β-七叶皂甙钠对大鼠缺血性脑水肿及水通道蛋白4表达的影响

目的研究β-七叶皂甙钠对大鼠脑梗死后脑水肿及水通道蛋白4（AQP4）表达的影响。方法取SD雄性大鼠150只,体质量240～300g。随机分为β-七叶皂甙钠治疗组60只,对照组60只,假手术组30只。各组大鼠又按缺血时间不同（6h和1、3、5、7d）分为5个亚组,其中治疗组和对照组每亚组各12只,假手术组每亚组6只。用线栓法制作大鼠大脑中动脉缺血模型,向治疗组大鼠腹腔注射β-七叶皂甙钠（5mg·kg-1.d-1）,对照组及假手术组在相应时间点注射等量等渗盐水。于造模后6h和1、3、5、7d断头取脑。采用干湿重法观察脑水肿的动态变化,通过HE染色观察脑梗死区的病理学变化,应用免疫组化方法检测脑组织AQP4的表达。结果①脑梗死后6h,病变侧大脑半球脑含水量开始增多,至3d时达高峰,以后逐渐下降,7d时仍高于假手术组。治疗组及对照组脑含水量组内不同时间点比较差异均有统计学意义（F=30.704,F=36.703,均P〈0.01）;各时间点治疗组及对照组梗死侧大脑半球脑含水量均明显高于假手术组（均P〈0.01）,治疗组均低于对照组（P〈0.05～0.01）。②脑梗死后6h,梗死侧AQP4表达水平即增高,3d时达高峰,以后逐渐下降,7d时仍高于假手术组;治疗组和对照组各组内各时间点比较差异均有统计学意义（F=20.878,F=28.489,均P〈0.01）。治疗组及对照组各时间点AQP4的表达均高于假手术组（均P〈0.01）,治疗组AQP4的表达均明显低于对照组（P〈0.05～0.01）。AQP4表达水平与脑含水量呈正相关（r=0.972,P〈0.001）。结论β-七叶皂甙钠能明显减轻脑梗死后脑水肿及降低AQP4的表达,β-七叶皂甙钠可能通过抑制AQP4的表达而减轻脑水肿。     

三七总皂苷分期治疗对脑出血大鼠微管相关蛋白-2及神经生长相关蛋白表达的影响

目的 探讨三七总皂苷(tPNS)分期治疗对脑出血大鼠微管相关蛋白-2(MAP-2)及神经生长相关蛋白(GAP-43)表达的影响.方法 大鼠随机分为 5 组:正常对照组、假手术组、脑出血模型组、脑出血后 12 h治疗 A 组、脑出血后 72 h 治疗 B 组.治疗 A 组于脑出血后 12 h、治疗 B 组于脑出血后 72 h开始以三七总皂苷70 mg/(kg·d)腹腔注射,连用 4 d.其余各组皆同时注射等量生理盐水.大鼠于造模后 7 d、14 d、21 d 每个时间点分别处死,干湿重法测脑含水量,脑组织切片行MAP-2、GAP-43 免疫组化.结果 治疗组大鼠肢体功能较脑出血模型组好,且治疗A组神经功能评分在7 d、14 d、21 d优于治疗B组(P<0.05);治疗组可明显改善脑水肿,7 d时治疗A组脑含水量低于治疗B组(P<0.05).治疗组血肿周边MAP-2及GAP-43表达皆高于脑出血模型组(P<0.05);且治疗A组在7 d、14 d、21 d 3个时间点表达高于治疗B组(P<0.05).结论 tPNS可以减轻大鼠脑出血后的脑水肿并积极上调MAP-2和GAP-43表达;早期给予tPNS治疗脑出血大鼠较晚期运用可能对大鼠康复更有利.     

银杏叶提取物(EGb761)对大鼠急性创伤性脑水肿的影响

目的探讨银杏叶提取物(EGb761)对大鼠急性创伤性脑水肿的疗效及其潜在机制。方法将60只SD大鼠随机分至假手术(A)组、创伤+等剂量生理盐水(B)组、创伤+低剂量EGb761(C)组、创伤+高剂量EGb761(D)组。采用Feeney自由落体致伤法制作大鼠顶叶局灶皮质挫裂伤模型,其中C组和D组,在创伤1 h后,分别给予EGb761 25 mg/kg和50 mg/kg治疗。采用干湿重法检测脑组织含水量,苏木素-伊红(HE)染色观察脑组织形态学变化,二氢乙锭(DHE)染色观察氧自由基表达水平,Western印迹检测AQP4蛋白表达水平。结果与A组相比,B、C、D组均出现脑水肿,氧自由基和AQP4表达水平上升,其中B组与A组有显著差异(P0.05);C组和D组脑水肿程度较B组有明显改善,脑组织中氧自由基,AQP4表达水平较B组显著降低(P0.05),且C组和D组之间无显著差异(P0.05)。结论 EGb761对创伤性脑水肿有保护作用,其机制可能与清除氧自由基,降低AQP4的表达有关。     

