Molecular Analysis of the Koala Reproductive Hormones and Their Receptors: Gonadotrophin‐Releasing Hormone (GnRH), Follicle‐Stimulating Hormone β and Luteinising Hormone β with Localisation of GnRH

During evolution, reproductive hormones and their receptors in the brain‐pituitary‐gonadal axis have been altered by genetic mechanisms. To understand how the neuroendocrine control of reproduction evolved in mammals, it is important to examine marsupials, the closest group to placental mammals. We hypothesised that at least some of the hormones and receptors found in placental mammals would be present in koala, a marsupial. We examined the expression of koala mRNA for the reproductive molecules. Koala cDNAs were cloned from brain for gonadotrophin‐releasing hormones (GnRH1 and GnRH2) or from pituitary for GnRH receptors, types I and II, follicle‐stimulating hormone (FSH)β and luteinising hormone (LH)β, and from gonads for FSH and LH receptors. Deduced proteins were compared by sequence alignment and phylogenetic analysis with those of other vertebrates. In conclusion, the koala expressed mRNA for these eight putative reproductive molecules, whereas at least one of these molecules is missing in some species in the amniote lineage, including humans. In addition, GnRH1 and 2 are shown by immunohistochemistry to be expressed as proteins in the brain.     

卵巢早衰临床特征探析

从卵巢早衰的病因病机、症状体征、性激素检查以及治疗探析本病的临床特征。结果表明,卵巢早衰的病因与家族遗传、性生活匮乏、过度操劳、不良生活习惯、失血性贫血、卵巢自身疾病以及医源性因素等相关;病机主要为肾亏脾虚,其次与心肾不交、肝脾失调、痰湿瘀浊等因素相关。症状以月经早闭为主,伴随绝经综合征的临床表现;体征为生殖器官的提前萎缩,促性腺激素水平升高,雌激素水平下降。治疗以补肾健脾为主,调理心肝为辅,配合理气活血,祛湿化痰,或中西医结合治疗。     

Ovarian Regulation of Kisspeptin Neurones in the Arcuate Nucleus of the Rhesus Monkey (Macaca mulatta)

Tonic gonadotrophin secretion throughout the menstrual cycle is regulated by the negative‐feedback actions of ovarian oestradiol (E₂) and progesterone. Although kisspeptin neurones in the arcuate nucleus (ARC) of the hypothalamus appear to play a major role in mediating these feedback actions of the steroids in nonprimate species, this issue has been less well studied in the monkey. In the present study, we used immunohistochemistry and in situ hybridisation to examine kisspeptin and KISS1 expression, respectively, in the mediobasal hypothalamus (MBH) of adult ovariectomised (OVX) rhesus monkeys. We also examined kisspeptin expression in the MBH of ovarian intact females, and the effect of E₂, progesterone and E₂ + progesterone replacement on KISS1 expression in OVX animals. Kisspeptin or KISS1 expressing neurones and pronounced kisspeptin fibres were readily identified throughout the ARC of ovariectomised monkeys but, on the other hand, in intact animals, kisspeptin cell bodies were small in size and number and only fine fibres were observed. Replacement of OVX monkeys with physiological levels of E₂, either alone or with luteal phase levels of progesterone, abolished KISS1 expression in the ARC. Interestingly, progesterone replacement alone for 14 days also resulted in a significant down‐regulation of KISS1 expression. These findings support the view that, in primates, as in rodents and sheep, kisspeptin signalling in ARC neurones appears to play an important role in mediating the negative‐feedback action of E₂ on gonadotrophin secretion, and also indicate the need to study further their regulation by progesterone.     

Seasonal Effect of Gonadotrophin Inhibitory Hormone on Gonadotrophin‐Releasing Hormone‐induced Gonadotroph Functions in the Goldfish Pituitary

We have shown that native goldfish gonadotrophin inhibitory hormone (gGnIH) differentially regulates luteinsing hormone (LH)‐β and follicle‐stimulating hormone (FSH)‐β expression. To further understand the functions of gGnIH, we examined its interactions with two native goldfish gonadotrophin‐releasing hormones, salmon gonadotrophin‐releasing hormone (sGnRH) and chicken (c)GnRH‐II in vivo and in vitro. Intraperitoneal injections of gGnIH alone reduced serum LH levels in fish in early and mid gonadal recrudescence; this inhibition was also seen in fish co‐injected with either sGnRH or cGnRH‐II during early recrudescence. Injection of gGnIH alone elevated pituitary LH‐β and FSH‐β mRNA levels at early and mid recrudescence, and FSH‐β mRNA at late recrudescence. Co‐injection of gGnIH attenuated the stimulatory influences of sGnRH on LH‐β in early recrudescence, and LH‐β and FSH‐β mRNA levels in mid and late recrudescence, as well as the cGnRH‐II‐elicited increase in LH‐β, but not FSH‐β, mRNA expression at mid and late recrudescence. sGnRH and cGnRH‐II injection increased pituitary gGnIH‐R mRNA expression in mid and late recrudescence but gGnIH reduced gGnIH‐R mRNA levels in late recrudescence. gGnIH did not affect basal LH release from perifused pituitary cells and continual exposure to gGnIH did not alter the LH responses to acute applications of GnRH. However, a short 5‐min GnIH treatment in the middle of a 60‐min GnRH perifusion selectively reduced the cGnRH‐II‐induced release of LH. These novel results indicate that, in goldfish, gGnIH and GnRH modulate pituitary GnIH‐R expression and gGnIH differentially affects sGnRH and cGnRH‐II regulation of LH secretion and gonadotrophin subunit mRNA levels. Furthermore, these actions are manifested in a reproductive stage‐dependent manner.     

