[Observation of effect of Wu Yun Liu Qi on reproductive hormones in varicocele patients].

Objective： To observe roproductive hormone levels in varicocele patients during a cycle (6 years) of Wu Yun Liu Qi, and explore whether the cycle had effect on the roproductive hormone levels. Methods： Data of roproductive hormone levels in varicocele patients from 2015 to 2020 were analyzed retrospectively. FSH、LH、T、PRL、E2 levels and T/E2 ratio were compared among the six years. According to Chinese sexagenary cycle heavenly stems and earthly branches of each year from 2015 to 2020 its yunqi characteristics were determined. Results: Totally data of 848 cases of varicocele patients were collected from 2015 to 2020. Among which, in 2015 (Yiwei year) there were 57 cases, in 2016 (Bingshen year) 83 cases, in 2017 (Dingyou year ) 133 cases, in 2018(Wuxu year) 156 cases, in 2019(Sihai year) 274 cases, and in 2020(Gengzi year) 145 cases. The levels of FSH、LH、PRL、T were not diferrent statistically from the six years except individual year. However, the level of E2 in 2016 when the Yunqi was Shao Yang Xiang Huo Si Tian and Jue Yin Feng Mu Zai Quan obviously higher than other years excpet 2018( All P< 0.05). And T/E2 ratio was lower in 2016 than other years except 2018 and 2020( All P< 0.05). Conclusions: Shi Xiang factors of Wu Yun Liu Qi had effect on roproductive hormone levels in varicocele patients, showing by higher E2 level in Yinshen year when the Shi Xiang factors may have bad effect on human fertility.

[Observation on effect of five evolutive phases and six climatic factors on pregnancy and sterility].

Objective： To verify statements about the effect of five evolutive phases and six climatic factors in 《huang di nei jing》 on pregnancy and sterility. Methods： Data of missed abortion from Nanjing Maternity and Child Health Care Hospital between 2006-2020 and Yangzhou Maternity and Child Health Care Hospital between 2012-2020 were collected. According to Chinese sexagenary cycle heavenly stems and earthly branches of each year between 2006-2020 was determined. And then based on the heavenly stems and earthly branches, evolutive phases and six climatic factors of each year were established. After that data of missed abortion in each year was compared. According to relative statements in 《huang di nei jing》 , it was observed whether the number of missed abortion cases in someone year increased significantly. Results: Totally there were 28059 cases of missed abortion from the two hospital were collected between 2006-2020 and 2012-2020 respectively. It was found that in the year of Shaoyang phase fire sitian and Jueyin wind wood in the spring (that is the year of 2010 and 2016) the number of missed abortion cases was higher obviously than the other year. When the year of Taiyang cold water in the spring (that is the year of 2009 and 2015), there was a relatively increased number of missed abortion cases. Conclusions: Five evolutive phases and six climatic factors have impacts on human pregnancy and sterility. It was an adverse condition for human reproduction in the year of Shaoyang phase fire sitian and Jueyin wind wood in the spring as well as Taiyang cold water in the spring. Until to now we can still verify statements about the effect of five evolutive phases and six climatic factors on pregnancy and sterility in 《huang di nei jing》.

[Effect of bilateral microsurgical subinguinal varicocelectomy on asthenozoospermia and oligozoospermia: A report of 109 cases].

OBJECTIVE: To investigate the effects of bilateral microsurgical subinguinal varicocelectomy (BMSV) in patients with asthenozoospermia or oligozoospermia. METHODS: Totally 147 patients with male infertility received BMSV from January 2018 to May 2019, of whom 109 had complete data recorded. We retrospectively analyzed the clinical data, including the total sperm count per ejeculate, sperm concentration and sperm motility before and after surgery, and the rate of natural conception achieved during the follow-up. RESULTS: BMSV achieved a total effectiveness rate of 79.00% in improving the percentage of progressively motile sperm (a rise of ≥20%) and a marked effectiveness rate of 70.00% (a rise of ≥50%) in the 100 cases of asthenozoospermia as compared with the baseline, with a mean recovery time of (110.13 ± 37.43) days. Besides, a total effectiveness rate of 74.29% (an increase of ≥20%) and a marked effectiveness rate of 71.43% (an increase of ≥50%) were attained in the improvement of sperm concentration in the 35 cases of oligozoospermia, with a mean recovery time of (117.00 ± 48.79 ) days. A natural conception rate of 30.30% was observed during the follow-up. No severe adverse events occurred postoperatively. CONCLUSIONS: BMSV is significantly effective for the treatment of asthenozoospermia and oligozoospermia.

