Morphological comparison and gonadotropins cell localization of mature female turbot and mouse pituitary

Reproduction is subtlety regulated by the hypothalamic-pituitary-gonad(HPG)axis in vertebrates.Pituitary gland is the center of the HPG axis,while pituitary gonadotropins follicle stimulating hormone(FSH)and luteinizing hormone(LH)were identified the key elements of the HPG axis in teleost and mammal.Morphology,cell lines,and gonadotropins cell localization of female turbot and mouse pituitary were determined at mature stage to illustrate the anatomical difference and cell characteristics in this study.Results show that turbot pituitary is chicken heart-shaped,dorsoventral,located on the ventral surface of the diencephalon.The mouse pituitary is oval,located in the pituitary fossa of the sella turcica at the skull base.Two well-distinguished areas adenohypophysis(AH)and neurohypophysis(NH)in pituitary were identified in turbot and mouse.Turbot AH comprised the rostral pars distalis(RPD),proximal pars distalis(PPD),and pars intermedia(PI).NH was not pronounced and with finger-like protrusions into PPD.However,mouse AH only comprised the pars distalis(PD)and PI.NH distribution was semicircular.Three main types of cells(acidophilic,basophilic,and chromophobic cells)were distributed in the mouse PD region,whereas appeared in the turbot PPD,RPD,and PI.Moreover,the percentage of mouse chromophobic and basophilic cells was higher and lower than that of turbot,respectively.The diameter of the aforementioned three cells in the mouse was significantly higher than turbot.fshβ-and lhβ-immunoreactive signals were identified in turbot-distinct pituitary cells that primarily occupied the peripheral and central regions of AH.However,mouse fsh-and lh-immunoreactive cells were expressed in the same cells and present in the PD.These results demonstrate the significantly difference of pituitary morphology,cell lines and gonadotropins(fshβand lhβ)location in female turbot and mouse.These differences help for fully understand the evolution and endocrinological functions of pituitary.

Live births from in vitro fertilization-embryo transfer following the administration of gonadotropin-releasing hormone agonist without gonadotropins:Two case reports

BACKGROUND The prevalence of female infertility between the ages of 25 and 44 is 3.5%to 16.7%in developed countries and 6.9%to 9.3%in developing countries.This means that infertility affects one in six couples and is recognized by the World Health Organization as the fifth most serious global disability.The International Committee for Monitoring Assisted Reproductive Technology reported that the global total of babies born as a result of assisted reproductive technology procedures and other advanced fertility treatments is more than 8 million.Advancements in controlled ovarian hyperstimulation procedures led to crucial accomplishments in human fertility treatments.The European Society for Human Reproduction and Embryology guideline on ovarian stimulation gave us valuable evidence-based recommendations to optimize ovarian stimulation in assisted reproductive technology.Conventional ovarian stimulation protocols for in vitro fertilization(IVF)–embryo transfer are based upon the administration of gonadotropins combined with gonadotropin-releasing hormone(GnRH)analogues,either GnRH agonists(GnRHa)or antagonists.The development of ovarian cysts requires the combination of GnRHa and gonadotropins for controlled ovarian hyperstimulation.However,in rare cases patients may develop an ovarian hyper response after administration of GnRHa alone.CASE SUMMARY Here,two case studies were conducted.In the first case,a 33-year-old female diagnosed with polycystic ovary syndrome presented for her first IVF cycle at our reproductive center.Fourteen days after triptorelin acetate was administrated(day 18 of her menstrual cycle),bilateral ovaries presented polycystic manifestations.The patient was given 5000 IU of human chorionic gonadotropin.Twenty-two oocytes were obtained,and eight embryos formed.Two blastospheres were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.In the second case,a 37-year-old woman presented to the reproductive center for her first donor IVF cycle.Fourteen days after GnRHa administration,the transvaginal ultrasound revealed six follicles measuring 17-26 mm in the bilateral ovaries.The patient was given 10000 IU of human chorionic gonadotropin.Three oocytes were obtained,and three embryos formed.Two high-grade embryos were transferred in the frozen-thawed embryo transfer cycle,and the patient was impregnated.CONCLUSION These two special cases provide valuable knowledge through our experience.We hypothesize that oocyte retrieval can be an alternative to cycle cancellation in these conditions.Considering the high progesterone level in most cases of this situation,we advocate freezing embryos after oocyte retrieval rather than fresh embryo transfer.

