肝细胞肝癌组织中miR-21表达水平及与预后的相关性

目的探讨肝细胞肝癌患者组织中miR-21的表达水平及其预后意义。方法共收集50例肝细胞肝癌的患者,采用免疫组化法检测肝细胞肝癌及配对癌旁组织中miR-21的表达水平,分析miR-21蛋白的表达与患者临床病理特征及预后生存的相关性。结果肝细胞肝癌组织中miR-21表达水平显著高于癌旁组织(58.0%vs 6.0%,χ~2=31.07,P0.000 1)。miR-21高表达与临床T分期相关(χ~2=8.043,P=0.018);但miR-21蛋白表达水平与患者年龄、性别、肿瘤大小、临床分期、N分期及分化程度均无显著关联(P0.05)。高表达miR-21的肝细胞肝癌患者总生存时间明显降低(P=0.032),但患者无疾病进展生存期差异无统计学意义(P=0.116)。结论肝细胞肝癌患者miR-21高表达可能作为肿瘤患者预后不良的重要标志物之一,而与肿瘤的发生及发展的机制性研究尚需进一步验证。     

ESPL1基因在肝细胞癌组织中的表达及其在预后评估中的作用

目的 探讨外纺锤体极样蛋白1(ESPL1)基因在肝细胞癌(HCC)中的表达及其对临床预后的影响，初步分析ESPL1变异情况及分子生物学功能。方法 从TCGA数据库获得HCC患者ESPL1表达数据和临床信息，比较癌、癌旁及正常组织中ESPL1的表达情况。采用Cox回归分析影响HCC患者预后的危险因素，Kaplan-Meier法分析ESPL1表达水平与HCC患者预后的关系。log rank检验ESPL1高表达组和低表达组的生存差异。cBioportal在线软件分析ESPL1突变情况，通过STRING数据库绘制ESPL1蛋白互作网络，并据此进行GO|KEGG富集分析其分子生物学功能。结果 肝癌组织中ESPL1表达水平高于癌旁及正常组织，分别为0.894±0.643比0.337±0.340、1.908±1.079比0.912±0.770(P<0.05);乙型肝炎相关肝癌ESPL1水平高于非乙型肝炎相关肝癌，为2.159±1.17比1.747±0.983(P<0.05)。ESPL1表达水平与性别、年龄、BMI、原发肿瘤分期、AFP水平有关(P<0.05)。原发肿瘤分期、ESPL...     

利用GEO数据库分析肝细胞癌组织中着丝粒蛋白F基因的表达及其与预后的相关性

目的探讨着丝粒蛋白F(CENPF)基因在肝细胞癌(HCC)患者组织中的表达情况及与临床特征和预后的相关性。方法利用在线工具Oncomine分析CENPF在HCC与正常肝组织的表达情况,从美国国立生物技术信息中心的GEO数据库下载相关芯片数据(GSE14520),收集215例HCC组织的基因表达谱及相关临床数据,将其分为CENPF低表达组(n=107)和CENPF高表达组(n=108)。对样本中的CENPF基因表达数据及其对应的临床信息进行分析。计量资料组间比较采用t检验;计数资料组间比较采用χ~2检验;生存分析采用log-rank(Mantel-Cox)检验;生存资料单因素及多因素分析采用Cox比例风险模型。结果数据集证实HCC组织中CENPF表达水平明显高于正常肝组织(t=12.217,P0.001)。CENPF基因表达水平与AFP水平、肿瘤大小、TNM分期、转移风险有关系(χ~2值分别为6.463、4.338、7.951、17.331,P值均0.05)。生存分析显示CENPF高表达患者预后明显差于CENPF低表达组(风险比=1.92,95%可信区间:1.24~2.96,P=0.005)。Cox单因素分析显示,HCC患者预后与AFP水平、肿瘤直径、合并肝硬化、肿瘤分期、转移风险及CENPF表达相关(P值均0.05);Cox多因素分析显示,合并肝硬化、肿瘤分期及CENPF高表达是HCC患者预后的独立危险因素(P值均0.05)。结论 CENPF基因高表达可能与HCC进展相关,可作为HCC患者的潜在预后生物标志物。     

D-双功能蛋白在肝癌组织中的表达及其与AFP和CA19-9的相关性

目的探讨D-双功能蛋白(DBP)在肝细胞癌(HCC)组织中的表达情况,以及与肿瘤血清学标志物AFP、CA19-9表达的相关性,进而分析DBP对HCC早期诊断和预后判断的临床价值。方法选择唐山市妇幼保健院和唐山市人民院2015年6月-2017年5月行肝癌切除术并随访的20例患者作为试验组,同期两所医院的20例健康体检者作为对照组,应用化学发光法检测AFP、CA19-9的表达水平,免疫组化法检测肝癌组织和癌旁组织DBP的表达水平,用积分光密度值(IOD)进行测量。计量资料2组间比较采用t检验,检测指标相关性检验采用Pearson相关分析。结果肝癌组织DBP的IOD值(220. 52±101. 30)明显高于癌旁组织(40. 49±19. 32),差异具有统计学意义(t=-7.00,P 0.01)。且DBP与AFP(r2=0. 868,P 0. 01)和CA19-9(r2=0. 814,P 0. 01)的表达水平均呈正相关。结论 HCC患者DBP表达升高,与HCC的发生发展密切相关,为肝癌的诊断提供新的靶点。     

