糖尿病并发弓形虫感染的初步研究

本文用ＩＨＡ、ＩＦＡ、Ｄｏｔ－ＥＬＩＳＡ三种血清学方法检测３２０例糖尿病病人血清的弓形虫抗体，阳性率分别是１０．３１％、１８．１３％和１９．０６％。均高于对照组（Ｐ＜０．００５），这是糖尿病患者并发弓形虫感染的首次报告。     

多项免疫吸附凝集试验检测孕妇弓形虫感染的实验研究

用弓形虫ＩｇＧ／ＩｇＡ／ＩｇＭ免疫吸附凝集试验检测５０７份孕妇血清，并与ＤＡ、ＳＦＴ、ＩＳＡＧＡ－ＩｇＭＥＩＡ－ＩｇＭ多项方法检测结果比较分析。多项检测结果阴性的２０１份血清中，该法检测出ＩｇＧ阳性２１份，ＩｇＡ、ＩｇＭ均为阴性，较ＤＡ敏感；多项检测结果为非活动性弓形虫感染的２０３份血清中，该法检出ＩｇＧ阳性１９７份、ＩｇＡ阳性１１份、ＩｇＭ阳性４份，阳性率分别为９７．０５％、５．４１％、１．９７％两者结果基本相符；多项检测结果为活动性感染的１０３份血清中，该法检出ＩｇＭ阳性８７份，ＩｇＡ阳性６１份，ＩｇＧ阳性１０３份，其中ＩｇＭ阳性与单项的ＩＳＡＧＡ－ＩｇＭ阳性结果相符率较高为８３．０２％。弓形虫ＩｇＧ／ＩｇＡ／ＩｇＭ免疫吸附凝集试验盒可作弓形虫感染的定性、定量检测，对判定孕妇的弓形虫感染时间是较好的方法。     

孕期弓形虫感染对胎儿的影响

目的为了解亚临床感染弓形虫的孕妇,母婴之间垂直传播情况。方法采用聚合酶链技术（ＰＣＲ）分别检测母婴血弓形虫ＤＮＡ（Ｔｏｘｏ－ＤＮＡ）及羊水Ｔｏｘｏ－ＤＮＡ．结果１．对３０例孕晚期孕妇检测羊水Ｔｏｘｏ－ＤＮＡ与其新生儿血Ｔｏｘｏ－ＤＮＡ,其检测结果无显著差异,Ｐ＞０．０５。２．孕中晚期组孕妇感染弓形虫后,其胎儿感染率明显高于孕早期组,Ｐ＜０．０５。结论孕中晚期孕妇感染弓形虫后,易于传染给胎儿且感染弓形虫为孕妇若能维持到足月分娩,出生的胎儿多为无临床症状的先天性弓形虫病（实验实诊断为阳性）,这组胎儿有可能于数年后发生远期并发症;由于羊水易于在孕中晚期获取,故检测羊水Ｔｏｘｏ－ＤＮＡ将有利于先天性弓形虫病的早期诊断及早期治疗,从而控制先天性弓形虫病儿的出生。     

弓形虫病IgM免疫吸附凝集试验（ISAGA）的建立

目的∶建立检测弓形虫 Ｉｇ Ｍ 抗体的免疫吸附凝集试验 （ Ｉ Ｓ Ａ Ｇ Ａ） 。方法∶ Ｕ 型微孔板以适宜浓度的羊抗人 Ｉｇ Ｍ 抗体包被， 用１ ％ 牛血清白蛋白封闭， 洗板后加入待测血清， 在３７ ℃孵育后洗涤， 加入弓形虫（ Ｒ Ｈ 株） 速殖子抗原悬液， 置３７ ℃过夜后观察结果。将其与丹麦的 Ｉ Ｓ Ａ Ｇ Ａ 和 Ｅ Ｌ Ｉ Ｓ Ａ 检测的结果以及玻片 Ｅ Ｉ Ａ 检测的结果作比较。结果∶本法与丹麦的试验检测丹麦孕妇４４ 份血清， 总符合率为９３２ ％ ；与玻片 Ｅ Ｉ Ａ 检测丹麦和上海孕妇６７ 份血清， 总符合率为９２５ ％ ， 两法的滴度之间明显相关 （γ＝ ０５８９ ， Ｐ＜ ０００１） ， Ｉ Ｓ Ａ Ｇ Ａ 滴度较玻片 Ｅ Ｉ Ａ 高１８ 倍； 用 Ｗ Ｈ Ｏ 国际生物标准化实验室提供的标准的抗弓形虫人血清定量测得其灵敏度为００８ Ｉ Ｕ／ ｍｌ。结论∶ Ｉｇ Ｍ Ｉ Ｓ Ａ Ｇ Ａ 具有敏感性、特异性高， 操作简便等优点， 不仅可作为弓形虫急性感染和慢性感染活动期的一种检测手段， 而且适用于弓形虫感染调查的大规模筛选。     

