Embryo–fetal exposure and developmental outcome of thalidomide following oral and intravaginal administration to pregnant rabbits

Studies in pregnant rabbits were conducted to evaluate if there are any differences in the uptake of thalidomide into the intrauterine compartment and developmental toxicity risk following oral and intravaginal administration. Thalidomide concentrations in maternal plasma, yolk sac cavity (YSC) fluid and embryo following intravaginal administration were 2- to 7-fold lower than their respective levels after oral administration. Ratios of thalidomide concentration in YSC fluid to maternal plasma were similar between these two routes, indicating no difference in uptake into the intrauterine compartment. A rabbit embryo–fetal development study using oral and intravaginal thalidomide administration at 2 mg/kg/day (a dose >10,000-fold higher than the expected amount of thalidomide in human semen) did not result in any developmental abnormalities. These data demonstrated no preferential transfer mechanism of thalidomide from vagina to conceptus, and no additional embryo–fetal developmental toxicity risks with thalidomide exposure via the vaginal route.     

体外受精和胚胎移植后妊娠合并急性粟粒性肺结核的临床特点

目的总结体外受精和胚胎移植（IVF-ET）后妊娠发生急性粟粒性肺结核的临床特点。方法回顾性分析2001年1月至2005年12月在浙江大学医学院附属妇产科医院生殖中心接受IVF-ET后妊娠并发粟粒性肺结核的6例临床资料。结果IVF-ET后妊娠并发粟粒性肺结核患者的临床表现多不典型，以发热为主要表现，呼吸道症状隐匿。胸部影像学表现以粟粒性结节和浸润性改变为主。平均于移植后53．2d发病，1例人工流产终止妊娠，其余5例均在发病后2～4周内发生自然流产。结论发热为急性粟粒性肺结核的主要临床表现，IVF-ET后妊娠发生粟粒性肺结核的妊娠结局差，对抗炎治疗无效的发热患者应警惕肺结核，尽早行结核病的相关检查。     

Changes in mRNA expression of MMP-2 in the Müllerian duct of chicken embryo

Although asymmetric development of the ovary and the oviduct is a unique characteristic in birds, the mechanism of asymmetric development still remains unclear. Recently, degradation of extracellular matrix has been suggested as an important factor related to the regression of the Müllerian duct in mammals. The present study was conducted to examine a possible role of metalloproteinase-2 (MMP-2) in the regression of the right Müllerian duct in the developing chicken embryo. Morphological changes in the Müllerian ducts were studied on day 15 of incubation and mRNA expresseion of MMP-2 was studied on days 12, 15, and 18 of incubation. Morphological observation demonstrated the disappearance of basement membrane in the right Müllerian duct which undergoes the regression. RT-PCR analysis showed that MMP-2 mRNA expression of the right Müllerian duct increased on days 15 and 18 of incubation coincidently with the time of regression. In the right Müllerian duct, regression was prevented by diethylstilbestrol treatment on day 4 of incubation and a coincident decrease in MMP-2 expression was observed when compared to the control group. These results suggest that MMP-2 may be involved in the regression of the right Müllerian duct in the female embryos of the chicken.     

Analysis of the functional integrity of cryopreserved human liver cells including xenografting in immunodeficient mice to address suitability for clinical applications.

BACKGROUND: The availability of well-characterized human liver cell populations that can be frozen and thawed will be critical for cell therapy. We addressed whether human hepatocytes can recover after cryopreservation and engraft in immunodeficient mice. METHODS: We isolated cells from discarded human livers and studied the properties of cryopreserved cells. The viability of thawed cells was established with multiple in vitro assays, including analysis of liver gene expression, ureagenesis, cytochrome P450 activity, and growth factor-induced cell proliferation. The fate of transplanted cells was analysed in immunodeficient NOD-SCID mice. RESULTS: After thawing, the viability of human hepatocytes exceeded 60%. Cells attached to culture dishes, proliferated following growth factor stimulation and exhibited liver-specific functions. After transplantation in NOD-SCID mice, cells engrafted in the peritoneal cavity, a heterologous site, as well as the liver itself, retained hepatic function and proliferated in response to liver injury. Transplanted hepatocytes were integrated in the liver parenchyma. Occasionally, transplanted cells were integrated in bile ducts. CONCLUSIONS: Cryopreserved human liver cell showed the ability to retain functional integrity and to reconstitute both hepatic and biliary lineages in mice. These studies offer suitable paradigms aimed at characterizing liver cells prior to transplantation in people.     

