A study of the distribution of 7-ethoxycoumarin O-de-ethylase activity in human placental subcellular fractions

Human placental homogenates from maternal smokers and non-smokers were fractionated using differential centrifugation techniques. Yields of the various subfractions were determined and their homogeneity assessed using electron microscopic procedures. The distribution and response of 7-ethoxycoumarin O-de-ethylase activity towards inhibition by dimethylsulphoxide, alpha-naphthoflavone and 9-hydroxyellipticine inhibitors in the placental subfractions were investigated. The low yield of microsomal protein obtained following differential centrifugation of placental homogenates (2.5 +/- 0.2 mg protein per g placenta) highlights the extremely refractory nature of human placental tissue towards homogenization. Enzymic studies showed that the majority (75 per cent) of the original O-de-ethylase activity in homogenates from smokers and non-smokers was to be found in the crude nuclear fraction. The 7-ethoxycoumarin O-de-ethylase activity present in both homogenate and crude nuclear preparations from a maternal smoker was found to be inhibited by both alpha-naphthoflavone and 9-hydroxyellipticine to a lesser extent than the O-de-ethylase activity which was present in both mitochondrial and microsomal fractions. While this observation suggests the existence of more than one induced O-de-ethylase activity in the human placenta, the possibility that such differences in inhibitory response may be due to other factors (e.g. inhibitor solubility effects) cannot be excluded. Studies using the above inhibitors also confirmed the results of earlier work by demonstrating that the O-de-ethylase activity in placental homogenates and subfractions from non-smokers is qualitatively different from the O-de-ethylase activities induced as a result of maternal smoking.     

Radiotherapy and/or chemotherapy as adjuvant treatment of uterine sarcomas

A retrospective evaluation of 87 cases of uterine sarcoma treated during 1965-1980 at two Swedish hospitals has been made. Adjuvant postoperative radiotherapy has been compared with adjuvant chemotherapy. Various types of combination treatments were also evaluated. Both types of adjuvant therapy seem to reduce the failure rate, both locally in the pelvis and at distant sites, although survival, studied by the life table technique, was unaffected. Radiotherapy appears to reduce the number of pelvic failures when used as combination therapy and chemotherapy tends to prevent distant recurrences when used together with surgery. Further prospective and randomized studies are needed, however, to answer the question of the long-term value of adjuvant radiotherapy and/or chemotherapy in the treatment of uterine sarcomas.     

C57BL/6小鼠上皮性卵巢癌模型的建立及其生物学特性

目的在免疫功能正常的C57BL/6小鼠体内建立上皮性卵巢癌腹腔转移瘤模型及皮下瘤模型,为卵巢癌的诊断、治疗及预防的相关研究提供基础。方法体外培养近交系C57BL/6小鼠卵巢上皮低分化腺癌细胞株ID-8,将对数生长期的ID-8细胞按1×10^7、5×10^6、1×10^6和1×10^5个细胞/只的剂量,分别接种于6～8周雌性SPF级C57BL/6小鼠腹腔及左侧肩部皮下,共8组,每组6只。观察腹腔瘤及皮下瘤的成瘤时间、成瘤率、腹水、腹腔肿瘤转移情况及小鼠生存期;处死小鼠时留取主要脏器、腹腔肿瘤及皮下肿瘤标本,行病理学检查。另外6只小鼠接种5×10^6个ID-8细胞,分别在4、8、16周后处死进行系统解剖,做常规病理学检查。结果将不同数量的ID-8细胞接种C57BL/6小鼠腹腔及皮下后,成瘤率均为100%,腹腔瘤模型组：腹腔注射1×10^5,1×10^6,5×10^6,1×10^7个ID-8细胞,平均生存时间分别为（141±6.7）d、（122.8±4.5）d、（83.4±7.2）d和（74.4±4.5）d,随着肿瘤细胞接种负荷增加,动物生存期明显缩短（P〈0.05）。皮下瘤组：1×10^7细胞组和5×10^6细胞组,1周左右成瘤;1×10^6细胞组,3周左右成瘤;1×10^5细胞组,6周左右成瘤。随着肿瘤接种负荷的增加,肿瘤直径和体积明显增大（P〈0.05）。结论 C57BL/6小鼠腹腔瘤模型类似人类Ⅲ、Ⅳ期卵巢上皮癌患者的临床特点。皮下瘤模型更易于观察免疫治疗或药物治疗的疗效。在免疫功能正常的C57BL/6小鼠建立的ID-8细胞卵巢癌肿瘤模型,是适合于卵巢癌分子和免疫治疗研究的模型。     

