Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells...Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells(SSCs)self-renew to maintain stem cell population within the testes and differentiate into mature spermatids.It is of great significance to uncover gene regulation and signaling pathways that are involved in the fate determinations of SSCs with aims to better understand molecular mechanisms underlying human spermatogenesis and identify novel targets for gene therapy of male infertility.Significant achievement has recently been made in demonstrating the signaling molecules and pathways mediating the fate decisions of mammalian SSCs.In this review,we address key gene regulation and crucial signaling transduction pathways in controlling the self-renewal,differentiation,and apoptosis of SSCs,and we illustrate the networks of genes and signaling pathways in SSC fate determinations.We also highlight perspectives and future directions in SSC regulation by genes and their signaling pathways.This review could provide novel insights into the genetic regulation of normal and abnormal spermatogenesis and offer molecular targets to develop new approaches for gene therapy of male infertility.展开更多
Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development re...Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development remain to be clarified.Here,we have demonstrated that Icariin promoted proliferation and DNA synthesis of mouse spermatogonial stem cells(SSCs).Furthermore,surface plasmon resonance iron(SPRi)and molecular docking(MOE)assays revealed that phosphodiesterase 5A(PDE5A)was an important target of Icariin in mouse SSCs.Mechanically,Icariin decreased the expression level of PDE5A.Interestingly,hydrogen peroxides(H2O2)enhanced the expression level of phosphorylation H2A.X(p-H2A.X),whereas Icariin diminished the expression level of p-H2A.X and DNA damage caused by H2O2 in mouse SSCs.Finally,our in vivo animal study indicated that Icariin protected male reproduction.Collectively,these results implicate that Icariin targets PDE5A to regulate mouse SSC viability and DNA damage and improves male reproductive capacity.This study thus sheds new insights into molecular mechanisms underlying the fate decisions of mammalian SSCs and offers a scientific basis for the clinical application of Icariin in male reproduction.展开更多
Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonucl...Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonuclease 1(PNLDCI)and male infertility,and we would like to contribute a commentary on this article.展开更多
Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production...Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.展开更多
Despite advancing therapeutic treatments,cancer remains the leading cause of death worldwide,with most of its patients developing drug resistance and recurrence after initial treatment.Therefore,incorporating preclini...Despite advancing therapeutic treatments,cancer remains the leading cause of death worldwide,with most of its patients developing drug resistance and recurrence after initial treatment.Therefore,incorporating preclinical models that mimic human cancer biology and drug responses is essential for improving treatment efficacy and prognosis.Patient-derived xenograft(PDX)models,as a promising and reliable preclinical trial platform,retain key features of the original tumor such as gene expression profiles,histopathological features,drug responses,and molecular signatures more faithfully compared with traditional tumor cell line models and cell line-derived xenograft models.Their significant advantages have been the preferred choice in cancer research,especially demonstrating remarkable potential in drug development,clinical combination therapy,and precision medicine.However,the successful construction and effective application of PDX models still face several challenges.In this review,we summarize the details of constructing PDX models and the drivers affecting their success rates,which will provide some theoretical basis for subsequent model optimization.In the meantime,we delineate the strengths and weaknesses of various mature PDX models and other developing preclinical models,including PDX-derived models,organoids,and genetically engineered models.Moreover,we highlight the challenges of newly developed technologies on the PDX models.Finally,we emphasize the innovative usage of PDX models in a variety of cancer studies and offer insights into their prospects.展开更多
Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs...Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs by key genes and signaling pathways remain elusive.Here,we report for the first time that Yes1-associated transcriptional regulator(YAP1)is required for fate determinations of SSCs and male fertility by interacting with RAD21 and targeting NEDD4 in humans and mice.YAP1 was mainly located at cell nuclei of human SSCs.YAP1 silencing resulted in the decreases in proliferation and DNA synthesis as well as an enhancement in apoptosis of human SSCs both in vivo and in vitro.RNA sequencing and real-time polymerase chain reaction assays identified NEDD4 as a target of YAP1,and NEDD4 knockdown inhibited the proliferation of human SSCs and increased their apoptosis.Furthermore,YAP1 interacted with RAD21 to regulate NEDD4 transcription in human SSCs.Importantly,YAP1 abnormalities were found to be associated with non-obstructive azoospermia(NOA)as manifested as lower expression level of YAP1 in testicular tissues of NOA patients and YAP1 single-nucleotide variants(SNVs)in 777 NOA patients.Finally,Yap1 germline conditional knockout(cKO)mice assumed mitotic arrest,low sperm count,and motility.Collectively,these results highlight a critical role of YAP1 in determining the fate determinations of human SSCs and male infertility through the YAP1/RAD21/NEDD4 pathway.This study provides new insights into the genetic regulatory mechanisms underlying human spermatogenesis and the pathogenesis of NOA,and it offers new targets for gene therapy of male infertility.展开更多
Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effec...Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effects of Pb2＋ on adult neural cells of humans or other mammals, only few of which have examined the effects of Pb2＋ on neural stem cells. The purpose of this study was to reveal the biological effects of Pb2＋from lead acetate [Pb （0H30OO）2] on viability, proliferation and differentiation of neural stem cells derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days, respectively. This study was carried out in three parts. In the first part, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay （MTT viability assay） was used to detect the effects of Pb2＋ on the cell viability of passage 2 hippocampal neural stem cells after 48-hour exposure to 0-200 pM Pb2＋. In the second part, 10 pM bromodeoxyuridine was added into the culture medium of passage 2 hippocampal neural stem cells after 48-hour exposure to 0- 200 pM Pb2＋, followed by immunocytochemical staining with anti-bromodeoxyuridine to demonstrate the effects of Pb2＋ on cell proliferation. In the last part, passage 2 hippocampal neural stem cells were allowed to grow in the differentiation medium with 0-200 pM Pb2＋. Immunocytochemical staining with anti-microtubule-associated protein 2 （a neuron marker）, anti-glial fibrillary acidic protein （an astrocyte marker）, and anti-RIP （an oligodendrocyte marker） was performed to detect the differentiation commitment of affected neural stem cells after 6 days. The data showed that Pb2~ inhibited not only the viability and proliferation of rat hippocampal neural stem cells, but also their neuronal and oligodendrocyte differentiation in vitro. Moreover, increased activity of astrocyte differentiation of hippocampal neural stem cells from both newborn and adult rats was observed after exposure to high concentration of lead ion in vitro. These findings suggest that hippocampal neural stem cells of newborn rats were more sensitive than those from adult rats to Pb2＋cytotoxicity.展开更多
Conflicting findings exist regarding the link between functional recovery and the regrowth of spinal tracts across the lesion leading to the restoration of functional contacts. In the present study, we investigated wh...Conflicting findings exist regarding the link between functional recovery and the regrowth of spinal tracts across the lesion leading to the restoration of functional contacts. In the present study, we investigated whether functional locomotor recovery was attributable to anatomical regeneration at postnatal day 1 （PN1）, PN7, PN14 and in adult rats two months after transection injury at the tenth thoracic segment of the spinal cord. The Basso, Beattie, and Bresnahan scores showed that transection led to a failure of hindlimb locomotor function in PN14 and adult rats. However, PN1 and PN7 rats showed a significant level of stepping function after complete spinal cord transection. Unexpectedly, unlike the transected PN14 and adult rats in which the spinal cord underwent limited secondary degeneration and showed a scar at the lesion site, the rats transected at PN1 and PN7 showed massive secondary degeneration both anterograde and retrograde, leaving a 〉5-mm gap between the two stumps. Furthermore, retrograde tracing with fluorogold （FG） also showed that FG did not cross the transection site in PN1 and PN7rats as in PN14 and adult rats, and re-transection of the cord caused no apparent loss in locomotor performance in the rats transected at PN1. Thus, these three lines of evidence strongly indicated that the functional recovery after transection in neonatal rats is independent of regrowth of spinal tracts across the lesion site. Our results support the notion that the recovery of locomotor function in developing rats may be due to intrinsic adaptations in the spinal circuitry below the lesion that control hindlimb locomotor activity rather than the regrowth of spinal tracts across the lesion. The difference in secondary degeneration between neonatal and adult rats remains to be explored.展开更多
The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by compari...The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by comparing immunoreactivity for p75 and phosphorylated c-Jun （p-c-Jun）, the production of JNK activation in axotomized motoneurons in postnatal day （PN）I, PN7, PN14 and adult rats. Intensive p-c-Jun was induced in axotomized motoneurons in PN1 and PN7. In PN14, p-c-Jun expression was sharply reduced after the same injury. The decreased expression of p-c-Jun at this age coincided with a developmental switch of re-expression of p75 in axotomized cells. In adult animals, no p-c-Jun but intensive p75 was detected in axotomized motoneurons. These results indicate differential expression or turnover of phosphorylation of c-Jun and p75 in immature versus mature spinal motoneurons in response to axonal injury. The non-co-occurrence of p75 and p-c-Jun in injured motoneurons indicated that p75 may not activate JNK pathway, suggesting that the p75 may not be involved in cell death in axotomized motoneurons.展开更多