应用PCR技术检测孕早期绒毛组织弓形虫DNA的研究

应用ＰＣＲ技术检测２７０例孕６－１２周妇女外周血及相应绒毛组织中的弓形虫ＤＮＡ，结果弓形虫ＤＮＡ阳性２６例，相应的绒毛组织中检出８例，垂直传播率为３０．７７％，同时用ＥＬＩＳ法检测血清中弓形虫循环抗原、ＩｇＭ和ＩｇＧ抗体，结果阳性率分别为３．７０％、７．０３％和１５．５％。ＩｇＭ和／或ＣＡｇ阳性者，外周血皆检出弓形虫ＤＮＡ。提示应用ＰＣＲ技术检测绒毛组织弓形虫ＤＮＡ结合ＥＬＩＳＡ进行筛选，可早期诊     

608例早孕绒毛宫内感染巨细胞病毒的免疫组织化学诊断

目的建立简便、快速的巨细胞病毒（ＣＭＶ）胚胎感染的诊断方法,早期诊断胚胎ＣＭＶ感染。方法应用免疫组化过氧化物酶抗过氧化物酶（ＰＡＰ）方法,检测与观察早孕绒毛组织的ＣＭＶ感染病灶。结果早孕绒毛６０８例,感染ＣＭＶ２２２例（３６．５％）。感染灶可分为三类,点状感染占７１％（１５８／２２２）,巢状感染占１７％（３７／２２２）,弥散状态感染占１２％（２７／２２２）。发现随胎龄增加巢状及弥散状感染增多,ＣＭＶ往往先侵入绒毛霍夫保尔细胞。经电镜观察验证了ＰＡＰ法。结论免疫组化ＰＡＰ法简便、灵敏、有效,可用于大量筛查宫内ＣＭＶ感染     

混合血清法检测献血员血清抗弓形虫抗体

将徐州市区义务献血员６人一组混合，用斑点免疫金银染色法（Ｄｏｔ－ＩＧＳＳ）进行检测，同时对所有血清用同样方法逐份进行检测，以研究混合血清法筛检抗弓形虫抗体的可能性。检测结果显示，混合血清法的敏感性为９２．３１％。特异性为９９．９６％，混合血清法与逐份血清法的一致率为９９．７３％，Ｋａｐｐａ值为０．９４７（Ｐ〈０．０１）。以全国弓形虫平均感染率（４．８６％）进行定量效益分析，用混合血清法筛检献血员，     

斑点免疫金银染色与Dot—ELISA检测抗弓形虫抗体的对比研究

为比较斑点免疫金银染色（Ｄｏｔ－ＩＧＳＳ）和Ｄｏｔ－ＥＬＩＳＡ检测弓形虫抗体的敏感性和特异性，本实验以弓形虫速殖子可溶性蛋白抗原为诊断用抗原，同时用Ｄｏｔ－ＩＧＳＳ和Ｄｏｔ－ＥＬＩＳＡ检测６５份弓形虫感染者血清、１７６份其他寄生虫感染者血清和７６份对照血清。结果显示，用Ｄｏｔ－ＩＧＳＳ及Ｄｏｔ－ＥＬＩＳＡ检测弓形虫感染者抗体阳性率分别为９８．４６％及９２．３１％，平均抗体滴度分别为１：３８４及１：     

先天性弓形虫病产前监测程序的研究

目的为减少官内感染和先天畸形，我们设计了弓形虫病的产前监测程序。方法采用ELISA法作为血清筛查的基本方法，绒毛、羊水和脐血用ELISA、PCR或小白鼠接种法进行监测。结果筛查出1192例监测对象，除271例拒绝监测外，无症状组509例，有先兆流产组113例，羊膜穿刺组299例。结论本研究结果提示，只要按程序监测，先天性弓形虫病是可以控制的。但卫生宣教也是需要的。     

COMPARATIVE STUDY ON DETECTION OF ANTI-TOXOPLASM A GONDII ANTIBODIES BY USING DOT-ELISA AND DOT-IMMUNOGOLD SILVER STAINING

