Deleterious variants in X-linked CFAP47 induce asthenoteratozoospermia and primary male infertility.

Asthenoteratozoospermia characterized by multiple morphological abnormalities of the flagella (MMAF) has been identified as a sub-type of male infertility. Recent progress has identified several MMAF-associated genes with an autosomal recessive inheritance in human affected individuals, but the etiology in approximately 40% of affected individuals remains unknown. Here, we conducted whole-exome sequencing (WES) and identified hemizygous missense variants in the X-linked CFAP47 in three unrelated Chinese individuals with MMAF. These three CFAP47 variants were absent in human control population genome databases and were predicted to be deleterious by multiple bioinformatic tools. CFAP47 encodes a cilia- and flagella-associated protein that is highly expressed in testis. Immunoblotting and immunofluorescence assays revealed obviously reduced levels of CFAP47 in spermatozoa from all three men harboring deleterious missense variants of CFAP47. Furthermore, WES data from an additional cohort of severe asthenoteratozoospermic men originating from Australia permitted the identification of a hemizygous Xp21.1 deletion removing the entire CFAP47 gene. All men harboring hemizygous CFAP47 variants displayed typical MMAF phenotypes. We also generated a Cfap47-mutated mouse model, the adult males of which were sterile and presented with reduced sperm motility and abnormal flagellar morphology and movement. However, fertility could be rescued by the use of intra-cytoplasmic sperm injections (ICSIs). Altogether, our experimental observations in humans and mice demonstrate that hemizygous mutations in CFAP47 can induce X-linked MMAF and asthenoteratozoospermia, for which good ICSI prognosis is suggested. These findings will provide important guidance for genetic counseling and assisted reproduction treatments.

CD137+ tumor infiltrating lymphocytes predicts ovarian cancer survival.

OBJECTIVE: Ovarian cancer (OC) is the leading cause of death from gynecologic malignancy in the United States, and biomarkers of patient outcomes are limited. Data using immunohistochemical (IHC) analysis are mixed regarding whether and which tumor infiltrating lymphocytes (TILs) impact survival, and IHC does not adequately quantify rare cell populations, including CD137+ (4-1BB) tumor-reactive TILs. Our study investigates if a higher percentage of CD3+ CD137+ TILs is associated with improved overall survival (OS) in OC. METHODS: Flow cytometry was performed on viably banked OC digests. Chart review and statistical analysis were performed. Forty-seven patients were included, 40 of whom were diagnosed with high-grade serous ovarian carcinoma (HGSOC), papillary serous carcinoma, or undifferentiated histology. RESULTS: A high percentage of CD3+ CD137+ TILs correlated with improved OS (n = 40, r = 0.48, P = 0.0016). Subjects were divided into CD3+ CD137+ TIL high and low groups by the median. Subjects with high CD3+CD137+ TIL frequencies (>9.6%) had longer OS (Wilcoxon rank-sum test; P = 0.0032) and improved OS (logrank test; P = 0.007). Differences in CD3+ or CD3+ CD8+ TILs did not impact survival. CD3+ CD137+ TILs were predictive of OS regardless of germline mutation or debulking status. Analysis of subgroups including late stage HGSOC and late stage HGSOC with primary optimal cytoreduction indicated CD3+ CD137+ TILs correlated with improved OS after adjusting for age and PARP inhibitor use (P = 0.034 and P = 0.016, respectively). CONCLUSIONS: Prevalence of CD3+ CD137+ TILs in digested OC specimens is associated with improved OS, while general TIL markers are not. CD137 has the potential to be a novel biomarker for survival in OC.

Homozygous mutations in CCDC34 cause male infertility with oligoasthenoteratozoospermia in humans and mice.

