Rectifying cow infertility under heat stress by immunization against inhibin and supplementation of progesterone.

This study was undertaken for the development of novel techniques that are based on immunoneutralization of inhibin bioactivity to improve Holstein cow fertility. A series of 4 experiments were carried out on 2 farms that were located in subtropical or temperate regions, to test the effects of immunization against inhibin alpha subunit on cow fertility under varying degrees of heat stress conditions. Though immunization against inhibin alone improved conception rate (CR) after TAI moderately in cows under mild heat stress conditions, the treatment plus progesterone supplementation substantially enhanced CR in the range of 25 to 35 percentages from severe heat stress to comfortable weather conditions. There existed an additive effect between immunization against inhibin and progesterone supplementation that maximally enhanced CR. Further, immunization against inhibin increased both FSH and activin A concentrations in blood during both follicular and luteal phases. It also significantly increased blood concentrations of E2 in the follicular phase but decreased P4 concentrations during the early pregnancy. However, interferon-tau concentrations in blood around the time of pregnancy recognition were doubled in the inhibin immunized cows. In conclusion, immunization against inhibin plus P4 treatment enhances ovarian follicle and the subsequent early embryo developments that help to greatly improve the fertility of Holstein dairy cows.

Calponin and MUC6 complement inhibin as diagnostic immunomarkers of serous cystadenoma in endoscopic ultrasound-guided aspiration/biopsy specimens.

AIMS: Because serous cystadenoma (SCA) does not usually require excision, it is critical to distinguish it from differential diagnoses which do, especially neuroendocrine tumour (NET). The gold standard for diagnosing SCA is assessment of endoscopic ultrasound-guided fine needle aspiration/biopsy (EUS-FNAB) material. Inhibin immunohistochemistry aids this assessment, but such positivity is not absolutely sensitive or specific to SCA. The following is the largest known study of SCA EUS-FNAB specimens and the first to compare four potential SCA immunomarkers between themselves and inhibin, compared against NET. METHODS AND RESULTS: Immunohistochemistry for calponin, mucin 6 (MUC6), glucose transporter 1 (GLUT1) and vascular endothelial growth factor A (VEGFA) was performed on 30 EUS-FNAB and three resection specimens of SCA and 32 EUS-FNAB specimens of NET. GLUT1 and VEGFA were suboptimal as diagnostic immunomarkers of SCA, being expressed by 10 and 44% of NETs, respectively. Further, their expression by cellular constituents of blood which often contaminate EUS-FNAB specimens hampered identification of neoplastic cells, especially in hypocellular samples. While 19% of NETs showed nuclear MUC6 positivity, cytoplasmic expression of the protein showed 100% specificity and sensitivity as an SCA marker. However, assessing MUC6 in EUS-FNAB specimens must also consider the protein's focal expression in physiological pancreatic, gastric or duodenal tissues, which can contaminate these specimens. Calponin was less sensitive (71% versus 100%) but more specific (100% versus 91%) than inhibin, although easier to assess in EUS-FNAB specimens than MUC6. CONCLUSIONS: Of the four potential immunomarkers of SCA suggested by the existing literature, calponin and MUC6 are useful complementary studies to inhibin for application to EUS-FNAB specimens.

Ethanolic extract of Moringa oleifera leaves alleviate cyclophosphamide-induced testicular toxicity by improving endocrine function and modulating cell specific gene expression in mouse testis.