Three-dimensional（3D） histology utilizes tissue clearing techniques to turn intact tissues transparent,allowing rapid interrogation of tissue architecture in three dimensions.In this article,we summarized the availa...Three-dimensional（3D） histology utilizes tissue clearing techniques to turn intact tissues transparent,allowing rapid interrogation of tissue architecture in three dimensions.In this article,we summarized the available tissue clearing methods and classified them according to their physicochemical principles of operation,which provided a framework for one to choose the best techniques for various research settings.Recent attempts in addressing various questions regarding the degenerating and regenerating nervous system have been promising with the use of 3D histological techniques.展开更多
Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.Ho...Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.However,the roles and molecular mechanisms of specific genes in mediating fate decisions of PCSCs and carcinogenesis of PCa remain to be elusive.In this study,we have explored the expression,function,and mechanism of AZGP1P2,a pseudogene of AZGP1,in regulating the stemness and apoptosis of PCSCs and treatment resistance of docetaxel in castration-resistant prostate cancer(CRPC).We revealed that AZGP1P2 was downregulated in CRPC cell lines and PCSCs,while it was positively associated with progression-free interval.Upregulation of the AZGP1P2 enhanced the sensitivity of docetaxel treatment in CRPCs via inhibiting their stemness.RNA pull-down associated with mass spectrometry analysis,co-immunoprecipitation assay,and RNA immunoprecipitation assay demonstrated that AZGP1P2 could bind to UBA1 and RBM15 as a“writer”of methyltransferase to form a compound.UBA1,an E1 ubiquitin-activating enzyme,contributed to RBM15 protein degradation via ubiquitination modification.Methylated RNA immunoprecipitation assay displayed that RBM15 controlled the mRNA decay of TPM1 in m6A methylation.Furthermore,a xenograft mouse model and patient-derived organoids showed that the therapeutic effect of docetaxel in CRPC was increased by AZGP1P2 in vivo.Collectively,these results imply that AZGP1P2 mediates the stemness and apoptosis of PCSCs and promotes docetaxel therapeutic effect by suppressing tumor growth and metastasis via UBA1/RBM15-mediated TPM1 mRNA decay in CRPC.展开更多
Early embryonic development is controlled by maternal factors originating from mature oocytes.The zygotic genome is activated from a transcriptionally quiescent state through a process called embryonic genome activati...Early embryonic development is controlled by maternal factors originating from mature oocytes.The zygotic genome is activated from a transcriptionally quiescent state through a process called embryonic genome activation(EGA),which involves the depletion and clearance of maternal factors.However,the mechanism by which maternal factors regulate EGA and embryonic development,particularly in humans,remains elusive.In this study,using tri-pronuclear(3PN) embryos and human embryonic stem cells(h ESCs),we demonstrated that the maternal transcription factor Orthodenticle Homeobox 2(OTX2),a paired-like homeobox gene,promotes EGA in human pre-implantation embryos.Knockdown of OTX2 through Trim-Away technology blocked embryonic development and minor EGA gene expression.Overexpression of OTX2(OTX2~(OE)) in h ESCs increased transcript products,primarily at the 2-cell embryo stage genes,including genes encoding methyltransferase of histone H3K4.OTX2~(OE) increased the level of H3K4me3 and increased the open chromatin region that co-occurs with the H3K4me3 region at the 4-cell stage in h ESCs.Based on these findings in h ESCs,we further verified that OTX2 directly induced the expression of SETD1A by binding to its promoter,leading to increased H3K4me3 levels in both h ESCs and 3PN embryos.These findings suggest that the maternal transcription factor OTX2 regulates EGA and early embryogenesis via epigenetic mechanisms.展开更多
Background： The worsening of semen quality, due to the application of Wi-Fi, can be ameliorated by Vitamin E. This study aimed to demonstrate whether a moderate dose of trolox,a new Vitamin E,inhibits oxidative damag...Background： The worsening of semen quality, due to the application of Wi-Fi, can be ameliorated by Vitamin E. This study aimed to demonstrate whether a moderate dose of trolox,a new Vitamin E,inhibits oxidative damage on sperms in vitro after exposure to Wi-Fi radiation. Methods： Each of the twenty qualified semen, gathered from June to October 2014 in eugenics clinic, was separated into four aliquots, including sham, Wi-Fi-exposed, Wi-Fi plus 5 mmol/L trolox, and Wi-Fi plus 10 mmol/L trolox groups. At 0 min, all baseline parameters of the 20 samples were measured in sequence. Reactive oxygen species, glutathione, and superoxide dismutase were evaluated in the four aliquots at 45 and 90 min, as were sperm DNA fragments, sperm mitochondrial potential, relative amplification of sperm mitochondrial DNA, sperm vitality, and progressive and immotility sperm. The parameters were analyzed by one-way analysis of variance and Tukey＇s posttest. Results： Among Wi-Fi plus 5 mmol/L trolox, Wi-Fi-exposed and Wi-Fi plus 10 mmol/L trolox groups, reactive oxygen species levels （45 min： 3.80 ± 0.41 RLU·10^-6·ml^-1. 7.50 ± 0.35 RLU·10^-6·ml^-1vs. 6.70 ± 0.47 RLU·10^-6·ml^-1P 〈 0.001; 90 min：5.40 ± 0.21 RLU·10^-6·ml^-1vs. 10.10 ± 0.31 RLU·10^-6·ml^-1. 