Effect of sodium arsenite on spermatogenesis, plasma gonadotrophins and testosterone in rats

<abstract> To investigate the effect of arsenic on spermatogenesis. Methods: Mature (4 months old) Wistar rats were intraperitoneally administered sodium arsenite at doses of 4, 5 or 6 mg-kg^-day1 for 26 days. Different varieties of germ cells at stage VII seminiferous epithelium cycle, namely, type A spermatogonia (ASg), preleptotene spermatocytes (pLSc), midpachytene spermatocytes (mPSc) and step 7 spermatids (7Sd) were quantitatively evaluated, along with radioimmunoassay of plasma follicle-stimulating hormone (FSH), lutuneizing hormone (LH), testosterone and assessment of the epididymal sperm count. Results: In the 5 and 6 mg/kg groups, there were significant dose-dependent decreases in the accessory sex organ weights, epididymal sperm count and plasma concentrations of LH, FSH and testosterone with massive degeneration of all the germ cells at stage VII. The changes were insignificant in the 4 mg/kg group. Conclusion: Arsenite has a suppressive influence on spermatogenesis and gonadotrophin and testosterone release in rats.

Constraints in the development of contraceptives for men

Considerable efforts have been made to develop a male contraceptive and the studies have provided very useful infor-mation in this field. At least five different strategies to develop a male contraceptive have been pursued, namely: inhi-bition of sperm production, interference with sperm function, interruption of sperm transport, prevention of sperm de-position, and prevention of sperm-egg interaction. Of all these approaches, inhibition of sperm production by using an-drogens either alone or in combination with progestins have given the most encouraging results. A nmnber of clinicaltrials substantiate that it is indeed possible to have a reversible, effective and safe hormonal method of contraception. Apostmeiotic and epididymal approach to interfere with sperm function or the secretory and metabolic processes of theepididymis is another attractive option of male contraceptive development. A number of chemical compounds have beenidentified which interfere with sperm function in the epididymis without affecting sperm production, however, the com-pounds evaluated so far were found to be toxic. Interruption of sperm transport through the vas either by vasectomy orpercutaneous intmvasal injection of liquids which form cure-in-place plugs is also an attractive option. However, re-versibility of the methods is of concern in their wide scale use. The major constraint in developing a long-acting male contraceptive seems to be the need for greater investment forproduct development. The clinical trials for evaluating the efficacy and safety of the new products and formulationsstretch over several years and require enormous financial commitment. Nevertheless, the long-term gain of having along-acting reversible contraceptive for men is far greater than the financial commitments over few years. Male attitudetowards using methods of family planning is much more favourable than originally believed. The pharmaceutical indus-try as well as the health care providers therefore have a greater responsibility. For early development of a contraceptivefor men, it is essential to increase investment and simplify the drug regulatory procedures. The advent of newer tech-nologies coupled with the convergent efforts of scientists will certainly make it possible to have an effective, safe andreversible male contraceptive in the near future.

Male reproductive hormones and semen quality

Male reproductive functions are mediated by different hormones whose orchestrations remain a major research interest. The 'master' regulator hormonal axis is the hypothalamo-pituitary-gonadal/testicular axis which is led by the pulsatile release of hypothalamic gonadotropin-releasing hormone. This, in turn, stimulates the anterior pituitary trophic hormones, the luteinizing hormone and follicle-stimulating hormone. Luteinizing hormone and follicle-stimulating hormone act upon the testicular cells, the Leydig cells for steroidogenesis and Sertoli cells to aid spermatogenesis, respectively. This primary axis is influenced by an array of other testicular hormones, metabolic hormones, and different regulatory factors. These hormonal crosstalks influence the intricate testicular functions, sexual behavior and semen quality in men. Given the growing concern in the last few decades over the increasing prevalence of male subfertility and/or infertility mostly in terms of deteriorating semen quality, it is required to find its underlying mechanisms. In this regard, the endocrine regulation of testicular functions is of prime importance in the determination of semen quality and sperm functions. This review article aims to present a concise updated overview on the mechanism by which the key hormones integrate the male reproductive functions and maintain the semen quality.

