川芎嗪对再灌注大鼠心肌细胞c-fos基因表达及凋亡的影响

目的 :观察川芎嗪 （TMP）对心肌缺血再灌注 （MIR）大鼠心肌 c- fos基因表达及凋亡细胞的作用。方法 :应用免疫组化法检测 MIR不同时期心肌 FOS蛋白的表达 ,并采用末端标记技术对心肌细胞凋亡数进行检测。结果 :1对照组心肌无 FOS蛋白表达 ,亦未见凋亡细胞出现。 2与 MIR组相比 ,TMP干预组心肌 FOS蛋白的表达明显减弱 （6 0 min分别为 5 7± 44个 /片和 6 .4± 3.2个 /片 ,90 m in分别为 46 7± 45 0个 /片和 171± 16 0个 /片 ,12 0 min分别为 2 46± 16 0个 /片和 130± 12 1个 /片 ,均 P<0 .0 5 ） ,TUNEL阳性细胞数均明显减少 （6 0 m in分别为 36 .3± 8.7个 /片和 2 4.7± 7.2个 /片 ,P<0 .0 5 ;12 0 m in分别为 48.0± 2 3.8个 /片和 10 .0± 8.1个 /片 ,P<0 .0 1）。结论 :MIR可诱导终末分化的心肌细胞高表达 FOS蛋白及发生细胞凋亡 ;应用 TMP可使缺血心肌细胞 c- fos基因表达减弱及凋亡减轻     

牛磺酸对新生大鼠心肌细胞模拟缺血/再灌注损伤中细胞凋亡的影响

目的 :研究牛磺酸对培养的心肌细胞模拟缺血 /再灌注 （I/R）过程中心肌细胞凋亡的影响。方法 :培养 SD大鼠乳鼠心肌细胞 ,建立模拟 I/R模型。实验分 3组 :1正常对照组 ;2模拟 I/R组 :缺氧 12 0 min,复氧 2 40 min;3牛磺酸组 :I/R前加用 2 0 mm ol/L 牛磺酸。计算台盼蓝摄取率 ,测定乳酸脱氢酶 （L DH）活性 ,以流式细胞仪 （FCM）和透射电镜观察细胞凋亡。结果 :在正常对照组 ,FCM未发现凋亡峰 ,电镜未发现凋亡细胞 ;I/R组 FCM检测的凋亡率为 15 .1%,电镜下可见到典型的凋亡细胞 ,台盼蓝摄取率和 L DH活性较对照组显著增加 [分别为 （35 .3± 1.7） %vs（2 .6± 0 .8） %和 （184.8± 4.6 ） U/m l vs（4 6 .3± 2 .4） U/ml;两者 P<0 .0 5 ]。牛磺酸组心肌细胞大部分存活 ,凋亡率明显降低 （平均 3.5 %,P<0 .0 5 ）。结论 :细胞凋亡参与了心肌 I/R损伤 ,牛磺酸可减少 I/R心肌细胞的凋亡。     

雄激素对离体大鼠心脏缺血/再灌注心功能及心肌凋亡的影响

目的观察雄激素对离体大鼠缺血/再灌注心脏功能和心肌细胞凋亡的影响。方法SD大鼠42只,随机分为雄激素组（TUD500mg/kg）、对照组、去势组,每组14只。制备大鼠离体缺血/再灌注模型（缺血30min,再灌注2h）,观察左室压力,再灌注结束后检测心肌梗死率和细胞凋亡指数,免疫组化技术检测心肌组织的Bcl-2、Bax和Caspase-3水平。结果①与对照组、去势组相比,雄激素组可以明显改善再灌注后的心脏左室收缩及舒张功能（P<0.05）;②雄激素组心肌梗死范围（31±11）%比对照组（39±8）%和去势组（44±7）%显著减少（均P<0.05）;③雄激素抑制心肌细胞凋亡的发生[（3.7±1.1）%vs（5.8±0.9）%、（6.5±2.3）%,均P<0.05],增加心肌组织中Bcl-2含量,Bax含量未见显著变化,Caspase-3激活减少,有统计学意义（P<0.05）。结论雄激素对离体心脏缺血/再灌注有保护作用,心肌细胞凋亡为其发挥保护作用的机制之一。     