7.00 ± 0.42 RLU·10^-6·ml^-1P 〈 0.001, respectively）, percentages of tail DNA （45 min： 16.8 ± 2.0% vs. 31.9 ± 2.5% vs. 61.3 ± 1.6%, P 〈 0.001; 90 min： 19.7 ± 1.5% vs. 73.7 ± 1.3% vs. 73.1 ± 1.1%, P 〈 0.001, respectively）, 8-hydroxy-2＇-deoxyguanosine （45 min： 51.89 ± 1.46 pg/ml vs. 104.89 ± 2.19 pg/ml vs. 106.11 ± 1.81 pg/ml , P = 0.012;90 min： 79.96 ± 1.73 pg/ml vs. 141.73 ± 2.90 pg/ml vs. 139.06 ± 2.79 pg/ml; P 〈 0.001）, and percentages of immotility sperm （45 min：27.7 ± 2.7% vs. 41.7 ± 2.2% vs. 41.7 ± 2.5%; 90 min： 29.9 ± 3.3% vs. 58.9 ± 4.0% vs. 63.1 ± 4.0%; all P 〈 0.001） were lowest, and glutathione peroxidase （45 min： 60.50 ± 1.54 U/ml vs. 37.09 ± 1.77 U/ml vs. 28.18 ± 1.06 U/ml; 90 min： 44.61 ± 1.23 U/ml vs. 16.86 ± 0.93 U/ml vs. 29.94 ± 1.56 U/ml; all P 〈 0.001）, percentages of head DNA （45 min： 83.2 ± 2.0% vs. 68.2 ± 2.5% vs. 38.8 ± 1.6%; 90 min： 80.3 ± 1.5% vs. 26.3 ± 1.3% vs. 26.9 ± 1.1%; all P 〈 0.001）, percentages of sperm vitality （45 min： 89.5 ± 1.6% vs. 70.7 ± 3.1% vs. 57.7 ± 2.4%;90 min： 80.8 ± 2.2% vs. 40.4 ± 4.0% vs. 34.7 ± 3.9%; all P 〈 0.001）, and progressive sperm （45 min： 69.3 ± 2.7% vs. 55.8 ± 2.2% vs. 55.4 ± 2.5%; 90 min： 67.2 ± 3.3% vs. 38.2 ± 4.0% vs. 33.9 ± 4.0%; all P 〈 0.001） were highest in Wi-Fi plus 5 mmol/L trolox group at 45 and 90 min, respectively. Other parameters were not affected, while the sham group maintained the baseline. Conclusion：This study found that 5 mmol/L trolox protected the Wi-Fi-exposed semen in vitro from the damage of electromagnetic radiation-induced oxidative stress.展开更多
Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remai...Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remain elusive.Here,we have demonstrated for the first time that OIP5(Opa interacting protein 5)controlled the self-renewal and apoptosis of human SSCs.RNA sequencing identified that NCK2 was a target for OIP5 in human SSCs,and interestingly,OIP5 could interact with NCK2 as shown by Co-IP(co-immunoprecipitation),IP-MS(mass spectrometry),and GST pulldown assays.NCK2 silencing decreased human SSC proliferation and DNA synthesis but enhanced their apoptosis.Notably,NCK2 knockdown reversed the influence of OIP5 overexpression on human SSCs.Moreover,OIP5 inhibition decreased the numbers of human SSCs at S and G2/M phases,while the levels of numerous cell cycle proteins,including cyclins A2,B1,D1,E1 and H,especially cyclin D1,were remarkably reduced.Significantly,whole-exome sequencing of 777 patients with nonobstructive azoospermia(NOA)revealed 54 singlenucleotide polymorphism mutations of the OIP5 gene(6.95%),while the level of OIP5 protein was obviously lower in testes of NOA patients compared to fertile men.Collectively,these results implicate that OIP5 interacts with NCK2 to modulate human SSC self-renewal and apoptosis via cell cyclins and cell cycle progression and that its mutation and/or lower expression is correlated with azoospermia.As such,this study offers novel insights into molecular mechanisms underlying the fate determinations of human SSCs and the pathogenesis of NOA,and it provides new targets for treating male infertility.展开更多
基金supported by the grants from the National Nature Science Foundation of China(No.32170862)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(No.2019SK1012)+1 种基金the Research Team for Reproduction Health and Translational Medicine of Hunan Normal University(No.2023JC101)Graduate Scientific Research Innovation Project of Hunan Province,China(No.CX2022520).
文摘Infertility has become one of the most serious diseases worldwide,and 50% of this disease can be attributed to male-related factors.Spermatogenesis,by definition,is a complex process by which spermatogonial stem cells(SSCs)self-renew to maintain stem cell population within the testes and differentiate into mature spermatids.It is of great significance to uncover gene regulation and signaling pathways that are involved in the fate determinations of SSCs with aims to better understand molecular mechanisms underlying human spermatogenesis and identify novel targets for gene therapy of male infertility.Significant achievement has recently been made in demonstrating the signaling molecules and pathways mediating the fate decisions of mammalian SSCs.In this review,we address key gene regulation and crucial signaling transduction pathways in controlling the self-renewal,differentiation,and apoptosis of SSCs,and we illustrate the networks of genes and signaling pathways in SSC fate determinations.We also highlight perspectives and future directions in SSC regulation by genes and their signaling pathways.This review could provide novel insights into the genetic regulation of normal and abnormal spermatogenesis and offer molecular targets to develop new approaches for gene therapy of male infertility.
基金supported by the grants from the National Nature Science Foundation of China(No.32170862)Developmental Biology and Breeding(No.2022XKQ0205)+2 种基金the Research Team for Reproduction Health and Translational Medicine of Hunan Normal University(No.2023JC101)Graduate Scientific Research Innovation Project of Hunan Province(No.CX2022520)Shanghai Key Laboratory of Reproductive Medicine(2022SKLRM01).
文摘Icariin is a pure compound derived from Epimedium brevicornu Maxim,and it helps the regulation of male reproduction.Nevertheless,the role and underlying mechanisms of Icariin in mediating male germ cell development remain to be clarified.Here,we have demonstrated that Icariin promoted proliferation and DNA synthesis of mouse spermatogonial stem cells(SSCs).Furthermore,surface plasmon resonance iron(SPRi)and molecular docking(MOE)assays revealed that phosphodiesterase 5A(PDE5A)was an important target of Icariin in mouse SSCs.Mechanically,Icariin decreased the expression level of PDE5A.Interestingly,hydrogen peroxides(H2O2)enhanced the expression level of phosphorylation H2A.X(p-H2A.X),whereas Icariin diminished the expression level of p-H2A.X and DNA damage caused by H2O2 in mouse SSCs.Finally,our in vivo animal study indicated that Icariin protected male reproduction.Collectively,these results implicate that Icariin targets PDE5A to regulate mouse SSC viability and DNA damage and improves male reproductive capacity.This study thus sheds new insights into molecular mechanisms underlying the fate decisions of mammalian SSCs and offers a scientific basis for the clinical application of Icariin in male reproduction.
