To report a case of a 46,XX SRY- negative man with a male phenotype and azoospermia. Case report. Molecular and Cytogenetic Unit in a University Hospital. A 35- year- old man with complete masculinization who referred...To report a case of a 46,XX SRY- negative man with a male phenotype and azoospermia. Case report. Molecular and Cytogenetic Unit in a University Hospital. A 35- year- old man with complete masculinization who referred to our institution because of a history of several years of infertility. Lymphocytic karyotype and genetic counseling. Peripheral blood metaphases were analyzed by standard G- banding and Q- banding. Fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) analyses were performed. Semen analysis showed azoospermia. Chromosome analysis revealed a 46,XX karyotype; molecular and cytogenetic analyses excluded the presence of SRY (the sex- determining region of the Y chromosome) gene. This case is one of the rare patients reported in the literature in whom testicular differentiation and a complete virilization in a 46,XX chromosomal constitution does not account for a translocation of the SRY gene to the X chromosome or to the autosomes. This finding suggests that other genes downstream from SRY, not yet identified, play an important role in sex determination.展开更多
Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding...Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding of eukaryotic transcriptomes to a new level, because it can resolve both gene expression level and alternative splicing events simultaneously. To gain a better understanding of cellular differentiation in gonads, we analyzed mRNA profiles from Drosophila testes and ovaries using RNA-seq. We identified a set of genes that have sex-specific isoforms in wild-type （WT） gonads, including several transcription factors. We found that differentiation of sperms from undifferentiated germ cells induced a dramatic downregulation of RNA splicing factors. Our data confirmed that RNA splicing events are significantly more frequent in the undifferentiated cell-enriched bag of marbles （barn） mutant testis, but downregulated upon differentiation in WT testis. Consistent with this, we showed that genes required for meiosis and terminal differentiation in WT testis were mainly regulated at the transcriptional level, but not by alternative splicing. Unexpectedly, we observed an increase in expression of all families of chromatin remodeling factors and histone modifying enzymes in the undifferentiated cell-enriched bam testis. More interestingly, chromatin regulators and histone modifying enzymes with opposite enzymatic activities are coenriched in undifferentiated cells in testis, suggesting that these cells may possess dynamic chromatin architecture. Finally, our data revealed many new features of the Drosophila gonadal transcriptomes, and will lead to a more comprehensive understanding of how differential gene expression and splicing regulate gametogenesis in Drosophila. Our data provided a foundation for the systematic study of gene expression and alternative splicing in many interesting areas of germ cell biology in Droso- phila, such as the molecular basis for sexual dimorphism and the regulation of the proliferation vs terminal differentiation programs in germline stem cell lineages. The GEO accession number for the raw and analyzed RNA-seq data is GSE16960.展开更多
Recent advances in the evolutionary genetics of sex determination indicate that the only molecular similarity in sex determination found so far among phyla is between the fly doublesex, worm mab-3 and vertebrate DMRTI...Recent advances in the evolutionary genetics of sex determination indicate that the only molecular similarity in sex determination found so far among phyla is between the fly doublesex, worm mab-3 and vertebrate DMRTI（dsx- and mab3-related transcription factor 1）/DMY genes. Each of these factors encodes a zinc-finger-like DNA-binding motif, DM domain. Insights into the evolution and functions of human DMRT1 gene could reveal evolutionary mechanisms of sexual development. Here we report the identification and characterization of multiple isoforms of human DMRT1 in the testis. These transcripts encode predicted proteins with 373,275 and 175 amino acids and they were generated by alternative splicing at 3＇ region. Expression level of DMRTla is higher than those of both DMRTlb and c, and the DMR Tlc expression was the lowest in testis, based on comparisons of mean values from real-time fluorescent quantitative RT-PCR analysis. Both DMRTlb and c result from exonization of intronic sequences, including the exonization of an Alu element. A further search for Alu elements within the DMRT1 gene demonstrated that all 99 Alu elements are non-randomly distributed among the non-coding regions on both directions. These new characteristics of DMRT1 would have an important impact on the evolution of sexual development mechanisms.展开更多
