山茱萸纳米水提液对衰老小鼠脑组织p66~(shc)表达的影响

目的 探讨山茱萸制剂对自然衰老小鼠脑组织的磷酸化酪氨酸信号适配蛋白(66kilodalton isoform of shcgene products,p66shc)表达的影响.方法 人工饲养自然衰老小鼠11～12月龄36只、青年鼠8只,分6组:青年对照组、衰老对照组和给药组,给药组又分为山茱萸纳米水提液和醇提液的大剂量、小剂量组;各组给药30 d,然后断头取小鼠肝组织做超氧化物岐化酶(SOD)、丙二醛(MDA)检测,RT-PCR法测定小鼠脑组织p66shc的表达.结果 山茱萸纳米水提和醇提液给药组SOD活性与衰老对照组相比显著升高(P<0.01),而MDA含量和脑p66shc的表达显著降低(P<0.01);山茱萸纳米制剂水提液小剂量、大剂量组和醇提液比较上述作用均更显著(P<0.01) .结论 山茱萸能够明显降低衰老小鼠脑p66shc活性亚基的表达,从而影响p66shc活性起到延缓衰老的作用.山茱萸纳米水提取液小剂量组的作用优于山茱萸纳米醇提液各组.     

缬沙坦对血管衰老中凋亡调控基因Bcl-2、Bax表达的影响

目的探讨缬沙坦对血管衰老中凋亡调控基因Bcl-2、Bax表达的影响。方法健康Wistar大鼠分为青年组、衰老组及缬沙坦组，测定血浆丙二醛（MDA）、超氧化物歧化酶（SOD）水平，同时采用恒速注人流体方法测定各组大鼠颈动脉血管的顺应性，利用免疫组织化学染色法、RT—PCR法和Western印迹法分析各组大鼠凋亡调控基因Bcl-2、Bax的mRNA及蛋白表达水平。结果与衰老组相比，缬沙坦组MDA浓度显著降低（P〈0．05），SOD浓度显著升高（P〈0．05），颈动脉血管的顺应性增高，其中弹性面积有显著性差异（P〈0．05），Bcl-2的mRNA及蛋白表达水平明显增高（P〈0．05），Bax的mRNA及蛋白表达水平降低（P〈0．05）。结论血管衰老有其特征性生理改变，Bcl-2、Bax的mRNA及蛋白表达的失衡可能是血管衰老的重要分子机制之一，缬沙坦有一定的逆转血管衰老的作用。     

衰老大鼠动脉顺应性与凋亡相关指标变化及缬纱坦对其影响

目的探讨衰老大鼠动脉顺应性与凋亡相关指标变化及缬纱坦对其影响,为阐明血管衰老对动脉硬化诊治的重要意义。方法Wistar大鼠分为青年组、衰老组及缬纱坦组,测定血浆丙二醛、超氧化物歧化酶水平,同时采用恒速注入流体方法测定大鼠颈动脉血管的顺应性,并利用免疫组织化学染色法、反转录—聚合酶链式反应和蛋白免疫印迹法分析各组凋亡相关基因Bcl-2 mRNA及蛋白表达、半胱氨酸天门冬氨酸蛋白酶3活性变化水平。结果与衰老组相比,缬纱坦组血浆丙二醛浓度明显降低(P<0.05),超氧化物歧化酶浓度明显升高(P<0.05);颈动脉血管的顺应性增高,其中弹性面积差异有显著性(P<0.05);主动脉血管Bcl-2阳性内皮细胞百分率增高(P<0.05),半胱氨酸天门冬氨酸蛋白酶3表达降低(P<0.05);Bcl-2 mRNA及蛋白表达水平明显增高(P<0.05),半胱氨酸天门冬氨酸蛋白酶3活性水平降低(P<0.01)。结论血管衰老有其特征性生理改变,凋亡相关基因Bcl-2表达下调、半胱氨酸天门冬氨酸蛋白酶3活性增高可能是血管衰老的重要分子机制之一,缬纱坦对血管衰老有一定保护作用,为延缓血管衰老以及防治动脉硬化开辟新途径。     

