胃癌组织RASSF1A甲基化对其mRNA及蛋白表达的影响

目的研究RASSF1A基因启动子区甲基化对胃癌组织中RASSF1AmRNA及蛋白表达的影响。方法RT-PCR方法检测54例胃癌及癌旁正常组织RASSF1AmRNA表达,甲基化特异性PCR方法检测RASSF1A启动子区CpG岛甲基化状态;Westernblot方法检测20例胃癌及癌旁正常组织中RASSF1A蛋白表达。结果(1)RASSF1AmRNA和蛋白表达水平在胃癌组织中明显低于癌旁正常组织(A值分别为0·2589±0·2407比0·5448±0·2971,P<0·0001;0·1874±0·0737比0·6654±0·2201,P<0·0001);(2)RASSF1A在胃癌组织和正常组织中的甲基化频率分别为66·7%和14·8%,差异有统计学意义(P<0·0001);(3)在胃癌组织中,甲基化组RASSF1AmRNA的表达明显低于非甲基化组(0·1384±0·1142比0·5018±0·2463,P<0·0001)。结论胃癌组织RASSF1AmRNA和蛋白表达缺失或低下,与其启动子甲基化程度增高显著相关。     

胃癌组织中血管内皮生长因子C表达及与淋巴结转移关系

目的探讨胃癌组织中血管内皮生长因子C(VEGFC)的表达及与肿瘤淋巴结转移的关系。方法取30例胃癌患者手术切除的新鲜标本和距癌灶5cm以外的胃组织各1块,采用半定量RTPCR法和免疫组化染色法检测胃癌组织及癌旁正常组织的VEGFC表达,图像分析仪检测VEGFC的免疫组化阳性指数(positiveindex,PI)。结果胃癌细胞呈高水平表达VEGFC,癌旁正常组织则未见表达。在胃癌组织中,有淋巴结转移(PI=1.345±0.079)、淋巴管浸润(PI=1.315±0.037)患者的VEGFC表达水平分别较无转移(PI=1.156±0.045)及无浸润者(PI=1.154±0.043)增高(P<0.05)。高分化胃癌患者的VEGFCmRNA[VEGFC/βactin比值(Ar)=0.846±0.042]和VEGFC表达(PI=1.372±0.023)均分别高于中低分化胃癌者(Ar=0.663±0.007,P<0.01和PI=1.126±0.078,P<0.05)。结论VEGFC阳性表达的胃癌患者可能更易发生淋巴结转移。     

食管癌中c-FLIP蛋白的表达及其与细胞增殖、凋亡的关系

目的探讨细胞型Fas相关死亡域样白介素-1β转换酶抑制蛋白（c—FLIP）在食管癌发生、发展中的作用。方法采用免疫组化SP法检测64份食管癌组织（观察组）和20份正常食管黏膜组织（对照组）中c—FLIP蛋白表达,并采用TNUEL法检测细胞增殖指数（1（I）和凋亡指数（AI）。结果观察组c—FLIP蛋白阳性率明显高于对照组,P〈0．05;c-FLIP蛋白表达与食管癌组织分化程度、TNM分期、淋巴结转移密切相关（P〈0．05）;随c-FLIP蛋白表达强度的增加,细胞增殖指数逐渐增强、凋亡指数逐渐降低（P〈0．05）。结论c—FLIP蛋白高表达可促进食管癌的发生、发展。     

胃癌组织中5-脂氧合酶的表达及其与磷酸化Akt相的关性

目的:检测5-脂氧合酶(5-lipoxygenase,5-LOX)在胃癌组织中的表达,并探讨5-LOX对磷酸化Akt(phosphorylatedAkt,p-Akt)表达的影响.方法:采用逆转录-聚合酶链反应(RT-PCR)检测30例新鲜胃癌及癌旁正常组织标本中5-LOXmRNA表达水平;免疫印迹法(Westernblot)检测5-LOX和p-Akt蛋白的表达水平.结果:5-LOX在胃癌组织中的表达显著高于癌旁正常组织(76.7%vs40%,P<0.05,mRNA水平;56.7%vs30%,P<0.05,蛋白水平).p-Akt蛋白在胃癌组织中的表达也显著高于癌旁正常组织(56.7%vs26.7%,P<0.05).相关性分析表明,胃癌组织中5-LOX蛋白表达和p-Akt水平间无明显相关性(r=0.186,P>0.05).结论:5-LOX蛋白对胃癌的发生、发展有一定促进作用,但与PI3-K/Akt信号通路无关.     

STAT3及相关生长调控基因在胃癌组织中的表达及意义

目的探讨转录信号传导子与激活子3（STAT3）及相关生长调控基因在胃癌发生发展中的作用及意义。方法应用RT—PCR检测STAT3基因及相关基因c—myc，p53，生存素（survivin），血管内皮生长因子（VEGF）的mRNA表达水平，应用Western印迹法及免疫组化法检测STAT3基因的蛋白水平及定位表达。结果胃癌组织及癌旁组织中STAT3 mRNA的表达均明显高于正常胃组织（P〈0．01），胃癌及癌旁组织中c—myc，surovivin，VEGF mRNA的表达上调（P〈0．01），p53 mRNA的表达下调（P〈0．01），胃癌组织及癌旁组织中STAT3蛋白水平的表达均高于正常胃组织（P〈0．01）。结论STAT3基因的持续激活在胃癌的发生发展中起重要作用，可作为早期诊断的指标及治疗的靶点。     