ETHNOPHARMACOLOGICAL RELEVANCE: Moringa oleifera Lam. is known for its nutritional and ethno medicinal values due to the presence of wide array of phytochemicals with multiple biological activities. We have previously reported that ethanolic extract of Moringa oleifera leaves (MOE) ameliorated cyclophosphamide (CP)-induced testicular toxicity and improved functional integrity of spermatozoa as well as spermatogenic cells. AIM OF THE STUDY: The present study was planned to investigate whether the mitigation of CP-induced testicular toxicity by MOE is mediated via modulation of endocrine profile, genes associated with function of different cell types and enhancement of DNA repair response in spermatogonial cells. MATERIALS AND METHODS: Adult Swiss albino mice (8 week) were injected with CP (100 mg/kg, one dose in a week for 3 weeks) and MOE (100 mg/kg, 5 doses in a week for 4 weeks) either alone or in combination intraperitoneally. At 35 day post CP injection (first dose), the functional characteristics such as count, motility, head morphology and DNA integrity were assessed in epididymal spermatozoa. Key reproductive hormones like testosterone, follicle stimulating hormone (FSH) and Inhibin B concentration were analyzed in serum and testis. In addition, mRNA expression of genes pertaining to the function of Leydig, Sertoli and spermatogonial cells as well as antioxidant enzymes were evaluated in the testis. To understand the DNA damage and repair process in germ cells, prepubertal (2 week) mice were administered with single dose of CP (200 mg/kg) and/or MOE (100 mg/kg) and analyzed for expression of DNA damage (γ-H2AX, P53 and Caspase3) and repair genes (Rad51 and Ku80) in isolated spermatogonial cells at various time points after treatment. RESULTS: CP administration resulted in decrease in count, motility and increase in morphological defects and DNA damage in spermatozoa. Testosterone level was marginally decreased while there was a significant increase in FSH (p < 0.001) and decrease in inhibin B (p < 0.05) observed in CP treated mice. Administration of MOE prior to CP, improved sperm functional characteristics, decreased FSH and increased inhibin B levels. Expression of Abp was down-regulated while Transferrin, Fshr and Gata4 (Sertoli cell specific genes) were up-regulated in testis treated with CP. Administration of CP down-regulated the expression of Oct4 and Ddx4 (Spermatogonia specific genes). MOE administration was shown to ameliorate CP-induced damage by modulating the expression of genes specific to Sertoli and spermatogenic cells. Furthermore, MOE treatment reduced CP-induced DNA damage as evident from lower percentage of γ-H2AX positive spermatogonial cells. CONCLUSION: Administration of MOE mitigated CP-induced testicular damage by improving blood and, intra-testicular hormonal milieu as well as modulating the expression of genes pertaining to Sertoli and spermatogonial cells.

Associations of Fish Oil Supplement Use With Testicular Function in Young Men.

Importance: Many young men have poor semen quality, and the causes are often unknown. Supplement intake of ω-3 polyunsaturated fatty acid has been found to improve semen quality among men with infertility, but the association with semen quality among healthy men is unknown. Objective: To determine if intake of ω-3 fatty acid supplements is associated with testicular function as measured by semen quality and reproductive hormone levels among healthy men. Design, Setting, and Participants: This cross-sectional study included young Danish men from the general population recruited between January 1, 2012, and December 31, 2017, at compulsory examinations to determine their fitness for military service. Young unselected men were approached after the examination and invited to participate in a study of reproductive function, regardless of their fitness for military service. Data analysis was conducted from September 1, 2018, to June 30, 2019. Exposures: Intake of supplements, including fish oil, during the past 3 months. Main Outcomes and Measures: Semen quality, measured as volume, concentration, total sperm count, percentage of morphologically normal spermatozoa, and motility, and serum reproductive hormone levels, measured as follicle-stimulating hormone, luteinizing hormone, testosterone, free testosterone, and inhibin B levels. Results: Among 1679 young Danish men (median [interquartile range] age, 18.9 [18.7-19.4] years) recruited to participate, 98 men (5.8%) reported use of fish oil supplements during the past 3 months, of whom 53 (54.1%) reported intake on 60 or more days. After adjustment and compared with men with no supplement intake, men with fish oil supplement intake on fewer than 60 days had semen volume that was 0.38 (95% CI, -0.03 to 0.80) mL higher, and men with fish oil supplement intake on 60 or more days had semen volume that was 0.64 (95% CI, 0.15 to 1.12) mL higher (P for trend < .001). Similarly, testicular size in men with supplement intake on fewer than 60 days was 0.8 (95% CI, -0.2 to 1.9) mL larger and in men with fish oil supplement intake on 60 or more days was 1.5 (95% CI, 0.2 to 2.8) mL larger compared with men with no supplement intake (P for trend = .007). After adjustment, men with fish oil supplement intake had a 20% (95% CI, 9%-31%) lower follicle-stimulating hormone level and 16% (95% CI, 8%-24%) lower luteinizing hormone level compared with men with no supplement intake. There were no associations of intake of other supplements with measures of testicular function. Conclusions and Relevance: These findings suggest that intake of fish oil supplements was associated with better testicular function, which is less likely to be due to confounding by indication, as no associations of intake of other supplements with testicular function were found. This cross-sectional study did not examine the actual content of ω-3 fatty acids in the supplements; therefore, these findings need confirmation in well-designed randomized clinical trials among unselected men.

