When hormone defects cannot explain it: Malformative disorders of sex development

The birth of a baby with malformations of the genitalia urges medical action. Even in cases where the condition is not life‐threatening, the identification of the external genitalia as male or female is emotionally essential for the family, and genital malformations represent one of the most stressful situations around a newborn. The female or male configuration of the genitalia normally evolves during fetal life according to the genetic, gonadal, and hormonal sex. Disorders of sex development occur when male hormone (androgens and anti‐Müllerian hormone) secretion or action is insufficient in the 46,XY fetus or when there is an androgen excess in the 46,XX fetus. However, sex hormone defects during fetal development cannot explain all congenital malformations of the reproductive tract. This review is focused on those congenital conditions in which gonadal function and sex hormone target organ sensitivity are normal and, therefore, not responsible for the genital malformation. Furthermore, because the reproductive and urinary systems share many common pathways in embryo‐fetal development, conditions associating urogenital malformations are discussed. Birth Defects Research (Part C) 102:359–373, 2014. © 2014 Wiley Periodicals, Inc.     

Site-specific Nitration of Apolipoprotein A-I at Tyrosine 166 Is Both Abundant within Human Atherosclerotic Plaque and Dysfunctional

We reported previously that apolipoprotein A-I (apoA-I) is oxidatively modified in the artery wall at tyrosine 166 (Tyr¹⁶⁶), serving as a preferred site for post-translational modification through nitration. Recent studies, however, question the extent and functional importance of apoA-I Tyr¹⁶⁶ nitration based upon studies of HDL-like particles recovered from atherosclerotic lesions. We developed a monoclonal antibody (mAb 4G11.2) that recognizes, in both free and HDL-bound forms, apoA-I harboring a 3-nitrotyrosine at position 166 apoA-I (NO₂-Tyr¹⁶⁶-apoA-I) to investigate the presence, distribution, and function of this modified apoA-I form in atherosclerotic and normal artery wall. We also developed recombinant apoA-I with site-specific 3-nitrotyrosine incorporation only at position 166 using an evolved orthogonal nitro-Tyr-aminoacyl-tRNA synthetase/tRNA_CUA pair for functional studies. Studies with mAb 4G11.2 showed that NO₂-Tyr¹⁶⁶-apoA-I was easily detected in atherosclerotic human coronary arteries and accounted for ∼8% of total apoA-I within the artery wall but was nearly undetectable (>100-fold less) in normal coronary arteries. Buoyant density ultracentrifugation analyses showed that NO₂-Tyr¹⁶⁶-apoA-I existed as a lipid-poor lipoprotein with <3% recovered within the HDL-like fraction (d = 1.063–1.21). NO₂-Tyr¹⁶⁶-apoA-I in plasma showed a similar distribution. Recovery of NO₂-Tyr¹⁶⁶-apoA-I using immobilized mAb 4G11.2 showed an apoA-I form with 88.1 ± 8.5% reduction in lecithin-cholesterol acyltransferase activity, a finding corroborated using a recombinant apoA-I specifically designed to include the unnatural amino acid exclusively at position 166. Thus, site-specific nitration of apoA-I at Tyr¹⁶⁶ is an abundant modification within the artery wall that results in selective functional impairments. Plasma levels of this modified apoA-I form may provide insights into a pathophysiological process within the diseased artery wall.     

Utero-tubal Embryo Transfer and Vasectomy in the Mouse Model

The transfer of preimplantation embryos to a surrogate female is a required step for the production of genetically modified mice or to study the effects of epigenetic alterations originated during preimplantation development on subsequent fetal development and adult health. The use of an effective and consistent embryo transfer technique is crucial to enhance the generation of genetically modified animals and to determine the effect of different treatments on implantation rates and survival to term. Embryos at the blastocyst stage are usually transferred by uterine transfer, performing a puncture in the uterine wall to introduce the embryo manipulation pipette. The orifice performed in the uterus does not close after the pipette has been withdrawn, and the embryos can outflow to the abdominal cavity due to the positive pressure of the uterus. The puncture can also produce a hemorrhage that impairs implantation, blocks the transfer pipette and may affect embryo development, especially when embryos without zona are transferred. Consequently, this technique often results in very variable and overall low embryo survival rates. Avoiding these negative effects, utero-tubal embryo transfer take advantage of the utero-tubal junction as a natural barrier that impedes embryo outflow and avoid the puncture of the uterine wall. Vasectomized males are required for obtaining pseudopregnant recipients. A technique to perform vasectomy is described as a complement to the utero-tubal embryo transfer.     

