Oxidative stress in granulosa cells contributes to poor oocyte quality and IVF-ET outcomes in women with polycystic ovary syndrome / 医学前沿

The increased levels of intracellular reactive oxygen species (ROS) in granulosa cells (GCs) may affect the pregnancy results in women with polycystic ovary syndrome (PCOS). In this study, we compared the in vitro fertilization and embryo transfer (IVF-ET) results of 22 patients with PCOS and 25 patients with tubal factor infertility and detected the ROS levels in the GCs of these two groups. Results showed that the PCOS group had significantly larger follicles on the administration day for human chorionic gonadotropin than the tubal factor group (P 0.05). PCOS group had slightly lower fertilization, cleavage, grade I/II embryo, clinical pregnancy, and implantation rates and higher miscarriage rate than the tubal factor group (P > 0.05). We further found a significantly higher ROS level of GCs in the PCOS group than in the tubal factor group (P < 0.05). The increased ROS levels in GCs caused GC apoptosis, whereas NADPH oxidase 2 (NOX2) specific inhibitors (diphenyleneiodonium and apocynin) significantly reduced the ROS production in the PCOS group. In conclusion, the increased ROS expression levels in PCOS GCs greatly induced cell apoptosis, which further affected the oocyte quality and reduced the positive IVF-ET pregnancy results of women with PCOS. NADPH oxidase pathway may be involved in the mechanism of ROS production in GCs of women with PCOS.

Relationship between obesity and serum testosterone level in 1 166 adult males / 重庆医学

Objective To explore the relationship between obesity and serum testosterone level among community adult males.Methods The stratified cluster sampling method was adopted to extract 1 166 male residents aged 20 years old and over in Zunyi City and conduct the questionnaire survey and physical examination.Fasting venous blood samples were collected for detecting the serum total testosterone(TT),sex hormone binding globulin(SHBG) and luteinizing hormone(LH).Then free testosterone index(cFT),free testosterone index(FTI) and testosterone secreting index(TSI) were calculated.Results The average levels of serum TT,SHBG,cFT,FTI,TSI and LH in 1 166 adult males were(16.83±4.90)nmol/L,(42.15 ± 20.21)nmol/L,(0.31 ±0.10)nmol/L,(0.46 ± 0.19) IU/L,(3.40 ± 2.04) IU/L and(6.77 ± 5.14) IU/L respectively;the Pearson correlation analysis showed that WHtR,BMI and WC were negatively correlated with TT,SHBG,TSI and LH,and were positively correlated with FTI,WHtR was negatively correlated with cFT,BMI and WC were negatively correlated with LH,the differences were statistically significant(P＜0.05).The multivariate linear regression analysis shown that WHtR was negatively correlated with TT,SHBG,TSI and LH;and was positively correlated with FTI,the differences were statistically significant(P＜0.01).Conclusion Adult male central obesity may be associated with the decrease of serum TT,SHBG,TSI and LH levels,and increase of FTI level.

Urgent need for contraceptive education and services in Chinese unmarried undergraduates: A multi-campus survey / 华中科技大学学报（医学）（英德文版）

In order to ascertain prevalence rate of premarital sexual intercourse, unintended pregnancy and abortion, and evaluate associated factors of unintended pregnancy among undergraduates from all over China, the representative sample of unmarried undergraduates was obtained by using a multi-stage, stratified, probability cluster design, and data were collected by using a survey questionnaire. 62 326 available responders were gained. 11.6% of them acknowledged having experiences of premarital sexual intercourse (standardized prevalence rate of sexual intercourse was 13.8%). 31.5% of students active in premarital sex acknowledged undergoing unintended pregnancy. 76.2% of pregnant students selected abortion to end it. Of students active in premarital sex, 46.2% used contraception at the first sexual intercourse, 28.2% replied "always" using contraception in sexual intercourse. The rate of using condoms, oral contraceptives (OCs), and withdrawal among students who had used contraception was 52.0%, 31.0%, and 27.2% respectively. "No preparation for sex" (40.3%), "pleasure decrement" (32.1%), "won't-be-pregnancy in occasional sexual intercourse" (30.2%) were their common excuses for using no contraception. The identified risk factors for unintended pregnancy among students active in premarital sex by multivariate analysis were as follows: having no steady lover [having no steady lover vs having a steady lover: odds ratio (OR), 1.875; 95% confidence interval (CI), 1.629-2.158], unaware of the course of conception (unaware vs aware: OR, 2.023; 95% CI, 1.811-2.260), considering abortion not endanger women's physical and mental health (no endangerment vs endangerment: OR, 2.659; 95% CI, 2.265-3.121), nonuse of contraception (never use vs always use: OR, 1.682; 95% CI, 1.295-2.185). Medical students were not less likely to experience an unintended pregnancy than nonmedical students (OR, 1.111; 95% CI, 0.906-1.287). The substantial proportion of unintended pregnancy among undergraduates indicates a need for convenient and targeted contraceptive education and services.

