大肠腺瘤与大肠腺癌肿瘤相关基因的差异表达

目的:探索大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因.方法:利用基因芯片技术筛选大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因,比较两组基因特点,寻找大肠腺癌重要致病基因,并以RT-PCR对部分差异表达基因进行检测来验证芯片结果.结果:(1)大肠腺瘤差异表达的肿瘤相关基因9个,均上调;(2)大肠腺癌差异表达的肿瘤相关基因47个...     

基因芯片筛选ghrelin miRNA转染胰腺炎细胞模型后的差异表达基因

背景:前期研究提示ghrelin在急性胰腺炎发病过程中发挥重要作用,但其机制尚不明确。目的:应用基因芯片技术筛选出ghrelin miRNA转染胰腺炎胰腺腺泡细胞后的差异表达基因。方法:以雨蛙肽构建胰腺炎胰腺腺泡细胞模型。将AR42J细胞分为ghrelin miRNA+雨蛙肽组、阴性对照+雨蛙肽组。提取细胞RNA,合成为ds-DNA,与大鼠全基因组基因芯片杂交、扫描、分析。筛选与炎症和钙通路相关的差异表达基因,并采用RT-PCR法对其中3个基因(Bcl-2、caspase-8、caspase-12)进行验证。结果:给予AR42J细胞ghrelin miRNA+雨蛙肽干预后,共筛选出2 938个差异表达基因,其中1 435个基因表达上调,1 503个基因表达下调。与CAMP/Ca2+信号通路相关的差异表达基因有60个,与炎症通路相关的差异表达基因有199个。RT-PCR结果表明,ghrelin miRNA+雨蛙肽组Bcl-2、caspase-12表达下调,caspase-8表达上调,与基因芯片筛查结果一致。结论:基因芯片技术可筛选出ghrelin miRNA转染胰腺炎胰腺腺泡细胞过程中发挥关键作用的基因,从而为研究内源性ghrelin对胰腺炎胰腺腺泡细胞的作用机制提供依据和参考。     

应用基因芯片筛选HBV 对肝癌细胞脂代谢相关基因影响的初步研究

目的通过基因芯片技术探测HBV感染对肝癌细胞脂代谢相关基因表达的影响,为深入研究HBV在肝癌脂代谢中的作用及机制提供新的依据。方法本研究以HepG2细胞作为对照,用稳定表达HBV的肝癌细胞HepG2.2.15来模拟HBV感染的肝癌细胞,利用基因芯片对肝癌细胞HepG2和HepG2.2.15进行分析,筛选出脂代谢相关的差异表达基因,对差异基因参与的信号传导通路进行分析。RT-PCR检测显著差异表达基因CYP1A1、CYP1B1和AHR。结果基因芯片共筛选出肝癌细胞HepG2和HepG2.2.15中差异表达基因共1263个,其中涉及脂代谢的基因有92个,具有表达上调的基因53个,表达下调的基因39个,涉及细胞生长、增殖、分化、运动和耐药等多种分子功能。RT-PCR检测脂代谢差异表达基因,其结果与芯片结果一致,确定芯片检测结果可靠。结论应用人类表达芯片成功筛选出HBV转染的人肝癌细胞后的脂代谢相关差异表达基因,为HBV感染在肝癌中的作用提供线索。     

重症肌无力患者胸腺自身免疫相关基因表达的研究

目的探讨MG患者和正常对照者的胸腺组织炎症反应与自身免疫性疾病相关基因的表达情况。方法取MG患者（MG组）和正常对照者（对照组）的胸腺组织标本，提取总RNA，反转录合成cDNA后，体外转录合成生物素标记的cRNA与基因芯片进行杂交，通过化学发光法检测基因表达，比较两组胸腺组织基因表达的差异。结果MG组与对照组比较，有61条基因存在显著性差异，其中表达上调33条，下调28条，这些基因包括炎症相关趋化因子基因、细胞因子及其受体基因、与细胞因子产生与代谢有关的基因、细胞因子及其受体相互作用相关的基因、炎症反应相关基因以及体液免疫相关基因等。结论芯片筛选MG患者胸腺自身免疫相关基因表达与正常对照组存在显著差异。     

应用基因芯片研究肝细胞癌组织差异基因表达谱

目的：应用基因芯片技术研究原发性肝细胞癌组织中的差异基因表达谱改变，以寻找肝细胞癌相关基因。方法：抽提正常肝组织和肝癌组织中的mRNA来制备探针，经杂交、洗涤后，通过计算机扫描分析正常肝组织和肝癌组织基因表达谱的差异情况。结果：在10000个候选基因中，筛选出102条差异表达基因，表达上调的有42条，表达下调的有60条。未知基因12条。结论：基于DNA微阵矩技术的肿瘤基因表达谱分析能够高通量筛选与肝癌发生发展相关的基因。     

采用人基因表达谱芯片筛选肺癌转移相关基因的初步研究

目的应用基因芯片技术分析人高、低转移肺巨细胞癌细胞株的基因差异表达谱，筛选与肺癌转移相关基因。方法提取人高转移肺巨细胞株95D和人低转移肺巨细胞株95C的mRNA，通过逆转录，制备分别用cy5-dUTP和cy3-dUTP进行标记的cDNA探针，并与芯片杂交。经过ScanArray3000扫描仪扫描，获取图象及对杂交信号进行数据分析，筛选出95D和95C细胞表达差异的基因谱，并对其中部分基因进行RT—PCR分析、验证。结果在所检测95C和95D细胞中，466条基因有表达差异，其中具有同-Genebank量的有108对，上调基因47对，下调基因61对。对其中KIAA1108、PGR1、JWA、S182、Jab1部分差异表达基因，经RT-PCR技术验证，结果与基因芯片基本符合。结论肺癌转移过程与多种基因作用有关，基因芯片技术可为肺癌转移相关基因的筛选，提供有效方法。     