基金This work was funded by the grant from the National Natural Science Foundation of China(No.32170862).
文摘Dear Editor,An interesting article from Zhao et al.recently published in Asian Journal of Andrology,has shown the association of a novel loss-of function(LOF)variant in PARN-like ribonuclease domain-containing exonuclease 1(PNLDCI)and male infertility,and we would like to contribute a commentary on this article.
文摘Objective Triple-negative breast cancer(TNBC)is the breast cancer subtype with the worst prognosis,and lacks effective therapeutic targets.Colony stimulating factors(CSFs)are cytokines that can regulate the production of blood cells and stimulate the growth and development of immune cells,playing an important role in the malignant progression of TNBC.This article aims to construct a novel prognostic model based on the expression of colony stimulating factors-related genes(CRGs),and analyze the sensitivity of TNBC patients to immunotherapy and drug therapy.Methods We downloaded CRGs from public databases and screened for differentially expressed CRGs between normal and TNBC tissues in the TCGA-BRCA database.Through LASSO Cox regression analysis,we constructed a prognostic model and stratified TNBC patients into high-risk and low-risk groups based on the colony stimulating factors-related genes risk score(CRRS).We further analyzed the correlation between CRRS and patient prognosis,clinical features,tumor microenvironment(TME)in both high-risk and low-risk groups,and evaluated the relationship between CRRS and sensitivity to immunotherapy and drug therapy.Results We identified 842 differentially expressed CRGs in breast cancer tissues of TNBC patients and selected 13 CRGs for constructing the prognostic model.Kaplan-Meier survival curves,time-dependent receiver operating characteristic curves,and other analyses confirmed that TNBC patients with high CRRS had shorter overall survival,and the predictive ability of CRRS prognostic model was further validated using the GEO dataset.Nomogram combining clinical features confirmed that CRRS was an independent factor for the prognosis of TNBC patients.Moreover,patients in the high-risk group had lower levels of immune infiltration in the TME and were sensitive to chemotherapeutic drugs such as 5-fluorouracil,ipatasertib,and paclitaxel.Conclusion We have developed a CRRS-based prognostic model composed of 13 differentially expressed CRGs,which may serve as a useful tool for predicting the prognosis of TNBC patients and guiding clinical treatment.Moreover,the key genes within this model may represent potential molecular targets for future therapies of TNBC.
基金supported by the National Natural Science Foundation of China(No.82172653)the Intra Institutional Open Fund of School of Medicine,Hunan Normal University(No.KF2022001)+1 种基金the Key Project of Developmental Biology and Breeding from Hunan Province,China(No.2022XKQ0205)The Research Team for Reproduction Health and Translational Medicine of Hunan Normal University(No.2023JC101).
文摘Despite advancing therapeutic treatments,cancer remains the leading cause of death worldwide,with most of its patients developing drug resistance and recurrence after initial treatment.Therefore,incorporating preclinical models that mimic human cancer biology and drug responses is essential for improving treatment efficacy and prognosis.Patient-derived xenograft(PDX)models,as a promising and reliable preclinical trial platform,retain key features of the original tumor such as gene expression profiles,histopathological features,drug responses,and molecular signatures more faithfully compared with traditional tumor cell line models and cell line-derived xenograft models.Their significant advantages have been the preferred choice in cancer research,especially demonstrating remarkable potential in drug development,clinical combination therapy,and precision medicine.However,the successful construction and effective application of PDX models still face several challenges.In this review,we summarize the details of constructing PDX models and the drivers affecting their success rates,which will provide some theoretical basis for subsequent model optimization.In the meantime,we delineate the strengths and weaknesses of various mature PDX models and other developing preclinical models,including PDX-derived models,organoids,and genetically engineered models.Moreover,we highlight the challenges of newly developed technologies on the PDX models.Finally,we emphasize the innovative usage of PDX models in a variety of cancer studies and offer insights into their prospects.
基金funded by the grants from National Nature Science Foundation of China(32470904 and 32170862)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)+3 种基金Key Grant of Research and Development in Hunan Province(2020DK2002)Developmental Biology and Breeding(2022XKQ0205)Shanghai Key Laboratory of Reproductive Medicine,Natural Science Foundation of Hunan Province of China(2024JJ5284,2023JJ30424,and 2024JJ5282)Research Foundation of Education Bureau of Hunan Province for Outstanding Young(23B0064).