E2F5在肝细胞肝癌中的表达及其与患者预后关系的研究

目的探讨E2F5在肝细胞肝癌(Hepatocellular carcinoma, HCC)组织中的表达水平;分析E2F5的表达水平与HCC患者的性别、年龄、血清AFP及ki-67浓度、肿瘤病理学分级、肿瘤TNM分期及患者预后的关系。方法通过癌症基因组图谱(TCGA)数据库获取HCC患者的E2F5基因组学数据、临床资料及预后信息,比较HCC患者癌组织和癌旁正常组织中的E2F5表达水平;比较E2F5低表达组和E2F5高表达组HCC患者的性别、年龄、肿瘤病理学分级、肿瘤TNM分期、血清AFP水平、ki-67水平、患者的整体生存率。结果 HCC患者癌组织的E2F5表达水平显著高于其癌旁正常组织的表达水平,差异有统计学意义(P0.05)。E2F5高表达组HCC患者的病理学分级、肿瘤TNM分期、血清AFP水平、血清ki-67水平均显著高于E2F5低表达组患者,差异有统计学意义(P0.05),但患者的性别、年龄比较,差异无统计学意义(P0.05)。Kaplan-Meier生存曲线显示,E2F5高表达组HCC患者的整体生存率显著低于E2F5低表达组患者,差异有统计学意义(P0.05)。结论 HCC患者癌组织中的E2F5呈高表达,E2F5高表达患者的肿瘤病理学分级及TNM分期、血清AFP水平、血清ki-67水平均显著高于E2F5低表达患者,HCC患者E2F5的高表达与其不良预后密切相关。     

MAGE-4基因在肝细胞肝癌中的表达及临床意义

目的通过研究MAGE-4基因在肝细胞肝癌组织中的表达,并与患者临床资料进行分析,探讨 MAGE-4基因与肝细胞肝癌(HCC)患者临床指标及转移与复发的关系,为MAGE-4基因编码蛋白用于HCC患者免疫治疗提供依据. 方法用RT-PCR的方法对31例HCC患者癌组织及相应癌旁组织MAGE-4基因表达进行测定,对全部RT-PCR扩增产物中目的基因片段进行DNA测序以证实其为 MAGE-4基因,患者均测定并统计AFP、AFU、抗HCV、HBsAg、AFP mRNA、肿瘤直径等临床指标. 结果 31例HCC患者肝癌组织中MAGE-4基因表达的阳性率38.7% (12/31)明显高于癌旁组织中MAGE-4基因表达的阳性率0% (0/31),P＜0.01.HCC患者肝癌组织中MAGE-4基因表达的阳性率与患者AFP、AFU、抗HCV、HBV标志物、AFP mRNA、肿瘤直径等临床指标均无关,P＞0.05. 结论 MAGE-4基因在HCC患者肝癌组织中特异高表达,可能作为HCC患者免疫治疗攻击的靶点,HCC患者肝癌组织中 MAGE-4基因表达的阳性率与HCC患者肿瘤标志物、转移、复发均无关.     

miR-92、MDK在乙型病毒性肝炎相关肝癌组织的表达及意义

目的:检测微小RNA-92(miR-92)、中期因子(MDK)在慢性乙型肝炎(CHB)相关肝癌患者癌组织中的表达水平,并分析二者与CHB相关肝癌患者预后的关系。方法:选取2015年6月至2017年6月由CHB导致肝细胞癌(HCC)并行肝癌切除术的患者66例为研究对象,取患者术中切除的癌组织及癌旁组织分别作为HCC组和对照组。采用实时荧光定量PCR(qRT-PCR)法检测受试者组织中miR-92、MDK mRNA水平。免疫组化法检测石蜡切片中MDK表达情况。分析miR-92、MDK表达水平与临床病理特征的关系。Kaplan-Meier(K-M)法分析miR-92、MDK表达水平与CHB相关HCC患者预后的关系。COX回归分析CHB相关HCC患者预后不良的影响因素。结果:HCC患者肝癌组织中miR-92、MDK mRNA及蛋白表达水平均高于对照组(P<0.05)。CHB相关HCC患者中miR-92、MDK表达水平与组织分化、TNM分期、门脉癌栓、肿瘤转移有关(P<0.05)。K-M生存分析显示,miR-92高表达HCC患者5年累积生存率低于miR-92低表达患者(P<0....     