BACKGROUND: Oligoasthenoteratozoospermia is a typical feature of sperm malformations leading to male infertility. Only a few genes have been clearly identified as pathogenic genes of oligoasthenoteratozoospermia. METHODS AND RESULTS: Here, we identified a homozygous frameshift variant (c.731dup, p.Asn244Lysfs*3) in CCDC34, which is preferentially expressed in the human testis, using whole-exome sequencing in a cohort of 100 Chinese men with multiple morphological abnormalities of the sperm flagella (MMAF). In an additional cohort of 167 MMAF-affected men from North Africa, Iran and France, we identified a second subject harbouring a homozygous CCDC34 frameshift variant (c.799_817del, p.Glu267Lysfs*72). Both affected men presented a typical MMAF phenotype with an abnormally low sperm concentration (ie, oligoasthenoteratozoospermia). Transmission electron microscopy analysis of the sperm flagella affected by CCDC34 deficiency further revealed dramatic disorganisation of the axoneme. Immunofluorescence assays of the spermatozoa showed that CCDC34 deficiency resulted in almost absent staining of CCDC34 and intraflagellar transport-B complex-associated proteins (such as IFT20 and IFT52). Furthermore, we generated a mouse Ccdc34 frameshift mutant using CRISPR-Cas9 technology. Ccdc34-mutated (Ccdc34mut/mut ) male mice were sterile and presented oligoasthenoteratozoospermia with typical MMAF anomalies. Intracytoplasmic sperm injection has good pregnancy outcomes in both humans and mice. CONCLUSIONS: Our findings support that CCDC34 is crucial to the formation of sperm flagella and that biallelic deleterious mutations in CCDC34/Ccdc34 cause male infertility with oligoasthenoteratozoospermia in humans and mice.

Integrating multiple vegetation indices via an artificial neural network model for estimating the leaf chlorophyll content of Spartina alterniflora under interspecies competition.

The invasive species Spartina alterniflora and native species Phragmites australis display a significant co-occurrence zonation pattern and this co-exist region exerts most competitive situations between these two species, competing for the limited space, directly influencing the co-exist distribution in the future. However, these two species have different growth ratios in this area, which increase the difficulty to detect the distribution situation directly by remote sensing. As chlorophyll content is a key indicator of plant growth and physiological status, the objective of this study was to reduce the effect of interspecies competition when estimating Cab content; we evaluated 79 published representative indices to determine the optimal indices for estimating the chlorophyll a and b (Cab) content. After performing a sensitivity analysis for all 79 spectral indices, five spectral indices were selected and integrated using an artificial neural network (ANN) to estimate the Cab content of different competition ratios: the Gitelson ratio green index, the transformed chlorophyll absorption ratio index/optimized soil-adjusted vegetation index, the modified normalized difference vegetation index, the chlorophyll fluorescence index, and the Vogelmann chlorophyll index. The ANN method yielded better results (R 2 = 0.7110 and RMSE = 8.3829 µg cm-2) on average than the best single spectral index (R 2 = 0.6319 and RMSE = 9.3535 µg cm-2), representing an increase of 10.78% in R 2 and a decrease of 10.38% in RMSE. Our results indicated that integrating multiple vegetation indices with an ANN can alleviate the impact of interspecies competition and achieve higher estimation accuracy than the traditional approach using a single index.

[Efficacy of Shugan Yiyang Capsules combined with sertraline on premature ejaculation].

OBJECTIVE: To observe the clinical effect of Shugan Yiyang Capsules combined with sertraline in the treatment of premature ejaculation (PE). METHODS: We randomly assigned 192 PE patients to receive sertraline hydrochloride 50 mg qd (control group, n = 96) or sertraline hydrochloride 50 mg qd plus Shugan Yiyang Capsules at the dose of 4 capsules tid ( combination therapy group, n = 96) , both for 6 weeks. We compared the intravaginal ejaculatory latency time (IELT) and Chinese Index of Premature Ejaculation ( CIPE) scores between the two groups of patients before and after medication and at 6 weeks after drug withdrawal. RESULTS: Compared with the baseline, the IELT was significantly increased after 6 weeks of medication in the combination therapy group ([1.41 ± 0.53] vs [6.69 ± 3.56] min, P < 0.05) and the control group ([1.43 ± 0.48] vs [5.37 ± 2.91] min, P < 0.05), and so was the CIPE score in the former (9. 80 ± 2.06 vs 21.62 ± 4.76, P < 0.05) and the latter group ([9.41 ± 1.97] vs [20.85 ± 4.83] , P < 0.05). In comparison with the pre-medication indexes, the IELT ([3.77 ± 1.63] min) and CIPE score (16.92 ± 3.37) of the combined therapy group were remarkably improved at 6 weeks after drug withdrawal (P < 0.05), but not those of the control ([1.19 ± 1.34] min and 10.59 ± 2.38, P > 0.05). CONCLUSION: Shugan Yiyang Capsules combined with sertraline have a definite and lasting effect on premature ejaculation.

