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Loss-of-function CFTR p.G970D missense mutation might cause congenital bilateral absence of the vas deferens and be associated with impaired spermatogenesis

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Asian Journal of Andrology 2023年第1期25卷 58-65页

作者： Jian-Wen Hou Xiao-Liang Li Li Wang Cong-Ling Dai Na Li Xiao-Hui Jiang Yue-Qiu Tan Er-Po Tian Qin-Tong Li Wen-Ming Xu Department of Obstetrics/Gynecology Key Laboratory of ObstetricGynecologic and Pediatric Diseases and Birth Defects of Ministry of EducationWest China Second University HospitalSichuan UniversityChengdu 610041China Reproductive Medical Center Chengdu Xi'nan Gynecology HospitalChengdu 610041China Reproductive Medical Center of West China Second University hospital Sichuan UniversityChengdu 610041China Reproductive Medical Center Gansu Provincial Maternal and Child-Care HospitalLanzhou 730050China Institute of Reproductive and Stem Cell Engineering School of Basic Medical ScienceCentral South UniversityChangsha 410000China Reproductive and Genetic hospital of CITIC-Xiangya Changsha 410000China Institute of Pediatrics Guangzhou Women and Children's Medical CenterGuangzhou Medical UniversityGuangzhou 510623China Human Sperm bank West China Second University HospitalSichuan UniversityChengdu 610041China Clinical Research Center for Reproduction and Genetics in Hunan Province Changsha 410000China Department of Obstetrics Gynecology and PediatricsWest China Second University HospitalKey Laboratory of Birth Defects and Related Diseases of Women and ChildrenMinistry of EducationSichuan UniversityChengdu 610041China

Congenital bilateral absence of the vas deferens(CBAVD)is observed in 1%–2%of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator(CFTR)*** is one of the most well-known genes related to male *** frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia(NOA).CFTR mutations are highly polymorphic and have established ethnic *** with F508Del in Caucasians,the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic ***,whether p.G970D participates in male infertility remains ***,a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and *** retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation(4.1%,5/122),excluding polymorphic ***,we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in *** Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli *** spermatocyte GC-2(spd)ts(GC2)Cftr p.G965D cells,RNA splicing variants were detected and CFTR expression decreased,which may contribute to the phenotypes associated with impaired ***,this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.

关键词： azoospermia congenital bilateral absence of the vas deferens CFTR mutation male infertility spermatogenesis

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A novel micro-straw for cryopreservation of sma number of human spermatozoon

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Asian Journal of Andrology 2017年第3期19卷 326-329页

作者： Feng Liu Sha-Sha Zou Yong Zhu Can Sun Yu-Fei Liu Shan-Shan Wang Wen-Bo Shi Jing-Jing Zhu Yong-Hua Huang Zheng Li Department of Andrology Center for Men's Health Shanghai China Urologic Medical Center Shanghai General Hospital Shanghai Jiao Tong University Shanghai China Center for Reproductive Medicine Ren Ji Hospital School of Medicine Shanghai Jiao Tong University Shanghai China Shanghai Human Sperm bank. Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics Shanghai China Institute of Refrigeration and Cryogenics School of Mechanical Engineering Shanghai Jiao Tong University Shanghai China Shanghai Key Laboratory of Reproductive Medicine Shanghai China

Cryopreservation of few spermatozoa is still a major challenge for male fertility preservation. This study reports use a new micro-straw （LSL straw） for freezing few spermatozoa for intracytoplasmic sperm injection （ICSI）. Semen samples from 22 fertile donors were collected, and each semen sample was diluted and mixed with cryoprotectant in a ratio of 1：1, and then frozen using three different straws such as LSL straw （50-100μl）, traditional 0.25 ml and 0.5 ml straws. For freezing, all straws were fumigated with liquid nitrogen, with temperature directly reducing to -130--140℃. Sperm concentration, progressive motility, morphology, acrosome integrity, and DNA fragmentation index were evaluated before and after freezing. After freezing-thawing, LSL straw group had significantly higher percentage of sperm motility than traditional 0.25 ml and 0.5 ml straw groups （38.5% vs 27.4% and 25.6%, P 〈 0.003）. Sperm motility and acrosomal integrity after freezing-thawing were significantly lower than that of before freezing. However, there was no significant difference in morphology, acrosome, and DNA integrity between the three types of straws （P 〉 0.05）. As LSL straws were thinner and hold very small volume, the freezing rate of LSL straw was obviously faster than 0.25 ml straw and 0.5 ml straws. In conclusion, LSL micro-straws may be useful to store few motile spermatozoa with good recovery of motility for patients undergoing ICSI treatment.