人工授精周期诱导排卵药物及方案的研究进展

诱导排卵联合宫腔内人工授精是广泛应用的一项辅助生育技术。诱导排卵目的是形成单一卵泡的发育成熟,尽可能地减少发生多胎妊娠和卵巢过度刺激综合征的风险。抗雌激素类和芳香化酶抑制剂因口服方便而广泛应用,单独使用妊娠率较低。促性腺激素类药物可以获得较高的临床妊娠率,采用小剂量递增的温和方案不但能够保证较高的单卵泡发育还能够明显的减少并发症的发生。关于促性腺激素促排卵治疗中卵巢反应预测因子还有待于进一步研究。     

Gonadotrophin‐Inhibitory Hormone in the Cichlid Fish Cichlasoma dimerus: Structure,Brain Distribution and Differential Effects on the Secretion of Gonadotrophins and Growth Hormone

The role of gonadotrophin‐inhibitory hormone (GnIH) in the inhibition of the reproductive axis has been well‐established in birds and mammals. However, its role in other vertebrates, such as the teleost fish, remains controversial. In this context, the present study aimed to evaluate whether GnIH modulates the release of gonadotrophins and growth hormone (GH) in the cichlid fish Cichlasoma dimerus. First, we partially sequenced the precursor polypeptide for GnIH and identified three putative GnIH peptides. Next, we analysed the expression of this precursor polypeptide via a polymerase chain reaction in the reproductive axis of both sexes. We found a high expression of the polypeptide in the hypothalamus and gonads of males. Immunocytochemistry allowed the observation of GnIH‐immunoreactive somata in the nucleus posterioris periventricularis and the nucleus olfacto‐retinalis, with no differences between the sexes. GnIH‐immunoreactive fibres were present in all brain regions, with a high density in the nucleus lateralis tuberis and at both sides of the third ventricle. Finally, we performed in vitro studies on intact pituitary cultures to evaluate the effect of two doses (10^?6 m and 10^?8 m ) of synthetic C. dimerus (cd‐) LPQRFa‐1 and LPQRFa‐2 on the release of gonadotrophins and GH. We observed that cd‐LPQRFa‐1 decreased β‐luteinising hormone (LH) and β‐follicle‐stimulating hormone (FSH) and also increased GH release to the culture medium. The release of β‐FSH was increased only when it was stimulated with the higher cd‐LPQRFa‐2 dose. The results of the present study indicate that cd‐LPQRFa‐1, the cichlid fish GnIH, inhibits β‐LH and β‐FSH release and stimulates GH release in intact pituitary cultures of C. dimerus. The results also show that cd‐LPQRF‐2 could act as an β‐FSH‐releasing factor in this fish species.     

男性肺原发性绒毛膜癌1例并文献复习

回顾性分析南京医科大学第一附属医院通过手术切除病理确诊的1例男性肺原发性绒毛膜癌(primary choriocarcinoma,PCC)的临床资料、影像学及组织病理学等特征,结合国内外文献复习,总结肺PCC的诊断及治疗进展.男性肺PCC是一种较为罕见高度恶性生殖细胞肿瘤,病因不明,临床表现多样,咯血最常见.早期诊断困难,易误诊为常见病,预后差,致死率高.手术联合化疗是目前较为提倡的治疗方案.男性肺PCC的诊断较难,需结合临床、影像学等综合分析,确诊需依赖组织病理学.     

The Roles of RFamide‐Related Peptide‐3 in Mammalian Reproductive Function and Behaviour

To maximise reproductive success, organisms restrict breeding to optimal times of the day or year, when internal physiology and external environmental conditions are suitable for the survival of both parent and offspring. To appropriately coordinate reproductive activity, internal and external standing is communicated to the hypothalamic‐pituitary‐gonadal axis via a coordinated balance of stimulatory and inhibitory neurochemical systems. The cumulative balance of these mediators ultimately drives the pattern of gonadotrophin‐releasing hormone secretion, a neurohormone that stimulates pituitary gonadotrophin secretion. Until 2000, a complementary inhibitor of pituitary gonadotrophin secretion had not been identified. At this time, a novel, avian hypothalamic peptide capable of inhibiting gonadotrophin secretion in cultured quail pituitary cells was uncovered and named gonadotrophin‐inhibitory hormone (GnIH). Subsequently, the presence and functional role for the mammalian orthologue of GnIH, RFamide‐related peptide, (RFRP‐3), was examined, confirming a conserved role for this peptide across several rodent species. To date, a similar distribution and functional role for RFRP‐3 have been observed across all mammals investigated, including humans. This overview summarises the role that RFRP‐3 plays in mammals and considers the implications and opportunities for further study with respect to reproductive physiology and the neural control of sexual behaviour and motivation.