阿托伐他汀对Livin、NF-κB与IL-1β在脑缺血再灌注大鼠脑组织中表达的影响

目的 检测脑缺血再灌注大鼠脑组织Livin、NF-κB和白介素(IL)-1β表达及观察阿托伐他汀对其表达的影响,探讨阿托伐他汀对脑缺血再灌注后的脑保护作用机制.方法 大脑中动脉线栓方法制作大鼠脑缺血再灌注模型;实验大鼠随机分为假手术组(S组)、模型组(M组)及阿托伐他汀治疗组(MT组);免疫印迹法和RT-PCR检测大鼠脑组织Livin、NF-κB和IL-1β的表达,TTC染色法测量脑梗死体积.结果 梗死灶体积MT组较M组显著降低(P<0.05), 较S组显著增高(P<0.01);MT组NF-κB和IL-1β表达较M组降低(P<0.05),MT组Livin表达较M组增高(P<0.05);MT组Livin、NF-κB和IL-1β表达较S组增高(P<0.05);M组Livin、NF-κB和IL-1β表达较S组显著增高(P<0.01).结论 阿托伐他汀可以通过降低脑缺血再灌注大鼠脑组织NF-κB和IL-1β、增强Livin表达抑制细胞凋亡和脑内炎症发挥脑保护作用.     

脑缺血再灌注后实验大鼠核因子kappaB的变化

目的研究大鼠局灶性脑缺血再灌注后核因子kappaB(NF-κB)在时间上表达的变化过程。方法线栓法建立右侧大脑中动脉闭塞大鼠模型,分别于缺血90min后再灌注2h,6h,12h,24h,72h,大鼠在各时间点断头取脑,测算脑梗死体积,用免疫印迹法检测脑组织胞核内NF-κB的表达。结果脑缺血再灌注后NF-κB的表达在6h出现明显升高(P<0.01),在24h达到高峰,72h略有下降,其脑梗死体积随着再灌注时间延长而增加,脑梗死体积与NF-κB表达水平之间有显著的相关性。结论脑缺血再灌注后,脑梗死区域出现胞核内NF-κB表达增加,NF-κB在再灌注损伤中起重要作用。     

眼针对急性脑缺血再灌注损伤大鼠海马组织核因子-κBp65、血管细胞黏附分子-1表达细胞黏附分子影响

目的观察眼针对脑缺血再灌注损伤大鼠海马组织核因子(NF)-κB p65、血管细胞黏附分子(VCAM)-1表达的影响,探讨眼针对急性脑缺血再灌注损伤的神经保护作用。方法将40只SD大鼠随机分为正常组、假手术组、模型组、眼针治疗组,采用改良线栓法制备局灶性脑缺血(MCAO)再灌模型,于再灌注后24 h进行神经功能缺损评分,运用免疫组化检测法,检测海马组织NF-κB p65、VCAM-1。结果与模型组比较,眼针组大鼠神经功能缺损评分明显下降;海马组织NF-κB p65、VCAM-1蛋白表达显著下降(P<0.05)。结论眼针能通过下调脑缺血再灌注损伤的NF-κB p65、VCAM-1蛋白水平,发挥神经保护作用。     

糖尿病通过炎症反应加重大鼠脑缺血再灌注损伤

目的：探讨肿瘤坏死因子-α（tumornecrosisfactor-α,TNF-α）和核因子-κB（nuclear factor-κB, NF-κB）在糖尿病大鼠缺血再灌注损伤中的作用。方法36只健康雄性Sprague-Daw ley大鼠按随机数字表法分为正常血糖假手术组、正常血糖脑缺血再灌注组和糖尿病脑缺血再灌注组,每组12只。采用腹腔注射链脲佐菌素制作大鼠糖尿病模型,然后应用栓线法建立局灶性脑缺血再灌注模型,再灌注24 h行神经功能缺损评分,然后2,3,5-氯化二苯四氮唑染色检测脑梗死面积,蛋白质印迹法检测缺血侧皮质NF-κB和TNF-α表达水平。结果正常血糖假手术组、正常血糖脑缺血再灌注组和糖尿病脑缺血再灌注组神经功能评分分别为（0.00±0.00）分、（2.50±1.08）分和（3.20±1.03）分,差异有统计学意义（F=38.015,P<0.001）,且糖尿病脑缺血再灌注组神经功能缺损评分较正常血糖脑缺血再灌注组显著性加重（ P<0.05）。正常血糖假手术组、正常血糖脑缺血再灌注组和糖尿病脑缺血再灌注组梗死面积分别为（0.00±0.00）％、（33.09±5.17）％和（55.45±9.29）％,各组间差异有统计学意义（F=206.614,P<0.001）,其中糖尿病脑缺血再灌注组梗死面积较正常血糖脑缺血再灌注组显著性增大（P<0.05）。再灌注后24 h,各组缺血侧皮质NF-κB（F=29.993,P<0.001）和TNF-α（F=28.722,P<0.001）表达水平差异有统计学意义,其中糖尿病脑缺血再灌注组NF-κB和TNF-α表达水平较正常血糖脑缺血再灌注组显著性增高（P均<0.05）。结论糖尿病会加重脑缺血再灌注损伤,TNF-α和NF-κB表达上调可能是糖尿病加重脑缺血再灌注损伤的机制之一。     