关键词： cryopreservation freezing rate LSL straw micro-straw motility

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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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Plasticity of male germline stem cells and their applications in reproductive and regenerative medicine

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Asian Journal of Andrology 2015年第3期17卷 367-372页

作者： Zheng Chen Zheng Li Zuping He State Key Laboratory of Oncogenes and Related Genes Renji-Med X Clinical Stem Cell Research Center Ren Ji Hospital School of Medicine Shanghai Jiao Tong University Shanghai Institute of Cancer Shanghai 200127 China Department of Urology Shanghai Human Sperm Bank Shanghai Institute of Andrology Ren Ji Hospital School of Medicine Shanghai Jiao Tong University Shanghai 200001 China Shanghai Key Laboratory of Assisted Reproduction and Reproductive Genetics Shanghai 200127 China Shanghai Key Laboratory of Reproductive Medicine Shanghai 200025 China

Spermatogonial stem cells （SSCs）, also known as male germline stem cells, are a small subpopulation of type A spermatogonia with the potential of self-renewal to maintain stem cell pool and differentiation into spermatids in mammalian testis. SSCs are previously regarded as the unipotent stem cells since they can only give rise to sperm within the seminiferous tubules. However, this concept has recently been challenged because numerous studies have demonstrated that SSCs cultured with growth factors can acquire pluripotency to become embryonic stem-like cells. The in vivo and in vitro studies from peers and us have clearly revealed that SSCs can directly transdifferentiate into morphologic, phenotypic, and functional cells of other lineages. Direct conversion to the cells of other tissues has important significance for regenerative medicine. SSCs from azoospermia patients could be induced to differentiate into spermatids with fertilization and developmental potentials. As such, SSCs could have significant applications in both reproductive and regenerative medicine due to their unique and great potentials. In this review, we address the important plasticity of SSCs, with focuses on their self-renewal, differentiation, dedifferentiation, transdifferentiation, and translational medicine studies.

关键词： differentiation direct transdifferentiation pluripotency self-renewal spermatogonial stem cells

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Nasopharyngeal brushing: a convenient and feasible sampling method for nucleic acid-based nasopharyngeal carcinoma research

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Cancer Communications 2018年第1期38卷 95-104页

作者： Pei-Fen Zhang Xiao-Hui Zheng Xi-Zhao Li Tian Tian Shao-Dan Zhang Ye-Zhu Hu Wei-Hua Jia Tumor Resource bank State Key Laboratory of Oncology in South ChinaCollaborative Innovation Center for Cancer MedicineSun Yat-sen University Cancer Center651 Dongfeng East RoadGuangzhou 510060Guang-dongP.R.China Affiliated Tumor hospital of Xinjiang Medical University Ürümqi 830000XinjiangP.R.China

Background:Tissue specimens for nasopharyngeal carcinoma(NPC)research are scarce because of sampling *** studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method for NPC *** present study aimed to evaluate the feasibility of nasopharyngeal brushing in the acquisition of NPC nucleic acids for ***:Nasopharyngeal brushing samples were acquired from 24 healthy individuals and 48 NPC *** from 48 NPC and 18 nasopharyngitis patients were collected by endoscopic *** expression levels of tumor suppressor genes(TSGs)and Epstein-Barr virus(EBV)-encoded microRNAs as well as EBV DNA copy number were measured by quantitative polymerase chain reaction in both types of ***:Among six TSGs examined,the expression levels of two genes were significantly decreased in nasopharyn-geal brushing and tissue samples from NPC patients as compared with those from healthy/nasopharyngitis *** EBV-encoded microRNAs,mir-bart1-5p,mir-bart5,mir-bart6-5p,and mir-bart17-5p,were significantly up-regulated in both NPC brushing and tissue samples compared with those from healthy/nasopharyngitis controls(Pal brushing samples(Palthy ***:Nasopharyngeal brushing can obtain sufficient tumoral materials for the analysis of viral nucleic acid,including EBV-encoded microRNAs and EBV *** the detection of TSG expression,nasopharyngeal brushings was feasible but inferior to tissue *** study confirms nasopharyngeal brushing as an applicable sampling method that can aid in nucleic acid-based NPC research.

