支气管哮喘患者外周血巨噬细胞移动抑制因子的表达及意义

目的 观察巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)在支气管哮喘(简称哮喘)患者外周血中的表达水平及其意义.方法 应用酶联免疫吸附试验法和荧光定量PCR技术,测定56例哮喘患者(哮喘组)和34名健康对照者(正常对照组)外周血MIF的表达水平.根据临床症状和肺功能检查将哮喘患者分为发作期组和缓解期组,其中发作期组32例,缓解期组24例,并对结果进行统计学处理.结果 血浆MIF的浓度和外周血单个核细胞MIF mRNA的表达水平:哮喘组高于正常对照组,差异有统计学意义(P<0.05);哮喘发作期组高于缓解期组,差异有统计学意义(P<0.05);但缓解期组与正常对照组之间差异无统计学意义.结论 MIF可能参与哮喘的病理过程并发挥重要作用,哮喘患者发作期组血MIF mRNA表达及血MIF水平显著增高,其变化可反映疾病的活动性.     

支气管哮喘急性发作期患者外周血Bcl-6mRNA、Blimp-1mRNA、IL-21水平变化及意义

目的通过观察支气管哮喘急性发作期患者外周血的Bcl-6mRNA、Blimp-1mRNA、IL-21的表达水平,探讨其在支气管哮喘发作、发展中的作用及价值,为临床的诊断及治疗提供参考及指导。方法搜集支气管哮喘急性发作期患者90例作为观察组,健康体检者60例作为对照组。采用酶联免疫吸附测定法(ELISA)检测各组的IL-21表达水平,实时荧光定量PCR法检测各组的Bcl-6mRNA、Blimp-1 mRNA表达水平。比较观察组与对照组之间Bcl-6mRNA、Blimp-1mRNA、IL-21的表达水平,明确Bcl-6mRNA、Blimp-1mRNA、IL-21在哮喘急性发作期所起的作用和在哮喘的诊断及治疗的价值。结果观察组与对照组的Bcl-6mRNA、Blimp-1mRNA、IL-21存在差异,两组的Blimp-1 mRNA表达水平分别为(0.621±0.216)和(1.166±0.661),两组的Bcl-6mRNA表达水平分别为(2.414±1.006)和(1.071±0.394),两组的IL-21表达水平分别为(80.917±22.764)和(17.612±6.079)。观察组Bcl-6mRNA、IL-21表达水平均显著高于对照组(P0.01),Blimp-1 mRNA表达水平显著低于对照组(P0.01)。结论 Bcl-6mRNA、Blimp-1mRNA、IL-21参与了哮喘的急性发作,其中Bcl-6mRNA、IL-21可能是哮喘发作的诱发因素,Blimp-1mRNA是哮喘发作的保护因素。Blimp-1mRNA、Bcl-6mRNA、IL-21可作为哮喘急性发作诊断过程中的重要参考指标,对其进行联合检测有利于哮喘的早期诊断及治疗。     

支气管哮喘患者呼出气一氧化氮与外周血嗜酸性粒细胞的相关性

目的观察支气管哮喘患者呼出气一氧化氮(Fe NO)的表达水平,分析其与外周血嗜酸性粒细胞(EOS)之间的相关性。方法收集2013年2月-2014年12月期间贵州省人民医院支气管哮喘患者58例,其中支气管哮喘急性发作期40例(急性发作期组),支气管哮喘缓解期18例(缓解期组),健康对照组30例。采用Fe NO分析仪测定Fe NO水平,并检测各组外周血嗜酸粒细胞计数,采用SPSS 11.5统计软件进行统计学处理,组间差异性比较采用单因素方差分析,相关性分析采用Pearson相关性分析。P0.05为差异有统计学意义。结果支气管哮喘急性发作期组Fe NO为101.44±32.87ppb,EOS为5.0±2.62×10~9/L;缓解期组Fe NO为32.83±11.42 ppb,EOS为1.32±0.59×10~9/L;健康对照组Fe NO为8.43±3.02ppb,EOS为0.22±0.09×10~9/L。发作期组及缓解期组Fe NO及EOS水平均高于健康对照组(P0.05)。急性发作期组与缓解期组比较,Fe NO及EOS水平差异有统计学意义(P0.05)。支气管哮喘患者Fe NO水平与EOS呈正相关性(P0.05)。结论 Fe NO能够一定程度上反应气道嗜酸细胞炎症,Fe NO检测可能有助于评估支气管哮喘的控制水平。     