NF-κB调节TNF-α表达减轻大鼠缺血性脑血管病脑组织损伤

目的研究缺血性脑血管病(ICVD)核因子(NF)-κB对肿瘤坏死因子(TNF)-α表达的影响及其对大鼠脑组织损伤的作用。方法将48只Wistar大鼠随机分成正常组(12只)、模型组(12只)、假手术组(12只)和吡咯烷二硫基甲酸酯(PDTC)+模型组,每组12只。采用线栓法制成大鼠局部缺血再灌注模型,用Ludmila Belayev 12分评分法评估大鼠神经功能,氯化-2,3,5-三苯基四氮唑(TTC)染色法观察脑梗死体积,免疫组化法检测脑组织NF-κB表达,酶联免疫吸附法(ELISA)测脑组织TNF-α含量。结果模型组大鼠Ludmila Belayev 12得分明显高于正常组与假手术组(P<0.05),PDTC+模型组得分显著低于模型组(P<0.01);模型组大鼠的脑梗死体积显著多于正常组和假手术组(P<0.01),与模型组比较,PDTC+模型组大鼠脑梗死体积显著减少(P<0.05);模型组大鼠脑组织NF-κB阳性细胞表达率显著高于正常组和假手术组(P<0.05),DPTC+模型组NF-κB表达明显低于模型组(P<0.05);模型组大鼠TNF-α的含量显著高于正常组和假手术组(P<0.01),PDTC+模型组TNF-α含量明显低于模型组(P<0.05)。结论 PDTC抑制NF-κB活性可以减少TNF-α含量,降低TNF-α的表达,所以可通过NF-κB调节TNF-α表达对缺血再灌注脑损伤起到保护作用,可能与其抑制炎症反应作用有关。     

丹红注射液对脑缺血再灌注损伤大鼠皮质区新生微血管的影响研究

背景新生血管形成是最终的侧支代偿途径,探究丹红注射液对新生血管的影响对其作用机制研究具有一定价值。目的探究丹红注射液对脑缺血再灌注损伤大鼠皮质区新生微血管的影响。方法2018年12月—2019年6月,将84只雄性SD大鼠随机分为假手术组(n=12)、造模组(n=36)及丹红组(n=36)。参照改良LONGA线栓法制备脑缺血再灌注损伤大鼠模型,其中造模组和丹红组大鼠制备脑缺血再灌注损伤模型,假手术组大鼠不插线栓;丹红组大鼠于拔出线栓后腹腔注射丹红注射液,造模组和假手术组大鼠均于相同时间点腹腔注射等量0.9%氯化钠溶液。比较三组大鼠造模后24 h、72 h、7 d神经行为学评分、脑梗死体积及脑缺血再灌注皮质区新生微血管数目。结果(1)假手术组大鼠造模后24 h、72 h及7 d神经行为学评分均为0。造模组和丹红组大鼠造模后24 h、72 h神经行为学评分比较,差异无统计学意义(P>0.05);丹红组大鼠造模后7 d神经行为学评分低于造模组(P<0.05)。(2)假手术组大鼠造模后24 h、72 h及7 d脑梗死体积均为0。丹红组大鼠造模后24 h、72 h及7 d脑梗死体积小于造模组(P<0.05)。(3)造模组和丹红组大鼠造模后24 h脑缺血再灌注皮质区新生微血管数目多于假手术组(P<0.05);造模组和丹红组大鼠造模后72 h及7 d脑缺血再灌注皮质区新生微血管数目多于假手术组,丹红组大鼠脑缺血再灌注皮质区新生微血管数目多于造模组(P<0.05)。结论丹红注射液可有效增加脑缺血再灌注损伤大鼠皮质区新生微血管数量,改善脑侧支循环,进而减轻神经功能缺损、缩小脑梗死体积。     

组织型激肽释放酶对脑缺血再灌注大鼠细胞凋亡和炎性反应的影响

目的探讨组织型激肽释放酶(TK)对脑缺血再灌注大鼠的治疗作用和对细胞凋亡、炎性反应的影响。方法将36只SD大鼠随机分为3组,A组:假手术;B组:生理盐水2ml·kg-1·d-1干预;C组:脑缺血再灌注后8h给予TK17.5×10-3U·kg-1·d-1治疗,每组12只。连续给药3天后分别进行神经功能缺损评分,脑梗死体积测定,梗死组织HE染色。对脑梗死组织进行TUNEL染色和细胞凋亡蛋白酶-3(caspase-3)、TNF-α、白细胞介素-1(IL-1)免疫组织化学检测。结果与A组和B组比较,C组能明显降低神经功能缺损,使脑梗死的体积变小(P<0.05);减轻脑梗死的病理改变;使缺血区内TUNEL、caspase-3、TNF-α、IL-1阳性细胞减少(P<0.05)。结论TK对脑缺血再灌注大鼠有治疗作用,能明显减轻缺血区内的细胞凋亡和炎性反应。     