Hepatitis C Prevention and Control: Implications for the Obstetrician/Gynaecologist

The hepatitis C virus, first isolated in 1989, is now prevalent throughout the world, and is estimated to affect approximately three percent of the entire population. The most efficient mode of transmission is through percutaneous exposure to contaminated blood. In a majority of infected patients, chronic hepatitis will ensue, potentially leading to the rare complications of cirrhosis and hepatocellular carcinoma. We aimed to review the natural history of the disease, and discuss screening and preventive strategies as they apply to women.     

Folate metabolism gene polymorphisms MTHFR C677T and A1298C and risk for Down syndrome offspring: a meta-analysis

Objectives

MTHFR C677T and A1298C have been associated with the risk of having an infant with Down syndrome (DS), but results were conflicting. We performed this meta-analysis to derive a more precise estimation of the association between maternal MTHFR polymorphisms and DS.

Study design

An electronic search of PubMed and Chinese Biomedicine database was conducted to select studies for meta-analysis. Twenty-eight case–control studies containing MTHFR C677T and A1298C gene polymorphisms were chosen, and odds ratio (OR) with confidence interval (CI) was used to assess the strength of this association.

Results

Case–control studies including 2806 cases and 4597controls for MTHFR C677T were identified. The overall results suggested that the variant genotypes MTHFR C677T were associated with DS risk (TT+CT vs. CC: OR = 1.305, 95% CI: 0.125–1.514, p = 0). In the stratified analysis, individuals with the T-carriers genotype in the dominant model had increased risk of DS (OR = 1.171, 95% CI: 0.976–1.405, p = 0.09) in Caucasian subjects and in Asian subjects (OR = 1.749, 95% CI: 1.084–2.824, p = 0.022). In addition, case–control studies including 1854 cases and 2364 controls for MTHFR A1298C were chosen. Associations between MTHFR A1298C and the risk of having a child with DS were not found. A symmetric funnel plot, the Egger's test (p = 0.126) suggested a lack of publication bias.

Conclusion

This meta-analysis supports the idea that MTHFR C677T genotype is associated with increased risk for DS offspring.     

HIV or human papillomavirus co-infection among Brazilian individuals infected with hepatitis B and/or hepatitis C

Objective

To investigate the incidence of co-infection with HIV and human papillomavirus (HPV) among Brazilian individuals with hepatitis B virus (HBV) or hepatitis C virus (HCV) infection.

Methods

A prospective study was conducted that enrolled 103 individuals from the Ceará region of Brazil between January 4, 2010, and August 9, 2012. Eligible participants were men (n = 45) or women (n = 58) infected with HBV and/or HCV. Pap smears were collected from female patients. Material from male (urethra) and female (cervix) patients was then collected via DNA-HPV test and visual inspection with acetic acid.

Results

In all, 48 participants tested positive for HBV and 55 tested positive for HCV; 2 had dual infection with HBV and HCV. Co-infection with HIV was detected among 15 participants, whereas 20 participants were found to be co-infected with HPV.

Conclusion

A high prevalence of co-infection with HIV and HPV was detected among Brazilian individuals with HBV and/or HCV.     

Vitamin-B12 deficiency following therapy in gynecologic oncology

Vitamin-B₁₂ deficiency results from inadequate absorption of the vitamin by the distal ileum and depletion of available stores. Both radiotherapy and intestinal resection can contribute to development of this condition. The significance of this problem in gynecologic oncology is discussed and two patients are described.     

