胆囊结石研碎镜检法对华支睾吸虫感染检出率的研究

收集2010年1~6月于广州市番禺区第二人民医院普外科实施内镜微创取石保胆术的179例患者的血清、粪便、胆汁和胆囊结石标本,分别采用胶体金免疫层析法检测华支睾吸虫IgG抗体,粪便直接涂片法、胆汁离心沉渣镜检法和胆囊结石研碎镜检法检查华支睾吸虫虫卵,并进行比较分析。结果显示,胶体金免疫层析法检测华支睾吸虫IgG抗体的阳性率为51.4%(92/179),粪便直接涂片法、胆汁离心沉渣镜检法和胆囊结石研碎镜检法华支睾吸虫卵阳性检出率分别为30.7%(55/179)、44.7%(80/179)和69.8%(125/179),粪便直接涂片法检出率最低,胆囊结石研碎镜检法检出率最高(均P<0.05)。其中粪便直接涂片法和胆汁离心沉渣镜检法检测结果为阳性的患者,其结石研碎镜检法的检测结果也均为阳性。     

基于胆结石患者胆汁、粪便和血清标本的4种诊断华支睾吸虫感染方法的评价

目的 比较并评价粪便改良加藤厚涂片法（Kato-Katz法）、胆汁离心镜检法、血清胶体金免疫层析法（colloid gold labeled immunochromatographic,CGIA）和酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）检测粪便、胆汁、血清诊断华支睾吸虫感染的价值. 方法 收集92例胆结石患者的胆汁、血清和粪便,分别采用镜检法检查胆汁华支睾吸虫卵、用Kato-Katz法检查粪便华支睾吸虫卵,用CGIA法和ELISA法检测血清华支睾吸虫IgG抗体,计算并分析各检测方法的阳性率、灵敏度、特异性、阳性预测值、阴性预测值和Youden指数. 结果 Kato-Katz法联合胆汁离心镜检法共确诊感染49例,阳性率为53.3％.与之相比,Kato-Katz法、胆汁离心镜检法、CGIA和ELISA的符合率分别为80.4％、84.8％、90.2％和96.7％.Kato-Katz法、胆汁离心镜检法、CGIA和ELISA法的灵敏度分别是63.3％、93.9％、98.0％和93.9％;特异性分别为100％、100％、69.8％和86.0％;阳性预测值则分别是100％、100％、78.7％和88.5％,阴性预测值分别为70.5％、93.5％、96.8％和92.5％,Youden指数为0.63、0.94、0.68和0.80. 结论 胆汁离心镜检是胆结石手术患者诊断华支睾吸虫感染的首选方法;但对于无法获得胆汁的受检者而言,血清IgG抗体检测在排除华支睾吸虫感染、粪便Kato-Katz法在确诊感染方面有较好的临床应用价值.     

实时荧光定量PCR检测胆囊结石中华支睾吸虫DNA的研究

目的运用基于Taqman探针实时荧光定量PCR技术检测胆囊结石中的华支睾吸虫DNA。方法选取2012年3～4月份40例实施内镜取石保胆手术患者的胆囊结石,通过傅里叶红外光谱法分析结石成分,对结石研磨涂片镜检观察,并提取其DNA,运用实时荧光定量PCR技术检测华支睾吸虫基因。结果 40例患者的胆囊结石按主要成分可分为胆色素性结石(25例),胆固醇性结石(6例),混合性结石(9例)。结石涂片镜检有23例发现华支睾吸虫虫卵(阳性率57.5%),其中胆色素性结石19例,混合性结石4例。实时荧光定量PCR检测华支睾吸虫DNA阳性率为65.0%(26/40),其中包括23例镜检虫卵阳性标本及3例镜检阴性标本。结论运用实时荧光定量PCR在胆囊结石中检测出华支睾吸虫DNA,敏感性高于镜检法查虫卵。这不仅为华支睾吸虫感染诱发胆囊结石形成提供了分子生物学证据,同时也提供了一种诊断华支睾吸虫感染的有效手段,对胆囊结石病人的病因诊断、临床治疗以及术后预防复发等具有重要的指导意义。     