为比较斑点免疫金银染色（Ｄｏｔ－ＩＧＳＳ）和Ｄｏｔ－ＥＬＩＳＡ检测弓形虫抗体的敏感性和特异性，本实验以弓形虫速殖子可溶性蛋白抗原为诊断用抗原，同时用Ｄｏｔ－ＩＧＳＳ和Ｄｏｔ－ＥＬＩＳＡ检测６５份弓形虫感染者血清、１７６份其他寄生虫感染者血清和７６份对照血清。结果显示，用Ｄｏｔ－ＩＧＳＳ及Ｄｏｔ－ＥＬＩＳＡ检测弓形虫感染者抗体阳性率分别为９８．４６％及９２．３１％，平均抗体滴度分别为１∶３８４及１∶２１８（１∶２０－１∶２５６０）。用Ｄｏｔ－ＩＧＳＳ检测，有２３份血清抗体滴度高于Ｄｏｔ－ＥＬＩＳＡ，有８份低于Ｄｏｔ－ＥＬＩＳＡ。二种方法所测抗体滴度呈直线正相关关系（ｒ＝０．６０８，Ｐ＜０．０１）。二种方法检测其他寄生虫感染者和对照血清，假阳性率分别为５．９５％及１３．８９％。Ｄｏｔ－ＩＧＳＳ操作流程和Ｄｏｔ－ＥＬＩＳＡ相似，但敏感性、特异性均优于Ｄｏｔ－ＥＬＩＳＡ，前者不使用对人体有害的底物，有较好的推广前景。     

绒毛、羊水弓形虫感染的研究

Aim In order to know well toxoplasma infection in intrauterine and to reduce congenital malformation.Methods 104 chorionic tissue and 494 amniotic fluid were determind by ELISA and mouse inoculation methods.Results Intrauterine infection was shown in choionic tissue(18.19%)and amniotic fluid(4.56%).After antitoxoplasma treatment,IgM and CAg positive was 3.86%(untreated 11.85%),and abnormal child birth was 2.56%(untreated 18.29%)Conclusion It indicated that seropositive pregnant women should avoid toxoplasma infection.The serveillance of amnioctic fluid is an important procedure of diagnosis and control for the maternalf vertical infection of the toxoplamosis,but the early prevention is the better.     

乙型肝炎病毒宫内感染的传播途径及早期诊断

目的分析羊水、脐带血、母血、胎盘等组织中乙型肝炎病毒标志物(HBV M)及HBV DNA与胎儿感染的关系，探讨HBV母婴传播机制。方法采用微粒子化学发光及核酸扩增杂交梳技术对65例血液乙型肝炎表面抗原(HBsAg)阳性不同孕龄孕妇的羊水、母血、脐带血进行HBV M和HBV DNA检测，对自然流产胎儿或死亡婴儿的胎盘、肝脏、心脏、肺脏进行免疫组织化学检测。结果65例血液HBsAg阳性，不同孕龄孕妇的羊水中HBsAg阳性率为21．50％，脐带血阳性率为20．00％；母血、羊水．脐带血HBsAg、乙型肝炎e抗原(HBeAg)、抗-HBc、HBV DNA均阳性者为6．15％；HBsAg、抗-HBc、抗-HBc阳性、HBV DNA阴性者占13．85％。对4例血液、羊水、脐带血HBsAg、HBeAg、抗-HBc、HBV DNA阳性孕妇分娩或自然流产后胎盘、胎儿及死亡婴儿的肝脏、肺脏、心脏进行免疫组织化学检测发现，胎盘各层组织镜下均可见到HBsAg、HBcAg阳性细胞；在肝脏、肺脏组织中可见到HBsAg、乙型肝炎核心抗原(HBcAg)阳性细胞，心肌组织内未见有HBsAg、HBcAg阳性细胞。结论胎儿感染HBV与羊水、胎盘中的病毒相平行；HBV在宫内可感染胎儿血液、肝脏、肺脏等组织；羊水检测HBV M及HBV DNA可作为胎儿早期HBV感染的诊断依据之一。     