关键词： Nasopharyngeal brushing Tissue samples Nasopharyngeal carcinoma Nucleic acid Cancer research

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Deciphering key genes involved in cisplatin resistance in kidney renal clear cell carcinoma through a combined in silico and in vitro approach

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Oncology Research 2023年第6期31卷 899-916页

作者： MUNEEBA MalIK MAMOONA MAQBOOL TOOBA NISAR TAZEEM AKHTER JAVED AHMED UJAN alANOOD SalGARNI FAKHRIA Aal JOUFI SULTAN SHAFI KalANAZI MOHAMMAD HADI alMOTARED MOUNIR MSalEM BEKHIT MUHAMMAD JAMIL Services hospital LahorePakistan Sabzazar Medical Center LahorePakistan BHU Pourmiana AttockPakistan Public Health Department University of Health SciencesLahorePakistan Department of Zoology Shah Abdul Latif UniversityKhairpurPakistan Department of Animal Sciences University of FloridaGainesvilleUSA Pharmacology and Toxicology Department College of Pharmacy Umm Al-Qura UniversityMakkahSaudi Arabia Department of Pharmacology College of PharmacyJouf UniversityAljoufSaudi Arabia Regional Laboratory and Blood bank(Total Laboratory Automation) RiyadhSaudi Arabia Health Affairs in Najran New Najran General HospitalNajranSaudi Arabia Department of Pharmaceutics College of PharmacyKing Saud UniversityRiyadhSaudi Arabia PARC Arid Zone Research Center Dera Ismail KhanPakistan

The low survival rate of Kidney renal clear cell carcinoma(KIRC)patients is largely attributed to cisplatin *** than focusing solely on individual proteins,exploring protein-protein interactions could offer greater insight into drug *** this end,a series of in silico and in vitro experiments were conducted to identify hub genes in the intricate network of cisplatin resistance-related genes in KIRC *** genes involved in cisplatin resistance across KIRC were retrieved from the National Center for Biotechnology Information(NCBI)database using search terms as“Kidney renal clear cell carcinoma”and“Cisplatin resistance”.The genes retrieved were analyzed for hub gene identification using the STRING database and Cytoscape *** and promoter methylation profiling of the hub genes was done using UalCAN,GEPIA,OncoDB,and HPA ***,survival,functional enrichment,immune cell infiltration,and drug prediction analyses of the hub genes were performed using the cBioPortal,GEPIA,GSEA,TIMER,and Drugbank ***,expression and methylation levels of the hub genes were validated on two cisplatin-resistant RCC cell lines(786-O and A-498)and a normal renal tubular epithelial cell line(HK-2)using two high throughput techniques,including targeted bisulfite sequencing(bisulfite-seq)and RT-qPCR.A total of 124 genes were identified as being associated with cisplatin resistance in *** of these genes,MCL1,IGF1R,CCND1,and PTEN were identified as hub genes and were found to have significant(pall survival(OS)of the KIRC ***,an aberrant promoter methylation pattern was found to be associated with the dysregulation of the hub *** addition to this,hub genes were also linked with different cisplatin resistance-causing ***,hub genes can be targeted with alvocidib,Estradiol,Tretinoin,Capsaicin,Dronabinol,Metribolone,Calcitriol

关键词： KIRC Cisplatin resistance Chemotherapy Overall survival

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Performance of Loop-Mediated Isothermal Amplification (LAMP)-Lateral Flow Detection and Xpert MTB/RIF Ultra for Diagnosis of Pulmonary Tuberculosis

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Journal of Tuberculosis Research 2021年第4期9卷 245-255页

作者： Siripan Kounmee Uraporn Phumisantiphong Yutthana Apichatbutr Pulmonology Division Department of Internal Medicine Faculty of Medicine Vajira Hospital Navamindradhiraj University Bangkok Thailand Department of Clinical Pathology Central Laboratory and Blood Bank Faculty of Medicine Vajira Hospital Navamindradhiraj Uni-versity Bangkok Thailand

