流行性出血热病毒垂直传播的实验研究

本文以Balb/c鼠为动物模型用IF技术进行EHFV垂直传播的研究。结果证实EHFV可由孕鼠经胎盘感染胎鼠,在胎鼠的脏器中不仅能查出和分离出EHFV,其血中也可检出EHF的IgG抗体。并对孕鼠不同时期感染EHFV进行观察证实母鼠在妊娠前、后接种EHFV均可引起胎鼠感染,但在妊娠早期和中期对胎鼠存活无影响,而在妊娠晚期感染EHFV,可造成死胎和流产。从而确切地证实了EHFV在鼠体内的垂直传播。     

研发的全液体和冻干剂型的三价B群链球菌疫苗在健康未孕妇女中的安全性和免疫原性:Ⅱ期随机对照研究

正背景:作者对研发的三价B群链球菌(GBS)的液体和冻干剂型疫苗在未孕妇女中的安全性和免疫原性进行了评价,并根据血清型特异性免疫应答评估了剂型的等效性。方法:在18～40岁的健康未孕妇女中进行了随机、可比较、观察者盲式Ⅱ期试验。妇女在第1天接种单剂量的全液体(n = 529)或冻干(n = 521)的三价GBS疫苗。安全性评估一直进行到第181天(研究终点)。     

早孕小鼠子宫内膜M2型丙酮酸激酶基因表达的研究

该研究分析了M2型丙酮酸激酶(pyruvate kinase M2,PKM2)基因在早孕小鼠子宫内膜的表达规律。通过建立正常妊娠小鼠模型,收集孕D1、D4、D5、D6、D7小鼠子宫内膜组织及孕D5小鼠着床点及着床旁子宫内膜组织。构建假孕小鼠模型,收集假孕PD1、PD4、PD5、PD6和PD7小鼠子宫内膜组织。用Real-time PCR和Western blot方法检测PKM2 m RNA和蛋白质表达水平;免疫组织化学方法检测PKM2蛋白质在孕D5着床点与着床旁子宫内膜的分布。研究结果显示,在正常妊娠小鼠子宫内膜,PKM2 m RNA表达从孕D5开始出现明显升高,孕D6达高峰,孕D7略有下降,孕D6、孕D7与孕D1相比有明显差异。PKM2蛋白质从孕D6开始出现明显升高,孕D7略有下降,孕D6、孕D7与孕D1相比有明显差异。假孕小鼠子宫内膜PKM2 m RNA水平从PD6开始有明显升高,PD7与PD6水平相当,PD6、PD7与PD1相比有明显差异。PKM2蛋白质水平每两组间无明显差异。孕D5小鼠子宫内膜组织中,PKM2 m RNA及蛋白质水平均呈现着床点明显高于着床旁趋势。该研究初步揭示了PKM2基因在早孕小鼠子宫内膜表达规律,为深入探讨PKM2在维持早孕小鼠子宫内膜正常功能的机制上的作用提供了重要线索。     

流行性出血热病毒在疫区鼠间垂直传播的首次证实

流行性出血热(EHF)病毒在鼠间垂直传播的问题一直未被证实。1985～1986年我们在实验研究的基础上进行了疫区现场研究,采用的基本方法是从感染的孕鼠子宫内剖取胎鼠,用免疫荧光法作抗原、抗体检查和病毒分离,结果从流行性出血热疫区捕获的4只黑线姬鼠孕鼠的胎鼠体内检出了特异性抗原和抗体(IgM和IgG)。从24只抗原阳性的褐家鼠孕鼠中检出抗原阳性胎鼠18只,垂直传播率达75％。从褐家鼠孕鼠和胎鼠中分离出病毒六株,经特异性试验和单克隆抗体鉴定确认为EHF病毒。上述事实证明,在自然条件下,流行性出血热病毒在疫区鼠类中存在着经胎盘感染或垂直传播。这一证实对于阐明疫源地形成、延续和扩散,以及人类发病等具有重要意义。     