华支睾吸虫病流行区胆石症患者胆囊结石类型及华支睾吸虫感染情况

目的探讨华支睾吸虫病流行区胆石症患者胆囊结石类型与华支睾吸虫感染的关系。方法选取2009年5月至2012年10月在广州市南沙区第六人民医院普外科实施内镜微创取石保胆手术的胆石症患者598例。胆石样品以红外光谱法分析结石成分,判断结石类型。胆石研碎后用光镜检查华支睾吸虫虫卵,计算不同类型结石的虫卵检出率。比较不同类型的结石患者,以及碳酸钙类结石患者中华支睾吸虫虫卵阳性和阴性患者的临床特点和生化特征。选取部分虫卵阳性的碳酸钙类结石行扫描电镜观察。结果 598例胆囊结石患者中,234例(39.1%)为胆固醇类结石,133例(22.2%)为胆色素类结石,112例(18.7%)为碳酸钙类结石,86例(14.4%)为混合类结石,33例(5.5%)为其他类结石。5类胆结石华支睾吸虫卵检出率分别为6.4%(15/234)、44.3%(59/133)、59.8%(67/112)、36.0%(31/86)和30.0%(10/33)。碳酸钙类结石的虫卵检出率最高,胆固醇类结石的虫卵检出率最低。碳酸钙类结石和混合类结石患者的血清CO2结合力高于胆固醇类结石患者(P0.05),碳酸钙类结石患者胆汁CO2结合力和p H值高于其他4组结石患者(P0.05)。碳酸钙类结石患者中华支睾吸虫虫卵阳性者的血清CO2结合力、胆汁CO2结合力和p H值均高于虫卵阴性者(P0.05)。光镜和扫描电镜观察均发现,碳酸钙类结石华支睾吸虫卵与碳酸钙结晶相互黏附。结论碳酸钙类胆囊结石患者的华支睾吸虫感染率高于其他结石类型的患者。     

广西藤县人群华支睾吸虫感染调查及其分子鉴定

目的了解广西藤县人群华支睾吸虫的感染状况及其分子鉴定,为当地华支睾吸虫病的防控提供理论依据。方法2016年11月,在广西藤县采用分层整群随机抽样抽取天平镇和金鸡镇各1个自然村作为调查点。收集3岁以上常驻居民的粪便,用改良加藤厚涂片法(一粪三检)检测华支睾吸虫虫卵并计数。通过问卷调查收集被调查者社会人口学等相关信息。从镜检阳性样本中随机选取部分新鲜粪样提取DNA, PCR扩增华支睾吸虫内转录间隔区2 (ITS2),测序后在NCBI核酸数据库作比对分析。结果共调查400人,检出华支睾吸虫虫卵阳性208人,感染检出率为52.0%(208/400),感染者以轻度感染为主(69.2%, 144/208)。天平镇华支睾吸虫感染检出率为66.5%(133/200),高于金鸡镇的37.5%(75/200)(χ2=33.69, P 0.01)。男性感染检出率为58.7%(131/223),高于女性的43.5%(77/177)(χ2=9.18, P 0.01); 30～39岁组感染检出率最高,为63.8%(60/94);高中及以上学历人群感染检出率最高,为68.0%(17/25)。不同年龄组、教育程度人群感染检出率差异有统计学意义(χ2=37.1, 23.02; P 0.01)。PCR扩增出156 bp的华支睾吸虫特异性条带,其序列与越南岘港分离株ITS2 (GenBank登录号:MF319655)的一致性为100%。结论广西藤县人群华支睾吸虫感染检出率为52.0%,属于超重流行区;男性、青壮年是华支睾吸虫高感染人群。广西藤县的华支睾吸虫ITS2序列与越南岘港分离株的基因型一致。     

广西146例华支睾吸虫疑似感染者粪样虫卵和血清IgG抗体的检测分析

检测广西华支睾吸虫疑似感染者粪样华支睾吸虫/扇棘单睾吸虫虫卵和血清华支睾吸虫IgG抗体,分析二者的相关性,为华支睾吸虫感染的监测和诊断提供参考。收集2017年3-9月于广西疾病预防控制中心预防性健康体检科就诊的疑似华支睾吸虫感染者粪样,采用改良加藤厚涂片法(一粪三检)检查华支睾吸虫/扇棘单睾吸虫虫卵并计算每克粪样虫卵数(EPG)。同步收集粪检阳性患者的血液标本,采用间接ELISA法检测血清华支睾吸虫IgG抗体。分析EPG值与IgG抗体A_(450)值的相关性,并根据华支睾吸虫感染度分类标准,分析轻度、中度和重度感染者EPG值与血清IgG抗体阳性率以及IgG抗体定性结果与EPG值的关系。在146份华支睾吸虫/扇棘单睾吸虫虫卵阳性粪样中,EPG最大值为21 502,最小值为8。血清华支睾吸虫IgG抗体阳性率为84.9%(124/146),男、女性IgG抗体阳性率分别为86.0%(111/129)和13/17,差异无统计学意义(P 0.05)。感染者EPG值和IgG抗体A_(450)值呈正相关性(r_s=0.436, P 0.01)。轻度、中度和重度感染者IgG抗体阳性率分别为78.4%(69/88)、 96.0%(48/50)和7/8,差异有统计学意义(P 0.05)。华支睾吸虫血清IgG抗体阳性组和阴性组粪样几何均数(GM EPG)值分别为736和204,差异有统计学意义(P 0.01)。提示在华支睾吸虫和扇棘单睾吸虫共感染的广西,粪样虫卵镜检和血清华支睾吸虫IgG抗体检测结果呈正相关;但单独粪便虫卵检查用于华支睾吸虫监测和诊断会存在误判,应辅助IgG抗体等血清学检测。     

