抗中性粒细胞胞浆抗体在肾炎综合征中的检测及其临床意义

目的　探讨检测抗中性粒细胞胞浆抗体 (ANCA)及其靶抗原在肾炎综合征中的临床意义。方法　应用间接免疫荧光 (IIF)法检测 10 0例肾炎综合征患者血清抗中性粒细胞胞浆抗体 ,对其阳性的 2 9例用酶联免疫吸附试验 (ELISA )检测靶抗原髓过氧化物酶 (MPO)和蛋白酶 3(PR3 )。结果　IIF检测肾炎综合征ANCA阳性率为 2 9% ,其中胞浆型 10 %、核周型 19%。急进型肾炎、狼疮性肾炎、紫癜性肾炎阳性率分别为 5 6%、2 0 %和 15 %。ELISA急进性肾炎和紫癜性肾炎大多数识别靶抗原MPO ,狼疮性肾炎ANCA不识别MPO或PR3。结论　ANCA在急进性肾炎和狼疮性肾炎中阳性率较高 ,检测ANCA对判断狼疮性肾炎活动及疗效具有参考价值     

丙基硫氧嘧啶导致的抗中性粒细胞胞质抗体阳性小血管炎及其靶抗原研究

目的 研究丙基硫氧嘧啶（PTU）引起的抗中性粒细胞胞质抗体（ANCA）阳性小血管炎的临床病理表现及其靶抗原。方法 对我院近年诊治的4例PTU引起的ANCA阳性小血管炎患者进行临床病理分析。以纯化的7种已知的ANCA靶抗原蛋白酶3（PR3）、髓过氧化物酶（MPO）、人白细胞弹力蛋白酶（HLE）、乳铁蛋白（LF）、组蛋白酶G（CG）、杀菌／通透性增高蛋白（BPI）和天青杀素（AZU）为固相抗原,采用ELISA法检测患者血清的靶抗原及治疗前后抗体滴度的变化。结果 4例病人中男女各2例,平均年龄30（11－57）岁,服PTU时间7－60个月。4例均有肾脏、肺脏、皮肤、关节肌肉和血液系统等受累,均为p-ANCA阳性,患者血清均识别MPO、LF和CG;3例识别HLE、AZU,2例识别PR3;无1例识别BPI。多数抗体滴度高,可大于1：25600;而服PTU无小血管炎临床表现的甲状腺功能亢进（甲亢）病人的30份血清均为阴性。肾活检2例为新月体性肾炎,2例为轻微病变,免疫荧光检查均为阴性。4例患者均立即停用PTU,3例应用免疫抑制剂,1例行血浆置换。4例小血管炎的临床症状均得以缓解,但1例晚期新月体肾炎患者发展为慢性肾衰竭而依赖透析。停药和治疗后各种抗体滴度均有所下降,但多未能短期阴转。结论 PTU可引起ANCA阳性小血管炎,其自身抗体可识别中性粒细胞胞质中多种已知的靶抗原;及时诊治,预后较好。     