孕妇孕前体重指数与妊娠期糖代谢异常发病相关性的研究

目的:探讨孕前体重指数与妊娠期糖代谢异常发病及其临床特点的关系。方法:采用病例-对照研究的方法,对孕期在我院进行健康保健确诊为糖代谢异常并已完成分娩的孕妇130例作为病例组,同时随机抽取同期分娩的260例糖代谢正常孕妇作为对照组。用Logistic回归模型对孕前体重指数进行单因素分析;计算其相对危险度。结果:病例组孕妇孕前BMI指数大于25者占80.77%,明显高于对照组的(19.23%),差别有统计学意义(X2=30.469,P<0.05)。logistic回归结果显示相对于BMI小于25者,BMI>25组发生糖代谢异常的相对危险度是3.90,95%CI为2.23-6.41。结论:孕妇孕前体重指数是妊娠期糖代谢异常的独立危险因素,因此孕前控制体重指数是降低孕期糖代谢异常发生的关键因素。     

不同胚龄大鼠胚胎组织切片制作体会

全胚切片标本是发育生物学教学及其相关研究的基础。本实验采用石蜡包埋和免疫组织化学技术对如何制作完整和连续的不同胚龄之全胚切片进行了探索,并研究了亨廷顿相关蛋白1在孕中期胚鼠的表达。     

炎症免疫刺激对孕鼠胚胎基因表达谱影响的研究

目的:探讨妊娠期大鼠经炎症免疫刺激后对胚胎基因表达谱的影响.方法:选取6只孕鼠,分别腹腔注射LPS及Zymosan后,取胚胎抽提总RNA,经纯化反转录等过程合成生物素标记的cRNA,经片断化后与Rat Genome 230 2.0 Array芯片杂交,读取芯片结果并进行分析.结果:LPS组有183个基因上调、270个基因表达下调,Zymosan组有144个基因表达上调、417个基因表达下调.有50个基因在上述两组中均表达上调,其中已知功能的有9个;有173个基因在上述两组中表达均下调,已知功能的有85个.结论:应用全基因组芯片筛选了孕鼠经炎症刺激后胚胎差异表达的基因,为进一步研究孕鼠经炎症刺激后仔鼠血压升高的机制提供了新的思路.     

待孕夫妇乙肝表面抗原及乙肝表面抗体阳性情况分析

目的分析与探讨待孕夫妇乙肝表面抗原及乙肝表面抗体检测结果,并研究其对临床孕前检查的影响及评价。方法随机选取2015-2017年度在我院进行孕前检查的夫妇2 440对(4 880例)为研究对象,按照年度将待孕夫妇分为两组,每组2 440例,两组均加强孕前检查中的乙肝表面抗原(HBsAg)及乙肝表面抗体(HBsAb)的检测。A组为2015年3月-2016年2月在我院进行乙肝表面抗原及乙肝表面抗体检查的待孕夫妇;B组为2016年3月-2017年2月在我院进行乙肝表面抗原及乙肝表面抗体检查的待孕夫妇。比较两组待孕夫妇乙肝表面抗原及乙肝表面抗体检测的阳性结果。结果 B组HBsAg阳性率、HBsAb阳性率明显高于A组(6.43%vs 4.63%;62.99%vs 58.44%),差异有统计学意义(P0.05)。B组、A组男性HBsAg阳性率明显高于同组女性(59.87%vs 40.13%;60.18%vs 39.82%),HBsAb阳性率低于同组女性(46.52%vs 53.48%;47.41%vs 52.59%),差异均有统计学意义(P0.05)。B组、A组高中及以上学历HBsAg阳性率明显低于同组高中以下学历(38.85%vs 61.95%;38.05%vs 61.15%),高中及以上学历HBsAb阳性率高于同组高中以下学历(53.15%vs 46.84%;51.75%vs 48.25%),差异均有统计学意义(P0.05)结论目前夫妇乙肝感染仍处于增高趋势,对于进行孕前检查的待孕夫妇加强乙肝表面抗原及乙肝表面抗体的检测,有助于疾病的早期诊断、干预及治疗,能够减少乙肝传播,可有效降低新生儿乙肝发病率,促进优生优育,提高出生人口整体素质。     

