替加色罗对结肠炎诱导的大鼠腰骶髓Fos、P物质和降钙素基因相关肽表达的影响

背景:临床研究发现,替加色罗可以明显改善肠易激综合征患者的腹部不适和腹痛,但其调节内脏感觉的机制目前尚不清楚。目的:观察替加色罗对结肠炎诱导的大鼠腰骶髓Fos、P物质(SP)和降钙素基因相关肽(CGRP)表达的影响,探讨替加色罗降低内脏敏感性的作用途径。方法:成年雄性Sprague-Dawley大鼠24只,以三硝基苯磺酸灌肠诱导结肠炎并随机分为实验组1:替加色罗灌胃,每天2mg/kg;实验组2:替加色罗灌胃,每天1mg/kg;对照组:生理盐水灌胃,2.0ml/d。连续灌胃7天后,采用免疫组化方法检测大鼠腰骶髓Fos、SP和CGRP的表达。结果:结肠炎可诱导对照组大鼠腰骶髓(L5～S1)背角深层Fos表达以及背角浅层SP和CGRP表达。实验组1大鼠腰骶髓背角Fos阳性神经元数(22.0±7.7)和SP密度(12.5%±1.4%)显著低于对照组(62.2±18.9和35.9%±8.9%,P<0.05),CGRP密度(1.2%±1.1%)与对照组(2.8%±2.4%)相比无显著差异。实验组2大鼠腰骶髓背角Fos、SP和CGRP的表达与对照组相比均无显著差异。结论:替加色罗可以明显减少结肠炎诱导的大鼠腰骶髓背角Fos和SP的表达,其降低内脏敏感性的作用可能与抑制脊髓背角SP的表达有关。     

伤害和非伤害性结直肠扩张时脊髓和部分脑干核团的c-fos表达

目的通过检测c—Fos表达研究伤害性和非伤害性结直肠扩张（CRD）对脊髓和脑干部分核团神经元活性的影响特点。方法分别建立20、40、80mmHg重复CRD的大鼠动物模型。扩张完毕取大鼠腰骶段脊髓和脑干组织，用免疫组化染色检测c-fos阳性神经元的表达。以插入气囊但不扩张（0mmHg）大鼠为对照组。结果①40、80mmHg扩张时c-Fos阳性神经元主要分布于脊髓后角Ⅰ—Ⅵ层，以及脊髓中间带Ⅶ层和脊髓中央管附近Ⅹ层；而20mmHg时主要在脊髓后角Ⅰ-Ⅳ层表达。②40或80mmHg扩张时腰骶段脊髓c-Fos阳性神经元表达显著高于20和0mmHg（P〈0．05）。③脑干延髓（NTS）的c—Fos阳性神经元表达随扩张压力增高而增高（P〈0.05），20、40、80mmHg扩张时均显著高于对照组（P〈0．05）。④中脑导水管周围灰质（PAG）的c-Fos阳性神经元在40、80mmHg扩张时显著高于20和0mmHg，20mmHg扩张时c—Fos表达和对照组无显著差异（P〉0．05）。结论伤害和非伤害性结直肠扩张刺激可以导致不同部位神经核团的不同反应。脊髓背角以及孤束核神经元对伤害性以及非伤害性内脏扩张刺激均产生反应，而脊髓中间带以及PAG神经元可能仅对伤害性内脏扩张刺激发生显著反应。     

