处理组)3组,每组8只。分别测定各组心肌缺血和梗死面积、细胞凋亡指数和Caspase12、 Grp78蛋白表达检测。结果 假手术组未见梗死心肌和缺血心肌,IPostC组心肌缺血区面积(46.46±2.13)%和梗死区面积(41.02±2.93)%均明显小于I/R组(53.31±3.87, 52.19±3.44)%,P值均<0.01。假手术组心肌细胞凋亡指数(6.70±2.25)%明显低于I/R组(26.92±1.91)% 、IPostC组(20.54±3.05)%,P值均<0.001。心肌组织Caspase12蛋白表达水平在假手术组(0.11±0.01)明显低于I/R组(0.41±0.06)和IPostC组(0.35±0.06),并且IPostC组低于I/R组,P值均<0.01;心肌组织Grp78蛋白水平在假手术组(0.13±0.03)亦明显低于I/R组(1.04±0.16)和IPostC组(1.17±0.14),并且IPostC组高于I/R组,P值均<0.01。结论 本研究从动物实验证实IPostC能减轻心肌细胞凋亡,而ERS激活参与了大鼠MIRI过程,推测IPostC在大鼠MIRI过程中可能通过调节ERS途径抑制细胞凋亡,改善MIRI。     

衰老大鼠心肌对缺血再灌注损伤的敏感性研究

目的观察衰老大鼠心肌对缺血再灌注损伤的敏感性,并探讨其可能的机制。方法成年和老年雄性Wistar大鼠各12只,分为4组：成年假手术组、成年缺血再灌注组、老年假手术组和老年缺血再灌注组,每组6只,进行缺血30 min再灌注3 h。脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测心肌细胞凋亡,比色法检测心肌组织半胱胺酸蛋白酶蛋白-3 (caspase-3)活性,化学发光法测定心肌组织总一氧化氮(NOx)含量,ELISA法检测心肌过氧亚硝基(ONOO~-)含量。结果老年组缺血再灌注引起心肌细胞凋亡的程度明显高于成年组,凋亡指数分别为(14．6±1.7)%、(19．0±2．1)%,差异有统计学意义(P＜0．05)；caspase-3活性：成年组为(340±32)μmol/mg,老年组为(436±35)μmol/mg,差异有统计学意义(P＜0．05)；心肌组织中NOx含量：成年缺血再灌注组、老年缺血再灌注组分别为成年假手术组的(2．1±0．2)、(4．4±0．5)倍,差异有统计学意义(P＜0．05)；ONOO~-含量：成年缺血再灌注组和老年缺血再灌注组分别为(4．68±0．15)nmol/g、(7．25±0．18)nmol/g,差异有统计学意义(P＜0．05)。结论老年大鼠对心肌缺血再灌注损伤的敏感性增加,可能的原因是老年鼠心脏中NO的毒性衍生物ONOO~-含量增加,从而导致功能蛋白的硝基化。     

倒卵叶五加总皂甙对大鼠心肌缺血/再灌注损伤后细胞凋亡的作用及机制

目的　探讨倒卵叶五加总皂甙 (SAOH)对大鼠缺血心肌再灌注后细胞凋亡的作用。方法　采用结扎左冠状动脉前降支再灌注复制大鼠心肌缺血再灌注损伤 (IRI)的模型 ,结扎前 10 min舌下静脉注射 SAOH,测定心肌缺血3 0 min后再灌注 6h心肌组织丙二醛 (MDA)和血浆超氧化歧化酶 (SOD)的含量 ,并用末端脱氧核苷酸转移酶介导的带生物素的 d UTP缺口末端标记 (TUNEL)的方法及免疫组化观察左心室心肌细胞凋亡及凋亡相关基因 Bcl- 2和 Bax的蛋白表达。结果　再灌注 6h后 ,SAOH1 组 (5 0 mg/ kg)及 SAOH2 组 (10 0 mg/ kg)较 IRI组 ,心肌组织 MDA含量明显减少 [(4.89± 0 .2 6)、(3 .5 7± 0 .2 1) nm ol/ mg· pro vs(6.97± 0 .3 4) nmol/ mg· pro,P值均 <0 .0 1];血浆 SOD活性明显提高 [(9.88± 0 .18)、(11.40± 0 .3 7)μIU / L vs (3 .3 9± 0 .2 4)μIU / L ,P值均 <0 .0 1];心肌细胞凋亡指数 (AI)显著减少 [(14.67± 3 .0 5 ) %、(7.49± 2 .62 ) % vs(2 1.96± 7.68) % ,P值均 <0 .0 1];Bax蛋白表达明显减少 [(7.73± 1.84) %、(5 .0 2± 1.0 1) % vs (18.5 9± 5 .68) % ,P值均 <0 .0 1];而心肌 Bcl- 2蛋白的表达显著增加 [(12 .5 1± 3 .42 ) %、(17.48± 4.72 ) % vs(7.3 2± 2 .0 5 ) % ,P值 <0 .0 5 ,<0     

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.     

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.     

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.     

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.