哮喘患者外周血单个核细胞NOD2 mRNA表达的研究

目的探讨外周单个核细胞NOD2 mRNA表达在哮喘发病机制中的作用以及与气道炎症的关系。方法将60例哮喘患者分为急性发作组和慢性持续期组,每组30人,经治疗症状体征好转后,作为临床缓解期组,采用实时荧光定量聚合酶链反应(PCR)法和酶联免疫吸附测定(ELISA)法分别检测哮喘各组及健康对照组30人外周血单个核细胞核内NOD2 mRNA的表达量及血清IL-6的含量。结果急性发作期哮喘患者的NOD2 mRNA的表达量及IL-6的含量均明显高于慢性持续期哮喘患者(P0.05)。急性发作期和慢性持续期哮喘患者的NOD2 mRNA的及IL-6的表达,均明显高于健康对照组(P0.05)。急性发作期和慢性持续期哮喘患者经治疗后的NOD2 mRNA及IL-6的表达较各自治疗前水平明显降低,且均高于健康对照组水平,具有统计学意义(P0.05)。哮喘患者外周血单个核细胞核内NOD2 mRNA表达随血清IL-6含量呈正相关,相关系数r=0.85,P0.05。结论 NOD2和IL-6都参与了哮喘的气道炎症反应,且与哮喘患者的严度程度呈正相关。     

支气管哮喘患者急性发作期外周血LncRNA NEAT1表达及与Th1/Th2平衡的关系

目的：探讨支气管哮喘（BA）患者急性发作期外周血长链非编码RNA（LncRNA）核富集转录本1（NEAT1）表达水平及其与辅助性T细胞1（Th1）/辅助性T细胞2（Th2）平衡的关系。方法：选取BA缓解期患者36例（缓解期组）、急性发作期患者44例（急性发作期组）,另选取同期体检健康者50例作为对照组。利用实时荧光定量PCR（qRT-PCR）测定外周血LncRNA NEAT1表达,应用流式细胞仪测定外周血单个核细胞（PBMC）中Th1、Th2细胞比例,应用ELISA法测定外周血干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）水平;采用Pearson检验分析BA急性发作期患者外周血LncRNA NEAT1表达与Th1/Th2、IFN-γ/IL-4比值的关系,并绘制ROC曲线分析LncRNA NEAT1表达水平对BA患者急性发作的预测价值。结果：急性发作期组患者外周血LncRNA NEAT1表达水平最高,其次为缓解期组,对照组最低,组间比较差异有统计学意义（均P＜0.05）。PBMC中Th1细胞比例及Th1/Th2比值高低依次为对照组、缓解期组、急性发作期组,Th2细胞比例高低依次为急性发作期组、缓解期组、对照组,组间比较差异有统计学意义（均P＜0.05）。外周血IFN-γ水平及IFN-γ/IL-4比值高低依次为对照组、缓解期组、急性发作期组,IL-4水平高低依次为急性发作期组、缓解期组、对照组,组间比较差异有统计学意义（均P＜0.05）。BA急性发作期患者外周血LncRNA NEAT1表达与Th1/Th2、IFN-γ/IL-4比值呈负相关（均P＜0.05）。外周血LncRNA NEAT1表达水平预测BA患者急性发作的曲线下面积为0.903,最佳截断值为1.73,敏感性为81.80%,特异性为88.90%。结论：BA患者急性发作期外周血LncRNA NEAT1高表达,可能与Th1/Th2平衡紊乱有关。     