大鼠结肠扩张性刺激引起内脏活动变化的实验研究

目的研究结肠扩张性牵张刺激导致的内脏活动变化的作用机制，为减少因结肠牵张性引起的并发症提供理论依据。方法采用气囊充气模拟结肠镜引起大鼠结肠扩张性牵张刺激的方法，利用电生理学方法观察大鼠支配内脏活动的神经中枢延髓内脏带（Medullary visceral zone，MVZ）中迷走复合体（vagus-solotary complex）放电频率的变化，并以胃的运动功能变化为指标，同步观察了大鼠胃电变化情况。应用免疫荧光技术，以原癌基因c—fos为标志，观察MVZ中迷走复合体的神经元被激活情况。结果给予气囊充气达0．8ml时，迷走复合体中神经元放电频率、Fos的表达以及胃电没有明显改变。给予气囊充气达1．0ml时，迷走复合体中神经元放电、Fos的表达以及胃电开始增加。当气囊充气达1．2ml时，迷走复合体中神经元放电、Fos的表达以及胃电开始有明显增加的改变。气囊充气达1．4ml时，迷走复合体中神经元放电、Fos的表达以及胃电的改变最为明显，而当气囊注入空气1．5ml时，迷走复合体中神经元放电、Fos的表达以及胃电的变化与气囊充气1．4ml时相比，差异不是很明显。结论结肠扩张性刺激可以对支配内脏活动的延髓内脏带中的神经元有明显的激活作用，进而导致内脏活动的相应变化。     

替加色罗对内脏高敏性大鼠直肠扩张后脊髓c-Fos表达的影响

目的　研究替加色罗对大鼠内脏敏感性的作用。方法　新生SD大鼠出生后第 8至 2 1天予醋酸灌肠 ,建立慢性内脏高敏模型为H组 ;生理盐水灌肠为C组。待大鼠成年后 ,将H组分为H病理 、H盐水 、H溶剂 、HTeg0 .1、HTeg0 .3 和HTeg1.0 组 ;C组分为C病理 、C盐水 和CTeg1.0 组。H病理 和C病理 组不扩张直肠行病理学检查和髓过氧化物酶活性测定 ;H盐水 和C盐水 组腹腔注射生理盐水 ,H溶剂 组注射溶剂 蒸馏水 ,HTeg0 .1、HTeg0 .3 和HTeg1.0 组分别注射替加色罗 0 .1、0 .3、1.0mg/kg ,CTeg1.0 组注射替加色罗 1.0mg/kg ,直肠球囊快速注水 (0 .4、0 .8、1.2ml)扩张观肠道 ,每次扩张持续 2 0s,间隔 5min ,察腹部撤离反射 (AWR)和脊髓 (L6～S1)c Fos表达。结果　H盐水 与H溶剂 组AWR积分明显高于其他组 (P <0 .0 5 ) ,腰骶髓c Fos阳性细胞总数明显增加 ,两组间无明显差异。HTeg0 .1、HTeg0 .3 和HTeg1.0 组比H盐水 组AWR积分明显降低 ,c Fos阳性细胞减少 (182± 14 ,16 6± 17,12 8± 15比 2 13± 13;P <0 .0 5 )。结论　替加色罗可明显降低内脏高敏大鼠的内脏敏感性 ,并剂量依赖性地减少脊髓c Fos的表达     