Bis(2-butoxyethyl) Phthalate Delays Puberty Onset by Increasing Oxidative Stress and Apoptosis in Leydig Cells in Rats

Objective This study investigated the effects of bis(2-butoxyethyl) phthalate(BBOP) on the onset of male puberty by affecting Leydig cell development in rats.Methods Thirty 35-day-old male Sprague-Dawley rats were randomly allocated to five groups mg/kg bw per day that were gavaged for 21 days with BBOP at 0, 10, 100, 250, or 500 mg/kg bw per day. The hormone profiles;Leydig cell morphological metrics;mRNA and protein levels;oxidative stress;and AKT,mTOR, ERK1/2, and GSK3β pathways were assessed.Results BBOP at 250 and/or 500 mg/kg bw per day decreased serum testosterone, luteinizing hormone, and follicle-stimulating hormone levels mg/kg bw per day(P < 0.05). BBOP at 500 mg/kg bw per day decreased Leydig cell number mg/kg bw per day and downregulated Cyp11a1, Insl3, Hsd11b1,and Dhh in the testes, and Lhb and Fshb mRNAs in the pituitary gland(P < 0.05). The malondialdehyde content in the testis significantly increased, while Sod1 and Sod2 mRNAs were markedly downregulated, by BBOP treatment at 250–500 mg/kg bw per day(P < 0.05). Furthermore, BBOP at 500mg/kg bw per day decreased AKT1/AKT2, mTOR, and ERK1/2 phosphorylation, and GSK3β and SIRT1levels mg/kg bw per day(P < 0.05). Finally, BBOP at 100 or 500 μmol/L induced ROS and apoptosis in Leydig cells after 24 h of treatment in vitro(P < 0.05).Conclusion BBOP delays puberty onset by increasing oxidative stress and apoptosis in Leydig cells in rats.The graphical abstract is available on the website www.besjournal.com.

高泌乳素血症妊娠相关治疗策略

高泌乳素血症（hyperprolactinemia,HPRL）在25~34岁妇女中发病率为每年23.9/10万,是育龄期女性生殖功能障碍的重要原因之一。接受不孕症检查的女性患者中HPRL的发病率达15%~20%,泌乳素（又称催乳激素,prolactin,PRL）、抑制促性腺激素释放激素（gonadotropin-releasing hormone,Gn RH）的过度分泌,影响卵泡发育,降低卵泡对促性腺激素（gonadotro-tropin，Gn）的敏感性。

排卵诱导药物应用的并发症及其防治

排卵诱导药物是指在排卵障碍或辅助生殖技术中用于诱发排卵的药物,常用的药物根据其作用机制不同主要有以下几类：（1）促性腺激素（gonadotropin,Gn）：包括尿促性素（HMG）、高纯度或重组卵泡刺激素（FSH）、重组黄体生成素（LH）,用于增加卵泡数量和促进卵泡发育;（2）绒促性素（HCG）：用于刺激排卵和黄体支持。

利血平和丘脑下部促黄体素释放激素类似物(LHRH-A)对大鳞副泥鳅促性腺激素细胞的分泌活动和排卵的促进作用

利血平能使神经分泌细胞的儿茶酚胺贮存量减少甚至消耗殆尽,导致多巴胺作为GRIF的作用减弱,从而显著增强LHRH-A促进GtH分泌和诱导排卵的效应。对大鳞副泥鳅,利血平和LHRH-A同时注射或者利血平比LHRH-A提前3小时注射,都能激活GtH细胞,显著增强其分泌活动并诱导排卵。利血平的最低有效剂量为1—5微克/克体重。利血平(25微克/克体重)+LHRH-A(0.05微克/克体重)的催产效果和多巴胺拮抗物pimozide(1微克/克体重)+LHRH-A(0.05微克/克体重)的催产效果一样,排卵率都达到了100％。