改良加藤厚涂片法和PCR法检测华支睾吸虫感染的 效果比较

目的　比较改良加藤厚涂片法（Kato?Katz法）和PCR法对现场人群粪便样本中华支睾吸虫感染的检出率,为华支睾吸虫感染检测方法的选择提供依据。方法　在2016年广西壮族自治区藤县人体华支睾吸虫感染流行病学调查的基础上,随机抽取133份粪便样本,-20 ℃保存。分别采用Kato?Katz法（1粪3检）和PCR法检测华支睾吸虫感染,比较两种方法对华支睾吸虫感染的检出率,采用Kappa分析评价两种检测方法结果的一致性。结果　133份人群粪便样本中,华支睾吸虫感染总检出率为77.44%（103/133）,其中Kato?Katz法检出率为57.14%（76/133）,PCR法检出率为70.68%（93/133）, PCR法检出率显著高于Kato?Katz法（[χ2] = 26.15,P <0.01）。76份Kato?Katz法阳性粪便样本中,88.16%（67/76）的样本PCR法扩增阳性;57份Kato?Katz法阴性样本中,47.37%（27/57）的样本PCR法扩增阳性。PCR法对每g粪便虫卵数（EPG）> 1 000的粪便样本中华支睾吸虫感染的检出率（94.7%,18/19）高于EPG < 1 000的样本（85.96%,49/57）,但差异无统计学意义([χ2] = 1.05,P = 0.436)。两种方法检出率一致性检验结果一般（Kappa = 0.73）。结论　PCR法对人群华支睾吸虫感染的检出率显著高于Kato?Katz法。建议在华支睾吸虫感染度较轻的地区,采用Kato?Katz法结合PCR法进行检测,以提高检出率。     

对流免疫电泳检测粪内华支睾吸虫抗原诊断华支睾吸虫病的试验研究

华支睾吸虫卵甚小,粪检时极易漏检。十二指肠及胆汁引流等方法,虽可提高检查效果,然操作不便,难于推广。血清学方法虽可提高检出率,但不易区分既往感染和现症病人。因此,我们初步探讨了对流免疫电泳法检测华支睾吸虫病人粪便内抗原的诊断价值。一、抗华支睾吸虫兔免疫球蛋白制备:     

2016—2020年福建省浦城县华支睾吸虫病国家监测点人群感染情况分析

2016—2020年根据《全国肝吸虫病和土源性线虫病监测方案》要求,将浦城县按地理方位划分东、南、西、北、中等5个片区,每个片区随机抽取1个乡镇的1个村作为人群华支睾吸虫感染情况的国家监测点。每个村随机调查200人以上,采集人群粪样采用改良加藤厚涂片法检测华支睾吸虫虫卵(一粪二检);根据村庄鱼塘溪渠分布情况,每年度采集各类淡水鱼类100尾,采用压片镜检法检测鱼鳃、鱼鳍及尾鳍部肌肉的华支睾吸虫囊蚴;各年度收集野外犬、猫粪样共25份,以水洗沉淀法检测华支睾吸虫虫卵;在监测点受检人群中随机抽查150人以上进行华支睾吸虫病防治知识、态度和行为等内容的问卷调查。2016—2020年共调查5 553人,吸虫总感染率为0.79%(44/5 553)。各年度感染率分别为0.60%、 1.02%、 1.16%、 0.97%和0.24%。其中,水洗沉淀检出华支睾吸虫感染者26人,感染率为0.47%(26/5 553);检出东方次睾吸虫感染者18人,感染率为0.32%(18/5 553)。检测3种淡水鱼类共639尾,感染囊蚴鱼类472尾,感染率为73.86%。其中,麦穗鱼的感染率2016年最高,达97.33%(73/75)。检测野外动物粪样共129份,检出华支睾吸虫卵5份,东方次睾吸虫卵2份,检出率分别为3.88%和1.55%。居民有效问卷共752份,总知晓率为37.10%(279/752)。男性与女性知晓率差异无统计学意义(χ2=0.73, P 0.05)。浦城县人群华支睾吸虫感染率低,居民对华支睾吸虫病知识的知晓率低。当地淡水鱼类特别是麦穗鱼的囊蚴感染率较高,应加强宣传教育与防治工作。     

华支睾吸虫病合并胆结石胆囊炎1例分析

目的了解1例患者出现华支睾吸虫病、胆结石、胆囊炎三联症的有关发病原因与临床表现,分析有关发病因素。方法直接涂片法检查患者T型管引流液中华支睾吸虫卵,kato-katz法检查粪便中华支睾吸虫卵,吡喹酮驱虫药物治疗。结果无论肉眼或镜下对患者T型管引流液中排出的虫体经鉴定结果一致,均为华支睾吸虫成虫。对患者采用吡喹酮总量120 mg/kg体重,3 d分服9次服完的驱虫治疗与对症处理的治疗方案,经治疗半月、1月、3月复查与回访,临床症状与体征消失,治后半月、1月及3月的粪便均未再检获华支睾吸虫卵。结论该患者出现的华支睾吸虫病、胆结石、胆囊炎三联症为有华支睾吸虫病的流行或有发病的地区所少见。     

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.     

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.     

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.     

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.