系统性红斑狼疮患者中抗中性粒细胞胞质抗体和细胞因子的关系

目的　探讨抗中性粒细胞胞质抗体 (ANCA)与系统性红斑狼疮 (SLE)血管炎临床特点的关系。检测SLE血管炎患者肿瘤坏死因子 (TNF) α、白细胞介素 (IL) 6等细胞因子的血清水平 ,探讨ANCA与SLE血管炎细胞因子的关系。方法　 6 0例SLE活动期患者和 30名正常对照组均通过间接免疫荧光法 (IIF)检测ANCA与酶联免疫吸附试验 (ELISA)检测抗髓过氧化物酶 (MPO)抗体 (即MPO ANCA) ,观察两组ANCA与抗MPO抗体的阳性率。通过ELISA法检测 30例SLE活动期患者和 30名健康对照者 ,外周血TNT α、白细胞介素 (IL) 6水平。结果　 6 0例SLE患者 1 5例ANCA阳性 ,均为核周型抗中性粒细胞胞质抗体 (pANCA) ,阳性率为 2 5 % ,抗MPO抗体 8例阳性(IIF法ANCA均阳性 ) ,阳性率为 1 3 3% ,健康对照组ANCA及抗MPO抗体均为阴性。病程超过 1年 ,伴有肾炎、浆膜炎、皮肤血管炎组 ,ANCA阳性率高 ,与无相应特点的对照组差异均有显著性 (P <0 0 5 ) ;有关节炎比无关节炎组ANCA阳性率高 (P =0 0 5 )。SLE活动期患者与对照组相比 :SLE组TNF α、IL 6水平较高 (P <0 0 5 )。抗MPO抗体阳性组与抗MPO抗体阴性组相比 :TNF α、IL 6水平升高 ,两组差异有显著性 (P <0 0 5 )。结论　部分SLE病人血清中可检测到 pANCA及抗MPO抗体 ,pANCA与SLE某些     

抗PR3抗体和抗MPO抗体的检测及临床研究

目的探讨抗蛋白酶-3(PR3)抗体、抗髓过氧化物酶(MPO)抗体和抗中性粒细胞胞质抗体(ANCA),在系统性血管炎患者中的检测及临床意义。方法对251例临床确诊为系统性血管炎和其他自身免疫性疾病患者血清,应用酶联免疫吸附试验(ELISA)检测抗PR3抗体和抗MPO抗体;用间接免疫荧光法(IIF)检测ANCA,并进行回顾性分析。结果①251例系统性血管炎患者与非血管炎病人的检测,经χ2检验,P<0.01,差异均有非常显著性。②33例韦格纳肉芽肿病(WG)患者主要表现为PR3和颗粒型抗中性粒细胞胞质抗体(cANCA)阳性均为22例,阳性率为67%;非血管炎病人,27例溃疡性结肠炎(UC)患者检测MPO和核周型抗中性粒细胞胞质抗体(pANCA)阳性分别为13和14例,阳性率分别是48%和52%;104例SLE患者检测MPO和pANCA阳性分别为19例、24例,阳性率分别为18%和23%。③ELISA法和IIF法阳性率,经χ2检验差异无显著性(P>0.05)。结论PR3、MPO抗体作为系统性血管炎的一种敏感标记抗体,有利于该疾病的早期治疗。     

抗中性粒细胞胞浆抗体对溃疡性结肠炎的诊断价值

目的　探讨抗中性粒细胞胞浆抗体 (ANCA)对溃疡性结肠炎 (UC)的诊断价值。方法　应用间接免疫荧光法 (IIF)、酶联免疫吸附法 (ELISA)和免疫印迹法 (Westernblot)分别对UC患者 ( 5 8例 )、非UC患者 ( 4 3例 )及健康献血员 ( 5 8例 )进行血清ANCA检测。结果　ANCA对UC诊断的敏感性为 3 7.93 % ,特异性为 10 0 %。UC患者中依据病情分为轻、中、重度组 ,ANCA的阳性率分别为17 65 %、4 1.67%和 5 2 .94 %。ANCA阳性肠黏膜炎症III～V级者占 78 95 % ,黏膜血管炎发生率为78 95 % ,而ANCA阴性者分别为 3 7.0 4 %和 4 4.4 4%。髓过氧化物酶 (MPO)、杀菌 /通透性增强蛋白(BPI)、乳铁蛋白 (LF)、组织蛋白酶G(CG)、蛋白酶 3 (PR 3 ) 5种ANCA抗原与UC患者血清的结合率分别为 13 .79%、13 .79%、10 .3 4%、10 .3 4%和 8.62 %。采用Westernblot法对UC患者血清进行检测 ,发现显示特异蛋白条带者占 4 8.2 8% ,其中显示分子量为 4 70 0 0条带者最多 ,占 2 2 .4 1%。结论　ANCA检测可作为UC的辅助诊断手段。目前 ,UC相关ANCA的靶抗原仍未明确 ,研究发现 4 70 0 0蛋白可能是UC相关ANCA的靶抗原之一。ANCA可能参与UC的致病机制。     

