白细胞介素-18在类风湿关节炎外周血单个核细胞中的表达

目的　探讨类风湿关节炎 (RA)外周血白细胞介素 18(IL 18)的表达及其与Th1/Th2平衡和疾病活动的关系。方法　以 β actin为内参照 ,用半定量反转录多聚酶链反应 (RT PCR)法测定 16例RA患者及 15名正常对照外周血单个核细胞 (PBMC)中IL 18、IL 4、IFN γmRNA水平 ,观察IL 18与IL 4、IFN γ及疾病活动指标红细胞沉降率 (ESR)、C反应蛋白 (CRP)的相关性。结果　①RA患者PBMC中IL 18mRNA表达与正常组相比增高非常显著 (1 82± 0 39比 0 45± 0 30 ,P <0 0 0 1)。②RA组IFN γ、IL 4均显著高于对照组 ,但IL 4/IFN γ显著低于对照组。③相关分析显示 :RA组IL 18mRNA与IFN γ、ESR显著正相关 (16例 ,r分别为 0 836 ,0 75 3,P均 <0 0 5 )。结论　RA患者外周血IL 18mRNA表达水平显著高于正常对照组 ,且与RA患者外周血IFN γ的产生及疾病活动性相关。     

系统性红斑狼疮外周血单个核细胞生长激素受体和mRNA表达的研究

目的　探讨SLE患者外周血单个核细胞 (PBMC)的生长激素受体 (GHR)的表达与系统性红斑狼疮 (SLE)的关系。方法　采用放射性受体配基结合试验 (RLBA)及反转录 聚合酶链反应(RT PCR)研究外周血淋巴细胞GHR变化。结果　SLE活动期患者PBMC的GHR的特异性结合率 (SB) (3 4± 2 0 ) %、总结合率 (TB) (13 8± 5 7) %和静止期患者SB (1 3± 1 2 ) % ,TB (11 4±4 6 ) %均高于正常对照组SB (1 0± 1 0 ) % ,TB (8 3± 0 9) % (P <0 0 1) ;SLE活动期患者PBMC的mRNA的表达水平 (0 77± 0 6 6 )高于静止期患者 (0 4 5± 0 2 8) (P <0 0 5 )和正常对照组PBMC的mRNA的表达 (0 31± 0 2 2 ) (P <0 0 1)。结论　SLE患者PBMC的GHR高表达与SLE的病情活动性相关。     

类风湿关节炎外周血单个核细胞和滑膜中人类软骨糖蛋白-39的研究

目的　通过观察人类软骨糖蛋白 39(HCgp 39)mRNA在类风湿关节炎 (RA)外周血单个核细胞 (PBMC)和滑膜中的表达水平 ,寻找反映RA早期骨破坏的敏感指标。方法　采集 31例RA、6例骨关节炎 (OA)、10例血清阴性脊柱关节病 (SpA)、5例系统性红斑狼疮 (SLE)以及 10例健康人新鲜抗凝静脉血 ,分离单个核细胞 ,并采集 7例RA、5例OA的滑膜 ,行半定量逆转录 (RT) PCR。结果　RA患者PBMC中HCgp 39mRNA行RT PCR与内对照tubulin共扩增后吸光度比值为0 86 90± 0 5 2 4 0 ,与OA(P =0 0 2 4 )、SpA(P =0 0 4 9)、SLE(P =0 0 4 3)以及健康对照 (P =0 0 33)相比差异均有显著性。而OA、SpA、SLE与健康对照之间差异无显著性。RA滑膜HCgp 39mRNA行RT PCR与tubulin共扩增后吸光度比值为 1 986 3± 1 9397,与OA相比差异有显著性 (P =0 0 4 )。在RA和OA中 ,同一患者的PBMC与滑膜HCgp 39mRNA表达差异无显著性。 结论　HCgp 39mRNA在RA患者PBMC和滑膜的表达明显高于其他炎症性关节病 ,提示HCgp 39可能是RA发病机制中重要的自身抗原 ,它的异常表达可能是造成T细胞介导自身免疫反应的机制之一。     