The effect of traditional Chinese medicine──Tian Gui Recipeon obesity and anovulation in androgen-sterilized rats

Objective: A traditional Chinese medicine Tian Gui Recipe (TGR) has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR). Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1. 25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n = 25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n = 15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno-and mRNA-containing neurons, and its relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER, NPY, gonadotrophin releasing hormone (GnRH) and gene expression levels on NPY, OB-Rb, POMC. Meanwhile, the criteria of energy state, including daily food intake, retroperitoneal fat depot pad and body weight, were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD). Results: Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0. 01, P<0. 01, P<0. 01). The mean serum leptin, T, E2 levels of group ASR were significantly higher than those of Group C and Group A+H (P< 0. 01,P<0. 01 ), while FSH, LH levels were lower (P<0. 01, P<0. 01 ). ERs were proved to be distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-syn- thesizing neurons, and both NPY and POMC mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions were increased (P<0. 01, P<0. 01, P<0. 01 ), OB-Rb mRNA, ER and GnRH immunoreactive expression were decreased significantly in the hypothalamus of ASR (P<0. 01, P<0. 01, P<0. 01 ). These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0. 05). Conclusion: The elevated peripheral E2 caused by high T and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY- containing neurons respectively, therefore challenge the NPY mRNA and NPY, POMC mRNA overexpressions. The overexpression, of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. TGR may reverse these abnormal neuroendocrine cascades by lowering the levels of T, E2 and leptin.

The effect of traditional Chinese medicine“Tian Gui Recipe (TGR)”on obesity and anovulation in androgen-sterilized rats(ASR)

Objectives: A traditional Chinese medicine “Tian Gui Recipe (TGR)” has been used to effectively treat clomiphene resistant anovulatory disease and obesity, especially in polycystic ovary syndrome (PCOS) cases with hyperinsulinemia. The effect of TGR on obesity and anovulation was investigated in androgen-sterilized rats (ASR).Methods: Female SD rats at the age of 9 days were divided into 3 groups. Group ASR (n=15): anovulation was demonstrated at the age of 70 days by vaginal smear in rats while 1.25 mg of testosterone propionate was injected subcutaneously on its 9th neonatal day. Group A+H (n=25): TGR were administered to ASR at the age of 80 days for 3 weeks. Rats with regular estrous cycle and ovulation after herbal treatment were included in this group. Group C (n=15): normal ovulated rats were recruited. Around the age of 112 days or on proestrous day, all rats were sacrificed under anesthesia. Serum leptin, testosterone (T), estrogen (E2), follicular stimulating hormone (FSH), luteinizing hormone (LH) levels were measured with radioimmunoassay (RIA). Double immunofluorescent staining combined with confocal laser scanning microscope and dual in situ hybridization were carried out on sections through areas of arcuate nucleus (ARC) to determine whether estrogen receptor (ER) immunoreactivity (IR) or long form of leptin receptor (OB-Rb) mRNA were expressed in neuropeptide Y (NPY) immuno- and mRNA-containing neurons, and their relationship to proopiomelanocortin (POMC) mRNA-containing neurons. Immunohistochemistry and in situ hybridization were used to observe the levels of ER-, NPY-,gonadotrophin releasing hormone (GnRH)-IR and gene expression levels of NPY, OB-Rb, and POMC in ARC. Meanwhile, the criteria of energy state,including daily food intake, retroperitoneal fat depot pad and body weight,were measured and evaluated. Values of immunohistochemistry and in situ hybridization were expressed in mean optic density (MOD).Results:Seventeen out of the 25 rats (68%) in Group A+H were ovulated after herbal treatment. ASR characterized with significantly high metabolic rate, energy imbalance and obesity (P<0.01,P<0.01,P<0.01).The mean serum leptin, T, E2 levels of ASR were significantly higher than that of Group C and Group A+H (P<0.01,P<0.01), while being contraryto that of FSH, LH levels (P<0.01,P<0.01). ERs were proved distributing in NPY-containing neurons, OB-Rb coexpressed in NPY-synthesizing neurons, and both NPY- and POMC- mRNA expressing neurons overlapped in ARC. Compared to Group C, levels of NPY mRNA and NPY, POMC mRNA expressions increased (P<0.01,P<0.01,P<0.01),OB-Rb mRNA, ER and GnRH immunoreactive expression decreased significantly in the hypothalamus of ASR (P<0.01,P<0.01,P<0.01).These dramatic neuroendocrine changes became normal in ovulated ASR after the herbal treatment (all P>0.05).Conclusion: The elevated peripheral estrogen caused by high testosterone and leptin levels in ASR may down-regulate their corresponding receptors oriented on hypothalamic NPY-containing neurons respectively, therefore challenge the NPY mRNA and NPY,POMC mRNA overexpression. The overexpression of NPY, POMC gene transcription and translation stimulate food intake, promote the deposit of adipocyte tissue, and inhibit GnRH expression and GnRH/FSH, LH release, which contribute to the occurrence of obesity and anovulation in ASR. “TGR” may reverse these abnormal neuroendocrine cascades by lowering levels of testosterone, estrogen and leptin.