实验性脑出血大鼠急性期脑组织含水量与ET、NO含量的关系

目的观察大鼠脑出血后血肿周围脑组织含水量与内皮素(ET)、一氧化氮(NO)含量之间的关系,探讨ET、NO在脑出血后脑水肿形成中的作用,为临床治疗提供新的理论依据和方法。方法选用健康雄性SD大鼠48只,体重250 g~300 g,随机分为脑出血组和假手术对照组,每组24只,两组动物再随机分为术后1 d、3 d、5 d、7 d四个时间点,每个时间点6只。通过脑内注射胶原酶建立大鼠脑出血模型。测定不同时间点脑组织含水量与ET、NO含量。结果脑出血组各时间点脑组织含水量及ET、NO含量均高于对照组(P0.05或P0.01)。脑组织含水量与ET、NO含量之间呈正相关关系(P0.05或P0.01)。结论脑出血后ET、NO过量生成,且与脑水肿程度呈正相关关系。脑出血后脑水肿的发生与ET和NO过量生成有关。     

依达拉奉联合甘油果糖对大鼠脑出血后水通道蛋白4的影响

目的 探讨脑出血依达拉奉和甘油果糖联合应用治疗大鼠脑出血的效果及机制的影响.方法 将100只健康SD大鼠随机均分为对照组、模型组、单药组及联合组各25只;后三组制作脑出血模型.成模后单药组注入依达拉奉3.0 mg/kg,联合组分别注入甘油果糖5 mL/kg和依达拉奉3.0 mg/kg.分别于术后6h、1d、3d、5d、7d各处死大鼠5只,干、湿重法测定脑含水量,免疫组化法测定脑组织水通道蛋4（AQP4）表达,观察血肿周围脑组织超氧化物歧化酶（SOD）和丙二醛（MDA）含量.结果 与对照组比较,各组AQP4于6h开始升高,3d达到高峰,7d仍高于正常,联合组各时点均明显低于单药组（P＜0.05）;模型组脑含水量6h开始增加,3d达高峰后缓慢下降,但7d仍高于正常（P＜0.05）;联合组及单药组各时间点含水量模型低于模型组,联合组均低于单药组（P＜0.05）,于6h降低最明显;各时点SOD活性其他各组均明显低于对照组,而MDA含量明显高于对照组,各时点AQP4表达与SOD活力呈明显负相关,与MDA含量呈明显正相关.联合组组各时点SOD活力显著高于模型组和单药组,MDA含量明显低于模型组和单药组;相关分析表明除对照组外,各组AQP4表达与脑组织含水量呈正相关.结论 脑出血脑水肿形成可能与APQ4表达上调有关,APQ4表达水平与脑水肿严重程度呈正相关;脑出血早期应用甘油果糖及依达拉奉可明显抑制病情进展.     

大鼠脑出血后血管内皮生长因子和基质金属蛋白酶9的表达及七叶皂苷钠的影响

目的探讨血管内皮生长因子和基质金属蛋白酶9在脑出血后的作用及七叶皂苷钠的影响。方法85只大鼠随机分为脑出血模型组(20)、治疗组A(20)、治疗组B(20)、假手术组(20)和正常对照组(5),采用胶原酶复制大鼠基底节区脑内血肿模型,通过免疫组织化学动态测定不同时间点(6 h、24 h、48 h和96 h)鼠脑内血肿周围脑组织中血管内皮生长因子和基质金属蛋白酶9的表达;并同时检测七叶皂苷钠干预后两者在相应时间点的动态变化。结果血管内皮生长因子和基质金属蛋白酶9在脑出血模型组出血后6 h表达明显增加,分别为12.67±1.50和9.27±1.28,24~96 h显著增加且均处于高峰(P<0.01);在出血后6~96 h两者的表达呈正相关,相关系数为0.479(P<0.01)。在七叶皂苷钠治疗组血管内皮生长因子、基质金属蛋白酶9的表达量明显受到抑制(特别是治疗B组抑制效果更明显),量效关系呈负相关其决定系数分别为0.107和0.083。结论1.脑出血后血管内皮生长因子的表达水平提高可能有协同诱导基质金属蛋白酶9的表达作用。2.七叶皂苷钠可能通过抑制基质金属蛋白酶9和血管内皮生长因子的相关途径发挥抗脑水肿作用。     