文摘Spermatogenesis is a sophisticated biological process by which spermatogonial stem cells(SSCs)undergo self-renewal and differentiation into spermatozoa.Molecular mechanisms underlying fate determinations of human SSCs by key genes and signaling pathways remain elusive.Here,we report for the first time that Yes1-associated transcriptional regulator(YAP1)is required for fate determinations of SSCs and male fertility by interacting with RAD21 and targeting NEDD4 in humans and mice.YAP1 was mainly located at cell nuclei of human SSCs.YAP1 silencing resulted in the decreases in proliferation and DNA synthesis as well as an enhancement in apoptosis of human SSCs both in vivo and in vitro.RNA sequencing and real-time polymerase chain reaction assays identified NEDD4 as a target of YAP1,and NEDD4 knockdown inhibited the proliferation of human SSCs and increased their apoptosis.Furthermore,YAP1 interacted with RAD21 to regulate NEDD4 transcription in human SSCs.Importantly,YAP1 abnormalities were found to be associated with non-obstructive azoospermia(NOA)as manifested as lower expression level of YAP1 in testicular tissues of NOA patients and YAP1 single-nucleotide variants(SNVs)in 777 NOA patients.Finally,Yap1 germline conditional knockout(cKO)mice assumed mitotic arrest,low sperm count,and motility.Collectively,these results highlight a critical role of YAP1 in determining the fate determinations of human SSCs and male infertility through the YAP1/RAD21/NEDD4 pathway.This study provides new insights into the genetic regulatory mechanisms underlying human spermatogenesis and the pathogenesis of NOA,and it offers new targets for gene therapy of male infertility.
基金supported by a grant from the University of Hong Kong, China
文摘Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effects of Pb2＋ on adult neural cells of humans or other mammals, only few of which have examined the effects of Pb2＋ on neural stem cells. The purpose of this study was to reveal the biological effects of Pb2＋from lead acetate [Pb （0H30OO）2] on viability, proliferation and differentiation of neural stem cells derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days, respectively. This study was carried out in three parts. In the first part, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay （MTT viability assay） was used to detect the effects of Pb2＋ on the cell viability of passage 2 hippocampal neural stem cells after 48-hour exposure to 0-200 pM Pb2＋. In the second part, 10 pM bromodeoxyuridine was added into the culture medium of passage 2 hippocampal neural stem cells after 48-hour exposure to 0- 200 pM Pb2＋, followed by immunocytochemical staining with anti-bromodeoxyuridine to demonstrate the effects of Pb2＋ on cell proliferation. In the last part, passage 2 hippocampal neural stem cells were allowed to grow in the differentiation medium with 0-200 pM Pb2＋. Immunocytochemical staining with anti-microtubule-associated protein 2 （a neuron marker）, anti-glial fibrillary acidic protein （an astrocyte marker）, and anti-RIP （an oligodendrocyte marker） was performed to detect the differentiation commitment of affected neural stem cells after 6 days. The data showed that Pb2~ inhibited not only the viability and proliferation of rat hippocampal neural stem cells, but also their neuronal and oligodendrocyte differentiation in vitro. Moreover, increased activity of astrocyte differentiation of hippocampal neural stem cells from both newborn and adult rats was observed after exposure to high concentration of lead ion in vitro. These findings suggest that hippocampal neural stem cells of newborn rats were more sensitive than those from adult rats to Pb2＋cytotoxicity.
基金supported by the Hong Kong SCI Fundthe National Basic Research Development Program (973 program) of China (2011CB504402)
文摘Conflicting findings exist regarding the link between functional recovery and the regrowth of spinal tracts across the lesion leading to the restoration of functional contacts. In the present study, we investigated whether functional locomotor recovery was attributable to anatomical regeneration at postnatal day 1 （PN1）, PN7, PN14 and in adult rats two months after transection injury at the tenth thoracic segment of the spinal cord. The Basso, Beattie, and Bresnahan scores showed that transection led to a failure of hindlimb locomotor function in PN14 and adult rats. However, PN1 and PN7 rats showed a significant level of stepping function after complete spinal cord transection. Unexpectedly, unlike the transected PN14 and adult rats in which the spinal cord underwent limited secondary degeneration and showed a scar at the lesion site, the rats transected at PN1 and PN7 showed massive secondary degeneration both anterograde and retrograde, leaving a 〉5-mm gap between the two stumps. Furthermore, retrograde tracing with fluorogold （FG） also showed that FG did not cross the transection site in PN1 and PN7rats as in PN14 and adult rats, and re-transection of the cord caused no apparent loss in locomotor performance in the rats transected at PN1. Thus, these three lines of evidence strongly indicated that the functional recovery after transection in neonatal rats is independent of regrowth of spinal tracts across the lesion site. Our results support the notion that the recovery of locomotor function in developing rats may be due to intrinsic adaptations in the spinal circuitry below the lesion that control hindlimb locomotor activity rather than the regrowth of spinal tracts across the lesion. The difference in secondary degeneration between neonatal and adult rats remains to be explored.
基金supported by Direct Grant (Project No.2030392) of the Chinese University of Hong KongHK Spinal Cord Injury FoundationNational Key Basic Research Support Foundation (973 Project: 2011CB504402)
文摘The neurotrophin receptor （p75） activates the c-Jun N-terminal kinase （JNK） pathway. Activation of JNK and its substrate c-Jun can cause apoptosis. Here we evaluate the role of p75 in spinal motoneurons by comparing immunoreactivity for p75 and phosphorylated c-Jun （p-c-Jun）, the production of JNK activation in axotomized motoneurons in postnatal day （PN）I, PN7, PN14 and adult rats. Intensive p-c-Jun was induced in axotomized motoneurons in PN1 and PN7. In PN14, p-c-Jun expression was sharply reduced after the same injury. The decreased expression of p-c-Jun at this age coincided with a developmental switch of re-expression of p75 in axotomized cells. In adult animals, no p-c-Jun but intensive p75 was detected in axotomized motoneurons. These results indicate differential expression or turnover of phosphorylation of c-Jun and p75 in immature versus mature spinal motoneurons in response to axonal injury. The non-co-occurrence of p75 and p-c-Jun in injured motoneurons indicated that p75 may not activate JNK pathway, suggesting that the p75 may not be involved in cell death in axotomized motoneurons.