Correlation of oxidation-reduction potential with hormones, semen parameters and testicular volume.

Seminal oxidative stress (OS) is a major cause of male factor infertility and can be measured as oxidation-reduction potential (ORP). Studies showed significant negative relationships of ORP with sperm count, motility or DNA integrity. Since these parameters are also positively or negatively associated with reproductive hormones follicle-stimulating hormone (FSH), luteinising hormone (LH), testosterone, testicular volume and the occurrence of varicocele, it is important to understand the mechanistic relationship between ORP and hormonal and/or testicular parameters. Therefore, we studied the relationship between ORP levels, standard hormone profiles and testicular volume in infertile men with and without varicocele. Results show a highly significant negative relationship of ORP with testicular volume and significantly positive correlations with FSH and LH. Yet, when adding varicocele as covariate, the relationship with FSH/LH became nonsignificant. Contrary, the presence of varicocele had only a contributing influence on the association of ORP with the testis volume. No association was found with estradiol. We propose that since OS causes degeneration of Sertoli cell with testicular shrinkage, such negative effect would result in a negative feedback on the hypothalamus with less inhibin secretion. This may result in increased secretion of LH and FSH. Thus, systemic and/or local OS may be responsible for smaller testis volumes.

The role of active immunization against inhibin α-subunit on testicular development, testosterone concentration and relevant genes expressions in testis, hypothalamus and pituitary glands in Yangzhou goose ganders.

The present study was designed to investigate the potential role of immunization against inhibin on testicular development, plasma testosterone concentration and expression of relevant genes in hypothalamus, pituitary, Leydig and Sertoli cells in Yangzhou ganders. For this purpose, Yangzhou ganders, n = 30 were divided into groups A and B. Group B ganders were actively immunized against inhibin α-subunit, while group A ganders were immunized with bovine serum albumin (BSA), which served as control. Immunization against inhibin elevated testes weights. In addition, immunization against inhibin elevated GnRH, StAR, CYP11A1 and AMH mRNA transcription expressions as depicted by qRT-PCR. Furthermore, hypothalamic GnRH-I mRNA expressions were up regulated, while GnIH mRNA transcription expression showed reciprocal expression on day 227. LH-ß mRNA transcription expression remained unaffected. In conclusion, our findings suggest that active immunization against inhibin affect spermatogenesis and testicular development through regulations of hypothalamic, pituitary and testicular genes expressions.

Identification of some novel pyrazolo[1,5-a]pyrimidine derivatives as InhA inhibitors through pharmacophore-based virtual screening and molecular docking.

The InhA inhibitors play key role in mycolic acid synthesis by preventing the fatty acid biosynthesis pathway. In this present article, Pharmacophore modelling and molecular docking study followed by in silico virtual screening could be considered as effective strategy to identify newer enoyl-ACP reductase inhibitors. Pyrrolidine carboxamide derivatives were opted to generate pharmacophore models using HypoGen algorithm in Discovery studio 2.1. Further it was employed to screen Zinc and Minimaybridge databases to identify and design newer potent hit molecules. The retrieved newer hits were further evaluated for their drug likeliness and docked against enoyl acyl carrier protein reductase. Here, novel pyrazolo[1,5-a]pyrimidine analogues were designed and synthesized with good yields. Structural elucidation of synthesized final molecules was perform through IR, MASS, 1H-NMR, 13C-NMR spectroscopy and further tested for its in vitro anti-tubercular activity against H37Rv strain using Microplate Alamar blue assay (MABA) method. Most of the synthesized compounds displayed strong anti-tubercular activities. Further, these potent compounds were gauged for MDR-TB, XDR-TB and cytotoxic study.

Melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, hormonal profile and total antioxidant capacity: A placebo-controlled, double-blind trial.