Male contraception:prospects for the new millennium

Effective regulation of human fertility has global consequences in terms of resource depletion, pollution and pover-ty. Current family planning services predominantly target a female clientele with few significant developments in malefertility regulation for over a century. The last two decades have witnessed a gathering interest, initially from the scien-tific community, and laterally from industry, in the development of safe, reliable, reversible methods of contraceptionfor men. This review summarises the methods of male fertility regulation which are currently available and critically ex-amines the published data on novel developments in male hormonal contraception which offer the potential of improvedcontraceptive choice for all in new millennium. (Asian J Androl 2000; 2: 3 - 12)

Evaluation of pig oocyte in vitro maturation and fertilization using three gonadotropin-based hormonal compounds

Objective:To evaluate the effect of human chorionic gonadotropins(hCG)and equine chorionic gonadotropins(eCG)on in vitro gilt oocyte maturation and embryonic development,using frozen semen for fertilization.Methods:Two independent experiments(6 replicates each)were carried out to evaluate gilt oocyte maturation,and fertilization and embryonic development by using ovaries from a local abattoir.Totally,712 oocytes were randomly distributed in four-well dishes to receive Novormon(eCG 5.0 IU),PG600(eCG 5.0 IU and hCG 2.5 IU),Chorulon(hCG 5.0 IU),or no hormones.Oocytes were incubated with 5%CO2,95%air and saturation humidity at 39℃for 44 h.Maturation of the oocytes to metaphaseⅡwas assessed by using the aceto-orcein technique.In addition,741 oocytes were used and randomly distributed in four-well dishes,and then oocyte maturation was carried out as mentioned,but matured oocytes were washed and placed in fertilization medium with frozen-thawed sperm.Gametes were co-incubated for 7 h,and then washed and placed in development medium,and incubated for further 7 days,at which time embryonic development was evaluated.Fertilization and embryo development media were not supplemented with the studied hormones.Results:Novormon(eCG)and PG600(eCG+hCG)treatments significantly improved the percentages of metaphaseⅡoocytes compared to the control group(P<0.05).Furthermore,a significant increase was also observed in the young blastocyst stage between the control group and the PG600 treatment group(P<0.05).Conclusions:Hormonal products Novormon(eCG)and PG600(eCG+hCG)can obtain the highest percentages of in vitro maturation in gilt oocytes;however,this effect is not transferred to fertilization rates.

GnRH-a治疗子宫内膜异位症用药方案的研究进展

1971年Shally和Guillenmin从猪下丘脑成功分离出了促性腺激素释放激素（gondaotropin-releasing hormone,GnRH）。随着人们对GnRH的不断研究,高效能的促性腺激素释放激素类似物/激动剂（gonadotropin-releasing hormone analog,GnRH-a）被合成出来。

男性迟发性性腺功能减退症的研究进展

男性迟发性性腺功能减退症是指性欲减退、晨勃减少、勃起功能障碍,其病因较多,如：下丘脑促性腺激素释放激素（gonadotropin releasing hormone,GnRH）分泌减少、紊乱;垂体对GnRH应答减少,导致黄体生成素（luteinizing hormone,LH）脉冲频率增加但不规则、振幅减小;睾丸自身功能损害;睾酮水平下降导致性激素结合球蛋白（sex hormone-binding globulin,SHBG）水平增加,从而导致血浆生物活性睾酮水平下降等。

Estrogens in males: what have we learned in the last 10 years?

This review focuses on the role of estrogen in men, mainly in male reproduction. The continuing increase in data obtained, and recent discoveries in this area will enable a better understanding of male physiology; these, in turn, will have important clinical implications.