文摘Three-dimensional（3D） histology utilizes tissue clearing techniques to turn intact tissues transparent,allowing rapid interrogation of tissue architecture in three dimensions.In this article,we summarized the available tissue clearing methods and classified them according to their physicochemical principles of operation,which provided a framework for one to choose the best techniques for various research settings.Recent attempts in addressing various questions regarding the degenerating and regenerating nervous system have been promising with the use of 3D histological techniques.
基金grants from the National Natural Science Foundation of China(81802554 and 32170862)the Experimental Animal Fund of Shanghai Science and Technology Commission(22140903800)a Project Funded by China Postdoctoral Science Foundation(2021M692432).
文摘Prostate cancer(PCa)is a common malignant tumor with high morbidity and mortality worldwide.The prostate cancer stem cell(PCSC)model provides novel insights into the pathogenesis of PCa and its therapeutic response.However,the roles and molecular mechanisms of specific genes in mediating fate decisions of PCSCs and carcinogenesis of PCa remain to be elusive.In this study,we have explored the expression,function,and mechanism of AZGP1P2,a pseudogene of AZGP1,in regulating the stemness and apoptosis of PCSCs and treatment resistance of docetaxel in castration-resistant prostate cancer(CRPC).We revealed that AZGP1P2 was downregulated in CRPC cell lines and PCSCs,while it was positively associated with progression-free interval.Upregulation of the AZGP1P2 enhanced the sensitivity of docetaxel treatment in CRPCs via inhibiting their stemness.RNA pull-down associated with mass spectrometry analysis,co-immunoprecipitation assay,and RNA immunoprecipitation assay demonstrated that AZGP1P2 could bind to UBA1 and RBM15 as a“writer”of methyltransferase to form a compound.UBA1,an E1 ubiquitin-activating enzyme,contributed to RBM15 protein degradation via ubiquitination modification.Methylated RNA immunoprecipitation assay displayed that RBM15 controlled the mRNA decay of TPM1 in m6A methylation.Furthermore,a xenograft mouse model and patient-derived organoids showed that the therapeutic effect of docetaxel in CRPC was increased by AZGP1P2 in vivo.Collectively,these results imply that AZGP1P2 mediates the stemness and apoptosis of PCSCs and promotes docetaxel therapeutic effect by suppressing tumor growth and metastasis via UBA1/RBM15-mediated TPM1 mRNA decay in CRPC.
基金supported by grants from the National Natural Science Foundation of China (32000564,31970504)the China Postdoctoral Science Foundation (2020M682574)+1 种基金the Natural Science Foundation of Hunan Province (S2021JJQNJJ0952)the Reproductive and Genetic Hospital of the CITIC-XIANGYA project (YNXM-201909)。
文摘Early embryonic development is controlled by maternal factors originating from mature oocytes.The zygotic genome is activated from a transcriptionally quiescent state through a process called embryonic genome activation(EGA),which involves the depletion and clearance of maternal factors.However,the mechanism by which maternal factors regulate EGA and embryonic development,particularly in humans,remains elusive.In this study,using tri-pronuclear(3PN) embryos and human embryonic stem cells(h ESCs),we demonstrated that the maternal transcription factor Orthodenticle Homeobox 2(OTX2),a paired-like homeobox gene,promotes EGA in human pre-implantation embryos.Knockdown of OTX2 through Trim-Away technology blocked embryonic development and minor EGA gene expression.Overexpression of OTX2(OTX2~(OE)) in h ESCs increased transcript products,primarily at the 2-cell embryo stage genes,including genes encoding methyltransferase of histone H3K4.OTX2~(OE) increased the level of H3K4me3 and increased the open chromatin region that co-occurs with the H3K4me3 region at the 4-cell stage in h ESCs.Based on these findings in h ESCs,we further verified that OTX2 directly induced the expression of SETD1A by binding to its promoter,leading to increased H3K4me3 levels in both h ESCs and 3PN embryos.These findings suggest that the maternal transcription factor OTX2 regulates EGA and early embryogenesis via epigenetic mechanisms.
