Suitability of antral follicle counts and computer-assisted analysis of ultrasonographic and magnetic resonance images for estimating follicular reserve in porcine,ovine and bovine ovaries ex situ

This study was conducted to determine if correlations exist between the numbers of microscopic follicles comprising ovarian follicular reserve (OFR) and antral follicle counts (AFCs), and to assess the usefulness of computerized analyses of ovarian ultrasonograms and magnetic resonance (MR) images for estimating OFR in excised porcine, ovine and bovine ovaries. As a pre-requisite to these analyses, we characterized and compared ovarian cortical histomorhpology and follicle populations in the three species varying in prolificacy and overall reproductive longevity, and hence the total number of microscopic and antral follicles. Ultrasonographic and MR images were obtained at the scanner settings optimized to provide opposing contrasts between antral follicles and the ovarian stroma. Commercially available ImageProPlus® analytical software was used to calculate numerical pixel values (NPVs) and pixel heterogeneity (standard deviation of the pixel values) along the computer-generated lines (4–6) placed in the area corresponding to the ovarian cortex. The numbers of primordial (r = 0.38, P < 0.01) and intermediate follicles (r = 0.37, P < 0.01) were correlated with the numbers of antral follicles in bovine ovarian sections. The numbers of primordial (r = 0.28, P < 0.05), intermediate (r = 0.31, P < 0.01) and primary follicles (r = 0.27, P < 0.05) correlated directly with mean NPVs of the ultrasonographic ovarian images in cattle. There was a negative correlation between primary follicle numbers and NPVs of MR images (3D FAST-SPOILED GRADIENT ECHO) of the porcine ovarian cortex (r = −0.31, P < 0.05). To summarize, the numbers of primordial and intermediate follicles could only be estimated from AFCs in cows. Using ultrasound NPVs, the numbers of primordial, intermediate and primary follicles could be directly estimated in bovine ovaries and the quantitative image attributes of MR images were useful for quantifying porcine primary follicles. The bovine ovarian model is compatible with human situation and hence future studies should be undertaken to ascertain the usefulness of AFCs and ultrasonographic image analyses for estimating OFR in women.     

An integrated assessment of histopathological changes of the enteric neuromuscular compartment in experimental colitis

Bowel inflammatory fibrosis has been largely investigated, but an integrated assessment of remodelling in inflamed colon is lacking. This study evaluated tissue and cellular changes occurring in colonic wall upon induction of colitis, with a focus on neuromuscular compartment. Colitis was elicited in rats by 2,4‐dinitrobenzenesulfonic acid (DNBS). After 6 and 21 days, the following parameters were assessed on paraffin sections from colonic samples: tissue injury and inflammatory infiltration by histology; collagen and elastic fibres by histochemistry; HuC/D, glial fibrillar acidic protein (GFAP), proliferating cell nuclear antigen (PCNA), nestin, substance P (SP), von Willebrand factor, c‐Kit and transmembrane 16A/Anoctamin1 (TMEM16A/ANO1) by immunohistochemistry. TMEM16A/ANO1 was also examined in isolated colonic smooth muscle cells (ICSMCs). On day 6, inflammatory alterations and fibrosis were present in DNBS‐treated rats; colonic wall thickening and fibrotic remodelling were evident on day 21. Colitis was associated with both an increase in collagen fibres and a decrease in elastic fibres. Moreover, the neuromuscular compartment of inflamed colon displayed a significant decrease in neuron density and increase in GFAP/PCNA‐positive glia of myenteric ganglia, enhanced expression of neural SP, blood vessel remodelling, reduced c‐Kit‐ and TMEM16A/ANO1‐positive interstitial cells of Cajal (ICCs), as well as an increase in TMEM16A/ANO1 expression in muscle tissues and ICSMCs. The present findings provide an integrated view of the inflammatory and fibrotic processes occurring in the colonic neuromuscular compartment of rats with DNBS‐induced colitis. These morphological alterations may represent a suitable basis for understanding early pathophysiological events related to bowel inflammatory fibrosis.     

Vasohibin-1基因在上皮性卵巢癌中的表达和临床病理意义

目的:探讨卵巢癌中Vasohibin-1的表达情况及临床病理学意义。方法:采用实时定量PCR和免疫组化方法检测在60例上皮性卵巢癌和12例正常卵巢组织中Vasohibin-1的表达情况,ELISA法检测卵巢癌组织中VEGF蛋白表达,分析vasohibin-1表达与VEGF之间的关系及其与卵巢癌分期,分级和预后之间的关系。结果:卵巢癌中vasohibin-1表达明显高于正常卵巢(P0.05)。Vasohibin-1表达水平与卵巢癌分期相关(P0.05),而与卵巢癌分级和淋巴转移无关。Vasohibin-1表达与VEGF蛋白水平呈正相关(P0.01)。高vasohibin-1表达卵巢癌患者三年生存率(50%)低于低vasohibin-1表达的患者(83%)。结论:vasohibin-1可以反映卵巢癌的血管生成潜能,是卵巢癌的不良预后因素。     

Oviposition and oogenesis in virgin fire ant females Solenopsis invicta are associated with a high level of dopamine in the brain

Abstract. In the fire ant Solenopsis invicta Buren, virgin females are capable of shedding their wings (dealation) and laying haploid eggs. However, dealation and reproduction are inhibited by a queen primer pheromone that depresses the release of Juvenile Hormone by the corpora allata. In an attempt to identify other neural signals that trigger the reproductive system, we measured the effect on brain biogenic amines of separation from the queen. Dopamine in the brain of virgin females increased from 552 ± 42 to 971 ± 65 fMol/brain when reproduction and dealation were stimulated by 15 days of separation. Octopamine and 5-hydroxytryptamine did not change significantly after the separation. Isolated virgin females fed with a tyrosine hydroxylase inhibitor 3-iodo- l -tyrosine mixed in sucrose for 15 days laid significantly fewer eggs and had fewer chorionated oocytes in their ovarioles than females fed with sucrose only. Restoring dopamine biosynthesis by adding l -dopa to the food also restored oogenesis and oviposition. Dealation was not affected by 3-iodo- l -tyrosine or l -dopa. The possible role of dopamine as the neural target of the queen pheromone regarding its potent allotoregulatory effect in other insects is discussed.     