早孕孕妇血清β-HCG及孕酮检测水平及临床应用

目的探讨早孕孕妇血清β-HCG及孕酮检测水平与胎儿生长发育状况的关系。方法用深圳产MAGLUMI1000全自动化学发光测定仪检测142例正常早孕孕妇和58例胎儿发育不良、停止发育等非正常早孕孕妇的血清β-HCG和孕酮水平,同时做彩色B超检查;48 h后再次检测血清β-HCG水平,对有流血等先兆流产孕妇进行保胎治疗并密切观察。结果非正常早孕孕妇的血清β-HCG与孕酮水平明显低于正常妊娠妇女且增长速度较慢(P<0.05),绝大部分正常妊娠妇女(94.6%)血清β-HCG水平的倍增<2 d,而大部分非正常妊娠妇女(85.7%)血清β-HCG水平的倍增>2 d。早孕孕妇的血清β-HCG与孕酮水平与胎儿生长发育状况成正相关,当血清β-HCG和孕酮平均水平分别达31 601 IU/L和20.8 ng/mL时,容易保胎成功。结论非正常妊娠妇女血清β-HCG与孕酮水平明显低于正常妊娠妇女且增长速度较慢。

Plasma β-Endorphin Levels in Women with Early Threatened Abortion before and after the Treatment of Integrated Chinese and Western Medicine

To observe plasm a β-endorphin (β-EP) and gonadotrophin releasing horm one (GnRH), hum an chorionic gonadotrophin (hCG), progesterone (P4) levelsin w om en w ith early threatened abortion and w ith a history of recurrent spontaneous abortions (RSA). Tw enty patientsw ith threatened abortion at7～8 w eeksof gestation w ere re- cruited, allof them had a history of 3 or m ore recurrentunexplained abortions. They w ere treated w ith psychologicalconsultation accompanied by traditionalChinese herbs. Blood samples w ere taken to m easure β-EP, GnRH, hCG and P4 levels by radioim - m unoassay (RIA). The treatm ents w ere continued till10～12 w eeks, blood w astaken during this period to compare changes in these peptides / horm ones. Tw enty norm al pregnantw om en at7～8 and 10～12 w eeksand 20 patientsw ith incompleteabortion at 10～12 w eeksw ererecruited for comparativestudies. Results: (1) In norm alpregnant w om en, plasm a β-EP, GnRH, hCGand P4 levelsat10～12 w eeksw ere significantly higher than thatat7～8 w eeks (P< 0.01). (2) In patients w ith threatened abortion and a history of RSA, plasm a β-EP levels at7～8 w eeks w ere significantly higher than those of norm al pregnantw om en (P< 0.01); on the contrary, plasm a GnRH, hCGand P4 levelsin these patientsw ere significantly low er than thosein norm alcases (P< 0.01). After treatm ent, 16 of the 20 patients succeeded in m aintaining their pregnancies, the levels of the four plasm a contents at10～12 w eeks w ere sim ilar to thosein norm alpregnantw om en (P> 0.05). (3) Plasm a β-EPlevelsin patientsw ith incomplete abortionsat10～12 w eeksw ere dram atically higher and GnRH, hCGand P4 levelsw ere low er than in norm alpregnantw om en (P< 0.01). β-EP m ightplay a role in the pathophysiology of spontaneousabortion.