哮喘患者外周血单个核细胞白细胞介素-13 mRNA表达的研究

目的 探讨哮喘患者外周血单个核细胞(PBMCs)白细胞介素(IL)-13 mRNA的表达和糖皮质类固醇激素治疗对其的影响.方法 采用逆转录聚合酶链反应(RT-PCR)法分别检测34例发作期哮喘患者(其中14例激素敏感型和10例激素抵抗型分别口服泼尼松治疗1周)、18例缓解期患者和30名健康对照者PBMCs IL-13 mRNA的表达.结果 52例哮喘患者和30名健康对照者PBMCs IL-13 mRNA的表达量分别为16.72±1.12和8.25±1.15,两组比较差异有统计学意义(P＜0.05).发作期和缓解期哮喘患者PBMCs IL-13 mRNA的表达量分别为20.72±1.02和13.67±3.15,两组比较差异有统计学意义(P＜0.05).在34例发作期哮喘患者中,轻、中、重度患者PBMCs IL-13 mRNA表达量分别为16.35±2.38、19.12±2.16、23.57±2.43,相互间比较差异有统计学意义(P＜0.05).激素敏感型哮喘患者经泼尼松治疗1周后PBMCs IL-13 mRNA表达量(21.03±3.77)较治疗前(15.67±2.67)明显下降,差异有统计学意义(P＜0.05);激素抵抗或依赖型哮喘患者经泼尼松治疗1周前后PBMCs IL-13 mRNA 表达量分别为20.35±3.58和19.57±3.34,差异无统计学意义(P＞0.05).结论 PBMCs IL-13参与哮喘发生的病理过程,并调节糖皮质类固醇激素的作用,可作为判断哮喘患者病变严重程度的指标之一.     

支气管哮喘患者外周血巨噬细胞移动抑制因子的表达及意义

目的观察巨噬细胞移动抑制因子（macrophagemigrationinhibitoryfactor,MIF）在支气管哮喘（简称哮喘）患者外周血中的表达水平及其意义。方法应用酶联免疫吸附试验法和荧光定量PCR技术,测定56例哮喘患者（哮喘组）和34名健康对照者（对照组）外周血MIF的表达水平。根据临床症状和肺功能检查将哮喘患者分为发作期、缓解期两组,其中发作期32例,缓解期24例,并对结果进行统计学处理。结果血浆MIF的浓度和外周血单个核细胞MIFmRNA的表达水平：哮喘组高于对照组,差异有统计学意义（P〈0．05）;哮喘发作期组高于缓解期组,差异有统计学意义（P〈0．05）;但缓解期组与正常之间差异无统计学意义。结论MIF可能参与哮喘的病理过程并发挥重要作用,哮喘患者发作期血MIFmRNA表达及血MIF水平显著增高,其变化可反映疾病的活动性。     