文摘Background： The worsening of semen quality, due to the application of Wi-Fi, can be ameliorated by Vitamin E. This study aimed to demonstrate whether a moderate dose of trolox,a new Vitamin E,inhibits oxidative damage on sperms in vitro after exposure to Wi-Fi radiation. Methods： Each of the twenty qualified semen, gathered from June to October 2014 in eugenics clinic, was separated into four aliquots, including sham, Wi-Fi-exposed, Wi-Fi plus 5 mmol/L trolox, and Wi-Fi plus 10 mmol/L trolox groups. At 0 min, all baseline parameters of the 20 samples were measured in sequence. Reactive oxygen species, glutathione, and superoxide dismutase were evaluated in the four aliquots at 45 and 90 min, as were sperm DNA fragments, sperm mitochondrial potential, relative amplification of sperm mitochondrial DNA, sperm vitality, and progressive and immotility sperm. The parameters were analyzed by one-way analysis of variance and Tukey＇s posttest. Results： Among Wi-Fi plus 5 mmol/L trolox, Wi-Fi-exposed and Wi-Fi plus 10 mmol/L trolox groups, reactive oxygen species levels （45 min： 3.80 ± 0.41 RLU·10^-6·ml^-1. 7.50 ± 0.35 RLU·10^-6·ml^-1vs. 6.70 ± 0.47 RLU·10^-6·ml^-1P 〈 0.001; 90 min：5.40 ± 0.21 RLU·10^-6·ml^-1vs. 10.10 ± 0.31 RLU·10^-6·ml^-1. 7.00 ± 0.42 RLU·10^-6·ml^-1P 〈 0.001, respectively）, percentages of tail DNA （45 min： 16.8 ± 2.0% vs. 31.9 ± 2.5% vs. 61.3 ± 1.6%, P 〈 0.001; 90 min： 19.7 ± 1.5% vs. 73.7 ± 1.3% vs. 73.1 ± 1.1%, P 〈 0.001, respectively）, 8-hydroxy-2＇-deoxyguanosine （45 min： 51.89 ± 1.46 pg/ml vs. 104.89 ± 2.19 pg/ml vs. 106.11 ± 1.81 pg/ml , P = 0.012;90 min： 79.96 ± 1.73 pg/ml vs. 141.73 ± 2.90 pg/ml vs. 139.06 ± 2.79 pg/ml; P 〈 0.001）, and percentages of immotility sperm （45 min：27.7 ± 2.7% vs. 41.7 ± 2.2% vs. 41.7 ± 2.5%; 90 min： 29.9 ± 3.3% vs. 58.9 ± 4.0% vs. 63.1 ± 4.0%; all P 〈 0.001） were lowest, and glutathione peroxidase （45 min： 60.50 ± 1.54 U/ml vs. 37.09 ± 1.77 U/ml vs. 28.18 ± 1.06 U/ml; 90 min： 44.61 ± 1.23 U/ml vs. 16.86 ± 0.93 U/ml vs. 29.94 ± 1.56 U/ml; all P 〈 0.001）, percentages of head DNA （45 min： 83.2 ± 2.0% vs. 68.2 ± 2.5% vs. 38.8 ± 1.6%; 90 min： 80.3 ± 1.5% vs. 26.3 ± 1.3% vs. 26.9 ± 1.1%; all P 〈 0.001）, percentages of sperm vitality （45 min： 89.5 ± 1.6% vs. 70.7 ± 3.1% vs. 57.7 ± 2.4%;90 min： 80.8 ± 2.2% vs. 40.4 ± 4.0% vs. 34.7 ± 3.9%; all P 〈 0.001）, and progressive sperm （45 min： 69.3 ± 2.7% vs. 55.8 ± 2.2% vs. 55.4 ± 2.5%; 90 min： 67.2 ± 3.3% vs. 38.2 ± 4.0% vs. 33.9 ± 4.0%; all P 〈 0.001） were highest in Wi-Fi plus 5 mmol/L trolox group at 45 and 90 min, respectively. Other parameters were not affected, while the sham group maintained the baseline. Conclusion：This study found that 5 mmol/L trolox protected the Wi-Fi-exposed semen in vitro from the damage of electromagnetic radiation-induced oxidative stress.
基金the grants from the National Nature Science Foundation of China(32170862 and 31872845)Major Scientific and Technological Projects for Collaborative Prevention and Control of Birth Defect in Hunan Province(2019SK1012)+3 种基金Key Grant of Research and Development in Hunan Province(2020DK2002)Developmental Biology and Breeding(2022XKQ0205)Natural Science Foundation of Hunan Province of China(2020JJ5380,2020JJ5383,and 2021JJ40365)a grant from the Shanghai Key Laboratory of Reproductive Medicine.
文摘Spermatogonial stem cells(SSCs)have important applications in both reproduction and regenerative medicine.Nevertheless,specific genes and signaling transduction pathways in mediating fate decisions of human SSCs remain elusive.Here,we have demonstrated for the first time that OIP5(Opa interacting protein 5)controlled the self-renewal and apoptosis of human SSCs.RNA sequencing identified that NCK2 was a target for OIP5 in human SSCs,and interestingly,OIP5 could interact with NCK2 as shown by Co-IP(co-immunoprecipitation),IP-MS(mass spectrometry),and GST pulldown assays.NCK2 silencing decreased human SSC proliferation and DNA synthesis but enhanced their apoptosis.Notably,NCK2 knockdown reversed the influence of OIP5 overexpression on human SSCs.Moreover,OIP5 inhibition decreased the numbers of human SSCs at S and G2/M phases,while the levels of numerous cell cycle proteins,including cyclins A2,B1,D1,E1 and H,especially cyclin D1,were remarkably reduced.Significantly,whole-exome sequencing of 777 patients with nonobstructive azoospermia(NOA)revealed 54 singlenucleotide polymorphism mutations of the OIP5 gene(6.95%),while the level of OIP5 protein was obviously lower in testes of NOA patients compared to fertile men.Collectively,these results implicate that OIP5 interacts with NCK2 to modulate human SSC self-renewal and apoptosis via cell cyclins and cell cycle progression and that its mutation and/or lower expression is correlated with azoospermia.As such,this study offers novel insights into molecular mechanisms underlying the fate determinations of human SSCs and the pathogenesis of NOA,and it provides new targets for treating male infertility.