The role of parathyroid hormone-related protein in intra-tracheal fluid

Parathyroid hormone-related protein (PTHrP), a multi-functional protein, is produced by many tissues in fetus. PTHrP concentration in amniotic fluid is reported to be significantly higher than in either fetal or maternal plasma. Other investigators have reported that PTHrP in amniotic fluid is derived mainly from amnion. The aim of this study was to investigate the contribution of fetus to PTHrP in amniotic fluid and the role of PTHrP in human fetal lung tissue. Samples of amniotic fluid, neonatal intra-tracheal fluid, gastric fluid, and the first urine of neonates were obtained at the time of elective cesarean section (n=11), and the concentrations of PTHrP were measured. The PTHrP level in intra-tracheal fluid (41.0+/-19.6 pmol/l, mean+/-SD) was significantly higher than the levels in amniotic fluid (22.1+/-0.8), neonatal gastric fluid (13.5+/-2.5), first urine (0.95+/-0.6), umbilical cord venous and arterial plasma (1.35+/-0.2, 1.63+/-0.3) and maternal plasma (1.05+/-0.1). PTHrP and PTH/PTHrP receptor mRNA were detected in human lung tissue obtained from a fetus stillborn at 36 weeks of gestation. The effects of PTHrP on fetal lung maturation were studied in H441 cells from a human lung epithelial cell line. PTHrP (10(-7)M) significantly suppressed cell proliferation (p<0.05) to approximately 80% of the control level, while administration of PTHrP significantly increased surfactant protein A production (p<0.01). We first demonstrated the high concentration of PTHrP in intra-tracheal fluid that may suggest the positive production of this protein from the fetal lung. The results obtained by in vitro study using a human lung epithelial cell line suggest that PTHrP derived from the fetal lung might modulate its own maturation.     

孕妇与胎儿弓形虫感染状况的研究

目的　探讨孕妇与胎儿弓形虫 (Toxoplasma,TOX)感染的状况与关系。　方法　用酶联免疫吸附法及荧光定量多聚酶链反应技术检测孕妇血中 TOX特异性抗体及 TOX DNA,对 TOX感染孕妇检测羊水或脐血诊断胎儿感染。　结果　 15 6 4例孕妇中血清 TOX- Ig M阳性者 4 1例 (2 .6 2 % ) ,TOX- Ig G阳性 5 9例 (3.77% ) ,TOX DNA阳性 4 5例(2 .88% ) ,4 1例 TOX- Ig M阳性者其 TOX- DNA均为阳性。 4 5例 TOX感染孕妇中羊水或脐血 TOX- DNA阳性 13例(2 8.89% )。 TOX感染胎儿中 2例自然流产 ,1例死胎、1例胎儿生长受限 (Fetal growth restriction,FGR) ,9例出生时无明显症状的新生儿中 ,有 1例生后 1个月患黄疸性肝炎 ,1例有单侧耳聋 ,7例生长发育正常。　结论　孕妇 TOX感染可危害胎儿 ,孕妇 TOX感染后取羊水或脐血检测 TOX DNA是诊断胎儿 TOX感染的有效方法。     

HBV宫内感染的传播途径及其机理的研究

目的 探讨HBV宫内感染的传播途径及其机理。方法 应用PCR技术检测HBV感染孕妇羊水、阴道分泌物、乳汁、脐血中HBV DNA；免疫组织化学技术检测胎盘组织中HBsAg和HBcAg的表达。结果 HBsAg、HBeAg、抗HBc阳性孕妇的羊水、阴道分泌物、乳汁、脐血中均检测到了HBVDNA，阳性率分别为48．28％（14／29）、27．59％（8／29）、37．93％（11／29）和24．14％（7／29）；健康对照组孕妇的上述样品中均未检出HBVDNA；HBV感染孕妇胎盘组织各层细胞均可表达两种抗原，但阳性细胞数目从母体面到胎儿面逐渐减少，阳性细胞的着色强度逐渐减弱。健康对照组孕妇胎盘组织中未发现HBsAg和HBcAg阳性染色细胞。结论 孕妇感染HBV后可通过多种途径传播，而羊水感染是导致胎儿感染的重要传播途径。     

Effect of Amniotic Fluid on Blood Coagulation

Amniotic fluids from, normal subjects and from patients with intrauterine death (IUD) were studied for their ability to shorten the clotting-time of blood in vitro . The IUD amniotic fluid was different in that it seemed to require the presence of factor VII for its action whereas normal amniotic fluid did not. In this it resembled tissue factor in its action, whereas normal amniotic fluid resembled Russell Viper venom.     

Mechanism of intrauterine infection of hepatitis B virus

AIM:To explore the possible mechanism of intrauterine infection of hepatitis B virus (HBV).METHODS: HBV DNA was detected in vaginal secretion and amniotic fluid from 59 HBsAg-positive mothers and in venous blood of their newborns by PCR. HBsAg and HBcAg in placenta were determined by ABC immunohistochemistry.RESULTS:The rate of HBV intrauterine infection was 40.1% (24/59). HBV DNA was detected in 47.5% of amniotic fluid samples and 52.5% of vaginal secretion samples respectively.HBsAg and HBcAg were detected in placentas from HBsAg-positive mothers. The concentration of the two antigens decreased from the mother‘s side to the fetus‘s side, in the following order:maternal decidual cells&gt;trophoblastic cells&gt; villous mesenchymal cells&gt;villous capillary endothelial cells. However, in 4 placentas the distribution was in the reverse order. HBsAg and HBcAg were detected in amniotic epithelial cells from 32 mothers.CONCLUSION:The main route of HBV transmission from mother to fetus is transplacental, from the mother side of placenta to the fetus side. However, HBV intrauterine infection may take place through other routes.     