系统性血管炎抗内皮细胞抗体与抗中性粒细胞胞质抗体的相关性初探

目的以来源于大小血管内皮细胞的两种永生细胞株为底物,检测系统性血管炎血清中抗内皮细胞抗体(AECA),分析其与抗中性粒细胞胞质抗体(ANCA)的相关性。方法细胞酶联免疫吸附试验(Cyto-ELISA)法检测血清中AECA;间接免疫荧光法(IIF)及抗抗体结合内皮细胞表面的蛋白酶3(PR3)、抗MPO-ELISA检测血清中ANCA;IIF及抗PR3、抗MPO-ELISA检测细胞株中PR3及MPO;反转录—聚合酶链反应(RT-PCR)法检测细胞株中PR3及MPOmRNA。结果AECAEA(EA.hy926为底物所测AECA)阳性率33.6%(41/122),AECAHMEC(HMEC-1为底物所测AECA)37.7%(46/122),ANCA35.3%(43/122),AECAEA或AECAHMEC与ANCA串联诊断系统性血管炎敏感性分别为59.8%(73/122)或60.7%(74/122)。AECAEA与ANCA,AECAHMEC与ANCA分别行配对字2检验,差异均无显著性(P>0.05),符合率仅分别为49.2%及51.6%。EA.hy926和HMEC-1中蛋白水平及mRNA水平均无PR3和MPO的表达。结论EA.hy926与HMEC-1中无PR3、MPO蛋白水平的表达,ANCA与AECA可能是两种相互独立的抗体,串联检测可提高诊断敏感性。     

抗蛋白酶-3抗体在韦格纳肉芽肿病和其他血管炎中的临床应用研究

目的探讨抗蛋白酶-3(PR3)抗体在韦格纳肉芽肿病(WG)和其他血管炎患者中的表达和临床意义。方法选取2001年3月至2006年7月山西医科大学第二医院确诊的住院患者576例。系统性血管炎组111例,其中9例WG(包含21份跟踪随访血清)；结缔组织病(CTD)组403例；各型肾小球疾病患者62例及健康对照30名,均采用酶联免疫吸附试验(ELISA)检测抗PR3抗体、抗髓过氧化物酶(MPO)抗体；采用间接免疫荧光法(IIF)检测抗中性粒细胞胞质抗体(ANCA),观察抗PR3与ANCA在WG和其他血管炎中的阳性率,且追踪WG治疗前后抗PR3吸光度值和ANCA的滴度变化。结果588例血清中抗PR3抗体阳性23例,分别为WG 15例(15/21,71．4％)；系统性红斑狼疮(SLE)6例(6/213,2．8％)：类风湿关节炎(RA)1例(1/135,0．7％)；混合结缔组织病(MCTD)1例。大动脉炎、白塞病,过敏性紫癜等常见的原发性系统性血管炎、肾病组、健康对照组未发现抗PR3抗体阳性。抗PR3抗体和胞质型ANCA(cANCA)在WG中阳性率最高,差异有统计学意义(P＜0．05)。抗PR3对WG诊断的敏感性71．42％,特异性98．58％。联合应用抗PR3与cANCA诊断WG的敏感性61．90％,特异性99．82％。抗PR3吸光度值、ANCA的滴度及伯明翰血管活动度评分(BVAS)随治疗好转下降。结论抗PR3抗体是诊断WG的一种敏感、特异标记抗体,抗PR3抗体和ANCA同时在临床应用,有利于WC和其他系统性血管炎的早期诊断和鉴别诊断。抗PR3抗体还可作为临床疗效观察指标。     