Evaluation of the vervet (Clorocebus aethiops) as a model for the assisted reproductive technologies

The vervet monkey was evaluated as a primate model for use in assisted reproductive technologies (ARTs). Eight adult female vervets were hormonally monitored for their potential use as egg donors and those six females displaying regular menstrual cycles were subjected to controlled ovarian stimulation with recombinant human gonadotropins. Three animals failed to respond while laparoscopic follicular aspiration was performed on the other three females at 27-30 h post-human chorionic gonadotropin administration. A total of 62, 40, and 18 oocytes was recovered from these three animals of which 30, 20, and 4, respectively, matured to the metaphase II stage and were subsequently inseminated using intracytoplasmic sperm injection. An average of 40+/-15% (SEM) of the inseminated oocytes were fertilized based on pronucleus formation and timely cleavage. One embryo from each of the two stimulated females developed into expanded blastocysts. Two adult male vervets were assessed as sperm donors. Neither adjusted well to the restraint and collection procedure required for penile electroejaculation. Samples collected via rectal electroejaculation were very low in sperm motility and concentration; however, cauda epididymal aspirations from one male yielded an adequate concentration of motile sperm. These results emphasize the need to establish species-specific ovarian stimulation protocols and semen collection techniques if vervets are to be considered for basic and applied (ARTs) research on primate gametes or embryos.     

兔卵巢组织石蜡切片技术的改良

针对常规石蜡切片方法步骤繁多,耗时长,所用的二甲苯试剂对人体健康有较大毒害性的缺陷,探讨提高制作兔卵巢组织石蜡切片质量的方法。试验从脱水和透明环节进行技术改良,用正丁醇与无水乙醇混合剂取代常规的脱水剂和透明剂。该混合剂具有良好的脱水和透明作用。实验结果表明：通过该技术改良,镜下观察组织结构清晰可辨、颜色鲜艳、对比度好、层次分明,组织切片效果良好。试验改良技术的工艺简单,具有质优、省时、快捷和实用性较好等优点,有推广应用价值。     

^125I标记白眉蝮蛇毒精氨酸酯酶代谢动力学研究

研究白眉蝮蛇(Agkistrodon halys ussuriensis)毒精氨酸酯酶的代谢动力学,为临床应用提供依据。125I标记的精氨酸酯酶对大鼠一侧颈静脉给药,不同时间从另一测颈静脉取血。5h后处死动物,取各组织、尿液、胆汁和粪便,对各样本的放射性进行测定并拟合时间-放射性关系曲线。代谢动力学拟合曲线符合一室模型,其中生物半衰期T1/2为55.9min,K值为0.0124min-1。125I标记的精氨酸酯酶在体内各组织广泛分布,但有血脑屏障存在,肝、肾和尿液中的放射性比其它组织要高很多,主要通过肝脏降解,肾脏排泄。     

125 I标记白眉蝮蛇毒精氨酸酯酶代谢动力学研究

研究白眉蝮蛇(Agkistrodon halys ussuriensis)毒精氨酸酯酶的代谢动力学,为临床应用提供依据。125I标记的精氨酸酯酶对大鼠一侧颈静脉给药,不同时间从另一测颈静脉取血。5h后处死动物,取各组织、尿液、胆汁和粪便,对各样本的放射性进行测定并拟合时间-放射性关系曲线。代谢动力学拟合曲线符合一室模型,其中生物半衰期T1/2为55.9min,K值为0.0124min-1。125I标记的精氨酸酯酶在体内各组织广泛分布,但有血脑屏障存在,肝、肾和尿液中的放射性比其它组织要高很多,主要通过肝脏降解,肾脏排泄。     

Extramuscular myofascial force transmission also occurs between synergistic muscles and antagonistic muscles

The purpose of the present study was to test the hypothesis that myofascial force transmission may not be limited by compartmental boundaries of a muscle group to synergists. Muscles of the anterior tibial compartment in rat hindlimb as well as of the neighbouring peroneal compartment (antagonistic muscles) were excited maximally. Length–force data, based on proximal lengthening, of EDL, as well as distal lengthening of the tibial muscles (TA + EHL) and the peroneal muscle group (PER) were collected independently, while keeping the other two muscle groups at a constant muscle–tendon complex length. Simultaneously measured, distal and proximal EDL active forces were found to differ significantly throughout the experiment. The magnitude of this difference and its sign was affected after proximal lengthening of EDL itself, but also of the tibial muscle complex and of the peroneal muscle complex. Proximal lengthening of EDL predominantly affected its synergistic muscles within the anterior crural compartment (force decrease <4%). Lengthening of either TA or PER caused a decrease in distal EDL isometric force (by 5–6% of initial force). It is concluded also that mechanisms for mechanical intermuscular interaction extend beyond the limits of muscle compartments in the rat hindlimb. Even antagonistic muscles should not be considered fully independent units of muscular function.

Particular, strong mechanical interaction was found between antagonistic tibial anterior muscle and peroneal muscle complexes: Lengthening of the peroneal complex caused tibial complex force to decrease by approximately 25%, whereas for the reverse a 30% force decrease was found.