A Randomized, Controlled Clinical Trial of Combining Therapy with Traditional Chinese Medicine-Based Psychotherapy and Chinese Herbal Medicine for Menopausal Women with Moderate to Serious Mood Disorder.

OBJECTIVE: To comparatively examine the effectiveness and safety of the combination therapy of traditional Chinese medicine formula Bushen-Shugan granule and psychotherapy (BSSG-P) and Chinese herbal medicine Bushen-Shugan granule (BSSG) alone in the treatment of moderate to serious mood disorder in menopausal women. METHODS: In our previous clinical studies, BSSG-P had been proved to be superior to BSSG, psychological treatment, and placebo in improving mild mood disorder in menopausal women. In this study, we analyzed the efficacy of BSSG-P and BSSG in the treatment of moderate to serious mood disorder. Eighty-five eligible participants, who were diagnosed as menopausal women with moderate to serious mood disorder and categorized as kidney deficiency and liver-qi stagnation pattern, were randomly assigned into two groups and treated with BSSG-P or BSSG. They were subjected to an 8-week treatment period and a 4-week follow-up study. The primary outcome instrument was the Greene Climacteric Scale, Self-Rating Depression Scale (SDS), and Self-Rating Anxiety Scale (SAS), respectively. RESULTS: When comparing all time points with baseline, both BSSG-P and BSSG markedly decreased the total score of Greene, SDS, and SAS and the score of each dimension, in which BSSG-P exerted superior effect after 8-week treatment and 4-week follow-up (P<0.05). Furthermore, BSSG-P also showed great advantage in reducing the score of Greene, SDS, and SAS for menopausal women with moderate mood disorder at the end of the 8th and 12th week when compared with BSSG (P < 0.05), whereas there was no significant difference between groups at any time point for patients with serious mood disorder (P>0.05). No serious event occurred in both groups, and no significant difference was found between groups in adverse event proportion. CONCLUSIONS: BSSG-P was superior to BSSG in improving the physical and psychological symptoms of menopausal women with mood disorder. For patients with moderate mood disorder, BSSG-P showed obvious advantages; however, no superiority was observed for serious mood disorder.

Adjuvant Therapy of Oral Chinese Herbal Medicine for Menopausal Depression: A Systematic Review and Meta-Analysis.

Objective. The aim of this meta-analysis was to evaluate the effectiveness of oral Chinese herbal medicine (OCHM) combined with pharmacotherapy for menopausal depression. Methods. The electronic databases were searched from their inception to December 25, 2016, comprising PubMed, Embase, Cochrane Central Register of Controlled Trials, Chinese National Knowledge Infrastructure (CNKI), Wanfang database, and Chinese Biomedical (CBM) database. Randomized controlled trials investigating the effectiveness of OCHM combined with pharmacotherapy for the people with menopausal depression were eligible. Risk of bias was evaluated according to the Cochrane handbook. Meta-analyses were performed to pool the effect size. Heterogeneity and publication bias were also examined. Results. Twenty-two RCTs with 1770 participants were included in the review. None of the studies used placebo as the control and the risk of bias was high in blinding the participants and personnel. Overall, the meta-analysis demonstrated that adjuvant therapy of OCHM was effective in reducing the Hamilton Rating Scale for Depression (HAMD) scores compared to pharmacotherapy (MD = -3.75; 95% CI = -5.22, -2.29; P < 0.00001). The meta-analysis also suggested that OCHM adjuvant therapy for menopausal depression was superior to pharmacotherapy in terms of response rate of reducing HAMD scores (RR = 1.17; 95% CI = 1.10, 1.25; I2 = 55%). Conclusions. OCHM may provide additional effectiveness to pharmacotherapy for the people with menopausal depression. RCTs including the placebo control were required to further determine the additional efficacy of OCHM for menopausal depression.

A randomized, double-blind, placebo-controlled trial of Chinese herbal medicine capsules for the treatment of premature ovarian insufficiency.

