胶体金免疫层析法与双单克隆抗体夹心ELISA检测鼠疫F1抗原的比较

目的比较胶体金免疫层析法（GICA）和双单克隆抗体夹心酶联免疫吸附试验（DMcAbS-ELISA）检测鼠疫F1抗原的敏感性和特异性。方法用GICA试纸条和DMcAbS-ELISA平行检测308份强毒鼠疫耶尔森菌感染鼠脏器标本和327份对照鼠标本,用RIHA微量法同时检测作为参照。结果327份对照鼠鼠疫细菌学检验及GICA,DMcAbS-ELISA和RIHA的F1抗原检测均为阴性,GICA,DMcAbS-ELISA和RIHA在108cfu/mL水平上未发现与近缘假结核耶尔森菌有交叉反应;308只感染鼠样本细菌培养阳性284只,24只样本未分离到鼠疫菌;F1抗原检测GICA阳性287份,DM-cAbS-ELISA阳性280份,RIHA阳性276份,GICA阳性检出率最高93.19%,与DMcAbS-ELISA和RIHA比较,差异均有统计学意义（χ^2=5.14,9.09;P=0.016,0.001）;GICA与DMcAbS-ELISA符合率97.73%（Kappa=0.845）;GICA和DMcAbS-ELISA与RIHA符合率分别是96.43%和98.70%（Kappa=0.774,0.926）;284份细菌培养阳性鼠标本F1抗原检测GICA阳性率是98.59%,高于DMcAbS-ELISA和RIHA,差异有统计学意义（χ^2=4.17,5.14;P=0.031,0.016）;24份细菌培养阴性实验鼠标本F1抗原检测：GICA检出7份阳性,阳性率29.17%,DMcAbS-ELISA检出6份阳性,阳性率25%,RIHA检出3份阳性,阳性率12.5%,GICA高于DMcAbS-ELISA和RIHA,但差异无统计学意义（χ^2=2.25,0;P=0.125,1.000）。结论GICA检测鼠疫F1抗原敏感特异,快速简便,优于DMcAbS-ELISA和RIHA,是鼠疫快速诊断中有应用价值的检测技术。     

胶体金免疫层析法检测鼠疫F1抗原的研究

目的 研究胶体金免疫层析试纸条检测鼠疫F1抗原的敏感性和特异性.方法通过鼠疫F1抗原单克隆抗体(F1MAb)捕捉F1抗原的胶体金免疫层析(GICA)试纸条,检测308只强毒鼠疫耶尔森菌(鼠疫菌)感染鼠脏器标本和225份对照鼠(达乌尔黄鼠217只,小白鼠5只,豚鼠3只)标本,同时用鼠疫反向间接血球凝集试验(RIHA)微量法和细菌培养做平行检测.结果 225只对照鼠鼠疫细菌学检验及GICA和RIHA的F1抗原检测均为阴性,GICA和RIHA在1×108 cfu/ml水平上未发现与近缘假结核耶尔森菌有交叉反应.GICA、RIHA对鼠疫菌检测灵敏性为2.5×105、2.0×105cfu/ml;检测F1抗原灵敏性为1、10 μg/L.GICA与细菌学检验符合率为97.94%(522/533),Kappa值=0.959,阳性检出率组间比较,差异无统计学意义(x2=0.36,P＞0.05);与RIHA符合率为97.94%(522/533),Kappa值=0.959,阳性检出率组间比较差异有统计学意义(x2=9.09,P＜0.05).308只实验鼠样本细菌培养阳性284只,284份细菌培养阳性标本中,GICA有280份检测阳性,阳性率为98.59%,高于RIHA[阳性率为96.13%(522/533)],两组比较差异有统计学意义(x2=5.14;P＜0.05).GICA的敏感度为98.59%(280/284),特异度为97.19%(242/249),阳性预测值为97.56%(280/287),阴性预测值为98.37%(242/246),Youden指数为0.9578.结论 GICA检测鼠疫F1抗原敏感特异,快速简便,是鼠疫早期快速诊断中有应用价值的检测技术.     

检测鼠疫F1抗原的三种免疫学方法比较

目的 比较三种检测鼠疫F1抗原方法用于诊断鼠疫的效果.方法在鼠疫疫源地,收集野生动物的血、肝、脾、淋巴组织标本,酶联免疫吸附试验(ELISA)、反向间接血凝试验(RIHA)、胶体金纸上色谱试验(GICA)三种方法检测鼠疫F1抗原.结果检测鼠疫疫源地内自毙和捕获野生动物脏器浸液标本414份,ELISA和GICA检出的阳性标本相同,阳性率均为5.31%(22/414),阳性符合率和阴性符合率均为100%.RIHA初试2孔以上凝集186份,复试仅18份阳性,非特异性凝集率高达40.57%(168/414),阳性率4.35%(18/414).RIHA与GICA、ELISA的阳性符合率均为81.82%(18/22),阴性符合率均为98.99%(392/396);阳性标本的平均滴度,ELISA为1:29.95,RIHA为1:28.68,组间比较差异有统计学意义(t=4.379,P＜0.01).结论三种方法检测鼠疫F1抗原诊断鼠疫,RIHA结果有定量的意义,非特异性凝集率高,复试工作量大;GICA和ELISA具有相同的特异性和敏感性,GICA结果只有定性意义,ELISA排除了RIHA、GICA两种方法的缺陷,综合了两种方法的优点.     

