大黄酸对肿瘤坏死因子α所致胰腺细胞损伤的保护作用

目的：探讨大黄酸是否对肿瘤坏死因子α（TNFα）作用下的胰腺细胞具有保护作用。方法：观察大黄酸对TNFα作用条件下体外培养的大鼠胰腺细胞的细胞活率、培养上清中的乳酸脱氢酶（LDH）、淀粉酶（AMY）和N－乙酰－D－氨基葡萄糖苷酶（NAG）的水平的影响,并进行了胰腺细胞形态学观察。结果：大黄酸可提高相应TNFα浓度下的胰腺细胞的细胞活率,降低培养液上清中LDH、AMY和NAG的水平。结论：大黄酸对TNFα所致胰腺细胞损伤有明显的保护作用。     

清胰汤对急性胰腺炎大鼠的保护作用

目的研究清胰汤对急性胰腺炎的治疗机制.方法采用逆行胆胰管注入牛磺胆酸钠制成大鼠急性胰腺炎模型,观察不同实验性急性胰腺炎治疗组(分别在造模2h后给予生理盐水、石蜡油和清胰汤治疗)大鼠血清肿瘤坏死因子(TNF)、内毒素(ET)、淀粉酶(AMY)改变.结果和其他治疗组比较,清胰汤可明显降低急性胰腺炎模型大鼠造模后24h和48h时的TNF和AMY的水平(和生理盐水、石蜡油组比较,P<0.05),造模后48h清胰汤治疗组大鼠血中ET含量显著低于其他治疗组(P<0.05).结论清胰汤的治疗作用可能是多方面的,包括保护胰腺细胞、调节细胞因子的释放、保护肠粘膜防止肠道细菌和内毒素的移位.     

清胰Ⅱ号对重症急性胰腺炎大鼠胰腺及肠组织中MCP-1表达的影响

目的探究清胰Ⅱ号对重症急性胰腺炎(SAP)大鼠胰腺及肠组织中单核细胞趋化蛋白(MCP)-1表达及细胞凋亡的影响。方法将36只大鼠随机分为对照组、模型组、模型+清胰Ⅱ号组,每组12只,对照组正常饲养,其余各组采用胆胰管逆行注射牛磺胆酸钠0.1 ml/100 g的方法建立SAP模型,模型+清胰Ⅱ号组造模后给予清胰Ⅱ号10 ml/kg。造模后12、24 h随机选取7只大鼠采样,比较各组腹水量、胰腺和回肠病理学变化,判断造模是否成功。酶联免疫吸附测定法(ELISA)检测血清MCP-1、白细胞介素(IL)-10、二胺氧化酶(DAO)、血清淀粉酶(AMY)活力。实时定量PCR(qRT-PCR)检测胰腺及肠组织中MCP-1 mRNA表达情况。流式细胞术检测胰腺细胞的凋亡率。结果模型组大鼠腹内血性腹水及胰腺坏死显著高于对照组,表明造模成功。造模后12、24 h,模型+清胰Ⅱ号组大鼠血性腹水、血清MCP-1、IL-10、DAO、AMY活力及胰腺、回肠组织中MCP-1mRNA的表达显著低于模型组(P<0.05);与对照组相比,模型组及模型+清胰Ⅱ号组胰腺细胞的凋亡率显著增加,且模型+清胰Ⅱ号组SAP显著高于模型组(P<0.05)。结论清胰Ⅱ号可下调炎症因子MCP-1、IL-10表达,降低DAO、AMY活力,促进胰腺细胞凋亡,对重症急性胰腺炎大鼠胰腺及肠组织具有良好的修复和保护作用。     