The role of brain-derived neurotrophic factor in mouse oocyte maturation in vitro / 华中科技大学学报（医学）（英德文版）

Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM), but the role of BDNF in oocyte maturation at cellular level is not still clear. In this study, mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage. Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy. The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs. 5.92%; P<0.05). Further, BDNF did not accelerate nuclear maturation of IVM oocytes. For the BDNF-treated oocytes at meiosis I, Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control, and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control. These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes. In conclusion, BDNF can promote the developmental competence of mouse IVM oocytes, by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis.

Clinicopathological features and prognosis of small cell carcinoma of the cervix / 华中科技大学学报（医学）（英德文版）

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis, treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage I b1, two stage I b2, one stage IIa, one stage IIb, and one stage IIIb. On the basis of their stages of condition, one subject with stage III b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively. It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.

The role of brain-derived neurotrophic factor in mouse oocyte maturation in vitro / 华中科技大学学报（医学）（英德文版）

Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM), but the role of BDNF in oocyte maturation at cellular level is not still clear. In this study, mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage. Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy. The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs. 5.92%; P<0.05). Further, BDNF did not accelerate nuclear maturation of IVM oocytes. For the BDNF-treated oocytes at meiosis I, Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control, and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control. These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes. In conclusion, BDNF can promote the developmental competence of mouse IVM oocytes, by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis 1.

Attractin gene deficiency contributes to testis vacuolization and sperm dysfunction in male mice / 华中科技大学学报（医学）（英德文版）

The effect of loss-of-function of Attractin (Atrn) on the male mouse reproduction system was examined in the study. The weights and pathological changes of testes and epididymes were compared between Atrn mutant (Atrn ( mg-3J )) mice and wild-type mice (C3HeB/FeJ) at different months of age. The number and motility of sperms were measured in the mutant and control mice. Furthermore, the testicular lactate dehydrogenase (LDH) and succinate dehydrogenase (SDH) in these animals were detected. The fertility potential of the sperms was observed in vivo and in vitro. The results showed that the testes of 3-month-old Atrn ( mg-3J ) mice experienced no significantly different pathological changes from the control mice at the same month of age but the SDH activity was substantially reduced. In the 5-month-old mutant mice, as compared with the control mice, mild vacuolation was found in the testes, the density and motility of sperms were decreased in the epididymes, the sperm fertility was impaired and the testicular enzyme activity was reduced. It is concluded that the age-related Atrn gene progressively loses its function and can cause testis vacuolation and impaired sperm function, which may be responsible for the impairment of male reproductive ability.

Efficacy and safety of drospirenone-ethinylestradiol on contraception in healthy Chinese women: a multicenter randomized controlled trial / 中华妇产科杂志

Objective To evaluate the contraception efficacy, mode of bleeding, side effects and other positive effects of drospirenone-ethinylestradiol (Yasmin) in healthy Chinese women. Methods This was a multicenter, randomized, control study of 768 healthy Chinese women who consulted about contraception. The subjects were randomized into Yasmin group (30 μg ethinylestradiol plus 3 mg drospirenone, 573 cases) or desogestrel group (30 μg ethinylestradiol plus 150 μg desogestrel, 195 cases) with the ratio of 3: 1. Each individual was treated for 13 cycles. Further visits were required at cycle 4, cycle 7, cycle 10 and cycle 13 of treatment. Weight, height, body mass index were evaluated at each visit. The menstrual distress questionnaire (MDQ) was given to the women at baseline, visit 3 (cycle 7) and visit 5 (after cycle 13). Results The values of basal features were similar between two groups (P> 0.05). The Pearl index (method failure) of Yasmin was 0. 208/hundred women year which was lower than that of desogestrel (0. 601/hundred women year). The mode of bleeding was similar between two groups after trial without showing any significant difference. According to MDQ subscale, the improvement of water retention and increasing appetite during inter-menstrual period and water retention and general well-being during menstrual period in the Yasmin group ( -0. 297, -0. 057, 0. 033, 0. 150 respectively) was more obvious than that in the desogestrel group ( - 0. 108, 0. 023, 0. 231, - 0. 023 respectively) with a significant difference (P < 0. 05 ). Some other values which improved in beth two groups, especially the improvement of breast tenderness and pain and skin abnormality in Yasmin group (18.0%, 89/494; 12. 6%, 62/494) was more distinct than that in desogestrel group (11.3%, 19/168; 5.4%, 9/168). The mean weight increased in desogestrel group (0. 57 kg) while it decreased in Yasmin group ( -0. 28 kg) with a significant difference (P < 0. 01 ). Conclusions Both Yasmin and desogestrel have good efficacy on contraception and similar modes of menstrual bleeding. Yasmin is better than desogestrel in terms of weight control and premenstrual syndrome of oral contraceptive.