p66shc基因表达与老年糖尿病患者早期肾损伤的相关性

目的研究p66shc基因在老年糖尿病患者外周血单核细胞中的表达及其与早期肾损伤指标的关系。方法采用实时PCR技术检测74例老年糖尿病患者和48例健康老年人(对照组)外周血单核细胞中p66shc基因表达水平,结合临床资料分析p66shc与早期肾损伤指标尿微量白蛋白(m Alb)、转铁蛋白(TRF)、免疫球蛋白(Ig G)、α1-微球蛋白(α1-MG)和N-乙酰-β-D-氨基葡萄糖苷酶(NAG)之间的相关性。结果 p66shc基因在老年糖尿病患者中的表达水平显著高于对照组(P<0.05);与对照组相比,老年糖尿病患者m Alb、TRF、Ig G、α1-MG、NAG均显著升高(均P<0.01),并均与p66shc基因的表达水平呈正相关。结论 p66shc与老年糖尿病患者早期肾损伤密切相关,可为老年糖尿病患者肾损伤的早期诊断提供新的依据。     

阿托伐他汀对大鼠动脉增龄性变化的影响

目的观察阿托伐他汀对大鼠动脉增龄性变化的影响。方法实验用Wistar大鼠分为12月龄组、18月龄组、阿托伐他汀组,每组8只。取胸主动脉,测定其超氧化物歧化酶(SOD)活性和丙二醛(MDA)的含量,Western印迹法测内皮型一氧化氮合酶(eNOS)蛋白的表达量。结果与12月龄组比较,18月龄组大鼠主动脉中SOD的活性降低,MDA含量升高,eNOS蛋白表达降低;与18月龄组比较,阿托伐他汀组大鼠主动脉中SOD的活性升高,MDA含量减少,eNOS蛋白表达增加。结论随着增龄的改变,大鼠主动脉抗氧化功能减弱,内皮功能出现异常,eNOS蛋白表达降低;阿托伐他汀可以通过改善衰老过程中的血管抗氧化能力和增加eNOS蛋白表达,改善大鼠血管内皮功能,从而延缓大鼠血管衰老的进程,发挥心血管保护作用。     

衰老大鼠心肌组织中活性氮簇的改变及可能意义

目的测定衰老大鼠心肌组织凋亡程度,探讨及测定衰老大鼠心肌组织中活性氮簇的改变及其与心肌细胞凋亡的关系。方法分别选取雄性成年SD大鼠（4月龄-6月龄）和衰老SD大鼠（22月龄-24月龄）。采用Caspase-3活性测定法检测大鼠心肌组织中细胞凋亡发生情况,用Western-blot蛋白印记法检测大鼠心肌组织中血管舒张剂刺激的磷蛋白（VASP）和磷酸化VASP（pVASP）的表达水平;ELISA法检测心肌组织硝基酪氨酸含量。结果与成年大鼠相比,衰老大鼠的Caspase-3比活性显著增高（P〈0.01）,心肌组织pVASP水平明显降低（P〈0.01）,硝基酪氨酸明显升高（P〈0.05）,硝基酪氨酸含量与Caspase-3活性呈正相关。结论衰老大鼠心肌组织中一氧化氮生物利用度下降,生成大量毒性的过氧亚硝基,这可能是衰老心脏凋亡增加的原因之一。     

雄激素缺乏小鼠体内活性氧和Rb蛋白表达的变化及对心脏衰老的影响

目的 探讨小鼠体内雄激素水平缺乏对心脏衰老的影响.方法 实验分为正常C57BL/ 6J雄鼠组(n=8)、去势组(n=8)、睾丸女性化鼠(tfm)组(n=10)、衰老组(n=8).测定各组血清睾酮浓度,分离心肌组织测定组织超氧化物歧化酶(SOD)和丙二醛(MDA)水平以及Western印迹测定去磷酸化Rb蛋白表达量.结果 与正常组相比,衰老组、去势组和tfm组血清睾酮浓度明显下降(P<0.01);心肌组织中SOD活性下降(P<0.05,P<0.01)、MDA含量增加(P<0.01);去磷酸化Rb蛋白表达量增加(P<0.01).与衰老组相比,去势组和tfm组SOD活性、MDA含量及去磷酸化Rb蛋白表达量差异无统计学意义.结论 体内雄激素水平缺乏可能通过增加活性氧水平(ROS)及Rb蛋白表达导致小鼠心肌组织衰老.     