血管性痴呆大鼠海马神经元核因子-κB与环氧合酶-2水平的动态变化

目的研究核因子(NF)-κB与环氧合酶(COX)-2在血管性痴呆(VD)大鼠海马的动态变化。方法将实验大鼠随机分为假手术组,全脑缺血再灌注30 min、2、6、24、48、7 d、15 d组。采用大鼠全脑缺血再灌注VD模型,Morris水迷宫检测空间学习记忆能力、HE染色观察大鼠海马神经元形态及数目、免疫组织化学染色法检测海马神经元NF-κB蛋白表达、Western印迹检测COX-2蛋白表达。结果与假手术组比较,VD组大鼠寻台潜伏期显著增加(P<0.05),海马神经元凋亡30 min开始出现(P<0.05),7 d达到峰值(P<0.01)。NF-κB蛋白表达30 min开始升高,24 h达到峰值,15 d仍明显高于假手术组(P<0.01)。COX-2蛋白表达2 h开始升高,7 d达到峰值,15 d仍高于假手术组(P<0.01)。结论 NF-κB活化可能上调VD大鼠海马神经元中COX-2表达水平,进而参与脑损伤病理生理过程的调节。     

兔脊髓缺血再灌注后小胶质细胞活化及炎性细胞因子变化的研究

目的:观察兔脊髓缺血再灌注后小胶质细胞活化及炎性细胞因子白细胞介素(IL)-6、IL-10、核转录因子(NF)-κB的变化规律,为后处理干预时机提供理论依据。方法:采用胸主动脉球囊阻断法建立兔脊髓缺血再灌注损伤模型。36周健康成年雄性新西兰大白兔54只,假手术组(n=6只)只置入球囊不阻断;48只脊髓缺血再灌注家兔按再灌注时间点分为8组:再灌注0h组、再灌注1h组、再灌注2h组、再灌注3h组、再灌注8h组、再灌注24h组、再灌注48h组、再灌注72h组,每组6只。分别于再灌注后0h、1h、2h、3h、8h、24h、48h和72h检测缺血段脊髓组织中正常神经元、凋亡神经元以及离子钙结合接头分子-1(Iba-1)、IL-6、IL-10、NF-κB的表达水平。结果:正常神经元数量随再灌注时间延长而减少;脊髓缺血再灌注损伤后8h原位末端转移酶标记(TUNEL)阳性神经元开始增多,再灌注24h组的TUNEL阳性神经元达高峰。再灌注2h组的Iba-1表达开始增多,再灌注8h组Iba-1表达达高峰;NF-κB于再灌注3h组开始增高,再灌注8h组NF-κB表达高峰;IL-6和IL-10表达均在再灌注24h组达高峰。脊髓缺血再灌注后NF-κB、IL-6、IL-10的表达水平与Iba-1呈相近的变化趋势。结论:脊髓缺血再灌注后小胶质细胞激活呈动态变化。NF-κB、IL-6、IL-10的表达水平与小胶质细胞激活显著正相关,在小胶质细胞激活前给予后处理可降低神经元损伤。     

吡格列酮通过下调Toll样受体/核因子-κB信号通路对大鼠局灶性脑缺血再灌注损伤的保护作用

目的研究过氧化物酶体增殖物激活受体(PPAR)γ激动剂吡格列酮对大鼠局灶性脑缺血再灌注损伤后脑组织中Toll样受体(TLR)4和核转录因子(NF)-κB及对血清中白细胞介素(IL)-6和肿瘤坏死因子(TNF)-α的影响,探讨吡格列酮对脑组织损伤的保护作用和机制。方法用线栓法制备大鼠中动脉脑缺血再灌注模型。随机将60只雄性SD大鼠分为假手术组、模型组、生理盐水干预组、吡格列酮(15 mg/kg)治疗组。脑缺血再灌注24 h后,采用神经功能缺损评分法,观察大鼠的行为学评分;氯化三苯基四氮唑(TTC)染色试验测定脑梗死体积;酶联免疫吸附(ELASA)技术测定血清中IL-6和TNF-α含量;免疫组化技术测定脑组织TLR4和NF-κB蛋白表达;逆转录聚合酶链反应(RT-PCR)技术测定脑组织TLR4和NF-κB mRNA表达。结果与生理盐水干预组相比,吡格列酮治疗组神经功能评分显著降低(P0.05),脑梗死体积显著减少(P0.05),血清中IL-6和TNF-α含量显著降低(P0.05),脑组织中TLR4和NF-κB蛋白和mRNA表达显著下调(P0.05)。结论吡格列酮对大鼠局灶性脑缺血再灌注损伤有一定的保护作用,作用机制可能与其下调TLR4/NF-κB信号转导通路并降低血中炎症反应相关因子IL-6和TNF-α的含量有关。     

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.     

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.     

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.