P2X3 receptor expression in DRG and bladder detrusor of an animal model of neurogenic bladder and ure-thra / 中华物理医学与康复杂志

Objective To investigate the P2X3 receptor expression in L6-S1 dorsal root ganglion (DRG)and bladder detrusor in a rat model of neurogenic bladder and urethra. Methods Eighty Sprague-Dawley rats wererecruited and randomly divided into a sacral injury group, a suprasacral injury group and a control group. Spinal tran-section was performed to establish the animal model of neurogenic bladder and urethra in rats of the sacral injurygroup and suprasacral injury group. Check the P2X3 receptor expression in DBG and bladder detrusor among thethree groups by Western blot test at 20 days after model establishment. Results P2X3 receptor expression in L6-S1DRG of sacral injury group was significantly less than that of the suprasacral injury group, which was in turn signifi-cantly higher than that of the control group. P2X3 receptor expression in bladder detrusor of sacral injury group wassignificantly lower than that of the suprasacral injury group, which was in turn significantly higher than that of thecontrol group. Conclusion There was close relationship between P2X3 receptor expression and dysfunction of blad-der and urethra.

Endogenous ouabain in hypertensive disorder complicating pregnancy / 华中科技大学学报（医学）（英德文版）

Expression of endogenous ouabain in placenta and the concentrations of serum ET-1 and NO were examined in 30 patients with hypertensive disorder complicating pregnancy (HDCP) and 30 healthy pregnant women to investigate the effect of endogenous ouabain on HDCP. Compared with the healthy pregnant group, the expression of endogenous ouabain dramatically increased in the HDCP groups (P<0.01). There was a significantly positive correlation between the expression of endogenous ouabain with ET-1 (r=0.5567, P<0.01), while the correlation of endogenous ouabain and NO was significantly negative (r=-0.6895, P<0.01). As expected, the correlation between ET-1 and NO was negative (r=-0.7796, P<0.01). ET-1 concentrations of maternal and cord sera in HDCP groups were significantly higher in comparison with healthy pregnant group (P<0.01). On the contrast, NO concentrations were much lower in the maternal and cord sera of HDCP groups as compared with healthy pregnant group (P<0.01). Our data suggest that endogenous ouabain is directly involved in the nosogenesis of HDCP, with accompanying decreased NO and the elevated of ET-1.

Endogenous Ouabain in Hypertensive Disorder Complicating Pregnancy / 华中科技大学学报（医学）（英德文版）

Expression of endogenous ouabain in placenta and the concentrations of serum ET-1 and NO were examined in 30 patients with hypertensive disorder complicating pregnancy (HDCP) and 30 healthy pregnant women to investigate the effect of endogenous ouabain on HDCP. Compared with the healthy pregnant group, the expression of endogenous ouabain dramatically increased in the HDCP groups (P<0.01). There was a significantly positive correlation between the expression of en- dogenous ouabain with ET-1 (r=0.5567, P<0.01), while the correlation of endogenous ouabain and NO was significantly negative (r=-0.6895, P<0.01). As expected, the correlation between ET-1 and NO was negative (r=-0.7796, P<0.01). ET-1 concentrations of maternal and cord sera in HDCP groups were significantly higher in comparison with healthy pregnant group (P<0.01). On the con- trast, NO concentrations were much lower in the maternal and cord sera of HDCP groups as com- pared with healthy pregnant group (P<0.01). Our data suggest that endogenous ouabain is directly involved in the nosogenesis of HDCP, with accompanying decreased NO and the elevated of ET-1.

