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scRNA-seq reveals that origin recognition complex subunit 6 regulates mouse spermatogonial cell proliferation and apoptosis via activation of Wnt/β-catenin signaling

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Asian Journal of Andrology 2024年第1期26卷 46-56页

作者： Shi-Wei Liu Jia-Qiang Luo Liang-Yu Zhao Ning-Jing Ou Chao-Yang Yu-Xiang Zhang Hao-Wei Bai Hong-Fang Sun Jian-Xiong Zhang Chen-Cheng Yao Peng Li ru-hui tian Zheng Li Zi-Jue Zhu Department of Andrology Center for Men's HealthUrologic Medical CenterShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China State Key Lab of Reproductive Medicine Nanjing Medical UniversityNanjing 211166China Shanghai Key Lab of Reproductive Medicine Shanghai Jiao Tong University School of MedicineShanghai 200080China Department of Urology Guangdong Provincial Key Laboratory of Biomedical ImagingThe Fifth Affiliated HospitalSun Yat-sen UniversityZhuhai 519000China Department of Urology First Affiliated Hospital of Nanjing Medical UniversityNanjing 210029China

The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining *** single-cell RNA sequencing(scRNA-seq)analysis of the testis was performed to identify genes upregulated in *** scRNAseq analysis,we identified the spermatogonia upregulated gene origin recognition complex subunit 6(Orc6),which is involved in DNA replication and cell cycle regulation;its protein expression in the human and mouse testis was detected by western blot and *** explore the potential function of Orc6 in spermatogonia,the C18-4 cell line was transfected with control or Orc6 ***,5-ethynyl-2-deoxyuridine(EdU)and terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assays,flow cytometry,and western blot were used to evaluate its effects on proliferation and *** was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 *** RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated(Wnt)/β-catenin *** blot revealed that the expression ofβ-catenin protein and its phosphorylation(Ser675)were significantly decreased when silencing the expression of *** findings indicated that Orc6 was upregulated in spermatogonia,whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

关键词： cell proliferation ORCG scRNA-seq analysis spermatogonia Wnt/β-catenin signaling

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Estradiol is an independent risk factor for organic erectile dysfunction in eugonadal young men

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Asian Journal of Andrology 2020年第6期22卷 636-641页

作者： hui-Rong Chen ru-hui tian Peng Li hui-Xing Chen Shu-Jie Xia Zheng Li Department of Andrology The Center for Men's HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong UniversityShanghai 200080China

Erectile dysfunctio n attributable to testosterone deficiency is less comm on in young males,and the effect of estradiol on erectile function in eug on adal young males is un *** an a lyzed data from 195 male participa nts,including 143 eug on adal patients with erectile dysfunction and 52 healthy *** distinguish psychogenic and organic erectile dysfunction,penile rigidity was measured using the nocturnal penile tumescenee rigidity *** levels of sexual hormones were quantified by electrochemiluminescence,and penile vascular status was assessed by penile color Doppler *** serum estradiol levels and the ratio of estradiol to testosterone were higher in patients with organic erectile dysfunction than in patients with psychogenic erectile dysfunction or healthy *** erectile dysfunction was negatively associated with estradiol levels and the ratio of estradiol to testosterone,and estradiol was the only sign ifica nt risk factor for orga nic erectile dysf un ction(odds ratio:1.094;95%con fide nee interval:1.042-1.149,P=0.000).Moreover,serum estradiol levels were negatively correlated with penile *** estradiol levels were higher and penile rigidity was lower in patients with venous erectile dysfunction than in patients with nonvascular erectile dysf *** con elude that elevated serum estradiol levels may impair erectile function and may be involved in the pathoge nesis of organic erectile dysfunction in eugonadal young men.