Serum β-hCG level on day 7 of frozen-thawed embryo transfer: association with the clinical pregnancy outcomes in artificial cycles

Objective:The relationship between serum beta human chorionic gonadotropin(β-hCG)levels of patients(7 days after the transplantation of frozen-thawed embryos)and the pregnancy outcomes was investigated.Methods:This study was designed as a retrospective clinical trial of 366 women who underwent frozen-thawed embryo transfers(FETs)in artificial cycles.Patients were divided into three groups:clinical pregnancy group,biochemical pregnancy group,and non-pregnant group according to their pregnancy outcomes.Serumβ-hCG levels were tested on day 4,7,9,11 and 14 after FET.Results:In the clinical pregnancy group,the serumβ-hCG levels after 7-day post-transplantation were significantly elevated(16.20 IU/L vs.3.07 vs.0.1 IU/L;P<0.05)compared with the other two groups.Furthermore,it was found that Area Under Curve(AUC=0.96)was significant with cut-off value higher than 4.26 IU/L(sensitivity=92.3%,specificity=90.2%)to predict the clinical pregnancy outcomes in the receiver operating characteristic(ROC)analysis ofβ-hCG concentrations on day 7 of post-transplantation.Conclusion:Our results suggested that the elevated serumβ-hCG levels on day 7 of post-transplantation could predict the positive clinical pregnancy outcomes in artificial FET cycles.

Pulmonary deportation of hydatidiform mole:a 12-year,single tertiary center experience in China

Background:Pulmonary deportation of hydatidiform mole is an exceedingly rare entity.The underlying mechanisms and proper management strategies remain unclear based on sporadic case reports over the past six decades.This study aimed to investigate the clinical features and rational treatment of patients with benign molar pregnancies with pulmonary deportation based on our experience.Methods:Medical records of 20 cases of hydatidiform mole with pulmonary deportation were retrospectively reviewed at Peking Union Medical College Hospital from November 2006 to May 2019.The detailed information of all patients was recorded and analyzed.Patients were divided into different groups according to their characteristics and Mann-Whitney U test was used to compare the duration to achieve a normal b-human chorionic gonadotrophin(b-hCG)level after the first evacuation among groups.Results:Initial pulmonary computed tomography scans showed suspected bilateral,left and right chest deportation of hydatidiform mole in 12,four,and four patients,respectively,with the maximum nodular diameter ranging from 0.6 to 1.2 cm.Ten patients achieved lesion resolution while the remaining ten patients achieved decreases in the size of their pulmonary lesions.The median duration to achieve a normal b-hCG level after the first evacuation was 15.5(13.0,21.9)weeks.There was no significant difference in the duration to achieve a normal b-hCG level after the first evacuation between two groups based on age(≥40 years vs.<40 years:15.8[12.2,21.5]weeks vs.15.5[12.9,23.0]weeks,Z=0.094,P=0.925),type of antecedent mole(partial mole vs.complete mole:15.2[12.5,27.4]weeks vs.15.9[12.9,21.5]weeks,Z=0.165,P=0.869),distribution of pulmonary nodules(bilateral lungs vs.unilateral lung:15.2[12.8,22.5]weeks vs.15.9[13.2,22.2]weeks,Z=0.386,P=0.700),maximum size of pulmonary nodules(>0.5 cm vs.0.5 cm:13.0[11.3,17.2]weeks vs.16.0[14.5,23.8]weeks,Z=1.815,P=0.070),and number of uterine evacuations(once vs.twice or three times:15.0[13.0,16.3]weeks vs.16.0[12.8,23.9]weeks,Z=0.832,P=0.405).The post-molar cohort was followed up for 17 to 139 months,and no gestational trophoblastic neoplasia was observed.Conclusions:No surgeries other than uterine evacuation and no chemotherapy regimens are recommended for such patients if they achieve satisfactory decreases in the level of hCG and gradual decrease or disappearance of pulmonary deportation nodules.Patients should be informed about the necessity of long-term follow-up.More collaborative international studies on this exceedingly rare condition may guide decisions regarding optimal management strategies.

Detection of Messenger RNA for Gonadotropin-Releasing Hormone (GnRH) but not for GnRH Receptors in Rat Pancreas

Although gonadotropin releasing hormone (GnRH), GnRH like molecule, and GnRH receptor (GnRH R) have been reported to exist in several tissues other than brain or anterior pituitary, there are no reports concerning GnRH or GnRH R gene expression in a normal pancreatic gland. In order to define the production of GnRH as well as GnRH R in the pancreatic gland, we examined their gene expression in various developmental stages of rat pancreas using the reverse transcriptase polymerase chain reaction (RT PCR). GnRH mRNA transcripts were found in pancreas of male and female rats at different ages, expressing at about the same level, whereas GnRH R mRNA transcripts could not be detected in any rat pancreatic gland samples. These results suggest a possible biological role of GnRH in rodent pancreas.