Effects on human heart valve immunogenicity in vitro by high concentration cryoprotectant treatment

It has been shown previously that cryopreservation, using an ice‐free cryopreservation method with the cryoprotectant formulation VS83, beneficially modulated immune reactions in vivo and in vitro when compared with conventionally frozen tissues. In this study, we assessed the impact of a VS83 post‐treatment of previously conventionally frozen human tissue on responses of human immune cells in vitro. Tissue punches of treated and non‐treated (control) aortic heart valve tissue (leaflets and associated aortic root) were co‐cultured for 7 days with peripheral blood mononuclear cells or enriched CD14⁺ monocytes. Effects on cellular activation markers, cytokine secretion and immune cell proliferation were analysed by flow cytometry. Flow cytometry studies showed that VS83 treatment of aortic root tissue promoted activation and differentiation of CD14⁺ monocytes, inducing both up‐regulation of CD16 and down‐regulation of CD14. Significantly enhanced expression levels for the C‐C chemokine receptor (CCR)7 and the human leukocyte antigen (HLA)‐DR on monocytes co‐cultured with VS83‐treated aortic root tissue were measured, while the interleukin (IL)‐6 and monocyte chemoattractant protein (MCP)‐1 release was suppressed. However, the levels of interferon (IFN)γ and tumour necrosis factor (TNF)α remained undetectable, indicating that complete activation into pro‐inflammatory macrophages did not occur. Similar, but non‐significant, changes occurred with VS83‐treated leaflets. Additionally, in co‐cultures with T cells, proliferation and cytokine secretion responses were minimal. In conclusion, post‐treatment of conventionally cryopreserved human heart valve tissue with the VS83 formulation induces changes in the activation and differentiation characteristics of human monocytes, and thereby may influence long‐term performance following implantation. Copyright © 2017 John Wiley & Sons, Ltd.     

宫腔镜输卵管近端栓塞术在体外受精-胚胎移植前预处理输卵管积水中的应用

输卵管积水发病率高,其不仅对育龄期妇女的生育力造成损害,还严重影响体外受精-胚胎移植(IVF-ET)的结果。如何在IVF-ET前妥善处理输卵管积水一直是临床治疗的难题。目前国际上尚缺乏相关的治疗指南。经典输卵管积水预处理方法主要包括经阴道输卵管积水抽吸术、输卵管切除术、输卵管近端结扎术、输卵管远端造口术以及输卵管栓塞术等。通过检索近年有关IVF-ET前输卵管积水预处理的相关文献,探讨临床常用输卵管积水预处理方法各自优缺点的同时,着重介绍宫腔镜下输卵管近端栓塞术的具体内容及作用机制,并分析该方法的可行性及安全性。近年国外多篇文献报道证实该方法效果良好。现行几种输卵管积水预处理方法的优劣目前尚无定论,宫腔镜下输卵管栓塞术作为一种新的方法,具有适应证广、有效、微创、简单经济、无放射损伤等优点,值得临床推广。     

自然周期冻融胚胎移植内膜准备中使用不同药物诱发排卵对妊娠结局的影响

目的:比较采用自然周期冻融胚胎移植内膜准备过程中自然排卵与人绒毛膜促性腺激素(h CG)诱发排卵及促性腺激素释放激素激动剂(Gn RHa)诱发排卵的不同临床结局。方法:回顾性分析湖南省郴州市第一人民医院生殖医学中心2011年10月—2013年10月采用自然周期准备子宫内膜的273例冻融胚胎移植周期。根据是否使用诱发排卵药物及药物的种类分为3组:A组为自然排卵组(n=181);B组为h CG诱发排卵组(n=47);C组为Gn RHa诱发排卵组(n=45)。比较3组患者的临床资料和妊娠结局。结果:13组年龄、移植日子宫内膜厚度、移植胚胎数、移植优胚率等比较差异无统计学意义(P0.05)。2A组临床妊娠率(56.4%)高于C组(33.3%,P0.01),而A组临床妊娠率与B组比较差异无统计学意义(P0.016 7)。结论:在自然周期冻融胚胎移植的内膜准备过程中,使用h CG诱发排卵对妊娠结局无不良影响,而使用Gn RHa诱发排卵则可能影响其临床结局。     