[Differential expression profiles of miRNAs in the vaginal tissue of the women with lubrication disorders].

OBJECTIVE: To explore the pathogenesis of lubrication disorder (LD), a most common type of female sexual dysfunction affecting women's physical health and conception, and find the therapeutic targets for its treatment and prevention. METHODS: We chose 3 LD patients and 3 healthy controls in Nanjing Maternal and Child Health Hospital, extracted their vaginal epithelial RNA for high-throughput miRNA sequencing, screened differentially expressed miRNAs for hierarchical cluster analysis, target gene prediction and gene ontology (GO) and KEGG pathway enrichment analyses. Finally, we verified the sequencing results by real-time fluorescent quantitative PCR. RESULTS: Totally 1 673 miRNAs were predicted by high-throughput sequencing and 64 likely to be the targets for the treatment of LD were screened, including 25 up-regulated more than 4 times and 39 down-regulated more than 4 times in the LD patients compared with the healthy controls. The neuron projection morphogenesis and AMPK signaling pathway were the most significant enrichment GO term and KEGG pathway. CONCLUSIONS: miRNAs are expressed differentially in LD patients. These miRNAs and target genes may be related to the occurrence of LD, and those that are expected to be the targets for the treatment of LD have important theoretical significance and potential application value.

[HPV infection of the external genitalia in men whose female partners have cervical HPV infection].

OBJECTIVE: Human papilloma virus (HPV) is a necessary cause of cervical cancer and is also closely related to penile cancer, oropharyngeal cancer, and anal cancer in males. However, few studies are reported on male HPV. This study aimed to investigate HPV infection of the external genitalia in men whose female partners have cervical HPV infection. METHODS: We collected the relevant data on the male outpatients whose partners had cervical HPV infection in our Department of Urology and Andrology from August to December 2016. We obtained samples with nylon swabs from the glans penis, corona, inner layer of the prepuce and penile body and detected different types of HPV infection using the Hybribio HPV typing kit, PCR and membrane hybridization. RESULTS: Valid data were collected from 140 males, which showed 83.5% of HPV infection of the external genitalia, including 60 cases of HPV6 (43.2%), 27 cases of HPV16 (19.4%), 14 cases of HPV39 (10.1%), 13 cases of HPV18 (9.4%), 13 cases of HPV58 (9.4%), and 13 cases of HPV52 (9.4%). Redundant prepuce was found in 75.5% of the males, but there was no statistically significant difference in the incidence rate of HPV infection between the normal and redundant prepuce groups (P > 0.05). CONCLUSIONS: Men who have the female partners with positive cervical HPV are at high risk of HPV infection and therefore need to be screened and treated so as to reduce HPV infection in both sexes.

[Relationship of abnormal sperm DNA methylation with early spontaneous abortion].