豆状带绦虫四川株的生物学研究

目的对豆状带绦虫四川株的生物学特性进行深入研究,为豆状囊尾蚴病的防控提供基础数据。方法用豆状带绦虫虫卵经口感染兔后,于第60天剖杀采集豆状囊尾蚴并记录其在兔体内的寄生部位。用豆状囊尾蚴经口感染实验犬,观察犬首次排出孕节的时间、每天排出孕节的数目及排孕节持续时间;选取55个孕节,记录孕节中虫卵的数目及大小;用1%次氯酸钠溶液激活虫卵,观察虫卵孵化过程。结果 5只犬在感染豆状囊尾蚴后分别在感染后39 d、43 d、45 d、50 d和57 d开始排出孕节;最多一天排出158片孕节;孕节中虫卵含量在60~26 480个之间,平均为3372个,虫卵大小为(41.32±4.53)μm×(36.23±4.92)μm;虫卵孵化需要4~5 min。豆状囊尾蚴在家兔体内主要寄生于胃大网膜和直肠浆膜。结论犬感染豆状囊尾蚴后,在犬体内发育成熟的豆状带绦虫排孕节持续时间长,孕节内虫卵量大,排出的虫卵可对环境造成较大的污染,极易引起中间宿主的感染。     

孕妇孕前体重指数与妊娠期糖代谢异常发病相关性的研究

目的：探讨孕前体重指数与妊娠期糖代谢异常发病及其临床特点的关系。方法：采用病例一对照研究的方法,对孕期在我院进行健康保健确诊为糖代谢异常并已完成分娩的孕妇130例作为病例组,同时随机抽取同期分娩的260例糖代谢正常孕妇作为对照组。用Logistic回归模型对孕前体重指数进行单因素分析;计算其相对危险度。结果：病例组孕妇孕前BMI指数大于25者占80．77％,明显高于对照组的（19．23％）,差别有统计学意义（X2=30．469,P〈0．05）。logistic回归结果显示相对于BMI小于25者。BMI〉25组发生糖代谢异常的相对危险度是3．90,95％CI为2．23—6．41。结论：孕妇孕前体重指数是妊娠期糖代谢异常的独立危险因素,因此孕前控制体重指数是降低孕期糖代谢异常发生的关键因素。     

Characterization of a trypsin inhibitor from equine urine.

A trypsin inhibitor was isolated from pregnant mares' urine by adsorption on bentonite and elution with aqueous pyridine followed by batch DEAE-cellulose treatment and column chromatography. Final purification to an electrophoretically homogenous glycoprotein was achieved by gel permeation chromatography. This equine urinary trypsin inhibitor (E-UTI) is acid- and heat-stable, has a molecular weight of 22 to 23 kDa, an isoelectric point of 4.55, forms a 1:1 molar complex with trypsin and has serine as its N-terminal amino acid. The N-terminal amino acid sequence of this protein is almost identical with that of EI-14, the inhibitor obtained from horse serum by tryptic treatment, except for two extra amino acid residues, Ser-Lys- on the N-terminal end of E-UTI. In its isoelectric point E-UTI differs from EI-14 and the inhibitor from human urine.     

Urine collection in the common marmoset (Callithrix jacchus) and its applicability to endocrinological studies

We investigated a new method of urine collection in the common marmoset. We entered the cage as soon as the light cycle started in the breeding room and collected urine from the animal directly without any restraint. We were able to take separate samples from completely different individuals housed together for behavioral studies in the same cage. Urine and blood samples were taken from individuals from late pregnancy through postpartum nursing period. Cortisol and prolactin concentrations measured in urine were compared to those measured in blood to evaluate this collection method. LH/CG level in the urine samples was also measured. Urine data in females indicated a tendency toward high cortisol values during late pregnancy, a sharp drop before parturition, and increase after delivery. In females cortisol levels measured in blood closely resembled concentrations measured in urine. Urine cortisol in males clearly indicated an increase postpartum, but the increase was not indicated in plasma. Plasma and urine prolactin concentrations in females made a similar increase during lactation. Male's plasma prolactin clearly indicated an increase directly proportional to strong behavioral contact with the infant. We also confirmed hormonal changes during pregnancy, and postpartum ovulation and subsequent pregnancies, from urine LH/CG data. We found this method extremely useful because of the high correlation between cortisol, prolactin and LH/CG data from blood and urine. Additionally, we collected urine samples with little stress to the animal from fear, irritation, pain, etc.     