针灸预处理对成年和老年甲醛致痛大鼠脊髓NO分布和c-fos表达的影响

目的比较成年和老年大鼠对炎症性疼痛反应的差别,以及针灸预处理对成年和老年甲醛致痛大鼠脊髓一氧化氮(NO)分布和cfos表达的影响。方法将4月龄和21月龄雄性SD大鼠分别随机分为对照组(不做任何处理)、甲醛组(仅注射甲醛)、针灸足三里穴+甲醛组(针灸足三里穴后注射甲醛)、针灸非穴位+甲醛组(针灸非穴位后注射甲醛),每组6只大鼠。针灸处理后,除对照组外,其余各组于针灸位点同侧足底注射甲醛。观察大鼠行为学反应,记录甲醛注射后1 h内的缩足次数和舔(咬)足时间,1.5 h后灌流取脊髓第4腰髓至第5腰髓节段,采用酶组织化学染色和免疫组织化学染色法检测脊髓NO的分布和c-fos的表达。结果对照组NO数量较低,且老年大鼠脊髓背角NO数量低于成年大鼠(P0.01);Fos免疫阳性神经元基础水平较低,且在成年和老年大鼠间无差异(P0.05);成年和老年大鼠脊髓未发现NO和Fos双标阳性神经元。甲醛组老年大鼠继发疼痛时相的缩足次数高于成年大鼠(P0.05);与对照组比较,NO阳性神经元、Fos免疫阳性神经元和二者双标阳性神经元数量在成年和老年大鼠脊髓均增加(P0.01),且老年大鼠的NO阳性神经元(脊髓背角,P0.05;中央管,P0.01)和Fos免疫阳性神经元数量(P0.01)高于成年大鼠。针灸足三里穴+甲醛组成年和老年大鼠继发疼痛时相的缩足次数(P0.01)和舔(咬)足时间(成年,P0.05;老年,P0.01)低于甲醛组;成年和老年大鼠脊髓NO阳性神经元、Fos免疫阳性神经元和二者双标阳性神经元数量均低于甲醛组(P0.01);且老年组NO阳性神经元和Fos免疫阳性神经元数量较甲醛组下降的幅度高于成年组大鼠(P0.01)。针灸非穴位+甲醛组成年和老年大鼠继发疼痛时相的缩足次数、舔(咬)足时间、NO阳性神经元、Fos免疫阳性神经元和二者双标阳性神经元数量与甲醛组比较无差异(P0.05)。结论老年大鼠对炎症性疼痛的反应强于成年大鼠,针灸预处理足三里穴可以提高大鼠痛阈,减轻疼痛反应,降低脊髓NO的数量和c-fos的表达,且对老年大鼠的作用优于成年大鼠。     

急性心肌损伤后大鼠孤束核的FOS核神经胶质原纤维酸性蛋白表达的变化

目的：观察急性心肌损伤后大鼠延髓孤束核(Nucleus Tractus Solitarii, NTS)神经元和星形胶质细胞的反应。方法：向心肌内注射福尔马林造成急性心肌损伤，免疫荧光染色观察NTS中Fos阳性和GFAP阳性细胞的反应情况。结果：急性心肌损伤后2 h，大鼠NTS中Fos阳性神经元显著升高，4 h达到峰值，然后开始下降。急性心肌损伤后4 h GFAP阳性星形胶质细胞荧光强度明显增强，24 h达到高峰然后下降。结论：急性心肌损伤后，大鼠延髓NTS中神经元和星形胶质细胞被心肌伤害性刺激激活，参与心肌损伤后疼痛的中枢调控。     

三黄汤灌肠对溃疡性结肠炎大鼠治疗作用研究

[目的]探讨三黄汤灌肠对三硝基苯磺酸(TNBS)诱导溃疡性结肠炎(UC)模型大鼠的疗效。[方法]采用TNBS诱导制备UC大鼠模型45只,随机分为模型组、阳性对照组及中药治疗组,各15只,并选未造模大鼠15只作为正常对照组。各组均灌肠给药,模型组和正常对照组予0.9%氯化钠注射液,阳性对照组予柳氮磺吡啶0.5 g/kg,中药治疗组予三黄汤浓煎剂60 g/kg,各组均按10 ml/kg每日灌肠1次。给药时间从造模后第3天开始,连续7 d。第11天观察各组大鼠结肠黏膜损伤指数(CMDI)及黏膜病理组织学情况。[结果]中药治疗组结肠黏膜病理损伤明显改善,CMDI为(1.5±0.535)分,与模型组的(3.55±1.214)分比较差异有统计学意义(P0.05);与阳性对照组的(2.63±1.188)分比较差异有统计学意义(P0.05)。在肉眼及电镜下观察发现,中药治疗组大鼠结肠黏膜较模型组、阳性对照组明显改善。[结论]三黄汤灌肠对TNBS诱导的大鼠UC疗效确切,能显著改善结肠黏膜的充血水肿、溃疡、糜烂等。     