OBJECTIVE: To investigate the relationship between the abnormal sperm DNA methylation level and early spontaneous abortion. METHODS: We randomly selected 98 males who met the inclusion criteria and whose wives suffered from unexplained abortion or embryo abortion, and included another 46 normal healthy men present for pre-pregnancy check-up as controls. We examined the semen quality and sperm morphology, obtained the sperm DNA fragmentation index (DFI) by modified sperm chromatin dispersion, and measured the sperm DNA methylation level using the methylated DNA quantification kit and the colorimetric method. RESULTS: Compared with the normal controls, the men in the unexplained abortion group showed a significantly lower rate of big-halo sperm (ï¼»45.50 ± 26.27ï¼½ vs ï¼»36.49 ± 23.06ï¼½%, P = 0.038), a higher rate of abnormal-head sperm (ï¼»77.08± 12.21ï¼½ vs ï¼»81.09± 10.89ï¼½%, P = 0.049), and a lower level of sperm DNA methylation (ï¼»0.47 ± 0.33ï¼½ vs ï¼»0.36 ± 0.26ï¼½ ng/µl, P = 0.035). The sperm DNA methylation level was positively correlated with the percentage of big-halo sperm (OR=0.546, P<0.01). Multivariate regression analysis manifested that sperm head abnormality was an independent risk factor of early spontaneous abortion or embryo abortion (OR=1.032, P = 0.049), while the high methylation level was protective factor against early spontaneous abortion or embryo abortion (OR=0.244, P = 0.03). CONCLUSIONS: The abnormal level of sperm DNA methylation may be one of the important reasons for early spontaneous abortion or embryo abortion.

MicroRNA-200a is up-regulated in aged rats with erectile dysfunction and could attenuate endothelial function via SIRT1 inhibition.

MiR-200a was shown to be upregulated in the corpus cavernosum (CC) of rats with aging-related erectile dysfunction (A-ED) in our previous study. Among its target genes, SIRT1 was also reported as a protective factor in erectile function by our groups previously. Thus, miR-200a might attenuate the erectile function in A-ED via SIRT1 inhibition. In the present study, three animal groups were included: aged rats with ED (group AE, n = 8), aged rats with normal erectile function (group AN, n = 8), and young rats as normal controls (group YN, n = 8). CCs from each group were collected for histological and molecular measurements to validate the dysregulation of miR-200a and SIRT1. After that, the cavernous endothelial cells (CECs) from CC of aged rats with normal erectile function were transfected with miR-200a in vitro. Then the expression of SIRT1 and molecules within the eNOS/NO/PKG pathway were measured to investigate whether the transfection could imitate the attenuated process of erectile function in the aged. As a result, miR-200a was upregulated while the SIRT1, the levels of eNOS and cGMP were all downregulated in the CCs from AE group. After transfection in vitro, the miR-200a was upregulated while the SIRT1 and levels of eNOS and cGMP were obviously downregulated. Finally, based on the results of our previous study, we further verify that up-regulation of miR-200a could participate in the mechanisms of A-ED via SIRT1 inhibition, and mainly attenuate endothelial function via influencing the eNOS/NO/PKGpathway.

[Semen quality and its influencing factors in preconception males in Nanjing area].

OBJECTIVE: To investigate the semen quality and its influencing factors in preconception males in Nanjing area so as to provide some evidence for working out effective intervention measures. METHODS: Totally 687 men receiving preconceptional physical examination were enrolled in this study. A questionnaire survey was conducted among the subjects along with an analysis of their semen quality. RESULTS: The median of sperm concentration was 63.3 x 10(6)/ml (95% CI [19.88-119] x 10(6)/ml). The median of grade a sperm was 33.03% (95% CI [19.38-55.05]%), that of grade a + b sperm was 52.08% (95% CI [39.53-69.37]%), and that of teratosperm was 91.75% (95% CI [69-100]%). The median concentration of seminal plasma PMN-elastase was 195.55 ng/ml (95% CI [76.16-3330.38] ng/ml) and that of seminal plasma zinc was 7.62 µmol/L (95% CI [1.5-23, 45] µmol/L). The positive rates of Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), and Gardnerella vaginalis (GV) were 42.4%, 0.3%, and 2.4%, respectively. The median of sperm DNA fragmentation index (DFI) of those whose wives had a history of adverse pregnancy was 20.25% (95% CI [2.15-68.25]%). Multivariate logistic regression analysis suggested that mental stress (OR 1.567, 95% CI [1.081-2.27]) and sedentariness (OR 1.772, 95% CI [1.211-2.592]) were independent risk factors for asthenospermia. CONCLUSION: The sperm quality of preconception males in Nanjing area is not encouraging, and it can be improved by changing undesirable lifestyle and reducing mental stress.

[Relationship between epigenetics of sperm and embryogenesis].