Effect of maternal tobacco smoking or exposure to second-hand smoke on the levels of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine of mother and the first urine of newborn

Tobacco smoking during pregnancy is associated with a variety of negative consequences not only for the mother, but also for the developing fetus. Many studies have shown that carcinogens contained in tobacco smoke permeate across the placenta, and are found in fetus. The aim of the study was to determine the prenatal exposure to tobacco-specific carcinogenic N-nitrosamines on the basis of measurements of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in urine of smoking and second-hand smoke (SHS) exposed women and in the first urine of their newborns. A questionnaire documenting demographics and socio-economical data, smoking habits and exposure to SHS was completed by 121 delivering women near or at term. Maternal concentrations of cotinine and NNAL were measured in urine of the mother and the first urine of her newborn infant by liquid chromatography tandem mass spectrometry (LC/MS/MS). The mean concentration of cotinine was 439.2 ng/mg creatinine and NNAL concentration in urine of smoking women was 74.0 pg/mg creatinine, and for her newborn 78.6 pg/mg creatinine. Among mothers exposed to SHS, cotinine and NNAL mean concentration were 23.1 ng/mg creatinine, and 26.4 pg/mg creatinine. In newborns of SHS exposed mothers during pregnancy the mean concentration of NNAL was 34.1 pg/mg creatinine, respectively. Active tobacco smoking as well as passive exposure to smoking during pregnancy is an important source of tobacco specific N-nitrosamines to the fetuses as evidenced by increased concentrations of this carcinogen. Determination of NNAL in maternal urine samples can be a useful biomarker of prenatal exposure of newborn to carcinogenic nitrosamines.     

A cytogenetical study of prenatal loss in the mare

The objective of this study was to investigate an hypothesis that chromosome anomalies are an important cause of prenatal loss in the mare. An attempt was made to analyse, cytogenetically, a series of 26 equine abortuses. Cell cultures were prepared from a range of tissues, but failed to grow, and chromosome analysis was therefore not possible for any of these specimens. Consequently, a study was made of the metaphase chromosomes prepared from 22 equine embryos after their surgical removal from mares' uteri. The karyotypes prepared for each specimen were normal. The current findings are discussed in the light of similar studies of induced abortions in women and embryonic loss in animals, and the potential for further investigation in this field is emphasized.     

子痫前期患者尿液中足细胞裂孔膜蛋白和标记蛋白的检测及意义

目的：检测子痫前期患者尿液中足细胞裂孔膜蛋白和足细胞标记蛋白的浓度并探讨其临床意义。方法：本实验以62例妊娠期妇女为研究对象,分为三组,其中正常妊娠期妇女25例为正常对照组,慢性高血压的妊娠期妇女17例为高血压组,子痫前期患者20例为子痫前期组。ELISA检测各组妊娠期妇女的尿液中足细胞裂孔膜蛋白和足细胞标记蛋白的表达;Bradford法检测各组妊娠期妇女的尿蛋白。结果：足细胞裂孔膜蛋白和足细胞标记蛋白在正常对照组尿液中含量极少,在高血压组中分泌增加,而子痫前期组患者中明显升高（P〈0．01）,且在子痫前期组尿液中足细胞裂孔膜蛋白和标记蛋白的分泌含量均成正相关（r2=0．79,P〈0．05）。子痫前期组患者中足细胞裂孔膜蛋白和足细胞标记蛋白与尿蛋白浓度成正相关（r2=0．58,P〈0．05;r2=0．79,P〈0．05）。结论：足细胞蛋白脱落主要发生于子痫前期患者,且足细胞蛋白脱落量与尿蛋白成正相关,能直接反映妊娠期患者的肾损伤程度,可作为预测罹患妊娠期高血压的指标。     

Pregnancy diagnosis in urine of Iberian lynx (Lynx pardinus)

Diagnosis of pregnancies is an important management tool for the Iberian lynx Conservation Breeding Program, a program geared to recover the world's most endangered felid. Non-invasive methods such as fecal hormone analyses are not applicable to the lynx, since fecal progestin does not follow the typical pregnancy pattern of felids. Therefore, we aimed to test whether urine can be used as an alternative substance for pregnancy diagnosis in the Iberian lynx.Progesterone immunoreactive metabolites were determined in urine samples of pregnant and non-pregnant females before and during breeding season. Additionally, we used the Witness^®Relaxin test to determine relaxin in blood and urine. No differences were found in progestin concentrations determined in urine samples collected from pregnant and non-pregnant animals between day 1 and 65 following mating. Although the Witness^®Relaxin test was positive in serum samples collected from animals between day 32 and 56 of pregnancy, it failed in both fresh and frozen urine samples collected from the same stage of pregnancy. A weak relaxin reaction in urine samples collected from animals between day 29 and 46 of pregnancy was detectable after urines were concentrated by ultrafiltration (>50×). Concentrated samples obtained from non-pregnant and early pregnant animals yielded negative test results. In conclusion, the Witness^®Relaxin test can be applied for pregnancy diagnosis in Iberian lynx in both serum and concentrated urine samples obtained during the second half of pregnancy. A positive relaxin test indicates an ongoing pregnancy, whereas negative tests must be judged carefully as hormone concentrations might be below detection thresholds.     