单克隆抗体的DOT-ELISA方法用于早期妊娠诊断

人绒毛促性腺激素(human chorionic gonadotropin,hCG)是由胎盘产生的糖蛋白,妇女尿或血清中的 hCG 是妊娠的最早指标。利用β-hCG 亚单位的特异性进行免疫分析,是测定体液 hCG 水平的最准确的方法。1985年国外首次报道应用单克隆抗体和酶免测定法测定 hCG。同年,陈捷平等在国内首次生产这一试剂盒并推广至全国。1987年国外报道了更简便的 DOT-ELISA 法检测 hCG,但无方法学详细的报道。

Challenges in the diagnosis and treatment of gestational trophoblastic neoplasia worldwide

Gestational trophoblastic neoplasia(GTN) is a rare tumor that originates from pregnancy that includes invasive mole, choriocarcinoma(CCA), placental site trophoblastic tumor and epithelioid trophoblastic tumor(PSTT/ETT). GTN presents different degrees of proliferation, invasion and dissemination, but, if treated in reference centers, has high cure rates, even in multi-metastatic cases.The diagnosis of GTN following a hydatidiform molar pregnancy is made according to the International Federation of Gynecology and Obstetrics(FIGO)2000 criteria: four or more plateaued human chorionic gonadotropin(hCG)concentrations over three weeks; rise in hCG for three consecutive weekly measurements over at least a period of 2 weeks or more; and an elevated but falling hCG concentrations six or more months after molar evacuation. However,the latter reason for treatment is no longer used by many centers. In addition,GTN is diagnosed with a pathological diagnosis of CCA or PSTT/ETT. For staging after a molar pregnancy, FIGO recommends pelvic-transvaginal Doppler ultrasound and chest X-ray. In cases of pulmonary metastases with more than 1cm, the screening should be complemented with chest computed tomography and brain magnetic resonance image. Single agent chemotherapy, usually Methotrexate(MTX) or Actinomycin-D(Act-D), can cure about 70% of patients with FIGO/World Health Organization(WHO) prognosis risk score ≤ 6(low risk), reserving multiple agent chemotherapy, such as EMA/CO(Etoposide,MTX, Act-D, Cyclophosphamide and Oncovin) for cases with FIGO/WHO prognosis risk score ≥ 7(high risk) that is often metastatic. Best overall cure rates for low and high risk disease is close to 100% and > 95%, respectively. The management of PSTT/ETT differs and cure rates tend to be a bit lower. The early diagnosis of this disease and the appropriate treatment avoid maternal death,allow the healing and maintenance of the reproductive potential of these women.

Complete response to multidisciplinary therapy in a patient with primary gastric choriocarcinoma

Primary gastric choriocarcinoma is a rapidly growing neoplasm with an average survival of several months in untreated patients.Gastrectomy with lymph node dissection followed by chemotherapy is the treatment of choice.Regimens used for gastric adenocarcinoma are usually selected.However,median survival remains less than six months.In this case report,we describe a case of primary gastric choriocarcinoma with a clinical complete response to multidisciplinary treatment including surgery,chemotherapy,and radiofrequency ablation(RFA).The patient was originally referred for general malaise.Esophagogastroduodenoscopy demonstrated a large tumor occupying the fornix,and total gastrectomy with lymph node dissection was performed.Seven days later,multiple liver metastatic recurrences with high serum levels of beta-human chorionic gonadotropin(β-hCG) were recognized.Chemotherapy with a gonadal choriocarcinoma regimen consisting of etoposide,methotrexate,actinomycin D,cyclophosphamide,and vincristine(EMA/CO),was initiated.After three cycles,serum β-hCG decreased markedly and the tumors disappeared.Six months later,multiple lung metastatic recurrences were found.After one cycle of EMA/CO,only one nodule remained.Computed tomography-guided RFA was performed for this oligometastatic tumor.The patient has been alive with no evidence of disease for 10 years after the initial diagnosis.To the best of our knowledge,this patient with recurrent primary gastric choriocarcinoma has achieved the longest survival.The present case is the first report of choriocarcinoma metastatic to the lung successfully treated with RFA.From our retrospective analysis of recurrent or unresectable primary gastric choriocarcinoma,we propose that gonadal choriocarcinoma regimens can be considered as first-line for primary gastric choriocarcinoma.