Epigenetics comprises the modifications made in gene expressions without changing the DNA sequence itself. Significant epigenetic changes take place during spermatogenesis and fertilization and exert direct influences on embryogenesis. This article provides an overview of the latest researches on epigenetics of male germ cells and a brief discussion on the correlation of sperm with embryogenesis in four aspects: DNA methylation, histone modification, regulation of non-coding RNAs, and genomic imprinting.

[Protective effect of pAdxsi-ERbeta adenovirus transfection on penile vascular endothelium in ERbetaKO mice].

OBJECTIVE: To investigate the effect of the overexpression of the ERbeta gene on the penile vascular endothelium of ERbeta knockout (ERbetaKO) mice and its molecular mechanisms. METHODS: We randomly divided 12 ERbetaKO male mice into groups A (ERbetaKO + TNFalpha + pAdxsi-ERbeta) and B (ERbetaKO + TNFalpha + empty virus), the former treated by pAdxsi-ERbeta adenovirus transfection, the latter with empty virus, and meanwhile both injected intraperitoneally with TNFalpha at 6 microg per kg body weight per d for 14 days. Then we observed the erectile function of the mice by APO, determined the changes of the endothelial markers CD34 and vWF by immunohistochemical staining, and detected the expressions of the relevant molecules in the eNOS-NO pathway by RT-PCR, Western blot and immunohistochemistry. RESULTS: Compared with group B, group A showed a significantly increased number of penile erections (0.50 +/- 0.55 vs 2.17 +/- 0.41, P < 0.05), shortened erectile latency ([28.83 +/- 1.33] min vs [24.00 +/- 1.27] min, P < 0.05), enriched CD34 and vWF markers (0.67 +/- 0.52 vs 1.50 +/- 0.55 and 0.50 +/- 0.55 vs 1.33 +/- 0.52, both P < 0.05), elevated expressions of eNOS and Cam (RT-PCR: 1.38 +/- 0.03 vs 1.62 +/- 0.05 and 1.02 +/- 0.09 vs 1.42 +/- 0.05, both P < 0.05; Western blot: 1.27 +/- 0.04 vs 1.55 +/- 0.07 and 0.76 +/- 0.05 vs 0.95 +/- 0.08, both P < 0.05), and reduced expression of caveolin-1 (RT-PCR: 2.13 +/- 0.13 vs 1.72 +/- 0.08, P < 0.05; Western blot: 3.99 +/- 0.16 vs 3.40 +/- 0.14, P < 0.05). The results of RT-PCR were consistent with those of Western blot. CONCLUSION: The ERbeta gene protects the penile vascular endothelium via the eNOS-NO pathway.

[Seminal parameters at different times of reanalysis of the normal semen samples detected in initial examination and their correlation with sperm DNA damage].

OBJECTIVE: To investigate the differences in semen quality at different times of reanalysis and the correlation of sperm DNA fragmentation index (DFI) with sperm motility alteration using semen samples completely liquefied and normal in initial examination. METHODS: We analyzed 127 semen samples up to the inclusion criteria with the computer-assisted semen analysis (CASA) system at 15, 30 and 60 min after semen collection, and obtained sperm morphology parameters and DFI by Shorr staining and acridine orange test (AOT) , respectively. RESULTS: Sperm concentration, and the percentages of grades a and b sperm showed no statistically significant differences at the three time points (P > 0.05). The percentages of grades a + b and a + b + c sperm were significantly higher at 15 min than at 30 and 60 min after semen collection (P < 0.05), but with no significant difference between the latter two time points (P > 0.05). The incidence of alternation from normal to abnormal in at least one index of sperm motility at different times was 25.2%, but there were no significant differences in sperm DFI and morphology between the normal and abnormal groups (P > 0.05). Among the altered parameters of sperm motility from 15 to 60 min, the percentages of grades a, a + b and a + b + c sperm were all positively correlated with sperm DFI (P < 0.05). CONCLUSION: Semen samples completely liquefied within 15 min after collection and normal in initial examination, when reanalyzed at 30 and 60 min, showed significant decreases in the percentages of grades a + b and a + b + c sperm, but not in the percentages of grades a and b sperm, and the parameters of sperm motility might be abnormal. Thus, at least 2 sperm analyses are required for a comprehensive evaluation of fertility. Significant difference between the results of the two analyses, and particularly a markedly reduced percentage of rapidly progressive sperm, might indicate sperm DNA damage, and thus the necessity of sperm DNA damage detection.