In this study, we aimed at investigating the impact of melatonin supplementation on semen parameters, hormonal profile and total antioxidant capacity after varicocelectomy. Infertile male patients who were diagnosed with varicocele and underwent subinguinal varicocelectomy were included in the study. After performing subinguinal varicocelectomy, the patients were randomised into two groups: 27 receiving melatonin for 3 months and 27 as the placebo-controlled group receiving placebo for 3 months. The pre-operative parameters of semen analyses, hormonal profile and seminal oxidative stress status of both groups were compared with those of post-operative parameters. There were statistically significant improvements in post-operative parameters of semen analyses (sperm concentration, motility and proportions of normally formed spermatozoa), peripheral blood inhibin B and total antioxidant capacity in melatonin group compared with placebo group. In conclusion, melatonin therapy adds extra benefit to varicecelectomy in terms of sperm parameters, peripheral blood inhibin B and total antioxidant capacity; however, further studies including large number of samples are needed to make a proper decision on melatonin supplementation after varicocelectomy.

Cancer Treatment and Bone Health.

Considerable advances in oncology over recent decades have led to improved survival, while raising concerns about long-term consequences of anticancer treatments. In patients with breast or prostate malignancies, bone health is a major issue due to the high risk of bone metastases and the frequent prolonged use of hormone therapies that alter physiological bone turnover, leading to increased fracture risk. Thus, the onset of cancer treatment-induced bone loss (CTIBL) should be considered by clinicians and recent guidelines should be routinely applied to these patients. In particular, baseline and periodic follow-up evaluations of bone health parameters enable the identification of patients at high risk of osteoporosis and fractures, which can be prevented by the use of bone-targeting agents (BTAs), calcium and vitamin D supplementation and modifications of lifestyle. This review will focus upon the pathophysiology of breast and prostate cancer treatment-induced bone loss and the most recent evidence about effective preventive and therapeutic strategies.

[Effects of moxibustion at Shenshu (BL 23) on level of sex hormones and AMH in sub-health peri-menopausal women].

OBJECTIVE: To compare the effects between moxibustion at Shenshu (BL 23) and oral administration of Vitamin E on quality of life and mechanism in sub-health perimenopausal women, aiming to provide clinical evidence of moxibustion for health care of sub-health female. METHODS: Sixty participants were randomly divided into a moxibustion group and a medication group, 30 cases in each one. The volunteers in the moxibustion group were treated with mild moxibustion at bilateral Shenshu (BL 23) for 15 min, once a day; five treatments were considered as a course of treatment, and totally 4 courses were given with an interval of 2 days between courses. The volunteers in the medication group were treated with oral administration of soft capsule of Vitamin E, once a day, continuously for 28 days. The scores of quality of life and serum levels of sex hormones, anti-Mullerian hormone (AMH) and inhibin B (INHB) were measured before and after treatment in the two groups. RESULTS: ① Compared before treatment, the scores of quality of life in the two groups were both significantly increased after treatment (both P<0.01), which was more superior in the moxibustion group (P<0.05). ② Compared before treatment, the serum levels of estradiol (E2) and progesterone in the moxibustion group were significantly increased after treatment, especially for premenopausal volunteers (both P<0.01). ③ After moxibustion, the serum level of AMH was significantly improved (P<0.01), however, the effect on INHB had no statistical difference (P>0.05). CONCLUSIONS: Moxibustion could effectively improve the quality of life in sub-health perimenopausal women, regulate the levels of sex hormones and AMH, improve ovarian reserve function, and delay ovarian aging.

Effect of Zinc and Melatonin on Oxidative Stress and Serum Inhibin-B Levels in a Rat Testicular Torsion-Detorsion Model.

The present study was aimed to examine the effects of 3-week zinc and melatonin administration on testicular tissue injury and serum Inhibin-B levels caused by unilateral testicular torsion-detorsion in rats. The study was performed on 60 Wistar Albino-type adult male rats. The animals were allocated to 6 groups in equal numbers. 1. Control; 2. Sham; 3. Ischemia-reperfusion; 4. Zinc + ischemia-reperfusion; 5. Melatonin + ischemia-reperfusion; 6. Zinc + melatonin + ischemia-reperfusion. Zinc and melatonin were administered before ischemia-reperfusion at doses of 5 and 3 mg/kg respectively, by intraperitoneal route for a period of 3 weeks. Testicular torsion-detorsion procedures consisted of ischemia for 1 h and then reperfusion for another hour of the left testis. Blood and testicular tissue samples were collected to analyze erythrocyte and tissue GSH and plasma and tissue MDA, Inhibin-B levels. The highest erythrocyte and testis GSH values were found in zinc, melatonin, and zinc + melatonin groups (p < 0.001). Torsion-detorsion group has significantly lower erythrocyte GSH levels and higher plasma MDA values (p < 0.001). Serum inhibin-B and spermatogenic activity levels in the torsion-detorsion group were also significantly lower than those in the other groups (p < 0.001). However, zinc-, melatonin-, and melatonin + zinc-supplemented groups have higher inhibin-B and spermatogenetic activity (p < 0.001). The results of the study show that zinc, melatonin, and melatonin + zinc administration partially restores the increased oxidative stress, as well as the reduced inhibin-B and spermatogenic activity levels in testes ischemia-reperfusion in rats. Suppressed inhibin-B levels in the testicular tissue may be a marker of oxidative stress.