关键词： erectile dysfunction estradiol organic etiology testosterone young

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Seminal plasma miR-192a： a biomarker predicting successful resolution of nonobstructive azoospermia following varicocele repair

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Asian Journal of Andrology 2018年第4期20卷 396-399页

作者： Er-Lei Zhi Guo-Qing Liang Peng Li hui-Xing Chen ru-hui tian Peng Xu Zheng Li Department of Andrology Center for Men's Health Department of ART Institute of Urologic Medical Center Shanghai General Hospital Shanghai Key Lab of Reproductive Medicine Shanghai Jiao Tong University 100 Haining Road Shanghai 200080 China Department of Andrology Seventh People's Hospital of Shangha University of TCM Shanghai 200000 China Oriental Jinghua Hospital of Shenyang Department of ART 156 Nanjing Road Shenyang 110005 China

This study was performed to investigate a potential marker for the presence of spermatozoa in the ejaculate following varicocelectomy in Chinese men with nonobstructive azoospermia and varicoceles. The micro-RNA （miR）-192a levels in seminal plasma and testicular tissue were evaluated by quantitative real-time polymerase chain reaction from 60 men with nonobstructive azoospermia and varicoceles （Group A： 27 men with spermatozoa found in the ejaculate after surgery; Group B： 33 men without spermatozoa found in the ejaculate after surgery） and 30 controls. The seminal plasma and testicular tissue miR-192a levels were higher in Group B than in Group A and the controls （P〈 0.001）, and there was no significant difference between Group A and the controls （P〉 0.05）. Apoptosis and proliferation assays with miR mimics and inhibitors showed that miR-192a induced GC-2 cell apoptosis through the activation of Caspase-3 protein. Thus, seminal plasma miR-192a appears to be a potential marker for successfully indicating spermatozoa in the ejaculate following microsurgical varicocelectomy in men with nonobstructive azoospermia and varicoceles. Seminal plasma miR-192a may be a useful clinical marker for prescreening to determine which patients with nonobstructive azoospermia and varicoceles would benefit from varicocelectomy.

关键词： miR-192a nonobstructive azoospermia seminal plasma varicocelectomy

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Successful microsurgical vasoepididymostomy for a case of cryptozoospermia

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Asian Journal of Andrology 2022年第4期24卷 436-437页

作者： ru-hui tian Yu-Hua Huang hui-Xing Chen Peng Li Er-Lei Zhi Chen-Cheng Yao Chao Yang Zheng Li Department of Andrology The Center for Men's HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China

Dear Editor,Cryptozoospermia is defined as the apparent absence of spermatozoa from fresh semen samples,but they *** found in centrifuged pellets.1 It is usually caused by a spermatogenic disorder,while cryptozoospermia associated with seminal duct obstruction is less common.2 Because of the extremely low sperm counts in semen samples,men with cryptozoospermia are often reliant on intracytoplasmic sperm injection(ICSI)for paternity.

关键词： injection crypt obstruction

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Low XIST expression in Sertoli cells of Klinefelter syndrome patients causes high susceptibility of these cells to an extra X chromosome

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Asian Journal of Andrology 2023年第6期25卷 662-673页

作者： Liang-Yu Zhao Peng Li Chen-Cheng Yao ru-hui tian Yu-Xin Tang Yu-Zhuo Chen Zhi Zhou Zheng Li Department of Andrology The Center for Men’s HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China Department of Urology The Fifth Affiliated Hospital of Sun Yat-sen UniversityZhuhai 519000China Department of Interventional Medicine Guangdong Provincial Key Laboratory of Biomedical ImagingThe Fifth Affiliated Hospital of Sun Yat-sen UniversityZhuhai 519000China School of Life Science and Technology ShanghaiTech UniversityShanghai 200120China