胰性脑病水通道蛋白-4的表达及意义

目的 观察水通道蛋白4(AQP4)在磷脂酶A2诱导的胰性脑病(PE)大鼠脑组织中的表达变化,探讨AQP4与胰性脑病的关系.方法 25只健康成年Wistar大鼠按数字表法随机分为空白组(5只)、对照组(10只)和PE组(10只),应用磷脂酶A2颈内动脉注射(0.1ml/100g体重)方法建立大鼠PE模型.对照组注射等容积生理盐水;空白组无任何处理.注射后1d处死大鼠,测脑湿/干重比值,常规行脑组织病理检查,采用免疫组化法和蛋白质印迹法检测AQP4的表达.结果 空白组、对照组大鼠脑组织无明显病理改变,PE组大鼠脑组织内神经元明显水肿,呈气球样变,炎细胞浸润、聚集,微血管内白细胞聚集及附壁.空白组、对照组、PE组大鼠脑组织含水量分别为(61.44±0.36)％、(63.20±0.32)％和(78.33±0.24)％,PE组大鼠脑组织含水量明显增加(P＜0.05);脑组织AQP4相对表达量分别为0.41±0.27、0.49±0.13、0.98±0.21,PE组大鼠脑组织AQP4表达明显上调(P＜0.05).结论 AQP4可能参与胰性脑病的发病机制.     

依达拉奉对创伤性脑水肿大鼠AQP4表达的影响

目的 探讨依达拉奉对创伤性脑水肿大鼠AQP4表达的影响。方法采用Wistar大鼠建立颅脑创伤模型,分成治疗组和模型组,并用假手术处理大鼠作为对照组。治疗组腹腔注射依达拉奉3mg／kg,每12h注射一次;模型组给予相同体积的生理盐水。分别于给药后6h、12h、1d,3d、5d处死各组大鼠,取出脑组织,测定脑含水量及AQP4mRNA表达水平。结果与模型组相比,12h后各个时间点治疗组脑组织含水量明显低于模型组（P均〈0．05）;模型组脑组织AQP4mRNA表达在3d达高峰,然后维持高水平表达,明显高于对照组（P〈0．05）;治疗组在6hAQP4mRNA表达水平最高,然后持续回落,5d时与对照组表达水平接近。结论依达拉奉可明显抑制脑水肿引发的AQP4表达上调,减轻脑水肿形成。     

地塞米松对脑出血大鼠模型脑组织AQP4表达的影响及意义

目的观察地塞米松对脑出血（ICH）大鼠模型脑组织水通道蛋白4（AQP4）表达的影响及意义。方法32只大鼠脑尾状核部位立体定向注入胶原酶建立ICH模型,随机分为观察1、2、3组和对照组各8只后单笼饲养,观察1、2、3组分别腹腔注射地塞米松1、15、30mg/kg,均为2次/d,连续3d;对照组常规饲养。检测各组脑含水量、血脑屏障通透性及脑组织AQP4蛋白及mRNA表达。结果用药3d后观察1～3组血脑屏障通透性、AQP4蛋白及AQP4 mRNA含量均低于对照组（P均〈0.05）,且AQP4蛋白及其mRNA表达与地塞米松呈剂量依赖性关系;观察1组脑含水量较对照组降低（P〈0.05）。结论地塞米松能降低ICH模型大鼠脑组织AQP4表达,此可能为其早期小量使用减轻脑水肿的机制之一。     

三七总皂甙与参麦注射液对大鼠脑出血后脑水肿、行为学变化的影响

目的 比较观察三七总皂苷及参麦注射液对脑出血大鼠脑组织损伤和脑水肿的作用.方法 采用胶原酶与肝素复合法造成大鼠脑出血模型,造模后3 h,各组按试验设计给药,在相应时间点先对大鼠进行神经行为学评分,测定脑组织含水量、细胞形态学的变化,TNF-α及HSP70表达.结果 三七、参麦均能够改善神经功能缺失症状,在第4天与模型组比较差异显著(P＜0.05).三七及参麦组能够减轻脑组织水肿,于第4天、第7天改善明显,与模型组比较差异显著(P＜0.05),与甘油作用相似,但起效较慢.三七和参麦均能够使大鼠脑出血后TNF-α表达降低、HSP70表达升高,以4 d、7 d与模型组比较有显著性差异(P＜0.05).结论 三七和参麦可以改善脑出血后神经功能缺陷症状;能减轻脑出血后脑水肿;可以抑制脑组织TNF-α的表达,增加HSP70表达,疗效优于甘油;综合各指标分析和比较,三七的作用较参麦好.     

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.