The Effect of Folate and Folate Plus Zinc Supplementation on Endocrine Parameters and Sperm Characteristics in Sub-Fertile Men: A Systematic Review and Meta-Analysis.

PURPOSE: To evaluate the effect of folate and folate plus zinc supplementation on endocrine parameters and sperm characteristics in sub fertile men. MATERIALS AND METHODS: We conducted a systematic review and meta-analysis. Electronic databases of Medline, Scopus , Google scholar and Persian databases (SID, Iran medex, Magiran, Medlib, Iran doc) were searched from 1966 to December 2016 using a set of relevant keywords including "folate or folic acid AND (infertility, infertile, sterility)".All available randomized controlled trials (RCTs), conducted on a sample of sub fertile men with semen analyses, who took oral folic acid or folate plus zinc, were included. Data collected included endocrine parameters and sperm characteristics. Statistical analyses were done by Comprehensive Meta-analysis Version 2. RESULTS: In total, seven studies were included. Six studies had sufficient data for meta-analysis. "Sperm concentration was statistically higher in men supplemented with folate than with placebo (P < .001)". However, folate supplementation alone did not seem to be more effective than the placebo on the morphology (P = .056) and motility of the sperms (P = .652). Folate plus zinc supplementation did not show any statistically different effect on serum testosterone (P = .86), inhibin B (P = .84), FSH (P = .054), and sperm motility (P = .169) as compared to the placebo. Yet, folate plus zinc showed statistically higher effect on the sperm concentration (P < .001), morphology (P < .001), and serum folate level (P < .001) as compared to placebo. CONCLUSION: Folate plus zinc supplementation has a positive effect on sperm characteristics in sub fertile men. However, these results should be interpreted with caution due to the important heterogeneity of the studies included in this meta-analysis. Further trials are still needed to confirm the current findings.

Ameliorative effect of omega-3 on spermatogenesis, testicular antioxidant status and preimplantation embryo development in streptozotocin-induced diabetes in rats.

PURPOSE: The present study was done to determine the ameliorative effect of polyunsaturated fatty acid omega-3 against experimental diabetes-induced damages on testicular tissue, sperm parameters and preimplantation embryo development in rat model. METHODS: Thirty-two mature male rats were divided into two control and test groups. The experimental diabetes (50 mg kg-1 streptozotocin, ip) was induced in test group and subdivided into non-treated diabetic, 300 and 600 mg kg-1 omega-3-treated (orally by gavage) groups. The rats in control group received 0.5 ml saline using intra-gastric gavage. Following 45 days, general histopathological changes, serum level of testosterone, inhibin B, glucose, and sperm parameters, testicular total antioxidant capacity (TAC) and malondialdehyde (MDA) content were analyzed. The mitochondria-dependent apoptosis was investigated by assessing the Bcl-2 and caspase-3 expression as well as DNA fragmentation. Finally, the in vitro fertilization (IVF) potential was examined by evaluating preimplantation embryo developing. RESULTS: The omega-3 significantly ameliorated the diabetes-induced histological damages, diminished serum level of glucose, testicular MDA content, and enhanced the serum testosterone, inhibin B and testicular TAC. The animals in omega-3-treated groups exhibited a significant (p < 0.05) up-regulation in Bcl-2, as well as remarkable (p < 0.05) down-regulation in caspase-3 expression compared to non-treated diabetic rats. Moreover, the omega-3 maintained DNA integrity, improved sperm quality as well as preimplantation embryo development. CONCLUSION: Our data showed that the omega-3 (especially at 600 mg kg-1 dose level) effectively ameliorates the experimental diabetes-induced infertility in rats by up-regulating the testicular endocrine and antioxidant statuses, preventing mitochondria-dependent apoptosis pathway and potentially improving the sperm quality.