[Treatment of varicocele by microsurgical varicocelectomy: report of 94 cases].

OBJECTIVE: To investigate the clinical effect of microsurgical varicocelectomy in the treatment of varicocele. METHODS: We retrospectively analyzed the clinical data of 94 varicocele patients treated by microsurgical varicocelectomy, of whom 36 complained of testicular pain or dragging and distending discomfort, and 58 infertility or oligoasthenospermia. Microsurgical varicocelectomy was performed through inguinal or subinguinal approach. Complications were observed and semen parameters detected through follow-up visits at 1, 3 and 6 months after the operation. RESULTS: Testicular pain or discomfort disappeared in 21 (65.6%) of the 32 followed-up patients who had complained of such symptoms, was relieved in 6 (18.8%), and remained unimproved in the other 5 (15.6%). The 56 followed-up patients with infertility or oligoasthenospermia all showed significantly improved sperm concentration and motility (grade a + b sperm), (15.47 +/- 3.21) x 10(6)/ml and (13.34 +/- 5.16)% at 3 months, and (18.39 +/- 4.05) x 10(6)/ml and (17.23 +/- 4.69)% at 6 months after operation, as compared with (8.26 +/- 1.68) x 10(6)/ml and (5.25 +/- 1.09)% preoperatively (P < 0.01). CONCLUSION: Microsurgical varicocelectomy can effectively improve symptoms and semen parameters in varicocele patients.

[Protective effect of ERbeta on penile vascular endothelium in mice].

OBJECTIVE: To investigate the protective effect of ERbeta on the penile vascular endothelium in mice. METHODS: We randomly selected 12 ERbeta knockout (ERbetaKO) and 12 C57BL/6 male mice, and divided them into four groups: normal control, ERbetaKO, ERbetaKO + TNFalpha, and wild-type + TNFalpha group. The former two were treated with normal saline, while the latter two by intraperitoneal injection of TNFalpha at 6 microg/(kg x d) for 14 days. Then we observed the spontaneous erectile response induced by APO and changes of the endothelial cells by immunohistochemical staining of CD34 and vWF, and detected cell apoptosis in the penile cavernous tissue by TUNEL. RESULTS: Compared with the normal control group, the ERbetaKO group showed significantly increased erectile latency (P<0.05), but no significant difference in the number of erections; the ERbetaKO + TNFalpha and wild-type + TNFalpha groups, too, exhibited remarkably longer erectile latency (P<0.05) but fewer erections (P<0.05), with even more obvious changes in the ERbetaKO + TNFalpha group. The expressions of CD34 and vWF were significantly reduced in the ERbetaKO group (2.25 +/- 0.50 and 2.00 +/- 0.00), ERbetaKO + TNFalpha group (0.25 +/- 0.50 and 0.33 +/- 0.58) and wild-type + TNFalpha group (1.50 +/- 0.58 and 1.25 +/- 0.50) as compared with those in the control (3.00 +/- 0.00 and 2.75 +/- 0.50) (P<0.05), even lower in the ERbetaKO + TNFalpha than in the wild-type + TNFalpha group (P<0.05). Apoptotic cells were found only in the ERbetaKO + TNFalpha group. CONCLUSION: After ERbeta knockout and especially after treated with the endothelial injury factor TNFalpha, endothelial cells are decreased in the penile vessels in mice, which suggests that ERbeta has a protective effect on the penile cavernous sinus endothelium.

[Human papillomavirus infection correlates with redundant prepuce or phimosis in the patients'sexual partners in Nanjing urban area].