葡萄糖调节蛋白78调控乙型肝炎相关性肝细胞癌患者线粒体生成蛋白的表达及临床分析

目的研究葡萄糖调节蛋白78(GRP78)在乙型肝炎相关性肝细胞癌组织中的表达及其与患者临床病理特征的关系,探索GRP78对肝癌细胞中线粒体生成相关蛋白分子的调控,为建立肝癌防治的新策略提供基础。方法收集乙型肝炎相关性肝细胞癌54例患者组织标本,用免疫组化和Western Blot检测肝癌及癌旁组织中GRP78、Lon、TFAM、COXⅣ的表达;用siRNA干涉肝癌细胞中GRP78的表达,检测细胞中GRP78、Lon、TFAM、COXⅣ的表达;用实时定量PCR(qRT-PCR)检测临床标本和干涉GRP78表达后肝癌细胞中线粒体DNA(mt DNA)的水平;对临床资料和实验数据进行统计分析。计量资料2组间比较比较采用t检验,计数资料2组间比较采用Fisher精确检验,患者生存分析采用Kaplan-Meier法。结果 GRP78及Lon在乙型肝炎相关性肝癌组织中的表达显著高于癌旁组织(t值分别为9.135、5.523,P值均0.001),而线粒体生成相关蛋白TFAM、COXⅣ的表达及mt DNA水平显著低于癌旁组织(t值分别为2.765、4.260、12.280,P值分别为0.011、0.001、0.001)。干涉肝癌细胞中GRP78的表达,可显著提高线粒体生成相关蛋白TFAM、COXⅣ的表达及mt DNA水平(P值均0.05)。GRP78在不同肿瘤数量、门静脉癌栓以及肿瘤分期的病例中,表达水平存在显著差异(P值分别为0.016、0.003、0.045);GRP78低表达患者术后总生存期及无复发生存期优于GRP78高表达的患者(χ2值分别为5.006、4.995,P值分别为0.025、0.026)。结论 GRP78对线粒体维持与生成具有潜在的调控作用,对建立潜在的肝癌防治新策略具有重要的临床指导意义。     

长链非编码RNA-lncRNA2在食管癌中高表达

目的探讨lncRNA2在食管癌中的表达情况,及lncRNA2异常表达作为食管癌的潜在诊断标志物和治疗靶标的可能。方法应用lncRNA表达芯片技术对14对配对的食管癌和癌旁组织进行分析,获得在癌组织和癌旁组织中差异表达>10倍的lncRNA 134个。应用RT-PCR技术在食管癌细胞系进行验证,并分析43对配对的癌及癌旁组织中的表达情况。结果在14个食管癌细胞系中,lncRNA2在10个食管癌细胞系中高表达,而在4个食管癌细胞系中缺失表达。在43对配对的食管癌及癌旁组织中,lncRNA2在26例癌组织中高表达,在癌旁组织中缺失表达或低表达;11例在癌组织中缺失表达或低表达,而在癌旁组织中高表达;另外6例在癌组织和癌旁组织中均缺失表达。lncRNA2在60.47%(26/43)的食管癌组织中高表达,而在25.58%(11/43)的癌旁组织中高表达,差异有统计学意义(P<0.01)。lncRNA2高表达与患者年龄、性别、TNM分期、肿瘤大小、吸烟史、饮酒史、家族史等临床因素无相关性(P均>0.05)。结论lncRNA2在食管癌中表达明显升高,是食管癌潜在的诊断标志物和治疗靶标。     

IQGAP2在肝细胞肝癌中的表达及其临床意义

目的:认识肝细胞肝癌中含IQ模体的GTP酶活化蛋白2(IQ motif containing GTPase activating protein 2,IQGAP2)的亚细胞定位和表达,及其在肝细胞肝癌(hepatocellular carcinoma,HCC)中的作用.方法:分别通过Western blot、免疫荧光和免疫组织化学染色分析IQGAP2在7种人肝癌细胞与正常肝细胞系中的表达、定位,及其在HCC组织样本中的表达与分布.结果:IOGAP2在Bel-7402、Bel-7404、SMMC-7721、SK-HEP-1、HLE和HL-7702等肝癌和正常成人肝细胞系中不表达,仅在HepG2和Hep3B等2种甲胎蛋白(AFP)表达阳性的人肝癌细胞系中表达,IQGAP2主要分布于细胞质,此外,在HepG2细胞中还具有明显的核膜和核仁定位,IQGAP2在HCC组织中表达降低(56.9%,29/51).其中,19例配对HCC组织的IQGAP2表达与肿瘤大小,AJCC分期和血清AFP水平相关(P=0.020,P=0.017,P=0.002);38例配对组织IQGAP2在肿瘤和癌旁正常肝组织中主要定位于胞质,部分细胞伴有胞核和胞膜定位,但是,IQGAP2的表达与肿瘤大小、分化程度、AJCC分期以及血清AFP表达水平无相关性.结论:IQGAP2蛋白可能参与细胞黏附、信号转导等过程,在肝癌的发生发展中发挥重要作用,是一种潜在的抑癌基因.     

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.