Klinefelter syndrome(KS)is the most common genetic cause of human male ***,the effect of the extra X chromosome on different testicular cell types remains poorly ***,we profiled testicular single-cell transcriptomes from three KS patients and normal karyotype control *** the different somatic cells,Sertoli cells showed the greatest transcriptome changes in KS *** analysis showed that X-inactive-specific transcript(XIST),a key factor that inactivates one X chromosome in female mammals,was widely expressed in each testicular somatic cell type but not in Sertoli *** loss of XIST in Sertoli cells leads to an increased level of X chromosome genes,and further disrupts their transcription pattern and cellular *** phenomenon was not detected in other somatic cells such as Leydig cells and vascular endothelial *** results proposed a new mechanism to explain why testicular atrophy in KS patients is heterogeneous with loss of seminiferous tubules but interstitial *** study provides a theoretical basis for subsequent research and related treatment of KS by identifying Sertoli cell-specific X chromosome inactivation failure.

关键词： Klinefelter syndrome nonobstructive azoospermia Sertoli cell spermatogenesis X chromosome inactivation

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Novel double-layer Silastic testicular prosthesis with controlled release of testosterone in vitro, and its effects on castrated rats

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Asian Journal of Andrology 2017年第4期19卷 433-438页

作者： hui-Xing Chen Shi Yang Ye Ning Hai-Hao Shao Meng Ma ru-hui tian Yu-Fei Liu Wei-Qiang Gao Zheng Li Wei-Liang Xia School of Biomedical Engineering & Med-X Research Institute Shanghai Jiao Tong University Shanghai 200030 China Department of Andrology Urologic Medical Center Shanghai General Hospital Shanghai Jiae Tong University Shanghai 200080 China Shanghai Human Sperm Bank Ren Ji Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200135 China Controlled Release Laboratory Shanghai Institute of Planned Parenthood Research Shanghai 200030 China

Testicular prostheses have been used to deal with anorchia for nearly 80 years. Here, we evaluated a novel testicular prosthesis that can controllably release hormones to maintain physiological levels of testosterone in vivo for a long time. Silastic testicular prostheses with controlled release of testosterone （STPT） with different dosages of testosterone undecanoate （TU） were prepared and implanted into castrated Sprague-Dawley rats. TU oil was applied by oral administration to a separate group of castrated rats. Castrated untreated and sham-operated groups were used as controls. Serum samples from every group were collected to measure the levels of testosterone （T）, follicle-stimulating hormone and luteinizing hormone （LH）. Maximum intracavernous penile pressure （ICPmax） was recorded. The prostates and seminal vesicles were weighed and subjected to histology, and a terminal dexynucleotidyl transferase-mediated UTP nick end labeling （TUNEL） assay was used to evaluate apoptosis. Our results revealed that the weights of these tissues and the levels of T and LH showed significant statistical differences in the oral administration and TU replacement groups compared with the castrated group （P 〈 0.05）. Compared with the sham-operated group, the ICPmax, histology and TUNEL staining for apoptosis, showed no significant differences in the hormone replacement groups implanted with medium and high doses of STPT. Our results suggested that this new STPT could release TU stably through its double semi-permeable membranes with excellent biocompatibility. The study provides a new approach for testosterone replacement therapy.

关键词： controlled-release preparations hormone replacement therapy hypogonadism prosthesis Silastic testosterone

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Microsurgical subinguinal varicocelectomy with spermatic cord double traction and vein stripping

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Asian Journal of Andrology 2020年第2期22卷 208-212页

作者： ru-hui tian Liang-Yu Zhao hui-Xing Chen Chao Yang Peng Li Yu-Hua Huang Zhong Wan Er-Lei Zhi Chen-Cheng Yao Zheng Li Department of Andrology Center for Men’s HealthDepartment of ARTUrologic Medical CenterShanghai General HospitalShanghai Key Lab of Reproductive MedicineShanghai Jiao Tong UniversityShanghai 200080China