胰岛素对人外周血单个核细胞肿瘤坏死因子α蛋白表达及分泌的影响以及普伐他汀的干预作用

探讨胰岛素对培养的人外周血单个核细胞肿瘤坏死因子α蛋白表达及分泌的影响，以及普伐他汀对其的作用。健康人静脉血加入淋巴细胞分离液后离心提取人外周血单个核细胞培养，加入不同浓度的胰岛素、普伐他汀及甲羟戊酸孵育48h，采用Western blot及酶联免疫吸附法等方法，观察人外周血单个核细胞中肿瘤坏死因子α蛋白表达水平及细胞培养上清中肿瘤坏死因子α蛋白水平。结果发现，胰岛素可诱导人外周血单个核细胞及其培养上清中肿瘤坏死因子α蛋白水平显著升高，且呈剂量依赖性，加入普伐他汀后可使胰岛素刺激的人外周血单个核细胞及其培养液上清中肿瘤坏死因子α蛋白水平显著下降，普伐他汀的这种作用可被甲羟戊酸所抑制。结果提示，普伐他汀对糖尿病患者心血管系统的保护作用可能部分与其抑制胰岛素介导的炎症反应有关。     

大黄酸抑制TGF—β1诱导的肾小管上皮细胞肥大及细胞外基质产生

目的观察大黄酸对TGF-β1诱导的近端肾小管上皮细胞细胞肥大及细胞外基质(ECM)的影响。方法在体外以TGF-β1（2μg／L）刺激LLC-PK1细胞,诱导细胞肥大和ECM合成增加。与此同时,以不同浓度的大黄酸处理细胞,检测细胞体积,蛋白质含量以及3H-亮氨酸掺入以观察细胞肥大的变化;检测细胞孵育24h后上清的纤维连接蛋白(FN)含量,3H-脯氨酸掺入以及细胞胶原Ⅳ及FNmRNA的表达以观察大黄酸对ECM的影响。结果TGF-β1（2μg／L）刺激可以导致LLC-PK1细胞出现细胞肥大,表现为细胞体积,细胞内蛋白量及3H-亮氨酸掺入量明显增加。大黄酸处理后细胞体积及细胞内蛋白量明显降低。TGF-β1也明显增加LLC-PK1细胞3H-脯氨酸掺入量、培养上清中FN含量、以及细胞内胶原Ⅳ和FNmRNA的表达。大黄酸则抑制上述ECM合成的增加,明显降低细胞内胶原Ⅳ,FNmRNA表达水平。结论大黄酸可以逆转TGF-β1诱导的近端肾小管上皮细胞肥大,抑制TGF-β1刺激的ECM合成。这可能是大黄酸预防或改善糖尿病肾脏病变,延缓糖尿病肾病进展的作用机制之一。     

阿司匹林对过氧化氢诱导肝癌细胞凋亡和坏死的抑制作用及其机制

目的：阿司芬林（AS）在胃肠道肿瘤，特别是结肠癌有抑制作用。但对胃肠道外肿瘤是否也有作用并不清楚。探讨AS对H2O2诱导的SMMC7721人肝癌细胞凋亡和坏死的作用及其机制，方法，应用细胞形态学、DNA片段电泳和流式细胞技术，测定乳酸脱氢酶（LDH）释放率，细胞存活率及胞浆铁蛋白浓度，观察AS对H2O2所致的肝癌细胞凋亡和坏死的作用。结果：0.8mmol/L的H2O2使肝癌细胞出现典型的亚二倍体Ap峰，“阶梯状”DNA电沪睛段及凋亡细胞形态学改变；而4mmol/L的H2O2则引起细胞明显坏死；但临床剂量范围内（≤5mmol/L）的AS不能诱导肝癌细胞凋亡，对H2O2所致的肝癌细胞凋亡和坏死有抑制作用，0．5mmol/L AS组可使Ap峰中细胞数（凋亡率）减少34％，LDH释放率减少40％，细胞存活率增加50％，随着AS剂量加大，对凋亡和坏死的抑制作用逐渐增强。5mmol/L的AS能完全对抗H2O2诱导的细胞凋亡。细胞坏死的情况也接近对照组（P＞0．05）；而去铁铁蛋白（apo-ferritin)也有类似的保护作用，检测发现，AS能使肝癌细胞铁蛋白表达显著增加，与其抗凋亡及坏死作用及明显的相关性。结论：AS可保护SMMC7721肝癌细胞对抗H2O2诱导的凋亡和坏死作用，其机制与AS增加肝癌细胞铁蛋白表达有关。     