子宫压迫缝合联合子宫动脉结扎术治疗产后出血的临床疗效

目的:探讨子宫压迫缝合联合子宫动脉结扎术治疗产后出血的临床疗效。方法:选取近年来60名产后出血患者,30名患者用子宫压迫缝合术(对照组),另外30名用子宫压迫缝合联合子宫动脉结扎术(实验组),观察手术时间、术后出血量及临床疗效。结果:实验组有效率为86.67%,对照组有效率为83.33%,两组患者的临床疗效差异无统计学意义(P0.05);两组手术时间,术中出血量差异无统计学意义(P0.05),但实验组的术后2 h出血量比对照组少,差异有统计学意义(P0.05)。结论:子宫压迫缝合联合子宫动脉结扎术治疗产后出血疗效显著,值得在临床上推广。     

消化道出血病因的临床分析

目的:回顾性分析消化道出血的病因构成、相关因素及治疗情况。方法:收集2008年7月至2013年9月因消化道出血在沈阳军区总医院消化内科住院的患者资料,包括患者的一般资料、入院日期、病因、出血部位、生命体征、疾病史。依据消化道出血程度分为轻度组(86例)、中度组(90例)、重度组(132例),分析消化道出血的病因特征。结果:与轻度组比较,重度组上消化道出血患者比例较高,中、重度组Blatchford评分、心率、尿素、血肌酐、凝血酶原时间、INR、血糖明显升高(P0.05);收缩压、舒张压、红细胞、血红蛋白、总蛋白、白蛋白、总胆固醇明显减低(P0.05)。重度组患有食管胃底静脉曲张的比例较高,行内镜下治疗的比例高(P0.05)。结论:根据患者出血程度可初步判断患者消化道出血的部位及病因,为临床诊治提供有价值的参考依据。     

An in vivo mouse model of primary dysmenorrhea

Primary dysmenorrhea (PD) is a common gynecological disorder. Hitherto, animal models which recapitulate clinical features of PD have not been fully established. We aimed to examine whether a pain model in mice could mimic the clinic features of PD. After pretreated with estradiol benzoate (1 mg/kg/day) intraperitoneally (i.p.) for 3 consecutive days, non-pregnant female Imprinting Control Region mice (6–8 weeks old) was injected with 0.4 U of oxytocin to induce the stretching or writhing response which was recorded for a time period of 30 min. During the writhing period, the uterine artery blood flow alterations were examined by Doppler ultrasound detection. After writhing test, the uterine morphological changes were observed by hematoxylin and eosin (H&E) staining histopathology. In addition, enzyme-linked immunosorbent assay kit was used to measure the levels of prostaglandins F_2α/prostaglandins E₂ (PGF_2α/PGE₂) and TXB₂ (a metabolite of TXA₂)/6-keto-PGF_1α (a metabolite of PGI₂) in the uterine tissue homogenates and plasma, respectively. Western blot analyses were performed to determine the expressions of oxytocin receptor (OTR), beta2-adrenergic receptor (beta2-AR), and cyclooxygenase-2 (COX-2) in uterine, which are responsible for the uterine contraction. The writhing response only occurred in the estrogen pretreated female mice. The area of uterine myometrium significantly decreased along with the increased thickness in the oxytocin-induced estrogen pretreated mice model. The uterine artery blood flow velocity dropped, while the pulsatility index and resistance index slightly increased after the injection of oxytocin. The PGF_2α/PGE₂ level significantly increased and the plasma TXB₂/6-keto-PGF_1α level significantly enhanced. Compared with the control group, the uterine histopathology demonstrated moderate to severe edema of endometrium lamina propria. In consistent with the uterine morphological changes, a significant reduction of beta2-AR and a significant increase of OTR and COX-2 in the uterine tissue were observed. The writhing response was caused by the abnormal contraction of uterus. The uterine spasm and ischemia changes of oxytocin-induced estrogen pretreated female mice model were similar to the pathology of human PD. We reported an in vivo mice model, which can be used to study PD and for clinical therapeutic evaluations.     

In vitro morphology,viability and cytokine secretion of uterine telocyte‐activated mouse peritoneal macrophages

Telocytes (TCs), a distinct interstitial cell population, have been identified in the uterus, oviduct and placenta, with multiple proposed potential biological functions. Their unique structure allows them to form intercellular junctions with various immunocytes, both in normal and diseased tissues, suggesting a potential functional relationship with the local immune response. It has been hypothesized that through direct heterocellular junctions or indirect paracrine effects, TCs influence the activity of local immunocytes that are involved in the inflammatory process and in immune‐mediated reproductive abnormalities. However, no reliable cytological evidence for this hypothesis is currently available. In this study, we cultured primary murine uterine TCs and collected TC conditioned media (TCM). Mouse peritoneal macrophages (pMACs) were co‐cultured for 48 hrs with TCM or with DMEM/F12 or lipopolysaccharide (LPS) as negative and positive controls, respectively. Normal uterine TCs with a typical structure and a CD‐34‐positive/vimentin‐positive/c‐kit‐negative immunophenotype were observed during culture. Morphologically, TCM‐treated pMACs displayed an obvious activation/immunoresponse, in contrast to over‐stimulation and cell death after LPS treatment and no sign of activation in the presence of DMEM/F12. Accordingly, a cell counting kit 8 (CCK‐8) assay indicated significant activation of pMACs by TCM and LPS compared to DMEM/F12, thus supporting the marked morphological differences among these groups of cells. Furthermore, within a panel of macrophage‐derived cytokines/enzymes, interleukin‐6 (IL‐6) and inducible nitric oxide synthase were significantly elevated in TCM‐treated pMACs; tumour necrosis factor α, IL1‐R1, and IL‐10 were slightly, but significantly, up‐regulated; and no changes were observed for transforming growth factor‐β1, IL‐1β, IL‐23α and IL‐18. Our results indicate that TCs are not simply innocent bystanders but are rather functional players in the activation of pMACs; they trigger and maintain the immune response, likely through indirect paracrine effects. Thus, we provide preliminary in vitro evidence of immunoregulatory and immunosurveillance roles for TCs.     

