石菖蒲有效成分对抑郁模型大鼠海马神经元的保护作用

目的探讨石菖蒲有效成分β-细辛醚对抑郁模型大鼠海马神经元细胞凋亡的保护作用。方法将大鼠随机分为正常对照组、模型组、西药对照组、中药组共四组;采用慢性不可预见性应激刺激结合孤养方式复制抑郁模型;通过敞箱实验与糖水偏爱度检测大鼠行为学改变;应用免疫组化与RT-PCR检测大鼠海马区转录因子环磷酸腺苷反应元件结合蛋白(CREB)及基因的表达水平。结果与正常对照组相比,模型组水平运动与垂直运动得分和糖水偏爱度明显降低(P<0.01⁰.001);与模型组比较,西药对照组和中药组的水平运动与垂直运动得分和糖水偏爱度均明显升高(P<0.050.001);与模型组比较,西药对照组和中药组的水平运动与垂直运动得分和糖水偏爱度均明显升高(P<0.05⁰.001);与正常对照组相比,模型组大鼠海马区CREB mRNA与蛋白表达明显下调(P<0.05);与模型组相比,西药对照组、中药组海马区CREB mRNA与蛋白表达明显上调(P<0.05)。结论β-细辛醚抗抑郁症的作用机制可能与其促进海马区CREB蛋白与mRNA的活性和表达,进而减少神经细胞凋亡有关。     

柴胡皂苷A对抑郁模型大鼠海马神经细胞凋亡的保护作用

目的探讨柴胡皂苷A对抑郁模型大鼠海马神经细胞凋亡的保护作用及机制。方法 60只SD雄性大鼠随机分为正常对照组、模型组、西药组、中药组;采用慢性不可预见性应激刺激结合孤养方式建立抑郁模型;利用敞箱实验与糖水偏爱度检测大鼠行为学改变;采用电镜观察大鼠脑海马区超微结构变化;应用免疫组化与RT-PCR检测大鼠海马区脑源性神经生长因子(BDNF)蛋白及基因的表达水平。结果与正常对照组相比,模型组水平运动得分、垂直运动得分和糖水偏爱度明显降低(P<0.01,P<0.001),与模型组比较,西药组和中药组水平运动得分、垂直运动得分和糖水偏爱度均明显升高(P<0.05,P<0.001);与正常对照组相比,模型组大鼠海马区BDNF mRNA与蛋白表达明显下调(P<0.05),与模型组相比,西药组、中药组海马区BDNF mRNA与蛋白表达明显上调(P<0.05)。结论柴胡皂苷A抗抑郁症的作用机制可能与其促进海马区BDNF蛋白与mRNA的活性和表达,进而减少神经细胞凋亡有关。     

柴胡疏肝散对抑郁症模型大鼠海马单胺氧化酶与乙酰胆碱酯酶活性的影响

目的 观察柴胡疏肝散对大鼠大脑海马乙酰胆碱酯酶(AChE)与单胺氧化酶(MAO)活性的影响.方法 选取60只4～5月龄的SD大鼠随机分为正常组、模型组、氟西汀组和中药组,共4组,每组15只.利用孤养结合慢性轻度不可预见性应激刺激制造抑郁症模型.21 d后,采用敞箱实验、糖水消耗量检测和体重检测等指标评定大鼠行为学改变,并用免疫组织化学染色法观察大鼠大脑海马区AChE的蛋白表达变化,利用比色法检测大鼠大脑海马区MAO活性.结果 与正常组相比,模型组水平运动得分、垂直运动得分、体重和糖水偏爱度均明显降低(P<0.01),与模型组比较,氟西汀组和中药组水平运动得分、垂直运动得分、糖水偏爱度与体重均明显升高(P<0.05～0.001);免疫组化结果显示,与正常组相比,模型组海马区AChE的蛋白表达均明显升高(P<0.05),与模型组相比,氟西汀组和中药组大鼠海马区AChE的蛋白表达均明显降低(P<0.05);比色法结果显示,与正常组相比,模型组海马区MAO活性明显升高(P<0.05),与模型组相比,氟西汀组、中药组海马区MAO活性明显下降(P<0.05).结论 柴胡疏肝散抗抑郁症的作用可能与其降低大脑组织海马区AChE蛋白表达和MAO活性有关.     