健康大鼠血管衰老性重塑与血管衰老相关基因的关系

目的通过研究健康大鼠血管衰老性重塑形态学变化及衰老相关基因表达,探讨血管衰老性重塑可能的分子调控机制,为临床有效干预血管衰老提供分子靶点。方法观察主动脉组织形态及内皮细胞显微结构变化,应用Western blotting分析4、10、16月和24月龄大鼠血管重塑p16INK4a和p21cip1蛋白表达变化。结果随增龄,大鼠主动脉管壁增厚,纤维化程度增高,内皮细胞形态呈现衰老改变,p16INK4a和p21cip1蛋白表达呈时间依赖性上调。结论血管衰老性重塑的分子机制之一可能与上调细胞周期蛋白p16INK4a和p21cip1的表达有关。进一步阐明其调控机制可为延缓血管衰老,防治动脉粥样硬化提供理论依据。     

原花青素对I型糖尿病大鼠血管内皮功能、氧化性损伤和凋亡相关蛋白表达的影响

目的研究原花青素(procyanidins,PC)对糖尿病大鼠血管内皮细胞的作用,并在三种凋亡相关蛋白水平探讨作用机制。方法雄性SD大鼠,尾静脉注射链脲佐菌素(STZ)30 mg/kg造成1型糖尿病动物模型,随机分为糖尿病模型组,50 mg/kg原花青素治疗组和100 mg/kg原花青素治疗组,每组8只,另选8只健康雄性大鼠为正常对照组。各组大鼠灌服给药12周后,用血糖仪及专用血糖试纸现场检测各组大鼠空腹血糖;采用放射免疫分析仪检测血清内皮素1(ET-1)和前列环素(PGI2)水平;采用比色法检测血管内皮组织MDA含量和SOD活性;采用常规免疫组织化学染色方法对血管内皮细胞三种凋亡相关蛋白Caspase-3、Bax和Bcl-2的表达进行检测;用Olympus成像分析系统高倍镜(×400)下观察和采集显微图像,采用美国显微镜图像分析测量软件Image-Pro Plus 6.0采集整个视野黄染灰度值并自动转化为光密度值,计算出单个内皮细胞平均光密度值(mean-density),分析三种凋亡相关蛋白的表达情况。结果糖尿病模型组大鼠,与正常对照组比较,大鼠空腹血糖水平明显增高,血清ET-1含量增高和PGI2含量降低(P0.01),血管内皮组织MDA含量提高(P0.01),血管内皮组织SOD活性降低(P0.01),血管内皮Caspase-3和Bax蛋白表达均增强(P0.01),Bcl-2蛋白表达及Bcl-2和Bax蛋白表达比值(Bcl-2/Bax)均减弱(P0.01);50 mg/kg原花青素治疗组和100 mg/kg原花青素治疗组,与糖尿病模型组比较,大鼠空腹血糖水平不同程度减低(P0.05和P0.01),血清ET-1含量降低和PGI2含量升高(P0.01),血管内皮组织MDA含量降低(P0.01),血管内皮组织SOD活性提高(P0.05和P0.01),血管内皮Caspase-3和Bax蛋白表达均减弱(P0.01),Bcl-2蛋白表达及Bcl-2和Bax蛋白表达比值(Bcl-2/Bax)均提高(P0.01),SOD含量均高于糖尿病模型组。结论原花青素对糖尿病大鼠血管内皮分泌功能和氧化性损伤有调节保护作用,同时可抑制血管内皮Caspase-3和Bcl-2蛋白表达水平,促进Bcl-2表达,提高Bcl-2和Bax蛋白表达比值。     

D-半乳糖诱发氧自由基升高对大鼠动脉衰老的影响

目的观察D半乳糖诱发氧自由基升高对大鼠颈动脉顺应性和腹主动脉脂褐素含量的影响。方法大鼠皮下注射D半乳糖140mg/kg连续7w,造成氧自由基升高动物模型。分别测定模型组与对照组大鼠血浆MDA、SOD水平。同时采用恒速注入流体方法测定大鼠颈动脉血管段的顺应性和荧光图像分析法测定腹主动脉脂褐质含量。结果模型组较对照组MDA含量显著升高(P<001),SOD略有下降。模型组血管顺应性低于对照组,其中弹性面积有显著性差异(P<005)。模型组腹主动脉脂褐质荧光强度显著高于对照组(P<005)。结论氧自由基是造成动脉衰老的重要因素。     

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.