Clomiphene Citrate as An Adjuvant to hMG Stimulation of the Ovaries in Mid-to-late Follicular Phase and Subsequently Pregnancy Outcome of Frozen-thawed Embryo Transfers

Objective To compare the results of a novel regimen of human menopausal gonadotrophin （hMG） in combination with clomiphene citrate （CC） in mid-to-late follicular phase with those of a short protocol of GnRH agonist （GnRHa） and hMG used for IVF. Methods In the retrospective study, 842 patients undergoing IVF were collected and classified into two groups： hMG in combination with CC in mid-to-late follicular phase （group A, n=319） and short protocol of GnRHa-hMG （group B, n=523）. The main outcome measures were ovarian responses in stimulation cycles and pregnancy outcomes in subsequent frozen-thawed embryo transfer （FET） cycles. Results In group A, the serum LH concentration on day 8 -10 was similar with that on the day of hCG administration （2.43 ± 1.92 IU vs 2.51 ±2.05 IU）. The number of mature follicles and oocytes retrieved was significantly lower in group A than in group B while the fertilization rate and the cleavage rate were comparable. The clinical pregnancy rate （47. 79% vs 48.04%）, the implantation rate （32.49% vs 33.11%） and the cumulative pregnancy rate （58.09% vs 60.22%） were respectively similar in group A and group B. Conclusion hMG in combination with CC in mid-to-late follicular phase results in the same pregnancy outcome as short protocol. The novel protocol may take the advantage of eliminating the occurrehce of a premature endogenous LH Surge.

Somatostatin 4 regulates growth and modulates gametogenesis in zebrafish

Somatostatin(SST)plays important roles in growth and development.In teleost fishes six SST encoding genes(sst1 to sst6)have been identified although few studies have addressed their function.Here we aim to determine the function of the teleost specific sst4 in the zebrafish.A CRISPR/Cas9 sst4 zebrafish mutant with loss of function(sst4−/−)was produced which grew significantly faster and was heavier at the onset of gonadal maturation than the wild type(WT).Consistent with their faster growth,liver igf1,igf2a and igf2b expression was significantly upregulated in the sst4−/−fish compared to the WT.Histological examination of the ovaries and testis indicated that sst4−/−fish had slightly delayed testicular gametogenesis compared to the WT.Significantly lower expression of igf3,amh,insl3,hsd17b3,hsd11b2,hsd20b,cyp11b and cyp17 was consistently observed in the sst4−/−testis.In contrast,the ovaries had lower expression of igf1,igf2a and cyp19a1a but increased expression of igf2b and hsd20b.The gonadotrophin beta subunits(fshb and lhb)in the brain were downregulated indicating the brain-pituitary-gonadal axis was downregulated in the sst4−/−fish and suggesting that the steroid production is compromised in the maturing gonads.In addition,analysis of sst1 and sst3 mRNA levels in sst4−/−fish suggests a dosage compensation effect of sst1 in the brain and liver.Altogether,the results from the zebrafish sst4−/−line support the idea that sst4 is involved in the regulation of igf signalling,somatic growth and reproduction since steroidogenesis and gametogenesis at pubertal onset were compromised.

Early detection of persistent trophoblastic tumour by serum human chorionic gonadotrophin monitoring after molar pregnancy

Objective To study the outcome of a multi centred post molar pregnancy serum human chorionic gonadotrophin (hCG) surveillance programme. Methods Patients recruited into the multi centred post molar serum hCG surveillance programme between 1988 and 1996 were studied. The clinical data were obtained from medical records and computer database. Results There were 616 patients in the study. Twenty five (11%) of 224 patients with molar pregnancy and 28 (7%) of 392 patients with partial molar pregnancy were diagnosed to have persistent trophoblastic tumour (PTT) requiring chemotherapy. Of the 53 patients treated for PTT, 58.5% received intravenous methotrexate (MTX), 22.6% received both MTX and actinomycin D, and 19% received CHAMOC, a multiple chemotherapeutic regimen. Four patients receiving single drug had to change the regimen because of poor response, and all recovered and remained well. There were 3 cases of recurrence, one in each group of patients receiving single, dual or multiple agents. One patient died because of treatment complication. The rest were well with a mean follow up of 42 months. Conclusions Post molar serum hCG surveillance is important to detecting persistent trophoblastic activity. Early treatment of PTT by the appropriate chemotherapy has a near hundred percent cure and prevents the progression of PTT into more advanced trophoblastic malignancy such as choriocarcinoma.