OBJECTIVE: To investigate the incidence of human papillomavirus (HPV) infection in women and its correlation with redundant prepuce or phimosis in the patients' sexual partners. METHODS: We conducted a questionnaire investigation among the women outpatients at the cervical disease clinic of Nanjing Maternity and Child Health Hospital from May to December 2011. We obtained information on their sexual life and determined whether their sexual partners had redundant prepuce or phimosis according to the schematic illustrations we offered. We used biology-hybridize HPV-type test kit, PCR and hybrid membrane methods for detection of different HPV genotypes in cervical exfoliated cells, taking any type of HPV detected as positive. We made between-group comparisons by chi-square test and analyzed independent risk factors by non-conditional logistic regression analysis. RESULTS: Of the 2 040 questionnaires, 1 568 were collected and 1 110 (71%) accepted as valid by inclusion criteria. Among the 1 110 subjects, 566 (50.9%) were infected with HPV, and 445 (78.6%) of the infected women admitted that their sexual partners had redundant prepuce or phimosis. The most frequent infection type was HPV16 (34.2%), followed by HPV58 (28.1%), HPV52 (20.2%) and HPV18 (10.8%). Multivariate logistic regression analysis suggested that redundant prepuce or phimosis in the women's sexual partners was an independent risk factor for HPV infection (OR 3.387, 95%CI [2.491-4.607]). CONCLUSION: In Nanjing urban area, the majority of the sexual partners of the HPV-infected women have redundant prepuce or phimosis, which is an independent risk factor for female cervical HPV infection. Male circumcision is necessitated in Nanjing to reduce the incidence of cervical HPV infection.

[Endothelial injury and erectile dysfunction].

The endothelium plays an important role in maintaining vascular homeostasis, regulating vascular tone and blood flow, and preserving a non-thrombogenic blood-tissue interface, and the normal function of the vascular endothelium is essential for penile erection. In most cases, erectile dysfunction (ED) is accompanied by endothelial dysfunction, and endothelial injury is a major pathological basis of ED, which can be induced by bad lifestyles, cardiovascular diseases, reactive oxygen species, and inflammatory mediators. The vascular endothelium is capable of self-repairing, and endothelial injury results from the unbalanced factors of injury and repair. This review focuses on the mechanism and repair of endothelial injury and the relationship of endothelial injury with ED.

[An investigation on female sexual dysfunction among urban Chinese women in Nanjing].

OBJECTIVE: Female sexual dysfunction (FSD) is a common problem affecting women's quality of life. However, reports are rarely seen on sexual problems in Chinese women. This study is to investigate the prevalence of FSD among urban Chinese women in Nanjing and offer some evidence for the establishment of preventative measures for FSD in China. METHODS: A cross-sectional hospital-based survey was conducted in Nanjing, China between August 2008 and March 2009. The sexual function of 609 women aged 20 -56 years from the urban area of Nanjing were investigated using the Female Sexual Function Index (FSFI). The total FSFI score of < 25 was used as the diagnostic criterion for FSD. RESULTS: The total FSFI score was 24.21 +/- 4.40 in this group of women, and it decreased with the increase of age. The overall prevalence of FSD was 56.8%, and it increased with the increase of age, 47.1%, 57.0%, 75.0% and 90.3% in the < 29 yr, 30 - 39 yr, 40 - 49 yr and > or = 50 yr groups, respectively. The most common problems were low sexual satisfaction (43.2%) and orgasm disorder (41.7%), followed by sexual pain (40.2%), hyposexuality (35.1%), vaginal dryness (31.4%) and sexual arousal disorder (29.6%). CONCLUSION: FSD is a common problem among urban Chinese women in Nanjing, and the most common sexual problems are low sexual satisfaction and orgasm disorder.

[Multivariate analysis of dyspareunia in women].