Fatty acid intake in relation to reproductive hormones and testicular volume among young healthy men.

Emerging evidence suggests that dietary fats may influence testicular function. However, most of the published literature on this field has used semen quality parameters as the only proxy for testicular function. We examined the association of fat intake with circulating reproductive hormone levels and testicular volume among healthy young Spanish men. This is a cross-sectional study among 209 healthy male volunteers conducted between October 2010 and November 2011 in Murcia Region of Spain. Participants completed questionnaires on lifestyle, diet, and smoking, and each underwent a physical examination, and provided a blood sample. Linear regression was used to examine the association between each fatty acid type and reproductive hormone levels and testicular volumes. Monounsaturated fatty acids intake was inversely associated with serum blood levels of calculated free testosterone, total testosterone, and inhibin B. A positive association was observed between the intake of polyunsaturated fatty acids, particularly of omega-6 polyunsaturated fatty acids, and luteinizing hormone concentrations. In addition, the intake of trans fatty acids was associated with lower total testosterone and calculated free testosterone concentrations (P trend = 0.01 and 0.02, respectively). The intake of omega-3 polyunsaturated fatty acids was positively related to testicular volume while the intake of omega-6 polyunsaturated fatty acids and trans fatty acids was inversely related to testicular volume. These data suggest that fat intake, and particularly intake of omega 3, omega 6, and trans fatty acids, may influence testicular function.

[Effect of Wenshen Yangxue Recipe on Inhibin-Activin-Follistatin System in Anovulatory Rats].

Objective To explore the effect of Wenshen Yangxue Recipe (WYR) on inhibin-ac- tivin-follistatin (INH-ACT-FS) system and gonadal hormone level in anovulatory rats. Methods Anovula- tory rat model was established in 76 rats (9 days old) by subcutaneous injecting testosterone propionate (1. 25 mg/0. 05 mL for each rat) from the nape. Totally 58 successfully modeled rats were divided into 5 groups according to random digit table, i.e., the model group (n =10), the Western medicine (WM) group (n =12), high, middle, and low dose WYR groups (n =12). Besides, another ten 22-day old rats were recruited as a normal group. Distilled water was daily administered to rats in the normal group and the model group by gastrogavage. Clomiphene citrate (0. 58 mg/100 g) was daily administered to rats in the WM group for 5 successive days. WYR at 5. 2, 2. 6, 1. 3 mg/100 g was daily administered to rats in high, middle, and low dose WYR groups for 21 successive days. Levels of follicular stimulating hormone (FSH) , luteinizing hormone (LH) , estradiol (E2) , progesterone (P) , and prolactin (PRL) were detected using radioimmunoassay. Contents of inhibin (INH) , activin (ACT) , and follistatin (FS) were measured using ELISA. Results Compared with the normal group, serum levels of FSH and LH increased, and P level decreased in the model group (P <0. 05) ; INH level decreased and FS level increased in the model group (P<0. 05). Compared with the model group, serum FSH level decreased in the WM group and 3 WYR groups, P level decreased in the WM group (P <0. 05); INH increased and FS levels decreased in the WM group and 3 WYR groups; ACT level increased in the high dose WYR group, with statistical differ- ence (P <0. 05). Conclusion WYR promoted follicular development possibly through regulating INH- ACT-FS system and gonadal hormone level.

[Treatment of Anovulatory Infertility with Shen Deficiency Syndrome by ZHU's Tiaojing Cuyun Recipe: a Clinical Evaluation].