文拉法辛对老年抑郁大鼠海马血管内皮生长因子表达的影响

目的 探讨抗文拉法辛对老年抑郁大鼠海马血管内皮生长因子(VEGF)表达的影响.方法 老年雄性SD大鼠,随机分为3组,模型组、文拉法辛组各12只均给予慢性不可预测的温和多相应激结合孤养共5 w,文拉法辛组于应激第3周末开始同时给予文拉法辛(5 mg.kg-1.d-1)治疗持续14 d,模型组同时给予生理盐水14 d.对照组12只不给予任何处理.采用敞箱实验和糖水消耗实验观察大鼠抑郁建模情况.采用免疫印迹检测海马VEGF蛋白表达,逆转录聚合酶链反应(RT-PCR)检测VEGF mRNA表达.结果 敞箱实验、液体消耗实验显示模型组及文拉法辛组抑郁模型建立成功.与模型组比较,文拉法辛组垂直得分增高,蔗糖消耗增加(P均＜0.05).与对照组比较,文拉法辛组海马VEGF蛋白及VEGFmRNA表达均增加(P ＜0.05,P＜0.01);模型组海马VEGF蛋白及VEGF mRNA表达均下降(P＜0.05).文拉法辛组与模型组比较,文拉法辛组VEGF蛋白及VEGF mRNA表达均明显增加(P＜0.01).结论 文拉法辛可提高老年抑郁大鼠VEGF表达,并对血管新生可能起到促进作用.     

小牛血清去蛋白注射液对大鼠局灶性脑缺血再灌注后Caspase-12表达的影响

目的 探讨小牛血清去蛋白注射液(DCSI)对脑缺血再灌注后大鼠海马Caspase-12表达的影响.方法 健康SD大鼠随机分为假手术组、模型组和DCSI组.DCSI组于术前1 w开始给药,每日大鼠尾静脉注射DCSI 80 mg/kg,末次给药5 h后建立脑缺血再灌注模型.分别在脑缺血再灌注后的5个时间点进行神经行为学观察,海马病理学、细胞凋亡、Caspase-12蛋白及mRNA表达指标的检测.结果 与假手术组相比,模型组大鼠脑缺血再灌注后的神经功能缺损明显,海马细胞呈明显凋亡表现,Caspase-12蛋白及mRNA表达上调,尤以24h时明显(P＜0.01);DCSI组与模型组相比,功能缺损评分和细胞凋亡相对较轻,Caspase-12蛋白及mRNA表达也相对下凋(P＜0.01,P＜0.05).结论 DCSI对缺血再灌注损伤大鼠大脑具有保护作用,其机制可能与其有效抑制脑组织Caspase-12 mRNA转录水平及蛋白表达、阻断内质网应激启动的凋亡通路有关.     

丁苯酞对大鼠海马神经元磷酸化环磷酸腺苷反应原件结合蛋白和Bcl-2表达的影响

目的探讨丁苯酞对氧糖剥夺/复氧的原代大鼠海马神经元磷酸化环磷酸腺苷反应原件结合蛋白(CREB)和Bcl-2表达的影响。方法以原代大鼠海马神经元设立对照组、模型组、丁苯酞组。丁苯酞组复氧8h后应用终浓度为0.1、1、10μmol/L的丁苯酞干预,通过Western blot法检测磷酸化CREB蛋白、RT-PCR法测定Bcl-2mRNA的表达。结果与对照组比较,模型组和不同浓度丁苯酞组大鼠海马神经元磷酸CREB蛋白、Bcl-2mRNA表达明显降低(P<0.05,P<0.01);与模型组比较,不同浓度丁苯酞组磷酸化CREB蛋白、Bcl-2mRNA表达明显升高,差异有统计学意义(P<0.01)。结论在实验剂量范围内,丁苯酞呈剂量依赖性地抑制海马神经元的凋亡,其作用机制可能是通过上调磷酸化CREB表达,提高Bcl-2活性,抑制海马神经元凋亡。     

下调PTEN对血管性痴呆大鼠海马神经元CREB的调节作用

目的探讨PTEN基因对血管性痴呆大鼠海马神经元CREB表达的影响机制。方法 30只大鼠随机分为正常组、AdR-siPTEN干预组、AdRFP阴性对照组,海马CA1区局部注射AdR-siPTEN腺病毒后2 d,采用双侧颈总动脉永久性结扎(2-VO)法建立血管性痴呆模型,4 w后应用HE染色检测神经元的损伤,RT-PCR和免疫组织化学检测CA1区Akt、CREB mRNA的表达。结果海马部位有PTEN基因红色荧光蛋白表达,并有PTEN基因的mRNA表达量明显下降(P0.01);HE染色显示,AdR-siPTEN组海马神经元损伤和变性程度明显轻于AdRFP阴性对照组大鼠;RT-PCR和免疫组化染色结果显示,与AdRFP组相比,AdR-siPTEN治疗组CA1区Akt、CREB的mRNA和蛋白表达水平显著升高(P0.05)。结论下调PTEN基因可通过Akt增加CREB的表达,从而改善血管性痴呆大鼠神经元突触可塑性,达到减轻神经元损伤、提高学习记忆和神经保护的目的 。     