OBJECTIVE: Dyspareunia is a common sexual trouble in women during the sexual intercourse. This study is to investigate the risk factors for dyspareunia in urban Chinese women and to supply some evidence for its preventive measures. METHODS: We conducted a hospital-based survey by distributing 2 658 copies of a questionnaire among the women in Nanjing urban area who came for regular physical examination in Nanjing Maternity and Child Health Hospital and their female companions aged over 20 years. The sexual function of the subjects was evaluated according to female sexual function indexes, dyspareunia indicated by sexual pain score < 4.4, and the results analyzed by multiple logistic regression analysis. RESULTS: Totally, 1 856 (69.8%) of the subjects completed the questionnaire, and 1 457 that met the criteria were included for analysis, of whom 43.0% (626/1457) admitted to dyspareunia during the sexual intercourse. Multiple logistic regression analysis showed that age (> or = 50 years) , smoking, hysterectomy, vaginal lubrication disorder, lack of sexual communication with partners were independent risk factors for dyspareunia (P < 0.05). CONCLUSION: Dyspareunia is associated with multiple factors including age (> or = 50 years), smoking, hysterectomy, vaginal lubrication disorder, lack of sexual communication with partners.

[Male factors in repeated spontaneous abortion].

Repeated spontaneous abortion (RSA), with very complicated pathogenesis, has an incidence of about 0.5% 2.0%. As for the paternal part which provides half of the genes for the embryo, current studies mainly focus on the genes of somatic cells or germ cells. Chromosome abnormality of somatic cells, chromosome disorder of sperm, defects in sperm quality, genetic mutation, senility, infection and any other paternal gene abnormalities that affect the embryo would induce RSA. This paper presents an update on the above mentioned paternal factors which might result in RSA.

[Evaluation of sperm mitochondrial membrane potential in varicocele patients using JC-1 fluorescent staining].

OBJECTIVE: To detect sperm mitochondrial membrane potential (MMP) of varicocele patients and investigate its clinical significance. METHODS: Sixty-seven varicocele patients were divided into a VC1 (grade 1, n = 26), a VC2 (grade 2, n = 21) and a VC3 group (grade 3, n = 20). And 29 normal fertile volunteers were included in a control group ( m = 29). Conventional semen analyses were performed by computer-assisted semen analysis (CASA). Semen samples were washed, followed by JC-1 staining to evaluate the sperm MMP (JC-1+ %) by flow cytometry. RESULTS: The sperm MMPs of the VC1, VC2 and VC3 groups were siginificantly lower ([56.29 +/- 16.32]%, P < 0.05; [45.04 +/- 13.21]%, P < 0.01; [31.63 +/- 12.91]%, P < 0.01) than that of the control ([76.21 +/- 13. 96]%). There was a significant positive correlation between the percentage of JC-1+ and that of grade (a + b) sperm (r =0.693, P=0.000). CONCLUSION: The decreased MMP in the sperm of varicocele men might be one of the important causes of male infertility.

Rapid molecular prenatal diagnosis of spondyloepiphyseal dysplasia congenita by PCR-SSP assay.

Heterozygous mutations of COL2A1 gene are responsible for type II collagenopathies. The common skeletal phenotypes include achondrogenesis type II, hypochondrogenesis, Stickler dysplasia, Kniest dysplasia, late onset spondyloepiphyseal dysplasia, and spondyloepiphyseal dysplasia congenita (SEDC). Prevention of SEDC can be achieved by prenatal diagnosis. This study reports the first rapid molecular prenatal diagnosis of SEDC performed in China by polymerase chain reaction sequence-specific primer (PCR-SSP) analysis. The pregnant woman we previously reported with SEDC carried the G to A substitution at nucleotide 1510 in exon 23 of COL2A1 gene, which caused a change from glycine to serine at codon 504 (G504S). By the time the woman got pregnant again, she had terminated two pregnancies and still had no child. In the first pregnancy, the molecular mutation of the family was not yet identified, and therefore prenatal diagnosis was unable to be performed by DNA analysis. In the second pregnancy, G504S mutation was found from fetal DNA. At the time of her third pregnancy, the woman and her husband became extremely worried about the potential SEDC for the fetus. For this reason, a quick and reliable molecular prenatal diagnosis of SEDC was performed by a PCR-SSP on an amniocyte sample collected at the 14th week of pregnancy. No mutation of the fetal DNA was identified. The result was obtained within 24 h after the sample was collected. The technique could be applied in confirmatory diagnosis and prenatal diagnosis for the affected family.