miR-126在大鼠心肌缺血再灌注早期的表达及抗凋亡作用

目的 探讨miR-126在大鼠心肌缺血再灌注（I/R）早期的表达及抗凋亡作用。方法 48只SD大鼠随机分为假手术（Sham）、I/R 2 h、I/R 4 h、I/R 6 h组,每组12只大鼠,比较各组缺血区心肌细胞凋亡蛋白水平及miR-126表达水平。构建重组腺病毒rAAV9-ZsGreen-pre-miR-126载体,24只SD大鼠随机分为：⑤I/R组：大鼠转染空白病毒后I/R 2 h;⑥I/R miR-126组：大鼠转染 rAAV9-ZsGreen-premiR-126 后 I/R 2 h,再次比较各组缺血区心肌细胞凋亡蛋白水平。结果 与Sham组相比,I/R 2 h、4 h、6 h组大鼠心肌缺血区miR-126表达进行性降低（P<0.05）,非缺血区miR-126表达进行性升高（P<0.05）;心肌凋亡蛋白BAX及CASPASE-3表达水平升高（P<0.05）,抗凋亡蛋白BCL-2表达水平降低（P<0.05）;与I/R组相比,I/R miR-126干预组可明显减少心肌凋亡蛋白BAX及CASPASE-3表达水平并增加抗凋亡蛋白BCL-2表达水平（P<0.05）。结论 在I/R早期大鼠心肌缺血区miR-126表达随再灌注时间延长进行性下降,并伴随心肌细胞凋亡进展;过表达miR-126可减少I/R导致的心肌细胞凋亡,发挥保护心功能的作用。     

缺氧与缺氧/复氧诱导心肌细胞凋亡的比较及意义

目的 :研究单纯缺氧与缺氧复氧对体外培养的新生大鼠心肌细胞凋亡的影响 ,探讨凋亡在心肌细胞缺氧 /复氧损伤中的作用。方法 :取体外培养的新生大鼠心肌细胞 ,分两组 ,均置于 95 0 ml/ L N2 ,5 0 ml/ L CO2 孵箱中培养16 ,32 ,4 8h,其中一组缺氧后再恢复正常条件培养 6 h,分别造成缺氧和缺氧 /复氧损伤的细胞模型 ,TUNEL 染色观察凋亡细胞形态学变化 ,流式细胞仪检测心肌细胞凋亡率。结果 :心肌细胞经历缺氧和缺氧 /复氧损伤后 ,TU NEL 染色可见凋亡阳性细胞 ;应用流式细胞仪定量检测 ,心肌细胞缺氧培养 16 ,32 ,4 8h后 ,其凋亡率分别为 :（2 .9± 0 .5 ） % ,（6 .2± 0 .8） %和 （2 6 .6± 3.0 ） % ;心肌细胞在缺氧 16 ,32和 4 8h后复氧 6 h,其凋亡率分别为 :（5 .5± 0 .7） % ,（11.0± 1.1） %和 （14 .2± 1.6 ） %。结论 :心肌细胞凋亡率随着缺氧时间的延长而增高 ;缺氧 /复氧较单纯缺氧可进一步加重心肌细胞的损伤     

缺血再灌注时心肌细胞凋亡情况及去甲肾上腺素预处理对其影响

目的　探讨微量去甲肾上腺素预处理对大鼠缺血再灌注心肌细胞凋亡的影响。方法　大鼠在体缺血再灌注 (I/ R)模型 ,分别以缺血前去甲肾上腺素预处理 (NE- P) ,缺血预处理 (IP) ;采用末端脱氧核苷酸转换酶介导的生物素平移缺口末端标记技术 (TUNEL )检测各组心肌细胞凋亡情况及心肌梗死范围。结果　 I/ R组细胞凋亡率 (43.33%± 4.92 % )较高 ,NE- P组及 IP组虽然也有一定的心肌细胞凋亡率 :2 5.2 4 %± 1 .56% ,2 4 .44%± 2 .96% ,但较 I/ R组明显降低 (P<0 .0 0 1 )。 IP组及 NE- P组心肌梗死范围较 I/ R组明显减少 ,IP组及 NE- P组两项指标无显著差异。结论　心肌缺血再灌注损伤可诱发心肌细胞凋亡 ,NE- P能明显减少缺血再灌注诱导的心肌细胞凋亡的发生率 ,能明显减少心肌梗死范围 ,减轻缺血再灌注损伤 ;NE- P减少心肌梗死范围、减轻缺血再灌注损伤的机制可能与其能明显减少心肌细胞凋亡有关。     