OBJECTIVE: To explore the molecular biological mechanism of ZHU's Tiaojing Cuyun Recipe (TCR) for treating anovulatory infertility patients with Shen deficiency syndrome (SDS) by observing its clinical efficacy. METHODS: Using randomized blocking methods, 80 patients were assigned to the treatment group (40 cases) and the control group (40 cases). Patients with regular menstrual cycle started medication from the 5th day of menstruation. Those with irregular menstrual cycle first took progesterone till withdrawal bleeding ,and then started medication from the 5th day of vaginal bleeding. Patients in the treatment group took ZHU's TCR, one dose per day, while those in the control group took Clomifene Citrate (CC), 50 mg per day. Three menstrual cycles consisted of one therapeutic course, a total of 2 courses. Clinical efficacy such as pregnancy rates and abortion rates were observed. Ovulation indices (the maximal diameter of mature follicles, luteinized follicles, ovulational follicles, and the endometrial thickness on the ovulation day), SDS, and integrals of menstrual symptoms were monitored before and after treatment. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) , and estradiol (E2) were determined using chemiluminescent immunoassay before treatment and after on therapeutic course. Serum levels of activin A (ACTA), inhibin B (INHB), and follistatin (FS) were detected using double antibody sandwich ELISA. RESULTS: Compared with the control group, the pregnancy rate was obviously elevated and the abortion rate was obviously lowered in the treatment group (P <0. 05). Ovulation rates of mature follicles and luteinizing follicles decreased more in the treatment group (P < 0.05). Compared with before treatment, integrals for SDS were lower, the maximal diameter of pre-ovulational follicles was increased, and integrals for menstrual symptoms in non-pregnant patients of the two groups were obviously lowered. Meanwhile, the endometrial thickness on the ovulation day was increased in the treatment group after treatment, but reduced in the control group (P < 0.05, P < 0.01). Compared with the control group, integrals for SDS were decreased, and the maximal diameter of pre-ovulational follicles was lowered in the treatment group after treatment (P < 0.05, P < 0.01). Integrals for SDS and the difference in the endometrial thickness on the ovulation day were increased, but the difference in the maximal diameter of pre-ovulational follicles were reduced (P < 0.05, P < 0.01). In the treatment group serum levels of E2 and ACTA increased more after one therapeutic course than before treatment (P < 0.01), but serum levels of INHB and FS decreased more after one therapeutic course than before treatment (P < 0.05). In the control group serum levels of FSH and ACTA increased more, and the serum level of FS decreased more after one therapeutic course than before treatment (P < 0.05, P < 0.01). Compared with the control group, serum levels of FSH and ACTA increased more, and serum levels of INHB decreased more in the treatment group after one therapeutic course than before treatment (P < 0.05, P < 0.01). CONCLUSIONS: ZHU'sTCR could improve SDS of anovulatory infertility patients, regulate the follicular development, and elevate the pregnancy rate. Its actions might be associated with regulating their sex hormones, expressions of ovary local factors such as INHB, ACTA, and FS.

Endocrinology of the Menopause.

In women, age-related changes in ovarian function begin in the mid-30s with decreased fertility and compensatory hormonal changes in the hypothalamus-pituitary-gonadal axis that maintain follicle development and estrogen secretion in the face of a waning pool of ovarian follicles. The menopause transition is characterized by marked variability in follicle development, ovulation, bleeding patterns, and symptoms of hyper- and hypoestrogenism. The menopause, which is clinically defined by the last menstrual period, is followed by the consistent absence of ovarian secretion of estradiol.

Fluoride exposure changed the structure and the expressions of reproductive related genes in the hypothalamus-pituitary-testicular axis of male mice.

Numerous studies have shown that fluoride exposure adversely affected the male reproductive function, while the molecular mechanism is not clear. The present study was to investigate the effects of fluoride exposure (60 days) on the expressions of reproductive related genes, serum sex hormone levels and structures of the hypothalamus-pituitary-testicular axis (HPTA), which plays a vital role in regulating the spermatogenesis in male mice. In this study, 48 male mice were administrated with 0, 25, 50, and 100 mg/L NaF through drinking water. Results showed that the malformation ratio of sperm was significantly increased (P<0.05). At transcriptional level, the expression levels of follicle-stimulating hormone receptor (FSHR), luteinizing hormone receptor (LHR), inhibin alpha (INHα), inhibin beta-B (INHßB), and sex hormone binding globulin (SHBG) mRNA in testis were significantly decreased (P<0.05). Moreover, histological lesions in testis and ultrastructural alterations in hypothalamus, pituitary and testis were obvious. However, the same fluoride exposure did not lead to significant changes of related mRNA expressions in hypothalamus and pituitary (P>0.05). Also, there were no marked changes in serum hormones. Taken together, we conclude that the mechanism of HPTA dysfunction is mainly elucidated through affecting testes, and its effect on hypothalamus and pituitary was secondary at exposure for 60 days.