SUBSTITUTION OF ACUPUNCTURE FOR HCG IN OVULATION INDUCTION

By using human menopausal gonadotropin (HMG) and human chorionic gonadotropin (HCG), fairly good clinical therapeutic efficacy has been obtained in the treatment of infertility. However, difficulties are brought about due to the ovarian hyperstimulation syndrome (OHSS) easily induced by these two drugs. Therefore,

Minimum Dose of hCG to Trigger Final Oocyte Maturation and Prevent OHSS in a Long GnRHa Protocol

This paper was aimed to study the minimum dose of human chorionic gonadotropin （hCG） to effectively trigger maturation of oocytes and prevent ovarian hyperstimulation syndrome （OHSS） in a series of hyper-responders treated with a long gonadotropin releasing hormone agonist （GnRHa） proto- col. Six women at high risk of developing severe OHSS in a long GnRHa protocol were enrolled into this study. Serum hormone levels on the day of and after hCO administration, antral follicle count, oo- cyte retrieval number and quality were determined. In total, 6 women aged between 29 and 36 years and at risk of developing severe OHSS, received 2000 U hCG Five of them were treated with coasting for 1 day and the rest one for 4 days. The mean number of oocytes collected was 19 （range 14-27） and the fertilization rate per collected oocyte was 72.81%. Of the 6 women in the study, only one cancelled em- bryos transfer and all embryos were frozen, and then she delivered two health boys on term in the sub- sequent frozen-thawed embryo transfer （FET） cycle. Pregnancies and births were achieved in 3 patients out of 5 in vitro fertilization-embryo transfer （IVF-ET） cycles. No woman developed moderate or severe OHSS. Triggering with 2000 U hCG is feasible to prevent OHSS in unpredicted hyper-responders undergoing IVF in a long GnRHa protocol.

Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary-testicular axis secretion in aging rats

An increasing amount of evidence demonstrates the anti-aging effect of Heshouwu in pill form. In this study, a subacute aging rat model was established by continuous intraperitoneal injection of D-galactose and treated with Heshouwu decoction （a Chinese herb for tonifying the kidney, comprising Heshouwu pill, Herba Epimedii, Radix Salviae Miltiorrhiae, and Poria）. Heshouwu pill treated rats were the positive control group. Radioimmunoassay, immunohistochemical staining, and western blot assay showed hypothalamic gonadotropin-releasing hormone, hypothalamic substance P, and serum gonadotropin levels to be significantly increased in the model rats; the concentrations of hypothalamic ~3-endorphin, and serum levels of insulin-like growth factor I and testosterone were significantly decreased. 1713- and 3[3-hydroxysteroid dehydrogenase expression in testicular tissue was also decreased. Intragastric administration of Heshouwu decoction at high （9.6 g/mL/100 g）, medium （4.8 g/mL/100 g）, and low （2.4 g/mlJ100 g） doses, Heshouwu decoction pretreatment at a medium dose （4.8 g/mL/100 g）, and Heshouwu pill （2.06 g/mL/100 g） significantly reversed these changes. Heshouwu decoction pretreatment and high-dose Heshouwu decoction had the greatest anti-aging effects. These experimental findings indicate that Heshouwu decoction can improve hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect superior to that of Heshouwu pill.

Predictive Value of Initial Serum Human Chorionic Gonadotropin Levels for Pregnancies after Single Fresh and Frozen Blastocyst Transfer