We retrospectively reviewed data for 286 patients with varicocele who underwent microsurgical subinguinal varicocelectomy from March 2015 to May 2017 in Shanghai General Hospital(Shanghai,China).In this surgical approach,the testis was delivered,and the gubernacular and external cremasteric veins were *** addition,the spermatic cord was delivered downward with continuous double traction away from the external *** remaining procedure was similar to the conventional *** followed patients for at least 3 months and evaluated postoperative semen parameters,pain symptoms,and *** excluded data for 32 men due to inadequate follow-up(ructive azoospermia,and 60 had symptomatic *** progressive sperm counts increased in the oligoasthenospermic patients from a median preoperative value of 9.15×10^6 ml^?1 to 25.33×10^6 ml^?1(n=34),and 35.6%(26/73)initially oligoasthenospermic men contributed to unassisted *** returned to the ejaculate in 12.4%(15/121)azoospermia *** patients with scrotal pain(n=60),43(71.7%)reported complete resolution of pain,16(26.7%)reported partial resolution,and 1(1.7%)reported no *** patients experienced varicocele *** doubletraction strategy avoids opening the external oblique aponeurosis,and results in less damage and faster *** addition,the stripping strategy eliminates potential damage to the testis caused by the varicose *** results showed that microsurgical subinguinal varicocelectomy using spermatic cord double traction in conjunction with testicular delivery for vein stripping is a safe and effective approach for varicocele repair.

关键词： azoospermia low sperm count sperm motility spermatic cord varicocele

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Generation of male germ cells from induced pluripotent stem cells （iPS cells）： an in vitro and in vivo study

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Asian Journal of Andrology 2012年第4期14卷 574-579页

作者： Yong Zhu Hong-Liang Hu Peng Li Shi Yang Wei Zhang hui Ding ru-hui tian Ye Ning Ling-Ling Zhang Xi-Zhi Guo Zhan-Ping Shi Zheng Li Zuping He Renji Hospital Sperm Development and Genetics LaboratoryShanghai Human Sperm BankShanghai Institute of AndrologyDepartment of UrologyShanghai Jiao Tong University School of MedicineShanghai 200001China Bio-X Institutes Key Laboratory for the Genetics of Developmental and Neuropsychiatric DisordersShanghai Jiao Tong UniversityShanghai 200240China Clinic Stem Cell Research Center Renji HospitalShanghai Jiao Tong University School of MedicineShanghai 200127China Department of Biochemistry and Molecular and Cellular Biology Georgetown University Medical CenterWashingtonDC 20057USA

Recent studies have reported that induced pluripotent stem （iPS） cells from mice and humans can differentiate into primordial germ cells. However, whether iPS cells are capable of producing male germ cells is not known. The objective of this study was to investigate the differentiation potential of mouse iPS cells into spermatogonial stem cells and late-stage male germ cells. We used an approach that combines in vitrodifferentiation and in vivotransplantation. Embryoid bodies （EBs） were obtained from iPS cells using leukaemia inhibitor factor （LIF）-free medium. Quantitative PCR revealed a decrease in Oct4 expression and an increase in StraSand Vasa mRNA in the EBs derived from iPS cells, iPS cell-derived EBs were induced by retinoic acid to differentiate into spermatogonial stem cells （SSCs）, as evidenced by their expression of VASA, as well as CDH1 and GFRal, which are markers of SSCs. Furthermore, these germ cells derived from iPS cells were transplanted into recipient testes of mice that had been pre-treated with busulfan. Notably, iPS cell-derived SSCs were able to differentiate into male germ cells ranging from spermatogonia to round spermatids, as shown by VASA and SCP3 expression. This study demonstrates that iPS cells have the potential to differentiate into late-stage male germ cells. The derivation of male germ cells from iPS cells has potential applications in the treatment of male infertility and provides a model for uncovering the molecular mechanisms underlying male germ cell development.