酶标鼠疫F1McAb在酶免疫染色法快速鉴定鼠疫菌中的应用

目的观察酶标鼠疫F1McAb在酶免疫染色法快速鉴定鼠疫菌中的应用效果.方法实验感染鼠疫强毒菌的病死小鼠和处死健康小鼠尸体,20℃以上室温放置,用鼠疫F1McAb酶免疫染色法检测,同时用细菌学检验方法分离鼠疫菌和反向血凝试验(RIHA)检测鼠疫F1抗原.结果直接培养仅从5d内的鼠尸分离检出鼠疫菌,阳性率为53.3%;动物实验检出鼠疫菌的最长时间为8d,阳性率为71.6%.存放13d内,酶免疫染色阳性率为99.1%,RIHA阳性率98.2%,用酶免疫染色法和RIHA检测健康动物腐败脏器全部阴性.以上方法检查鼠疫疫区自毙动物标本12份,酶免疫染色法阳性7份,RIHA阳性6份,动物实验阳性5份,直接培养未获得阳性结果.结论酶免疫染色法检查动物腐败脏器中的鼠疫菌具有较高的敏感性和特异性.     

2015─2017年新疆青河县山区动物鼠疫血清学和病原学调查

目的通过鼠疫病原学和血清学的实验检测结果,了解新疆青河县山区存在鼠疫自然疫源地的潜在状况。方法采用鼠疫"四步检验"法分离青河县山区捕获的长尾黄鼠及灰旱獭脏器、自毙动物脏器及骨骼和寄生蚤中的鼠疫菌;间接血凝试验(HA)和酶联免疫吸附试验(ELISA)检测动物脏器和寄生蚤悬液中的鼠疫F1抗原;长尾黄鼠、灰旱獭以及牧犬血清中的鼠疫F1抗体。结果经鼠疫"四步检验"和反向间接血凝(RIHA)检验长尾黄鼠脏器552份、灰旱獭脏器4份、自毙长尾黄鼠和灰旱獭脏器及骨骼13份、长尾黄鼠体外寄生蚤583只,均未检出鼠疫菌及F1抗原;检测长尾黄鼠血清552份,IHA法检出阳性1份、滴度1∶2~9,ELISA法检出阳性3份、平均滴度1∶2~8;检测牧犬血清129份,IHA法检出阳性6份、平均滴度1∶2^(3.83),ELISA法检出阳性8份、平均滴度1:26.75;其中牧犬鼠疫F1抗体阳性标本中检出鼠疫IgM抗体阳性2份,平均滴度1∶2^(5.5)。结论新疆青河县山区啮齿类动物间有鼠疫流行迹象,流行区域为花海子地区,存在潜在鼠疫自然疫源地,建议对该区域开展进一步鼠疫疫源性调查。     

胶体金免疫层析法检测鼠疫F1抗原的研究

Objective To study the sensitivity and specificity of gold-immunochromatography assay (GICA) for detection of Yersiniapestis(Y. pestis ) F1 antigen. Methods Viscera organ(liver and spleen) specimens of 308 mice with virulent Y. pestis infection and 225 control specimens of rats(217 Spermophilus dauricus, 5 mice,3 guinea pigs) were detected by GICA dipstick with monoclonal antibody against plague F1 antigen (F1MAb).Meanwhile, micro-method of reverse indirect hemagglutination assay(RIHA) and bacteria culture were carried out for parallel testing. Results Bacteriological examination of 225 control specimens, and F1 antigen detected with GICA and RIHA were all negative. No cross-reaction with related Yersinia pseudotuberculosis at 1 x 108 cfu/ml level was found in GICA and RIHA. Detection sensitivity of Y. pestis by GICA and RIHA were 2.5 × 105 cfu/ml and 2.0 × 105 cfu/ml, respectively, and of F1 antigen were 1μg/L and 10 μg/L, respectively. Coincidence was 97.94% (522/533) between GICA and bacteriological test, Kappa = 0.959, and the difference was statistically insignificant(x2 = 0.36, P ＞ 0.05); and 97.94%(522/533) between GICA and RIHA, Kappa = 0.959, with statistically significant difference in the positive detection rates(x2 = 9.09, P ＜ 0.05). Out of the 308 infected mice, 284 were positive of plague bacterial cultured, In 284 samples with positive bacterial culture, there were 280 of positive detected by GICA for F1 antigen, positive rate of F1 antigen was 98.59%, higher than that by RIHA[the positive rate of 96.13%(522/533)], with statistically significant difference(x2 = 5.14, P ＜ 0.05). Sensitivity of GICA was 98.59% (280/284), specificity was 97.19% (242/249), positive predictive value (PPV) was 97.56% (280/287),negative predictive value ( NPV ) was 98.37% (242/246), and Youden index was 0.9578. Conclusions GICA is sensitive and specific, fast and simple in detection of F1 antigen of the plague. It's a valuable detection technique for early and rapid diagnosis of plague.     