胚胎移植术前指导患者憋尿对移植结局影响的研究

摘要 目的：探讨胚胎移植术前患者憋尿准备的细化指导方法,并分析其对患者移植结局的影响。方法：选取2019年9月至2020年12月期间我院收治的194例胚胎移植患者（包括新鲜周期胚胎移植和冻融周期胚胎移植）,按随机数字表法分为指导组（101例）和对照组（93例）。对照组患者给予常规指导憋尿,指导组患者在对照组的基础上给予更细化的憋尿指导方法。比较两组总妊娠率、不同年龄段患者的移植妊娠率,分析提前指导憋尿对瘢痕子宫和无子宫手术史患者移植妊娠率的影响。结果：指导组总妊娠率为59.41%,对照组总妊娠率为53.76%,指导组总妊娠率高于对照组,但是未见显著性差异（P>0.05）;在<35岁的患者中,指导组妊娠率（76.47%）高于对照组（58.33%）,差异有统计学意义（P<0.05）;在瘢痕子宫和无子宫手术史的患者中,两组移植妊娠率比较未见显著性差异（P>0.05）。结论：胚胎移植术前给予患者细化的憋尿指导方法能提高<35岁患者的移植妊娠率,对瘢痕子宫和无子宫手术史患者的移植妊娠率无明显影响。     

THE USE OF THE MALE CALIFORNIA TOAD IN THE DIAGNOSIS OF PREGNANCY

The California male toad, readily available in most areas of the state, and for many other reasons a better subject for test purposes than the frog, was used in 237 tests to determine the presence or absence of pregnancy in humans. Human urine was injected into the toads, and the cloacal discharge then was examined for the presence of sperm. There were only two false reactions in the series—both false negative. In subsequent tests in both these cases, the result was positive.     

The effect of weight on intravaginal pressure and vulval conformation of mares

Pressures were recorded monthly at two sites in the vaginas of nine horse mares with normal vulval conformation. The pressures were lowest when the mares' weights were minimal, and there was evidence of a descending pressure gradient from the vestibule to the cervix. Loss of body weight was also associated with shortening and angulation of the vulva, although it was not displaced above the pelvic brim.     

Increased urinary excretion of nephrin, podocalyxin, and βig-h3 in women with preeclampsia

Emerging evidence has shown that podocyte injury and reduced specific podocyte protein expressions contribute to proteinuria in preeclampsia. We collected urine specimens from women with preeclampsia to study whether podocyte-specific protein shedding is associated with renal barrier dysfunction. Urine specimens from women with normal pregnancies and from pregnant women complicated by chronic hypertension were used for comparison. We determined soluble podocyte slit protein nephrin levels in the urine specimens. Podocalyxin, βig-h3, and VEGF concentrations were also measured. We found that nephrin and podocalyxin were barely detectable in the urine specimens from normal pregnant women and from women with chronic hypertension. In preeclampsia, urinary nephrin and podocalyxin concentrations were significantly increased and highly correlated to each other, r(2) = 0.595. Nephrin and podocalyxin were also correlated with urine protein concentrations. βig-h3 was detected in the urine specimens from women with preeclampsia, and it is highly correlated with nephrin and podocalyxin concentrations in preeclampsia. βig-h3 was undetectable in normal pregnancy and pregnancy complicated by chronic hypertension. Elevated VEGF levels were also found in women with preeclampsia compared with those of normal pregnancy and pregnancy complicated by chronic hypertension. These results provide strong evidence that podocyte protein shedding occurs in preeclampsia, and their levels are associated with proteinuria. The finding of urinary βig-h3 excretion in preeclampsia suggests that increased transforming growth factor activity might also be involved in the kidney lesion in this pregnancy disorder.