具生殖系嵌合能力的C57BL/6J小鼠胚胎干细胞系的建立

目的:建立能实现种系传递的C57BL/6J小鼠胚胎干细胞系。方法:从C57BL/6J小鼠3.5d的囊胚中分离培养内细胞团。经体外适宜培养建系后,将C57BL/6J胚胎干细胞(ES)注入ICR小鼠受体囊胚腔,制备嵌合体小鼠。结果:成功地建立了3个C57BL/6J小鼠胚胎干细胞系,该C57BL/6JES细胞呈集落状生长,正常稳定的核型率>80%,具高水平的磷酸酶活性,并表达ES细胞特殊性细胞表面标记SSEA-1,不表达SSEA-3和SSEA-4;体内外的分化实验也证实mC57ES1具向三胚层组织分化的能力。经显微注入ICR小鼠囊胚腔中,产生4只嵌合体小鼠,经证实其中1只为种系嵌合小鼠。结论:建立了能实现种系传递的C57BL/6J小鼠胚胎干细胞系,该系的ES细胞可用于今后制备转基因动物和基因敲除动物。     

hCGβ-C3d3融合蛋白免疫BALB/c小鼠及其抗血清对hCG诱导鼠子宫增重的抑制效应

目的:通过分子佐剂C3d3增强hCGb避孕疫苗的免疫原性。方法:采用分子生物学技术以phCMV1为载体分别构建分泌型、带有6个组氨酸纯化标签的真核表达质粒phCMV1-6His-hCGb-C3d3和phCMV1-6His-hCGb,在CHO细胞中获得稳定、高效表达的重组蛋白,并用镍柱和凝胶过滤层析对其进行分离、纯化。分别用hCGb-C3d3融合蛋白和单用hCGb间隔4周两次免疫生育期雌性BALB/c小鼠,ELISA测定血清中抗hCGb抗体滴度,并对各组小鼠产生的抗血清拮抗hCG诱导的小鼠子宫增重效应进行比较。结果:C3d3使hCGb蛋白疫苗的免疫原性增强1 995倍,hCGb-C3d3融合蛋白免疫小鼠产生的抗血清具有很强的抑制小鼠子宫增重作用。结论:通过分子佐剂C3d3可以大幅提高机体对hCGb的体液免疫应答能力,hCGb-C3d3融合蛋白免疫小鼠产生的抗血清对hCG的生物学作用具有更强的抑制效应。     

Caspase-3活性改变与人卵巢癌细胞系COC1/DDP耐药的关系

目的 探讨人卵巢癌顺铂耐药细胞株COC1/DDP中抗凋亡蛋白Bcl-XL、Bcl-2、细胞色素C的表达及半胱天冬氨酸蛋白酶-3(caspase-3)活性与人卵巢癌顺铂耐药的关系。方法:采用RT-PCR和Western Blot分析人卵巢癌顺铂敏感细胞株COC1和顺铂耐药株COC1/DDP中Bcl-XL、Bcl-2、细胞色素C的表达和caspase-3的活性。并用流式细胞术测定顺铂作用后COC1和COC1/DDP细胞株的凋亡率。结果:在COC1/DDP细胞中,Bcl-XL和Bcl-2的表达明显高于COC1细胞;顺铂作用后,在COC1/DDP细胞株中细胞色素C的表达明显减少,caspase-3活性明显降低(P<0.05);其凋亡率也明显低于COC1细胞株(P<0.05)。结论:人卵巢癌细胞株COC1/DDP对顺铂产生耐药可能与细胞内Bcl-XL、Bcl-2过度表达抑制了线粒体细胞色素C的释放及caspase-3活性有关。     

Decreased expression of complement 3a receptor (C3aR) in human placentas from severe preeclamptic pregnancies

Objectives

The aim of this study was to determine the expression of the anaphylatoxin receptors complement C3a receptor (C3aR) and C5a receptor (C5aR) in the placentas of pregnancies complicated by severe early onset preeclampsia.

Study design

We recruited women with pregnancies complicated by severe early-onset preeclampsia (n = 19, 11 of which were further complicated with IUGR) and women with preterm pregnancies not affected by preeclampsia (n = 8). Gene and protein expression of C3aR and C5aR was analysed by quantitative RT-PCR and Western blotting, respectively.

Results

C3aR was detected in the Hofbauer cells in the villous stroma of the placenta. C5aR staining was detected in the syncytiotrophoblast and endothelial cells. We found significantly decreased expression of C3aR mRNA and protein expression in placentas with preeclampsia compared to controls. However, C5aR expression was not significantly different between preeclamptic and control placentas at either the mRNA or protein level.

Conclusions

Decreased C3aR expression indicates a dysregulation of the complement system in the placentas of preeclamptic women. Further studies would elucidate the exact mechanisms that complement has in preeclampsia.