活动性狼疮肾炎患者外周血单个核细胞钙调磷酸神经酶活性的检测及其意义

目的　检测活动性狼疮肾炎 (LN)患者外周血单个核细胞 (PBMC)钙调磷酸神经酶(calcineurin ,CaN)活性及其与PBMC分泌免疫球蛋白、自身抗体的关系 ,探讨FK5 0 6治疗狼疮肾炎的作用机制。方法　体外培养活动性LN患者PBMC ,应用发色底物法检测胞质CaN活性 ,酶联免疫吸附法 (ELISA)测定细胞培养上清免疫球蛋白和抗dsDNA抗体。结果　①单纯培养时 ,LN组CaN活性显著高于对照组 [(4 8 6± 4 7)nmol/mg蛋白vs(8 9± 2 7)nmol/mg蛋白 ,P <0 0 0 1 ];在PMA +Ionomycin刺激下 ,各组CaN活性均升高 ,LN组CaN活性明显高于正常对照组 [(71 2± 1 2 9)nmol/mg蛋白vs (34 2± 8 4 )nmol/mg蛋白 ,P <0 0 0 1 ];②单纯培养时 ,LN组PBMC培养上清中IgG浓度显著高于对照组 [(2 1 0 8± 6 0 0 )mg/Lvs (1 4 97± 5 1 6 )mg/L ,P <0 0 5 ];刺激条件下 ,LN组IgG水平显著高于对照组 [(4 991± 1 2 0 2 )mg/Lvs (3374± 1 1 6 6 )mg/L ,P <0 0 0 1 ];③单纯培养时 ,LN组PBMC培养上清中抗dsDNA抗体水平显著高于对照组 [(1 37± 0 1 6 )BIvs (0 71± 0 0 5 )BI,P <0 0 5 ];刺激条件下 ,LN组抗dsDNA抗体水平显著高于对照组 [(2 38± 1 1 7)vs (1 0 9±0 0 2 )BI ,P <0 0 0 1 ];④CaN特异的拮抗剂FK5 0 6显著抑制LNPBM     

血管内皮生长因子的表达与胃癌浸润和转移的关系

目的　研究血管内皮生长因子165(VEGF)mRNA在胃癌中的表达 ,探讨VEGF与胃癌浸润和转移的关系。方法　采用RT PCR方法 ,对 31例胃癌及非癌组织手术标本中VEGF165mRNA的表达进行相对定量研究。结果　胃癌组织中VEGF165mRNA表达的平均相对量 (1.12 5± 0 .35 6 )明显高于非癌组织的表达量 (0 .76 0± 0 .2 78,P <0 .0 5 ) ,其中淋巴结转移组 (1.2 19± 0 .377)和Ⅲ、Ⅳ期组 (1.2 6 2±0 .386 )分别高于无淋巴结转移组 (0 .92 7± 0 .2 0 5 )和Ⅰ、Ⅱ期组 (0 .934± 0 .194 ,P均 <0 .0 5 )。VEGF高表达者中淋巴结转移率为 83.3% ,Ⅲ和Ⅳ期占 77.8% ,均明显高于VEGF低表达者的 4 6 .2 %和 33.8%(P <0 .0 5 )。结论　胃癌组织中有VEGF的高表达 ,VEGF的表达在胃癌浸润和转移过程中发挥重要作用。     

系统性红斑狼疮患者外周血单个核细胞趋化因子受体4、5 mRNA的表达

目的探讨趋化因子受体CCR4及CCR5在系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的表达,与疾病的相关性.方法收集93例确诊的SLE患者和30名正常对照,通过外周血单个核细胞(PBMC),提取RNA.应用反转录-聚合酶链反应(RT-PCR)方法检测研究对象CCR4、CCR5 mRNA表达水平.结果趋化因子受体CCR4 mRNA在SLE患者PBMC中的表达水平,患者组(1.57±0.70)与正常对照组(0.19±0.18)比较,两者差异有显著性(t=3.012,P=0.003);活动期(2.03±1.04)非活动期(0.26±0.19)及对照组比较,两者差异有显著性;非活动期与对照组比较,差异无显著性(P＞0.05).趋化因子受体CCR5 mRNA在PBMC中表达水平,患者组(0.56±0.44)与对照组(0.37±0.14)比较,两者差异有显著性(t=3.262,P=0.002);活动期(0.53±0.51)与非活动期(0.59±0.34)比较,差异无显著性(P＞0.05);活动期与对照组比较(t=2.039,P=0.045)及非活动期与对照组比较(t=3.461,P=0.001),两者差异有显著性.SLE患者组CCR4及CCR5 mRNA水平与疾病活动度评分关系CCR4 mRNA水平与SLEDAI(r=0.376,t=3.851,P=0.000)呈正相关.CCR5 mRNA水平与疾病的活动性(SLEDAI)(r=0.062,t=-0,589,P=0.557)不相关.结论CCR4 mRNA表达水平在患者组比对照组表达增高,活动期比对照组表达水平增高,与疾病的活动性(SLEDAI)呈正相关.CCR5 mRNA表达在患者组比对照组增高,差异有显著性,与SLEDAI不相关.     