As one of the earliest markers for predicting pregnancy outcomes, human chorionic gonadotropin(h CG) values have been inconclusive on reliability of the prediction after frozen and fresh embryo transfer(ET). In this retrospective study, patients with positive h CG(day 12 after transfer) were included to examine the h CG levels and their predictive value for pregnancy outcomes following 214 fresh and 1513 vitrified-warmed single-blastocyst transfer cycles. For patients who got clinical pregnancy, the mean initial h CG value was significantly higher after frozen cycles than fresh cycles, and the similar result was demonstrated for patients with live births(LB). The difference in h CG value existed even after adjusting for the potential covariates. The area under curves(AUC) and threshold values calculated by receiver operator characteristic curves were 0.944 and 213.05 m IU/m L for clinical pregnancy after fresh ET, 0.894 and 399.50 m IU/m L for clinical pregnancy after frozen ET, 0.812 and 222.86 m IU/m L for LB after fresh ET, and 0.808 and 410.80 m IU/mL for LB after frozen ET with acceptable sensitivity and specificity, respectively. In conclusion, single frozen blastocyst transfer leads to higher initial h CG values than single fresh blastocyst transfer, and the initial h CG level is a reliable predictive factor for predicting IVF outcomes.

Bowel endometriosis:Recent insights and unsolved problems

Bowel endometriosis affects between 3.8% and 37% of women with endometriosis.The evaluation of symptoms and clinical examination are inadequate for an accurate diagnosis of intestinal endometriosis.Transvaginal ultrasonography is the first line investigation in patients with suspected bowel endometriosis and allows accurate determination of the presence of the disease.Radiological techniques (such as magnetic resonance imaging and multidetector computerized tomography enteroclysis) are useful for estimating the extent of bowel endometriosis.Hormonal therapies (progestins,gonadotropin releasing hormone analogues and aromatase inhibitors) significantly improve pain and intestinal symptoms in patients with bowel stenosis less than 60% and who do not wish to conceive.However,hormonal therapies may not prevent the progression of bowel endometriosis and,therefore,patients receiving long-term treatment should be periodically monitored.Surgical excision of bowel endometriosis should be offered to symptomatic patients with bowel stenosis greater than 60%.Intestinal endometriotic nodules may be excised by nodulectomy or segmental resection.Both surgical procedures improve pain,intestinal symptoms and fertility.Nodulectomy may be associated with a lower rate of complications.

Studies of GnRH-A Active Immunization Effects on LH and FSH Secretion and Histostructure of the Ovary and Uterus in Rabbits

The objective of the study is to investigate the effects of active immunization against gonadotropin releasing hormone agonist （GnRH-A） on secretion of luteinizing hormone （LH） and follicle-stimulating hormone （FSH） in the pituitary, and to observe the histological structures and development about ovaries and uteri in female rabbits. 24 female rabbits （Oryctolagus cuniculus） were divided randomly into 4 groups （n=6）, namely, experimental group I （EG-I）, experimental II （EG-II）, experimental III （EG-III）, and control group （CG）. Rabbits were subcutaneously injected with 1.0 mL GnRH-A （alarelin） antigen respectively at concentrations of 100, 100 and 50 μg mL-1 respectively, in EG-I, EG-II and EG-III. Alarelin antigen was re-injected in EG-II and EG-III with the same dosage on 20 d. CG was a blank. The ovarian and uterine samples were collected aseptically at the end of the experiment of 70 d. The tissue slices were observed under light and electron microscopes. Serum concentrations of LH and FSH were measured with ELISA. The results showed that serum LH concentrations in EG-II and EG-III reached the peak levels on 50 and 40 d respectively, and LH level in EG-II exceeded other 3 groups on 50 d （P0.05）. FSH level in EG-II was higher than those in EG-I, CG （P0.01） and EG-III （P0.05） on 40 d. GnRH-A could increase the number of primary follicles, enlarge the primary follicle vertical diameter （PFV） and primary follicle transverse diameter （PFT）, and promote growth and maturation of follicles. The endometrial epithelium thickness （EET） and uterine wall thickness （UWT） in three EGs were less than that in CG （P0.05）. GnRH-A can increase the quantities of mitochondrial cristaes, cortex granules in cytoplasm, broaden and lengthen zona pellucidas and microvilli of oocytes. It also enlarged nuclei of ooxytes and mitochondria, thereby it promoted the development of oocytes. Re-injection of 100 μg alarelin antigen enhanced the secretion of LH and FSH. GnRH-A promoted the growth and maturation of ovaries and follicles, suppressed uterine development, and also influenced histostructure of ovaries and uteri in female rabbits.