OBJECTIVE: This study was conducted to evaluate the treatment effectiveness of Chinese herbal medicine capsules containing the Yangyin Shugan formula (YYSG) in premature ovarian insufficiency (POI). METHODS: One-hundred forty-six women with POI participated in this stratified, randomized, double-blind, placebo-controlled clinical trial. Participants in two groups (n = 73 in each)-the YYSG group and control group-underwent treatment for 12 weeks. Outcome measures included the Chinese version Menopause-Specific Quality of Life questionnaire (CMS), serum levels of basal follicle-stimulating hormone (bFSH), basal estradiol, and anti-Mullerian hormone (AMH), the antral follicle count (AFC), and ovarian peak systolic velocity (PSV; cm/s). RESULTS: Treatment with YYSG significantly reduced the total scores of the CMS at the end of the 12th week with statistical significance (P < 0.01); the vasomotor, psychosocial, physical, and sexual domains significantly improvement after treatment (P < 0.01). Compared with the baseline hormone levels, YYSG markedly decreased the bFSH level with statistical significance (P < 0.01) and improved the AMH level (P < 0.01). Furthermore, YYSG greatly improved the participants' AFC and ovarian PSV, compared with placebo (P < 0.01). There were no serious adverse events, and the safety indices of whole blood counts, renal function, and liver function were within the normal range, both before and after treatment. CONCLUSIONS: Treatment with YYSG was more effective than placebo for improving menopausal symptoms, basal hormone levels, and ovarian function in women with POI in Guangdong, China.

Effects of Jiawei Shaoyao-Gancao Decoction and Its Drug-Containing Serum on Proliferation, Apoptosis, and Ultrastructure of Human Adenomyosis Foci Cells.

OBJECTIVE: The present study aimed to investigate the effects of Jiawei Shaoyao-Gancao Decoction (JSGD) and its drug-containing serum (CDS) on the proliferation, apoptosis, and ultrastructure of human adenomyosis foci cells. METHODS: Primary cultures of human adenomyosis foci cells were prepared from hard uterine lesions of adenomyosis patients. The cells were treated with JSGD (10 and 20 mg/ml), CDS, and mifepristone (MIF) for 24 or 48 h. Cell proliferation was detected using CCK-8 assay, cell apoptosis was measured by flow cytometry, and the cell ultrastructure was observed by transmission electron microscopy (TEM). RESULTS: JSGD and CDS significantly induced cell apoptosis and inhibited cell proliferation for 24 h or 48 h, in which the effects of JSGD were in a dose-dependent manner. The effect of CDS for 24 h was higher than that of CDS for 48 h. Moreover, JSGD and CDS treatments induced marked apoptosis in adenomyosis foci cells, characterized by nucleus chromatin, condensation, fragmentation, mitochondria and endoplasmic swelling, and autophagy-lysosome. CONCLUSIONS: JSGD and CDS can suppress proliferation and induce apoptosis in adenomyosis foci cells, through altering their ultrastructure. The results provided support for JSGD and CDS in the treatment of adenomyosis and gained further insight into the effect of this prescription.

[Role of estrogen, estrogen receptors, and aromatase in the pathogenesis of uterine adenomyosis].

OBJECTIVE: To study the role of estrogen (E2), estrogen receptor (ER) and aromatase (P450arom) in the pathogenesis of uterine adenomyosis. METHODS: Paraffin-embedded specimens of the uterine tissue from patients with uterine adenomyosis and patients with cervical lesions (CIN; control) were examined for expressions of E2, ER and P450arom by immunohistochemistry and ELISA. The cells isolated from the lesions of patients with adenomyosis were cultured in vitro, and the changes in cell growth in response to treatments with E2, ER inhibitor, ER inhibitor + E2, estrogen deprivation, and estrogen deprivation+ ICI182780 were assessed using CCK-8 method. RESULTS: The expression levels of E2, ER, and P450arom were significantly higher in adenomyosis ectopic lesions and eutopic endometrium than in the myometrium and endometrium in the control group (P<0.05); no significant difference in E2 and P450arom expressions was found between adenomyosis ectopic lesions and eutopic endometrium (P>0.05), while the expression levels of ER in ectopic lesions was significantly higher than that in eutopic endometrium. The cell inhibition rates were similar between ER inhibitor group and ER inhibitor + Estrogen activation group (P>0.05), and was significantly higher in estrogen deprivation+ ER inhibitor group than in estrogen deprivation group (P<0.05). CONCLUSION: The high expression levels of E2, ER, and P450arom in adenomyosis ectopic lesions and eutopic endometrium promote uterine adenomyosis cell proliferation, in which process E2 combines with ER to execute its biological effect; ER also promotes the occurrence and development of uterine adenomyosis through other pathways.