The Effect of Vitis vinifera L. Juice on Serum Levels of Inhibin B, Sperm Count in Adult Male Rats

PURPOSE: Vitis vinifera is a species of Vitis that is native to the Mediterranean region, central Europe, and southwestern Asia, and has been used as a drug in traditional medicine. Traditional medicinal plants have been used for medical purposes with increasing effectiveness. It is important to identify drugs that inhibit spermatogenesis. Therefore, the present study aimed to investigate the effect of grape juice (GJ) on serum levels of inhibin B and sperm count in normal male rats. MATERIALS AND METHODS: Thirty-five adult male rats were randomly divided into five groups, each containing seven rats. Rats in the control group received 1 mL of normal saline over the course of the study. The experimental groups received GJ (100, 200, 400, and 1,600 mg/kg, orally, for 35 days consecutively). At the end of the treatment period, fertility indices were measured, including body weight difference, sex organ weight, sperm motility and count, epididymal sperm reserve, daily sperm production (DSP), and serum inhibin B levels. RESULTS: We found that GJ reduces body weight difference, was associated with decreased sperm motility and count in all treatment groups (p< or =0.05 and p< or =0.001, respectively). Moreover, DSP was significantly decreased in all treatment groups compared to the control group (p< or =0.05), except in the group receiving 100 mg/kg of GJ. Inhibin B levels were significantly decreased in all treatment groups (p< or =0.05). CONCLUSIONS: The results of our study suggest that GJ in all doses, but especially in higher doses, may decrease fertility in male rats.

Treatment of Anovulatory Infertility with Shen Deficiency Syndrome by ZHU's Tiaojing Cuyun Recipe: a Clinical Evaluation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the molecular biological mechanism of ZHU's Tiaojing Cuyun Recipe (TCR) for treating anovulatory infertility patients with Shen deficiency syndrome (SDS) by observing its clinical efficacy.</p><p><b>METHODS</b>Using randomized blocking methods, 80 patients were assigned to the treatment group (40 cases) and the control group (40 cases). Patients with regular menstrual cycle started medication from the 5th day of menstruation. Those with irregular menstrual cycle first took progesterone till withdrawal bleeding ,and then started medication from the 5th day of vaginal bleeding. Patients in the treatment group took ZHU's TCR, one dose per day, while those in the control group took Clomifene Citrate (CC), 50 mg per day. Three menstrual cycles consisted of one therapeutic course, a total of 2 courses. Clinical efficacy such as pregnancy rates and abortion rates were observed. Ovulation indices (the maximal diameter of mature follicles, luteinized follicles, ovulational follicles, and the endometrial thickness on the ovulation day), SDS, and integrals of menstrual symptoms were monitored before and after treatment. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) , and estradiol (E2) were determined using chemiluminescent immunoassay before treatment and after on therapeutic course. Serum levels of activin A (ACTA), inhibin B (INHB), and follistatin (FS) were detected using double antibody sandwich ELISA.</p><p><b>RESULTS</b>Compared with the control group, the pregnancy rate was obviously elevated and the abortion rate was obviously lowered in the treatment group (P <0. 05). Ovulation rates of mature follicles and luteinizing follicles decreased more in the treatment group (P < 0.05). Compared with before treatment, integrals for SDS were lower, the maximal diameter of pre-ovulational follicles was increased, and integrals for menstrual symptoms in non-pregnant patients of the two groups were obviously lowered. Meanwhile, the endometrial thickness on the ovulation day was increased in the treatment group after treatment, but reduced in the control group (P < 0.05, P < 0.01). Compared with the control group, integrals for SDS were decreased, and the maximal diameter of pre-ovulational follicles was lowered in the treatment group after treatment (P < 0.05, P < 0.01). Integrals for SDS and the difference in the endometrial thickness on the ovulation day were increased, but the difference in the maximal diameter of pre-ovulational follicles were reduced (P < 0.05, P < 0.01). In the treatment group serum levels of E2 and ACTA increased more after one therapeutic course than before treatment (P < 0.01), but serum levels of INHB and FS decreased more after one therapeutic course than before treatment (P < 0.05). In the control group serum levels of FSH and ACTA increased more, and the serum level of FS decreased more after one therapeutic course than before treatment (P < 0.05, P < 0.01). Compared with the control group, serum levels of FSH and ACTA increased more, and serum levels of INHB decreased more in the treatment group after one therapeutic course than before treatment (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>ZHU'sTCR could improve SDS of anovulatory infertility patients, regulate the follicular development, and elevate the pregnancy rate. Its actions might be associated with regulating their sex hormones, expressions of ovary local factors such as INHB, ACTA, and FS.</p>