极化液给予时机对缺血/再灌注犬心脏功能及心肌损伤的影响

目的:探讨不同时间开始给予极化液（葡萄糖-胰岛素-钾液,G IK）对犬心肌缺血/再灌注（M I/R）后心脏功能及心肌细胞损伤的影响。方法:制备犬M I/R模型,心肌定量缺血（左前降支血流量降低80%）50 m in,再灌注4 h。分别于:①再灌注前30 m in（再灌注前）、②再灌注即刻（再灌注时）、③再灌注后1 h（再灌注后）给予G IK。观察对动脉血压、心率、左室压的影响及测定心大静脉血清乳酸脱氢酶（LDH）、肌酸激酶（CK）活性,再灌注结束后检测心肌梗死率或细胞凋亡指数。结果:与再灌注后组相比,再灌注前组和再灌注时组可明显改善再灌注后左室收缩及舒张功能,降低血清CK、LDH活性,减少心肌梗死范围[分别为（5.9±1.1）%和（5.2±0.8）%vs（9.1±1.2）%,均P<0.01]、抑制心肌细胞凋亡的发生[（4.6±0.9）%和（3.7±1.1）%vs（8.9±2.3）%,均P<0.05]。结论:再灌注早期给予G IK可降低M I/R引起的心肌细胞损伤,促进再灌注后心脏功能的恢复,在再灌注后1 h给予G IK对心脏的上述保护作用则明显减弱。     

依达拉奉对大鼠心肌再灌注抗凋亡作用的机制

目的：观察大鼠心肌缺血再灌注时依达拉奉抗凋亡作用的机制。方法：采用体外结扎大鼠左前降支制备大鼠心肌缺血再灌注模型。将Wistar大鼠随机分为对照组、缺血再灌注组、依达拉奉组（再灌注即刻经尾静脉注射依达拉奉3mg／kg）。分别用TUNEL染色法及免疫组化法检测心肌细胞凋亡、心肌凋亡蛋白Bcl-2、Bax的表达。结果：（1）凋亡检测结果示,对照组无细胞凋亡,依达拉奉组较缺血再灌注组凋亡细胞数明品减少[（13．11±1．58）比（24．33±1．38）,P〈0．01];（2）免疫组化结果示,依达拉奉组与缺血再灌注组Bcl-2、Bax蛋白表达均明显高于对照组（P〈0．01）;依达拉奉组较缺血再灌注组Bcl-2蛋白表达明显升高[（0．2300±0．0218）比（0．0924±0．0152）],而Bax蛋白表达明显降低[（0．0249±0．0042）比（O．1312±0．0173）],P均〈0．01;缺血再灌注组Bcl-2／Bax比值较对照组明显降低[（0．6906±0．0035）比（1．1632±0．0212）],依达拉奉组Bcl-2／Bax比值（7．4752±0．5573）较缺血再灌注组和对照组明显升高（P〈0．01）。结论：依达拉奉能够减少心肌细胞凋亡,心肌细胞凋亡的减轻与下调Bax蛋白表达,上调Bcl-2蛋白表达以及Bcl-2／Bax比值有关。     

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     

灯盏花素对大鼠心肌缺血再灌注损伤诱导细胞凋亡的影响

目的观察灯盏花素（BRE）对大鼠心肌缺血再灌注（I/R）诱导细胞凋亡及凋亡相关基因Bcl-2、Bax表达的影响,并对其作用机制进行初步探讨。方法30只SD大鼠随机分为3组（n=10）:①假手术（Sham）组:前降支穿线不结扎,观察1.5 h;②缺血再灌注（I/R）组:结扎前降支30 m in,再灌注前5 m in生理盐水（4 mg/kg）iv,再灌注1h;③BRE组:再灌注前5 m in应用BRE（4 mg/kg）iv,余同I/R组。常规方法检测丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性,免疫组化法测定Bc l-2、Bax蛋白的表达,通过细胞图像分析系统测量细胞阳性染色面积百分率,应用透射电镜观察心肌超微结构的变化。结果同I/R组相比,BRE组心肌组织MDA含量显著降低[（7.56±1.67）μmol/gvs（16.58±2.59）μmol/g,P<0.05],SOD活性显著升高[（2.37±0.22）mkat/gvs1.38±0.14 mkat/g,P<0.05],Bax蛋白表达显著降低（5.02±1.01vs18.59±5.68,P<0.01）,而Bc l-2蛋白表达显著增高（17.48±4.72vs7.32±2.05,P<0.01）,Bax/Bc l-2显著降低（0.29±0.57vs3.32±0.10,P<0.01）,细胞阳性染色面积百分率显著减少（0.42±0.06vs13.84±1.97,P<0.01）,细胞超微结构损伤明显减轻。结论BRE通过调控Bax/Bc l-2表达抑制心肌I/R诱导的细胞凋亡,对缺血再灌注心肌具有保护作用。     

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.