关键词： differentiation induced pluripotent stem cells male germ cells retinoic acid transplantation

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Ultrasonographic evaluation of the rete testis thickness:a promising approach to differentiate obstructive from nonobstructive azoospermia

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Asian Journal of Andrology 2023年第6期25卷 725-730页

作者： Xin Li ru-hui tian Peng Li Chun-Xiao Li Ming-Hua Yao Chen-Cheng Yao Xiao-Bo Wang Li-Ren Jiang Zheng Li Rong Wu Department of Ultrasound Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China Department of Andrology The Center for Men’s HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China Department of Pathology Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China

This study aimed to evaluate the ability of rete testis thickness(RTT)and testicular shear wave elastography(SWE)to differentiate obstructive azoospermia(OA)from nonobstructive azoospermia(NOA).We assessed 290 testes of 145 infertile males with azoospermia and 94 testes of 47 healthy volunteers at Shanghai General Hospital(Shanghai,China)between August 2019 and October *** testicular volume(TV),SWE,and RTT were compared among patients with OA and NOA and healthy *** diagnostic performances of the three variables were evaluated using the receiver operating characteristic *** TV,SWE,and RTT in OA differed significantly from those in NOA(all P≤0.001)but were similar to those in healthy *** with OA and NOA were similar at TVs of 9–11 cm^(3)(P=0.838),with sensitivity,specificity,Youden index,and area under the curve of 50.0%,84.2%,0.34,and 0.662(95%confidence interval[CI]:0.502–0.799),respectively,for SWE cut-off of 3.1 kPa;and 94.1%,79.2%,0.74,and 0.904(95%CI:0.811–0.996),respectively,for RTT cut-off of 1.6 *** results showed that RTT performed significantly better than SWE in differentiating OA from NOA in the TV overlap *** conclusion,ultrasonographic RTT evaluation proved a promising diagnostic approach to differentiate OA from NOA,particularly in the TV overlap range.

关键词： azoospermia differential diagnosis rete testis thickness shear wave elastography

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Efficacy of stepwise mini-incision microdissection testicular sperm extraction for nonobstructive azoospermia with varied etiologies

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Asian Journal of Andrology 2023年第5期25卷 621-626页

作者： Yu-Xiang Zhang Chen-Cheng Yao Yu-Hua Huang Peng Li Er-Lei Zhi Zi-Jue Zhu Jian-Xiong Zhang Fu-Jun Zhao Zheng Li ru-hui tian Department of Andrology The Center for Men's HealthUrologic Medical CenterShanghai Key Laboratory of Reproductive MedicineShanghai General HospitalShanghai Jiao Tong University School of MedicineShanghai 200080China

Stepwise mini-incision microdissection testicular sperm extraction(mTESE)is a procedure that attempts to minimize testicular ***,the mini-incision approach may vary in patients with different ***,we performed a retrospective analysis of 665 men with nonobstructive azoospermia(NOA)who underwent stepwise mini-incision mTESE(Group 1)and 365 men who underwent standard mTESE(Group 2).The results showed that the operation time(mean±standard deviation)for patients with successful sperm retrieval in Group1(64.0±26.6min)was significantly shorter than that in Group2(80.2±31.3min),with P0.05),even when the etiologies of NOA were taken into *** results of consecutive multivariate logistic regression analysis(odds ratio[0R]:0.57;95%confidence interval[Cl]:0.38-0.87;P=0.009)and receiver operating characteristic(ROC)analysis(area under the ROC curve[AUC]=O.628)showed that preoperative anti-Mullerian hormone(AMH)level in idiopathic NOA patients was a potential predictor for surgical outcomes after initial three small incisions made in the equatorial region without sperm examined under an operating microscope(Steps 2-4).In conclusion,stepwise mini-incision mTESE is a useful technique for NOA patients,with comparable SRR,less surgical invasiveness,and shorter operation time compared with the standard *** AMH levels may predict successful sperm retrieval in idiopathic patients even after a failed initial mini-incision procedure.

关键词： etiology male infertility nonobstructive azoospermia standard mTESE stepwise mini-incision mTESE

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