Incubation at 32.5 °C and above causes malformations in the zebrafish embryo

Zebrafish embryos are increasingly used for developmental toxicity screening of candidate drugs and are occasionally co-incubated with a metabolic activation system at 32 °C for 1, 2 or 4 h, depending on their developmental stage. As this temperature is higher than the optimal temperature for zebrafish embryonic development (26–28.5 °C), we investigated whether continuous incubation of zebrafish embryos from 2.5 until 96 h post fertilization (hpf) at high temperatures (30.5–36.5 °C) causes malformations. At 32.5 °C tail malformations were observed as early as 24 hpf, and these became even more prominent at 34.5 and 36.5 °C. Cardiovascular and head malformations, edema and blood accumulations throughout the body were present at 36.5 °C. Finally, temperatures higher than 28.5 °C accelerated embryonic development except for 36.5 °C, at which a lower hatching rate and hatching enzyme activity were observed. In conclusion, incubation of zebrafish embryos at 32.5 °C and above from 2.5 until 96 hpf causes malformations as early as 24 hpf.     

Helicobacter pylori is a fragile bacteria when stored at low and ultra-low temperatures

BACKGROUND AND AIM: Usually, bacteria are cryopreserved for short-term storage at low and ultra-low temperatures. There are no reports as to whether Helicobacter pylori is a fragile bacteria when stored at low and ultra-low temperatures as compared with other intestinal bacteria. A study was done on seven H. pylori strains and other intestinal bacteria to compare different temperatures for storage of organisms in saline solution. METHODS: Seven H. pylori strains, specifically American Type Culture Collection (ATCC) strains 43504 and TN2GF4, and five strains isolated from the present patients were grown on a modified Skirrow's agar for H. pylori. Escherichia coli and Bacteroides distasonis, both representing isolates from the present patients, were grown on trypticase soy blood agar for E. coli, and EG agar for B. distasonis. Culture was for 4-5 days under microaerobic, aerobic and anaerobic conditions at 37 degrees C. Cells were harvested by scraping growth from the solid medium and into sterile saline. The cells were adjusted to concentrations of 109 viable cells/mL in saline and preserved at 4 degrees C, -20 degrees C, or -80 degrees C for 3 weeks before reculture under microaerobic, aerobic and anaerobic conditions at 37 degrees C for 7 days. After incubation, morphologically distinct colonies were counted, isolated, and identified by standard bacteriologic techniques. The H. pylori were morphologically analyzed by electronic microscopy before and after preservation. Mongolian gerbils were inoculated with the cryopreserved H. pylori to evaluate the bacterial infectivity. RESULTS: Six of the seven H. pylori strains failed to culture after being preserved at 4 degrees C, -20 degrees C, or -80 degrees C. Only ATCC 43504 could be cultured after freezing at -80 degrees C. The number of H. pylori ATCC 43504 before preservation was 9.0 +/- 0.5 (log10 no. organisms/mL) and decreased to 5.7 +/- 0.6 after preservation. Morphologically, all H. pylori except ATCC 43504 strains transformed from a bacillary to a coccoid form after preservation. In addition, none of the H. pylori strains could infect Mongolian gerbils after preservation. Escherichia coli and B. distasonis were recovered. Titers before and after 4 degrees C, -20 degrees C, and -80 degrees C, respectively, were 9.1 +/- 0.2, 8.9 +/- 0.5, 8.6 +/- 0.3, and 8.7 +/- 0.3 for E. coli and 9.1 +/- 0.4, 8.7 +/- 0.6, 8.6 +/- 0.5, and 8.8 +/- 0.3 for B. distasonis. CONCLUSIONS: Helicobacter pylori is a fragile bacteria for storage at low and ultra-low temperatures in comparison with other intestinal bacteria.