西藏尼木县一起动物间鼠疫的实验室判定

目的对西藏自治区拉萨市尼木县一起动物间鼠疫进行实验室判定。方法取病死喜马拉雅旱獭脏器标本进行细菌学检测和抗原检测,采用实时荧光定量PCR检测鼠疫杆菌核酸(chro392基因)。结果病死喜马拉雅旱獭脏器标本鼠疫杆菌检测阳性、鼠疫F1抗原检测阳性、实时荧光定量PCR检测鼠疫杆菌核酸阳性。结论该病死喜马拉雅旱獭实验室检测为鼠疫杆菌感染所致,据此判定尼木县此次事件为动物间鼠疫疫情。     

鼠疫检验新技术在乌鲁木齐县灰旱獭鼠疫疫源地中的应用

目的比较分析酶联免疫吸附试验(ELISA)、红细胞凝集试验(IHA和RIHA)、胶体金纸上层析试验(GICA)3种免疫学检测技术的敏感性、特异性和稳定性。方法按照《鼠疫检测新技术在灰旱獭鼠疫监测和应急处置中的应用》课题设计要求,以Spss 13.0对2007—2015年乌鲁木齐县鼠疫监测数据整理并进行分析。结果 2007—2015年检测血清样本3 260份,其中灰旱獭血清2 753份、牧犬血清507份,ELISA检出阳性36份、阳性率1.10%,IHA检出阳性13份、阳性率0.40%,两种方法检出阳性率差异有统计学意义(χ~2=10.877 7,P0.01),检出鼠疫抗体滴度差异无统计学意义(t=0.036 6,P0.01);共检测以自毙灰旱獭脏器及骨骼为主的各种自毙动物材料322份,检出鼠疫F1抗原分别为ELISA8份、GICA 8份和RIHA 6份,阳性率分别为2.48%、2.48%和1.86%,四步检验法分离出鼠疫菌2株、检菌阳性率0.62%,ELISA与RIHA检出阳性率(χ~2=0.292 1)和检菌率(χ~2=3.465 2)差异无统计学意义(P0.01),RIHA与四步法检菌率差异亦无统计学意义(χ~2=1.925 5,P0.01);检测自毙动物脏器标本鼠疫F1抗原ELISA和GICA阳性率一致,二者略高于RIHA和检菌,差异无统计学意义(χ~2=0.292 1,P0.01)。结论在鼠疫监测和应对突发鼠疫疫情处置工作中,3种方法各有其优缺点,联合使用才能发挥各自特长,依据不同情况采用不同的方法,可提高鼠疫监测质量和快速应对鼠疫突发事件能力。     

酶免疫染色技术快速检测鼠疫耶尔森菌

目的研究酶免疫染色技术检测鼠疫耶尔森菌(鼠疫菌)的敏感性及特异性,建立鼠疫免疫快速检测技术。方法用酶免疫染色技术对细菌标本和动物标本检测鼠疫菌,动物标本同时用细菌学、RIHA、RGICA和ELISA法平行检测鼠疫菌。结果细菌标本酶免疫染色法检测结果与实际相符,酶免疫染色技术检测鼠疫菌的灵敏性为1 000个/ml;动物标本酶免疫染色法检测鼠疫菌结果和细菌学、RIHA、RGICA和ELISA一致。结论酶免疫染色技术检测鼠疫菌敏感、特异、简便、快速,可用于鼠疫检测工作。     

云南省洱源县鼠疫阳性线索实验室检测分析

目的了解云南省洱源县三营镇永乐村驻地是否处于鼠疫流行期,为做好鼠疫防控工作提供依据。方法2014年5月采用现场调查和实验室检测相结合的方法开展鼠疫调查工作,采用间接血凝试验(IHA)、酶联免疫吸附试验(ELISA)、胶体金免疫层析(GICA)方法对采集的样本进行鼠疫F1抗原和F1抗体的实验室检测,对死鼠脏器进行鼠疫菌分离培养。结果 3份死鼠脏器未分离出鼠疫菌,F1抗原检测均阴性;128份犬血清、3份鼠血清进行F1抗体检测,IHA均阴性、ELISA阳性3份、GICA阳性5份,两种方法均阳性2份。结论洱源县可能存在鼠疫F1抗体阳性犬,需进一步加强该地区的鼠疫监测工作。     

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.