MSCs-肝细胞共培养上清对体外培养L02细胞的影响

目的以体外培养L02细胞为实验对象,了解骨髓间充质干细胞(MSCs)与肝细胞共培养上清的生物活性。方法采用两步酶分离法、全骨髓贴壁筛选法分别分离肝细胞、MSCs,按比例直接共培养,收集不同时段的共培养上清,用于人肝细胞系-L02细胞的培养。分别以单独培养的肝细胞上清、MSCs上清为对照,观察共培养上清对L02细胞活力和总蛋白合成的影响;同时观察其对L02细胞损伤模型的AST、LDH及细胞凋亡的影响。结果肝细胞上清、MSCs上清以及共培养上清对L02细胞的活力和增殖均有显著促进作用,表现为L02增殖活跃,总蛋白合成量增加;加入上述细胞上清的酒精损伤L02细胞AST、LDH释放和细胞凋亡显著低于未加细胞上清的对照组,其中共培养上清效果最为显著(P<0.05)。结论共培养上清对L02有明显的刺激增殖和损伤保护作用。     

ICOS联合IFN-γ单抗对活化T细胞诱导的RINm5F细胞凋亡的保护作用

目的 探讨可诱导性协同刺激分子(ICOS)单抗和γ-干扰素(IFN-γ)单抗联合应用对T细胞活化诱导的胰岛细胞凋亡及其相关凋亡机制.方法 植物血凝素(PHA)活化的T细胞与胰岛细胞株RINm5F共同培养,联合应用ICOS单抗和IFN-γ单抗48 h后,AnnexinV/PI法观察RINmSF细胞凋亡率,RT-PCR法观察凋亡相关和胰岛功能相关基因表达,放射免疫法检测观察胰岛素分泌水平.结果 ICOS单抗与IFN-γ单抗联用,T细胞活化诱导的RINm5F细胞株凋亡率(%)显著下降(IgG对照71.89±3.29,ICOS单抗39.66±2.05,IFN-γ单抗45.65±1.59,ICOS单抗+IFN-γ单抗20.49±0.95),这可能与死亡受体途径FAS/FAS配体,肿瘤坏死因子及其受体和肿瘤坏死因子相关凋亡诱导配体等相关的基因表达下调有关;同时两者联合应用后,RINmSF细胞株INS1、INS2及PDX-1基因表达量显著上调,胰岛素分泌水平也显著升高.结论 ICOS单抗和IFN-γ单抗联合应用降低胰岛细胞凋亡的同时,对胰岛细胞的胰岛素分泌功能具有一定的保护作用,是一种有效的诱导T细胞对胰岛细胞无反应性的手段.     

百令提取液对环孢素A导致肾小管损伤的防治作用

目的：从细胞水平观察百令提取液对环他素A(CsA)肾毒性的影响。方法：体外培养肾小管上皮细胞，通过^3H-TdR掺入、MTT比色法、PI染色法和细胞LDH释放等观察百令提取液对CsA引起的肾小管上皮细胞毒性的防治作用。结果：^3H-TdR掺入和MTT比色法显示在一定的剂量范围百令提取液可明显促进培养的肾小管上皮细胞增生，5mg／L以上的CsA抑制细胞增殖；细胞周期的石研究显示15mg／L以上的CsA使小管上皮细胞细胞周期5且滞在G2期；此外，更大剂量的CsA(25mg／L)对细胞有直接的毒性作用，使培养上清LDH值升高。百令提取液能部分逆转CsA引起的细胞增殖受抑和细胞周期阻滞，但足对CsA导致的细胞培养上清LDH的升高没有明显的影响。结论：本研究证实了百令提取液可以促进体外培养的肾小管上皮细胞增生，对CsA的肾小管细胞毒性具有一定的拈抗作用。     

NOD小鼠胰岛内TNF/TNFR1凋亡通路的动态变化

随着病程的进展和胰岛炎的加重,NOD小鼠胰腺γ干扰素(IFNγ)、肿瘤坏死因子α(TNFα)、肿瘤坏死因子受体(TNFR)1和半胱氨酸蛋白32(CPP32)mRNA表达增加,胰岛细胞及浸润的炎症细胞TNFα、TNFR1、CPP32蛋白表达增加,提示Th1细胞因子和TNF/TNFR1凋亡通路在1型糖尿病发病中起重要作用。     

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.     

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.     

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.     

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.