应用基因芯片技术筛选胰腺癌相关基因

目的应用基因芯片技术筛选胰腺癌相关基因。方法将14000种人类基因PCR产物按微矩阵排列点样于化学涂层的载玻片上，制成基因芯片。按一步法抽提4例胰腺癌和癌旁正常胰腺组织的总RNA，将等量的RNA分别逆转录合成荧光分子掺人的cDNA一链作探针，混合后杂交上述基因芯片。经严格洗片后用ScanArray 4000扫描仪扫描芯片荧光信号图像，每点上两种荧光信号的强度分别代表Cy5-dCTP和Cy3-dCTP的量，获得的荧光信号图像用计算机分析。结果按差异显著性标准，从14000个基因中筛选出在胰腺癌组织中共同差异表达基因189条，其中已知基因101条，新基因88条。在筛选出的已知基因中，有50条表达上调，51条表达下调。结论基因芯片技术的肿瘤基因表达谱分析能够高通量筛选胰腺癌相关基因。并高效对基因功能进行研究。胰腺癌基因表达谱的分析有助于认识肿瘤发病机制。     

结直肠癌相关差异表达基因的生物信息学分析

背景:结直肠癌(CRC)是消化系统常见肿瘤,发病率和死亡率较高。目的:应用生物信息学方法对CRC差异表达基因进行分析,筛选与CRC发生、发展相关的基因。方法:从公共基因表达数据库(GEO)下载CRC基因芯片GSE32323、GSE21510、GSE9348数据集,使用R语言筛选差异表达基因。在DAVID数据库中对差异表达基因行GO和KEGG分析。应用STRING、Cytoscape构建蛋白质-蛋白质相互作用(PPI)网络,筛选出CRC的核心基因。结果:在三个数据集中共筛选出834个CRC共同差异表达基因,包括376个上调基因和456个下调基因。GO分析表明差异表达基因主要参与细胞分裂、增殖、代谢等过程。KEGG分析显示差异表达基因主要富集于p53通路和细胞外基质蛋白通路。PPI网络共筛选出20个核心基因。结论:运用生物信息学方法对CRC基因芯片进行分析可为CRC发病机制、肿瘤标记物的筛选和治疗药物靶点的选择提供理论基础。     

心肌淀粉样变人血清对培养心肌细胞基因表达谱的影响

目的:利用基因表达谱芯片技术筛选正常人血清和心肌淀粉样变患者血清干预培养心肌细胞的差异表达基因,以探讨淀粉样变心肌病发病的可能机制。方法:通过将分离的血清稀释10倍后干预培养SD乳鼠心肌细胞建立模型,采用CCK-8检测各组细胞活性抑制程度,基因表达谱芯片技术筛选两组大鼠差异表达基因,采用实时定量PCR验证相关基因表达量。结果:CCK-8显示与正常control组相比,阴性对照组细胞存活率略有下降,但无统计学意义。与control组相比,实验组细胞存活率明显下降至约70%(P0.05)。基因芯片分析显示,患者血清实验组及正常血清对照组两组相比,差异表达的基因有869条,其中上调基因710条,下调基因159条。实验组中胶原形成相关基因Col1a1、Col3a1、Col5a2、Col6a1、Col6a2、Col6a3,促纤维形成相关基因Lama1、Itga4、Vwf、Mmp14的表达均明显上调(P0.05);影响血管重建相关基因Angptl4、Asb4、Cited1表达下调(P0.05)。经RT-PCR验证,结果一致。结论:心肌淀粉样变患者血清干预影响培养心肌细胞的活性,促进胶原纤维形成并影响血管重建。淀粉样变心肌病的病理机制涉及多个基因,其中基质纤维化、血管重建受阻对淀粉样变心肌病的发生发展起着至关重要的作用,其具体机制还有待进一步研究。     

Iprl对巨噬细胞抗结核分枝杆菌感染免疫相关基因表达的影响

目的应用基因芯片技术检测Iprl的表达对巨噬细胞感染分枝杆菌H37Ra后参与免疫应答的相关基因的表达差异。方法实验组与对照组细胞分别感染分枝杆菌H37Ra,基N芯片检测实验组和对照组细胞感染H37Ra96h后免疫相关基因表达差异。定量PCR反应检测芯片结果中上调的3个基因的表达差异用以验证芯片结果的可靠性。结果基因4心4-片检测结果显示Iprl基因的表达上调了11个固有免疫机制中相关基因表达,其中与M中抗Mtb感染关系密切的基因有：TI．R2、TLR4、Irakl、Traf6、Ifngrl、Tnfrsfla,而参与调节适应性免疫应答的相关基N如：IL1,II．一12无明显表达差异。结论Ip。1增强M西抗Mtb感染的机制可能主要是通过上调M西抗感染固有免疫机制的有关基因的表达从而发挥抗菌作用。本实验为进一步研究Iprl促进M西的活化及杀伤胞内吞噬的Mtb的作用机制奠定了基础。     

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.