凋亡抑制基因Bcl-2表达与支气管哮喘关系的研究

目的 支气管哮喘的发病机制非常复杂,至今尚未完全阐明.目前认为细胞凋亡与支气管哮喘发病相关.Bcl-2基因是近年来广泛重视的一种凋亡抑制基因,可抑制多种细胞凋亡,与多种疾病的发生存在密切的关系.本研究测定支气管哮喘患者外周血白细胞Bcl-2基因mRNA的表达量及其血清中蛋白的含量,研究Bcl-2在支气管哮喘中的表达及其相互关系,初步探讨Bcl-2基因在支气管哮喘的发生、发展中的生物学意义. 方法 20例支气管哮喘中重度发作患者为实验组,20例无支气管哮喘、变态反应性疾病的健康人为对照组.采取抗凝静脉血标本5ml,立即用Dextron-PBS分离出血白细胞,采用TristarTM试剂按试剂盒说明提取总RNA,-80°超低温冰箱保存,采用TaKaRa试剂盒检测Bcl-2基因mRNA表达量.两步法第一步用ExScriptTM反转录酶,对Bcl-2mRNA反转录成cDNA;第二步用Premix Ex TaqTM酶对反转录后的cDNA进行扩增,10倍梯度稀释cDNA模板,分别为100、10-1、10-2、10-3、10-4倍梯度.用美国PE公司5700型实时荧光定量PCR仪进行检测,以GAPDH基因为对照基因,用仪器附带SDS软件自动算出Bcl-2基因与对照基因比值,确定Bcl-2基因的相对含量.同时抽取抗凝静脉血2ml,伊红染色,行外周血嗜酸性粒细胞计数.抽取不抗凝静脉血2ml,分离血清,采用澳大利亚Bender Medsystems公司人Bcl-2蛋白检测试剂盒,按试剂盒说明操作,通过酶联免疫吸附法(ELISA)检测Bcl-2蛋白含量.应用SAS 6.12计算机软件对实验结果进行统计学分析. 结果 支气管哮喘患者外周血嗜酸性粒细胞计数较正常对照组比较有非常显著性增加(P＜0.01),20例支气管哮喘患者外周血细胞Bcl-2基因mRNA表达量高于正常对照组,两组有非常显著性差异(P＜0.01),外周血清Bcl-2蛋白含量与正常对照组差异无显著性(P＞0.05). 结论 本研究证实支气管哮喘患者外周血嗜酸性粒细胞显著增多.支气管哮喘患者外周血细胞Bcl-2基因mRNA表达显著增多,揭示Bcl-2基因在支气管哮喘发病中具有一定的作用和意义,Bcl-2基因与支气管哮喘密切相关.而外周血清中Bcl-2蛋白含量与支气管哮喘的发病无直接相关.     

SORL1基因表达与阿尔茨海默病关系的研究

目的研究中国汉族人迟发散发型阿尔茨海默病(sporadic Alzheimer’s disease,SAD)外周血SORL1 mRNA的表达,为临床诊断提供依据。方法选择迟发SAD患者23例(SAD组),根据WHO疾病分类第10修订版临床诊断标准为迟发SAD,另选健康体检者12例(对照组)。分离2组外周血白细胞,提取总RNA,反转录合成cDNA,采用实时荧光定量PCR检测,以SORL1和磷酸甘油醛脱氢酶的mRNA含量比值作为评价SORL1 mRNA表达水平的指标,并计算出待测样本中SORL1 mRNA表达的准确含量。结果 SAD组外周血SORL1 mRNA表达明显高于对照组,差异有统计学意义(1.17±0.33 vs 0.39±0.29,P0.01),与性别、年龄无明显相关性。结论汉族人外周血SORL1 mRNA表达水平与迟发SAD密切相关;通过实时荧光定量检测外周血SORL1 mRNA表达,对迟发SAD早期诊断可起到辅助的指导作用。     

Aiolos基因mRNA表达与支气管哮喘发病的相关性研究

目的探讨淋巴细胞转录因子Aiolos基因mRNA的表达水平与支气管哮喘(简称哮喘)发病的关系,为哮喘发病和治疗研究寻找新的突破点。方法选取33例哮喘患者及38位健康对照者,采用SYBR Green荧光实时定量PCR方法,以磷酸甘油醛脱氢酶(GAPDH)基因作为内参,检测两组Aiolos基因mRNA的表达水平的差异。结果利用比较平均循环数值(Ct)法测定mRNA的相对表达程度,哮喘患者组Aiolos基因与内参基因GAPDH的△Ct值为5.29±2.29,健康对照组△Ct值3.45±4.99,Aiolos基因在哮喘患者外周血中的表达水平显著低于正常人(P=0.046)。结论Aiolos基因的低水平表达可能是导致哮喘发病的原因之一。     

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.