胃扩张对大鼠脑、脊髓和肌间神经丛Fos蛋白和降钙素基因相关肽表达的影响

背景:功能性消化不良是一种常见疾病,越来越多的证据显示其发病与内脏神经敏感性增高有关。目的:通过测定大鼠伤害性胃扩张后脑、脊髓和肌间神经丛Fos蛋白和降钙素基因相关肽(CGRP)的表达,探索内脏刺激传入的途径和方式。方法:24只成年Sprague-Dawley雄性大鼠随机分为3组:实验组(12只)、手术对照组(6只)和空白对照组(6只)。实验组和手术对照组先植入胃内气囊。48h后,实验组接受反复的气囊扩张[80mmHg(1mmHg=0.133kPa)],2h后处死全部实验动物,立即取材。采用免疫组化法观察脑、脊髓和肌间神经丛Fos蛋白和CGRP的表达。结果:实验组杏仁核、延髓和胸髓Fos蛋白的表达较其他两组显著增强(P均<0.01),肌间神经丛的Fos蛋白表达3组间无显著差异。实验组延髓、胸髓CGRP表达较其他两组显著增强(P<0.05和P<0.01)。延髓和胸髓中Fos蛋白与CGRP的表达显著相关(rs分别为0.794和0.728,P均<0.01)。结论:胃扩张刺激可以兴奋皮层下中枢,CGRP在内脏刺激信号的传入过程中起重要作用。     

七氟醚对甲醛致痛诱导Fos蛋白在大鼠脊髓内表达的影响

15只SD大鼠,随机分为对照组、吗啡组、七氟醚组,各5只。七氟醚组吸入3%七氟醚5 min后腹腔注射生理盐水,右侧后肢跖部皮下注射4%甲醛150μl,继续吸入七氟醚;对照组、吗啡组腹腔分别注射生理盐水、10 mg/kg吗啡5 min后,右侧后肢跖部皮下注射甲醛。2 h后处死大鼠,取脊髓L3~L5节段,用免疫组化法观察脊髓内Fos蛋白的表达。结果:甲醛致痛仅诱导Fos在右侧脊髓表达,与对照组比较,吗啡组、七氟醚组脊髓各层Fos蛋白免疫反应阳性神经元(FLIN)数量显著降低(P<0.01),且七氟醚组FLIN量显著多于吗啡组(P<0.01)。认为七氟醚能明显抑制甲醛致痛诱导Fos蛋白在大鼠脊髓内的表达,脊髓背角是七氟醚发挥抗伤害作用的重要部位。     

电针对糖尿病胃轻瘫大鼠胃电活动与延髓神经元和星状胶质细胞可塑性的影响

[目的]观察电针对糖尿病胃轻瘫（DGP）大鼠胃电活动与延髓迷走孤束复合体（VSC）内神经元和星状胶质细胞可塑性变化的影响。[方法]实验大鼠分为空白对照（空白）组、DGP模型（模型）组、模型加电针足三里穴（足三里）组和模型加电针三阴交穴（三阴交）组。模型制备采用腹腔注射5％四氧嘧啶和熟地灌胃诱导的方法。实验3周后记录大鼠的胃电活动,取延髓进行抗Fos蛋白和抗胶质原纤维酸性蛋白（GFAP）的免疫组化染色。[结果]与空白组比较,模型组的胃电平均频率和振幅明显降低。而足三里组和三阴交组的平均频率和振幅较模型组明显升高（P〈O．01,〈0．05）;以足三里组升高更明显,与三阴交组比较差异有统计学意义（P〈0．05）。除空白组外,Fos阳性神经元、GFAP阳性星状胶质细胞集中表达于VSC。以模型组的Fos和GFAP表达最高,而足三里组与三阴交组的表达较模型组明显减少,且它们之间差异有统计学意义（P〈0．01）。[结论]针刺可改善糖尿病胃运动功能障碍,延髓VSC内免疫阳性神经元和胶质细胞可能参与了此调节作用。     

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.     

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.