激光心肌血管重建术结合转染血管内皮生长因子基因治疗心肌缺血的实验研究

目的　探讨激光心肌血管重建术 (transmyocardiallaserrevascularization ,TMLR)与转血管内皮生长因子 (vascularendothelialgrowthfactor,VEGF)基因相结合能否更有效地改善心肌缺血。方法在猪的急性心肌缺血模型上行TMLR ,实验组在孔道边缘心肌内注射VEGF裸质粒 2mg,对照组注射等体积灭菌水。术后 4周、8周取材。RT PCR、免疫组化检测VEGFmRNA、蛋白表达 ;碱性磷酸酶组化法显示新生血管。结果　术后 4周VEGFmRNA、蛋白水平实验组高于对照组 ;8周时两组表达均显著下降。血管计数实验组 4周 (4 9 6± 11 5 ) mm2 ,对照组 4周 (2 6 4± 4 1) mm2 ;实验组 8周 (5 9 7±8 0 ) mm2 ;,对照组 8周 (2 8 7± 2 9) mm2 。实验组新生血管多于对照组。结论　与TMLR相比 ,TMLR结合转染VEGF基因能提高VEGF表达 ,促进新生血管形成。     

系统性红斑狼疮凋亡相关基因Fas、sFas mRNA表达研究

目的　从转录水平研究系统性红斑狼疮患者凋亡相关基因Fas、sFas表达情况 ,探索凋亡在SLE发病中的作用。方法　利用荧光标记RT PCR半定量方法 ,以 β actin为内参照 ,对 5 0例SLE、2 4例RA和 2 4名正常人外周血白细胞凋亡相关基因Fas、sFasmRNA表达水平进行了检测。结果　SLEFas、sFas表达均高于正常对照 ( 3 2± 1 8vs 2 1± 0 6,P <0 0 0 1和 1 2± 0 7vs 0 5 8±0 2 7,P <0 0 0 0 1) ,并且表达量与病情活动度相关 (相关系数分别为 0 5 2 18,P =0 0 0 7和 0 4 188,P =0 0 3 7) ,而RA仅sFas表达增高 ( 0 9± 0 5vs 0 5 8± 0 2 7,P <0 0 1)。结论　SLE患者外周血白细胞凋亡相关基因表达存在异常 ,提示凋亡在SLE发病中可能具有一定作用和意义。     

急性白血病患者血浆VEGF测定及其临床意义

目的 :探讨急性白血病 (AL)患者血浆血管内皮细胞生长因子 (VEGF)含量变化与临床病情的关系。方法 :用双抗体夹心酶联免疫吸附法 (ELISA)检测 5 4例AL患者血浆VEGF含量 ,同时检测骨髓涂片有核细胞中原始及幼稚白细胞数。结果 :①AL患者血浆VEGF含量 [(6 4 .76± 34.86 )ng/L]明显高于对照组 [(33.13± 11.97)ng/L](P <0 .0 1) ,且AL患者血浆VEGF升高与骨髓中原始及幼稚白细胞百分数增加呈正相关 (r =0 .4 37,P <0 .0 1) ;②初治组AL患者血浆VEGF含量 [(76 .0 1± 4 0 .18)ng/L]明显高于缓解组 [(42 .0 7± 17.75 )ng/L](P <0 .0 1) ,难治 /复发组患者血浆VEGF含量 [(78.2 1± 31.77)ng/L]又明显上升 ,与缓解组比较差异有显著性意义 (P<0 .0 1) ,而与初治组比较差异无显著性意义 (P >0 .0 5 )。结论 :①AL患者血浆VEGF含量升高与白血病细胞生长、增殖有关 ;②AL患者血浆VEGF含量与临床病情变化密切相关 ,可作为了解病情、观察疗效、判断预后的指标之一。     

冠心病患者血小板膜糖蛋白CD_(62P)表达和血粘度的临床研究

目的　探讨血小板膜糖蛋白CD62P表达及血粘度与冠心病的关系。方法　 82例冠心病患者分为A组 :稳定型心绞痛 2 6例 ;B组 :不稳定型心绞痛 2 8例 ;C组 :急性心肌梗死 2 8例 ,并选健康人 2 8例为D组。分别利用流式细胞仪技术测定CD62P活性及血液流变学指标。结果A组、B组、C组和D组的CD62P阳性表达分别为 3 6 2± 1 2 4 %、5 2 6± 1 4 9%、6 0 8± 1 75 %和 2 4 6± 0 81%。A、B、C组与健康人D组比较 ,有显著性差异(P <0 0 5 )。血浆粘度分别为 1 31± 0 30、1 6 4± 0 2 2、2 19± 0 35及 1 30± 0 18mPa .s。B、C组与对照组相比差异显著 (P <0 0 5 )。CD62P与血浆粘度和纤维蛋白原浓度呈正相关 (r =0 872 ,P <0 0 5 ;r =0 793,P <0 0 5 )。结论　CD62P及血浆粘度均可能成为判断冠心病患者病情进展程度的良好指标。     

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.