老年人抗中性粒细胞胞浆抗体相关性小血管炎的临床表现

目的　为了解老年人抗中性粒细胞胞浆抗体 (ANCA)相关性小血管炎所引起的多器官损害的临床表现。方法　分析总结近 5年来检出的 1 4 5例 60岁以上老年ANCA相关性小血管炎患者的临床病理资料。结果　1 4 5例患者 ,男 74例 ,女 71例 ,平均年龄 68.2岁。胞浆型ANCA(c ANCA)阳性 1 7例 ,均识别蛋白酶 3(PR3) ;环核型ANCA (p ANCA)阳性 1 2 8例 ,其中 1 2 0 1 2 8例单独识别髓过氧化物酶 (MPO) ,8 1 2 8例同时识别PR3和MPO。不足 1 2的患者在 3个月内确诊。确诊病例均呈多器官损害 ,其中肾脏受累占 93 .8% ,且 75 %出现肾功能不全 ;肺脏受累占 74.5 % ,半数以上表现为肺出血和 (或 )肺部阴影 ,此外还有不同程度的消化道、神经系统、肌肉关节、眼、耳器官或组织损害及发热、乏力、消瘦等非特异症状。实验室检查 ,90 %以上的患者有中重度贫血、血沉增快和C反应蛋白升高。结论　老年人ANCA阳性小血管炎较为多见 ,临床表现为多器官损害 ,ANCA检测有助于诊断     

狼疮肾炎中识别新抗原的抗中性粒细胞胞质抗体及其与光过敏和口腔溃疡的关系

目的　寻找狼疮肾炎 (LN)患者中抗中性粒细胞胞质抗体 (ANCA)的未知靶抗原 ,并分析其相应抗体与临床和病理损害的关系。方法　收集 72例有完整临床病理资料的狼疮肾炎患者的血清 ,分离健康人外周血中性粒细胞的初级颗粒和次级颗粒制备成酸性可溶性颗粒蛋白抗原液 ,以此为抗原在非还原状态下进行Western blot分析 ,检测狼疮肾炎患者血清中的ANCA。结果　部分血清可识别一些蛋白条带 ,其中 1 4 / 72 (1 9 4% )例识别分子量为 690 0 0的蛋白条带 ,后者不同于ANCA其他已知的靶抗原如组织蛋白酶G和乳铁蛋白等 ;1 0 / 72 (1 3 9% )例识别 55 0 0 0的蛋白条带 ,后者可能是杀菌 /通透性增高蛋白。进一步的研究发现 ,在抗 690 0 0蛋白抗体阳性的病人中 ,光过敏和口腔溃疡的发生率均显著高于抗 690 0 0蛋白抗体阴性者 (分别为 57 1 %、1 2 1 % ,P <0 0 0 5 ;50 0 %、1 7 2 % ,P <0 0 5)。结论　中性粒细胞混合颗粒中的 690 0 0蛋白可能为狼疮肾炎ANCA新的特异性靶抗原 ,抗 690 0 0蛋白抗体可能与狼疮肾炎患者光过敏和口腔溃疡相关。     

抗中性粒细胞抗体相关性血管炎31例临床分析

目的分析抗中性粒细胞抗体(ANCA)相关性血管炎(AAV)伴肾脏损害患者临床特征。方法收集2002年9月至2007年9月四川大学华西医院收治的31例临床诊断为AAV患者。间接免疫荧光及免疫酶联吸附试验(ELISA)均为ANCA阳性。逐项分析ANCA抗原谱,肾脏和肾外临床表现。结果本组AAV患者男16例,女15例。年龄18~84岁,平均(54.19±20.00)岁。显微镜下多血管炎(MPA)27例,韦格纳肉芽肿(WG)4例。肾脏症状首发16例,肾外症状首发15例,以呼吸系统症状首发者8例,8例伴咯血。27例MPA患者就诊时血肌酐平均(460.42±354.55)μmol/L。全组患者P-ANCA阳性24例,C-ANCA阳性7例,抗原识别抗MPO阳性25例,抗PR3阳性6例,抗BPI阳性3例。结论AAV临床表现多样,肺、肾是最常见的受累器官。本病晚期治疗方法十分有限,预后甚差,重视ANCA筛查,早期诊断是改善预后的关键。     

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.