Background: Due to the limitations of diagnosis by Xpert MTB/RIF assay, WHO suggests Loop—Mediated Isothermal Amplification (TB-LAMP) instead of sputum-smear microscopy for pulmonary TB diagnosis in patients. Dr. Thongchai Kaewphinit et al. invented a fast TB detection kit called TB d-tect (LAMP-LFD assay). There was no clinical trial to estimate the performance of TB d-tect. Objective: This study was aimed to find the performance of LFD assay and Xpert MTB/RIF ultra for pulmonary TB. Material and methods: A cross-sectional study was conducted. Suggestive pulmonary TB patients were enrolled from June 2020-28 February 2021. Respiratory specimens were collected from each patient and sent for AFB smear, LAMP-LFD assay, Xpert MTB/RIF ultra and culture TB. Result:

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In situ cornea harvesting through the Red Cross Organization:a new approach to relieving severe cornea donor shortage in Chinese eye banks

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International Journal of Ophthalmology(English edition) 2017年第10期10卷 1611-1613页

作者： Su-Xia Li Fu-Hua Wang Ting Wang Sha-Sha Han Wei-Yun Shi Shandong Eye hospital Shandong Eye InstituteShandong Academy of Medical Sciences The People hospital of Dongying

Corneal diseases are currently the second main cause of blindness in *** most of the corneal blindness could be treated by corneal transplantation,only about 10 000 operations were carried out each year owing to the severe shortage of corneal donors and limited eye bank programs.A feasible cornea donation program was established through the organization of the Red Cross,and in situ corneal removal techniques were developed to avoid conflicts with Chinese traditions of keeping the deceased *** number of donated corneas,which had a safe and secure quality,increased significantly year by year.

关键词： cornea donor transplantation eye bank

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Women's health and Chinese integrative medicine

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2017年第3期18卷 183-185页

作者： Fan QU Fang-fang WANG Nicola ROBINSON Paul J.HARDIMAN Women's hospital School of Medicine Zhejiang University School of Health and Social Care London South Bank University Institute for Women's Health University College London Medical School

Researchers in the fields of obstetrics, gynecology, and Chinese integrative medicine （CIM） were invited to share their perspectives on the most recent studies.

关键词：女性健康中西医结合医学妇产科学

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Basic Phenotyping of Male Fertility from 2019 to 2020 at the Human Sperm bank of Fudan University

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Phenomics 2022年第3期2卷 211-218页

作者： Yadong Li Can Sun Haitao Ma Hong Zhu Feng Zhang Feng Jiang Human Sperm bank of Fudan University Obstetrics and Gynecology Hospital of Fudan UniversityShanghai 200011China Obstetrics and Gynecology hospital of Fudan University NHC Key Laboratory of Reproduction Regulation(Shanghai Institute of Planned Parenthood Research)Shanghai Key Laboratory of Female Reproductive Endocrine Related DiseasesState Key Laboratory of Genetic Engineering at School of Life SciencesFudan UniversityShanghai 200011China Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology Hospital of Fudan UniversityFudan UniversityShanghai 200011China

The aim of this cross-sectional survey was to analyze the semen parameters of volunteers from the Human Sperm bank of Fudan University,as well as the related factors influencing these *** January 2019 to December 2020,semen parameters from a total of 5214 men were included in this *** Kruskal-Wallis test was used to detect differences associated with several independent variables.A total of 5214 volunteers were *** volunteers were registered in 33 provinces,autonomous regions,municipalities(including Macao and Taiwan)and 294 prefecture-level *** average age of volunteers was 27.40 ***,76.50%of the volunteers had a college education or *** with BMI values of 18.5-23.9 kg/m2 accounted for 60.70%of *** parameters were significantly different according to season,education level,duration of abstinence,age group and *** basic male fertility phenotypes(semen parameters)showed new trends in the study period,and accurate long-term tracking of male semen parameters will help researchers to better understand the changes in male fertility phenotypes(semen).

关键词： Male fertility phenotypes Semen parameters Sperm quality Human sperm bank

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