心肌梗死后抑郁大鼠心肌细胞Bax/Bcl-2的变化及意义

目的 研究抑郁对心肌梗死大鼠心肌细胞Bax/Bcl-2的影响及意义.方法 将SD雄性大鼠随机分为4组:正常组、抑郁组、心肌梗死组(心梗组)和心肌梗死后抑郁组(模型组).通过结扎冠状动脉和28天慢性不可预测性温和应激分别建立心肌梗死(以下简称心梗)和抑郁模型,造模成功后行糖水消耗实验及旷场实验评价大鼠行为学改变,将大鼠心脏取材后应用Western blot技术检测心肌Bax、Bcl-2蛋白表达.使用Bax/Bcl-2比值评估心肌细胞凋亡.结果 心梗组、抑郁组及模型组大鼠糖水偏爱百分比、旷场实验中总行程、运动速度、直立次数较正常组比均明显降低(P＜0.05),其中模型组与正常组各项指标的比较分别为(41.04 ±3.25)％与(72.32 ±8.33)％,(18.17 ±5.94)m与(70.85±6.64) m,(3.03±0.99) cm/s与(11.81 ±1.11)cm/s,(15.13 ±1.73)次与(7.13±1.73)次,差异均有统计学意义(P＜0.05).心梗组、抑郁组及模型组大鼠心脏Bax/Bcl-2比值均明显高于正常组(P＜0.05),且模型组大鼠最明显.结论 心梗和抑郁均可致大鼠心肌细胞凋亡,抑郁加重心梗后心肌细胞凋亡,凋亡机制可能为心梗与抑郁的共病机制之一.     

丹星通络汤治疗大鼠脑缺血再灌注Bax蛋白表达的影响

目的 观察丹星通络汤对大鼠脑缺血再灌注损伤海马区细胞凋亡调控基因Bax蛋白表达的影响,探讨丹星通络汤预防与治疗大鼠脑缺血再灌注损伤的作用.方法 SD大鼠40只,随机分成假手术组、模型组、尼莫地平组、丹星通络汤小剂量组、丹星通络汤大剂量组,每组8只.采用线栓法制备大鼠大脑中动脉闭塞(MCAO)再灌注模型.HE染色检测大鼠脑组织海马区的病理学变化,免疫组织化学法和医学图像分析法检测大鼠脑组织海马区Bax蛋白的平均光密度(OD)值.结果 与模型组比较,丹星通络汤大、小剂量组大鼠脑组织海马区Bax蛋白的OD值明显减低(P＜0.05),并且丹星通络汤大剂量组与尼莫地平组相比有统计学意义(P＜0.05).结论 丹星通络汤可能通过下调Bax蛋白的表达,从而抑制脑组织的凋亡机制发挥对脑组织的保护作用.     

参芎注射液对大鼠脑缺血再灌注后神经细胞凋亡及Fas死亡结构域蛋白表达的影响

目的:探讨参芎注射液对大鼠脑缺血再灌注后神经细胞凋亡及Fas相关死亡结构域蛋白(FADD)及其mRNA表达的影响.方法:100只SD大鼠随机分为正常组(n=10)、假手术组(n=10)、脑缺血再灌注组(n=40)和参芎注射液组(n40).线栓法制备大脑中动脉闭塞模型,TUNEL法检测神经细胞凋亡,分别应用免疫组织化学染色和逆转录-聚合酶链式反应检测FADD蛋白和mRNA表达.结果:与正常组和假手术组相比,脑缺血再灌注组凋亡神经细胞计数显著增加(P＜0.01),FADD蛋白和mRNA表达均显著增强(P均＜0.01).与脑缺血再灌注组比较,参芎注射液组凋亡神经细胞显著减少(P＜0.05,P＜0.01),FADD蛋白和mRNA表达显著减弱(P＜0.05,P＜0.01).结论:参芎注射液能通过下调FADD表达抑制大鼠脑缺血再灌注后神经细胞凋亡.     

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.