采用组织芯片技术研究c-FLIP在胃癌中的表达及其意义

目的 检测C-FLIP(细胞型fas相关死亡区域蛋白样白介素1-β转化酶抑制蛋白,FADD-like interleukin-1-β converting enzyme inhibitory protein,c-FLIP)在胃癌中的表达并提示其在胃癌发生、发展中的意义.方法 收集60例胃癌和27例正常胃黏膜标本,用组织芯片(tissue micro-array)和免疫组织化学法检测c-FLIP基因的表达.结果 c-FLIP在60例胃癌组织中有59例呈阳性表达(98.03%),在27例正常胃黏膜组织中有1例呈阳性表达(3.7%),胃癌组织中c-FILP基因表达显著高于正常胃黏膜(P<0.05).结论 c-FLIP基因高表达于胃癌组织,这为揭示胃癌的发生、发展提供了新的理论依据,并为胃癌的有效治疗提供了新思路.应用组织芯片大规模高效检测临床样本是可行的,具有快速、准确、方便、经济的特点.     

胃癌组织碱性成纤维细胞生长因子表达及对血管新生和肿瘤进展的影响

目的　探讨碱性成纤维细胞生长因子 (bFGF)在胃癌组织中表达及其对血管新生和肿瘤生物学行为的影响。方法　应用免疫组化SP法检测 74例胃癌 ,17例癌旁组织bFGF表达及间质微血管密度 (MVD)。结果　胃癌组织中肿瘤细胞、间质新生血管高度表达bFGF。癌组织bFGF表达(77.0 3% )明显高于癌旁组织 (2 9.4 1% ,P <0 .0 1)。癌旁胃黏膜及伴有肠上皮化生的胃黏膜表达bFGF较弱。bFGF高表达组的平均MVD值 (79.3± 11.2 )明显高于bFGF低表达组 (71.2± 11.9,P <0 .0 5 )。此外bFGF表达程度与胃癌淋巴结转移和癌浸润深度密切相关。结论　bFGF可促进肿瘤间质微血管生成 ,加速肿瘤浸润和转移。     

RhoC基因及蛋白在胃癌肝转移中的表达及意义

目的:探讨胃癌肝转移中胃镜组织及肝转移癌组织中RhoC mRNA及蛋白的表达及意义。方法:采用逆转录聚合酶链式反应技术(RT-PCR)分析28例胃癌患者的癌组织(GC)及癌旁胃粘膜组织(PGMT)、肝转移癌(ML)组织中RhoC mRNA及蛋白的表达;采用免疫组织化学染色分析Rhoe蛋白的表达情况。结果:GC组织中RhoC mRNA光密度相对值为0.135±0.065;PGMT的光密度相对值为0.118±0.074,两者无显著性差异(P>0.05);ML中RhoC mRNA光密度相对值为0.641±0.113,与GC相比,差异有显著性意义(P<0.05)。RhoC蛋白的表达中,癌旁胃粘膜组织、胃癌组织、肝转移癌灶组织的灰度值分别为(128.523±11.486)U,(146.694±7.079)U,(162.351±14.612)U。经Student's t检验,差异有显著性意久(P<0.05);ML中RhoC mRNA及蛋白光密度相 对值明显高于GC(P<0.05)。结论:RhoC mRNA及蛋白的表达与胃癌转移有关,可用作判定胃癌转移及预后的指标。     

胃癌组织中CTGF蛋白表达及其临床意义

目的探讨结缔组织生长因子（CTGF）蛋自在胃癌发生、发展中的作用。方法采用免疫组化法检测10份正常胃黏膜组织、50份胃癌组织及其癌旁组织中CTGF蛋白表达情况。结果胃癌组织中CTGF阳性表达高于癌旁组织及正常胃黏膜组织,P〈0．05;癌组织CTGF阳性表达率明显高于正常黏膜组织（P〈0．01）,CTGF的高表达与胃癌分化程度、淋巴结转移密切相关（P均〈0．05）;与肿瘤浸润的深度无明显关系。结论CTGF蛋白可作为胃癌前病变及胃癌早期诊断和预后判断的指标。     

胃癌组织中c-FLIP、NF-κB p65的表达变化及意义

目的 探讨细胞型Fas相关的死亡区域蛋白样白介素-1转换酶抑制蛋白(c-FLIP)、细胞核因子κB(NF-κB)在胃癌发生、发展中的作用.方法 选择胃癌组织标本80份(胃癌组)和癌旁组织80份(癌旁组),采用免疫组化法检测两组的c-FLIP、NF-κB p65,分析其与胃癌临床病理特征的关系.结果 胃癌组c-FLIP、NF-κB p65阳性率明显高于癌旁组(P均＜0.05);c-FLIP的表达与胃癌临床分期、分化程度、淋巴结转移、术后复发相关(P均＜0.05),NF-κB p65的表达与浸润深度、分化程度、淋巴结转移、术后复发相关(P均＜0.05);c-FLIP和NF-κBp65在胃癌组织中的表达呈正相关(r =0.303,P=0.006).结论 c-FLIP、NF-κB p65的高表达可能在胃癌的发生、发展中起重要作用.     

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.     

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.