Adjusting Nitrogen Application in Accordance with Soil Water Availability Enhances Yield and Water Use by Regulating Physiological Traits of Maize under Drip Fertigation

Knowledge of the interactive effects of water and nitrogen (N) on physio-chemical traits of maize (Zea mays L.) helps to optimize water and N management and improve productivity. A split-plot experiment was conducted with three soil water conditions (severe drought, moderate drought, and fully water supply referring to 45%–55%, 65%–75%, and 85%–95% field capacity, respectively) and four N application rates (N₀, N₁₅₀, N₂₄₀, and N₃₃₀ referring to 0, 150, 240, 330 kg N ha^–1 respectively) under drip fertigation in 2014 and 2015 in the Huang-Huai-Hai Plain of China. The results indicated that drought stress inhibited physiological activity of plants (leaf relative water content, root bleeding sap, and net photosynthetic rate), resulting in low dry matter accumulation after silking, yield, and N uptake, whereas increased WUE and NUE. N application rates over than 150 kg ha^–1 aggravated the inhibition of physiological activity under severe drought condition, while it was offset under moderate drought condition. High N application rates (N₃₃₀) still revealed negative effects under moderate drought condition, as it did not consistently enhance plant physiological activity and significantly reduced N uptake as compared to the N₂₄₀ treatment. With fully water supply, increasing N application rates synergistically enhanced physiological activity, promoted dry matter accumulation after silking, and increased yield, WUE, and N uptake. Although the N₂₄₀ treatment reduced yield by 5.4% in average, it saved 27.3% N under full water supply condition as compared with N₃₃₀ treatment. The results indicated that N regulated growth of maize in aspects of physiological traits, dry matter accumulation, and yield as well as water and N use was depended on soil water status. The appropriate N application rates for maize production was 150 kg ha^–1 under moderate drought or 240 kg ha^–1 under fully water supply under drip fertigation, and high N supply (>150 kg ha^–1) should be avoided under severe drought condition.     

种植密度对不同耐密性春玉米基部茎节维管束及根系伤流的影响

选用不同耐密性玉米品种‘郑单958’(密植品种)和‘辽单526’(稀植品种)为材料,设置4个不同的密度水平,分别在拔节期和乳熟期采用徒手切片显微观测基部茎秆内部维管束结构变化,采集茎基部伤流液测定其主要成份含量并计算伤流强度,以期为春玉米的密植高产提供理论依据.结果显示:(1)在拔节期,‘郑单958’维管束结构对密度变化反应相对敏感,而‘辽单526’维管束结构在各密度间并无显著差异;当密度最大时,‘辽单526’伤流强度的下降程度要大于‘郑单958’,但两品种伤流液成分对密度反应无明显变化.(2)在乳熟期,密度对‘郑单958’维管束结构无显著影响,而‘辽单526’在密度增加后维管束结构各指标明显下降且与对照(适宜密度)差异显著;各密度‘郑单958’的伤流强度均大于‘辽单526’,增加密度会提高两品种伤流液中可溶性糖的含量,尤其以‘辽单526’增加更显著.(3)在高于对照密度条件下,两品种的籽粒产量都不同程度低于对照,但‘辽单526’产量降低程度要大于‘郑单958’.研究表明,密植性玉米对密度压力的反应要早于稀植性玉米,其在高密度表现出的适应性也要优于稀植性玉米,这从茎基部的结构和功能上揭示了玉米品种耐密性差异的生理基础.     

胸窗萤的防卫行为：反射性出血及翻缩腺反应

为了探讨胸窗萤Pyrocoelia pectoralis Oliver, 1883的防卫行为, 对成虫的反射性出血行为及幼虫的翻缩腺体形态进行了研究。结果表明： 胸窗萤具有复杂的多重防卫行为策略。成虫利用闪烁、 假死、 反射性流血进行防卫, 幼虫则利用闪烁、 假死和微小的翻缩腺体进行防卫。反射性流血发生在雄成虫的前胸背板和鞘翅边缘, 而雌成虫仅在前胸背板边缘观察到有反射性流血行为。扫描电镜观察到直径大约32 μm的圆坑状结构环绕前胸背板或鞘翅边缘一圈。雌虫鞘翅牙也具有相似的圆坑状结构。幼虫具有9对乳头状的翻缩腺体, 位于中胸、 后胸和第1～7节侧板上。超薄切片发现, 腺体表面的棒状结构内部中空, 棒状结构连接一个发达的分泌细胞。细胞内连接棒状结构的部位着生致密的管状内质网。行为学实验发现, 胸窗萤血液对蚂蚁具有非常有效的拒避作用。