Unusual pulmonary embolism: septic pulmonary embolism and amniotic fluid embolism.

BACKGROUND: Septic and amniotic fluid emboli are rare sources of pulmonary embolism (PE), so the present study sought to elucidate the background of these cases. METHODS AND RESULTS: A total of 11,367 PE cases were identified from 396,982 postmortem examinations. The incidence of septic PE was 247 (2.2%) of the total. The origin of infection was found in 85.6% of the cases. Fungal embolus was detected more often than bacterial embolus. The most frequently detected fungus was aspergillus (20.8%). The primary disease associated with fungal embolus was leukemia (43.2%). The incidence of PE cases associated with pregnancy and/or delivery was 89 (0.8%) of the total PE cases. Among them, amniotic fluid embolism was found in 33 (73.3%) of 45 PE cases with vaginal delivery, and in 7 (21.2%) of 33 PE cases with cesarean delivery (p<0.0001). CONCLUSION: Fungal embolus was more frequent than bacterial embolus, and leukemia was most frequent as the primary disease in cases of fungal embolus. The main cause of PE in cesarean section cases was thrombotic embolism, and the main cause in vaginal delivery cases was amniotic fluid embolism.     

Cytomegalovirus infection during pregnancy with maternofetal transmission induces a proinflammatory cytokine bias in placenta and amniotic fluid

Congenital infection with cytomegalovirus (CMV) can induce immune responses and placental damage. By use of immunoassay panels, 27 cytokines were assessed in midtrimester amniotic fluid from 8 patients with congenital CMV, in midtrimester sera from 12 pregnant women with primary CMV infection, and in amniotic fluid and serum from uninfected maternal controls. Levels of the cytokines tumor necrosis factor α, interleukin 1β, interleukin 12, and interleukin 17; the chemokines CCL2, CCL4, and CXCL10; and the growth factors granulocyte-macrophage colony-stimulating factor and platelet-derived growth factor bb were significantly elevated in amniotic fluid from congenital CMV patients (P < .01). Only CXCL10 was significantly higher in sera from CMV-infected pregnant women. CMV infection during pregnancy is associated with a shift in cytokine expression toward a proinflammatory state.     

Ursodeoxycholic acid administration in patients with cholestasis of pregnancy: effects on primary bile acids in babies and mothers

Little is known about the effects on the fetus of ursodeoxycholic acid (UDCA) treatment for intrahepatic cholestasis of pregnancy (ICP). Twenty ICP patients were given UDCA at 1.5 to 2 g/d, to our knowledge the highest dosage yet reported. Effects were evaluated on conjugated bile acids (BA) in amniotic fluid (15 of 20 patients) and umbilical cord serum obtained at delivery (20 of 22 newborns), as compared with 10 untreated patients (amniotic fluid, 9 of 10 patients; cord serum, 9 of 10 newborns). Liver function tests, serum BA and UDCA were evaluated on enrollment and then weekly until 1 week after delivery. Maternal serum conjugated cholic (CCA) and chenodeoxycholic (CCDCA) acids levels fell (18.5 +/- 1.9 to 10.5 +/- 1.9 micromol/L, and 5.8 +/- 0.8 to 2.97 +/- 0.7 micromol/L, respectively [P <.01]) in treated patients, and remained unaffected (20.0 +/- 3.1 vs. 20.3 +/- 2.3, and 5.6 +/- 0.6 vs. 5.4 +/- 0.5, respectively [P = not significant]) in untreated ones. Serum conjugated UDCA levels rose to 16.5 +/- 1.8 micromol/L (P<.001). Median values of CCA and CCDCA in amniotic fluid around delivery were 4.9 +/- 12.4 and 4.8 +/- 7.7 micromol/L, respectively, in treated patients, as against 17.9 +/- 27.5 and 18.5 +/- 20.9 micromol/L in untreated ones. In treated mothers, CCA and CCDCA concentrations in cord blood were 6.0 +/- 0.9 and 5.2 +/- 0.95 micromol/L, respectively, as against 21.9 +/- 5.6 and 18.9 +/- 2.1 micromol/L in untreated ones. In treated patients, median UDCA values in amniotic fluid and cord blood were 0.8 +/- 2.4 and 0.9 +/- 0.14 micromol/L, respectively. We conclude that increasing the dose of UDCA more effectively controls ICP and improves maternal clinical outcome after delivery.