Effects of murine cytomegalovirus infection on sperm viability in mice / 华中科技大学学报（医学）（英德文版）

In order to explore the effects of testicular infection of murine cytomegalovirus (MCMV) on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups: an experimental group (n = 56) and a control group (n = 35). The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1, 1. 5, 2, 4, 6, 9 and 14 post-inoculation (D1) 1. 5, 2, 4, 6, 9 and 14 PI). The MCMV M83 mRNA gene was detected in the testis by in situ hybridization (ISH) with MCMV late-mRNA probe labeled with digoxin. Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5 PI (P 0.05). This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently.

Effects of Murine Cytomegalovirus Infection on Sperm Viability in Mice / 华中科技大学学报（医学）（英德文版）

In order to explore the effects of testicular infection of murine cytomegalovirus (MCMV) on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups: an experimental group (n=56) and a control group (n= 35). The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1,1.5, 2, 4, 6, 9 and 14 post-inoculation (D1, 1.5,2, 4, 6, 9 and 14 PI). The MCMV M83 mRNA gene was detected in the testis by in situ hybridization (ISH) with MCMV late-mRNA probe labeled with digoxin.Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5PI (P＜ 0.05). No statistically significant difference in the sperm viability was found after D2 PI between two groups (P＞0.05). This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently.

Experimental study of the effect of rhTNF-alpha on human sperm mitochondrial function and motility in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effect of rhTNF-alpha on human sperm mitochondrial function and motility in vitro.</p><p><b>METHODS</b>Fifty-six semen samples collected by masturbation were analyzed according to WHO protocols. Semen samples from 40 healthy men were prepared using Percoll centrifugation. Sperm suspension was diluted to a concentration of 10 x 10(6)/ml in Ham's F10 medium. Sperm samples were incubated with rhTNF-alpha solution (final concentration 0.03 microg/L, 0.06 microg/L, 0.09 microg/L and 0.27 microg/L, respectively) for 0.5 h, 1 h, 2 h, 3 h and 4 h at 37 degrees C in 5% CO2, and comparative studies were made with a control group. Ten microl sperm samples were examined with CASA technique, 250 microl stained in the presence of 10 microg/ml Rh123 and PI, and mitochondrial function analyzed by flow cytometry.</p><p><b>RESULTS</b>Significant differences were found between the experimental groups (final concentration 0.06 microg/L, 0.09 microg/L and 0.27 microg/L) and the control group in viability, straight line velocity, curvilinear velocity, average path velocity, progressive motility of human sperm and the number of spermatozoa with normal mitochondrial function (P < 0.01) except the final concentration 0.03 microg/L group (P > 0.05). Motility of human sperm lowered with the increase of rhTNF-alpha concentration and incubation time, and r values were 0.675, 0.691, 0.762, 0.693, 0.724 and 0.571, 0.594, 0.752, 0.791, 0.816, respectively (P < 0.01). The number of spermatozoa with normal mitochondrial function decreased with the increased rhTNF-alpha concentration and incubation time, and r values were 0.615, 0.643, 0.752, 0.691, 0.754 and 0.532, 0.567, 0.782, 0.692, 0.854, respectively (P < 0.01).</p><p><b>CONCLUSION</b>rhTNF-alpha can reduce human sperm motility function in vitro, possibly by interfering with human sperm mitochondrial function.</p>

Immunohistochemical localization of attractin in the testis and epididymis of mature male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To localize the attractin protein in the testis and epididymis of mature male rats.</p><p><b>METHODS</b>Testes and epididymides were obtained from mature male Sprague dawley rats (n = 20). Tissues were fixed and prepared for immunohistochemical (IHC) and indirect immunofluorescent (IIF) assay, carried out with antiserum against rat attractin.</p><p><b>RESULTS</b>In the testis of the male rat, there was distinct immunopositive staining on cell membrane and cytoplasm within Leydig cells, primitive spermatogonia, primary spermatocytes, spermatids, Sertoli cells, and peritubular myoid cells. In the epididymis, including caput, corpus, cauda, there was no definitive immunopositive staining within the efferent ductule and epididymal duct.</p><p><b>CONCLUSION</b>Attractin is expressed in the male rat reproductive system and localized within Leydig cells and germ cells. It may be invoved in acting on the reproductive system. And its physiological function has yet to be further studied.</p>

Progress in Attractin gene / 中华男科学杂志

Attractin, the research of which has flourished in recent years, is an autosomal recessive gene. The gene is widely expressed, and involved in a number of physiological and pathological events. This paper reviews the research development of the Attractin gene, including the characterization, expression, mutation, function and molecular mechanism of this gene. Besides, further research on the gene is also suggested.