Objective： To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. Method： Both carotid arteries and arteries of lower extremity were subjected to ultra...Objective： To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. Method： Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppier＇s method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein （CRP） was assayed by nepheloturbidity. Tumor necrosis factor-α（TNF-α）, lnterleukin-6 （IL-6）, lnterleukin-8 （IL-8）, lnterleukin-10 （IL-10）, Interleukin-18 （IL-18）, soluble intercellular adhesion molecule-1 （sICAM-1） and soluble vascular cell adhesion molecule-1 （sVCAM-1） were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. Results： Free testosterone was （6.337±3.371） pg/L in case group and （11.375±4.733）pg/L in control group, P〈0.01. No differences were found in total testosterone and estradiol. CRP was （27.294±10.238）mg/L in case group and （12.843±6.318） mg/L in control group, P〈0.01. IL-6 was （41.700±31.385）pg/L in case group and （25.396±20.772）pg/L in control group, P〈0.05. IL-8 was （89.249±58.357）pg/L in case group and （67.873±31.227）pg/L in control group, P〈0.05. sICAM-1 was （470.491±134.078）pg/L in case group and （368.487±97.183）pg/L in control group, P〈0.01. sVCAM-1 was （537.808±213.172）pg/L in case group and （457.275±157.273）pg/L in control group, P〈0.05. There were no differences in TNF-α, IL-10 and IL-18. Correlation analysis showed that FT （free testosterone） had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was -0.678. Conclusion： Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low tree testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male.展开更多
Gonad development requires a coordinated soma-germline interaction that ensures renewal and differentiation of germline and somatic stem cells to ultimately produce mature gametes. The Drosophila tumour suppressor gen...Gonad development requires a coordinated soma-germline interaction that ensures renewal and differentiation of germline and somatic stem cells to ultimately produce mature gametes. The Drosophila tumour suppressor gene discs large （dig） encodes a septate junction protein functioning during epithelial polarization, asymmetric neuroblast division, and formation of neuromuscular junctions. Here, we report the role of dig in testis development and its critical function in somatic cyst cells （SCCs）. In these cells dig is primarily required for their survival and expansion, and contributes to spermatocyte cyst differentiation. Cell death primarily occurred in SCCs at the end of spermatogonial amplification at a time when Dig becomes restricted in wild-type （wt） testes to the distal somatic cells capping the growing spermatocyte cysts. RNAi depletion of dig transcripts in early SCCs fully prevented testis development, whereas depletion in late SCCs resulted in a breakdown of spermatocyte cyst structure and germ cell individualization. Specific dig expression in SCCs resulted in developmental rescue of dig mutant testes, whereas its expression in germ cells exerted no such effect, dig overexpression in wt testes led to spermatocyte cyst expansion at the expense of spermatogonial cysts. Our data demonstrate that dig is essentially required in SCCs for their survival, expansion, and differentiation, and for the encapsulation of the germline cells.展开更多
文摘To report a case of a 46,XX SRY- negative man with a male phenotype and azoospermia. Case report. Molecular and Cytogenetic Unit in a University Hospital. A 35- year- old man with complete masculinization who referred to our institution because of a history of several years of infertility. Lymphocytic karyotype and genetic counseling. Peripheral blood metaphases were analyzed by standard G- banding and Q- banding. Fluorescent in situ hybridization (FISH) and polymerase chain reaction (PCR) analyses were performed. Semen analysis showed azoospermia. Chromosome analysis revealed a 46,XX karyotype; molecular and cytogenetic analyses excluded the presence of SRY (the sex- determining region of the Y chromosome) gene. This case is one of the rare patients reported in the literature in whom testicular differentiation and a complete virilization in a 46,XX chromosomal constitution does not account for a translocation of the SRY gene to the X chromosome or to the autosomes. This finding suggests that other genes downstream from SRY, not yet identified, play an important role in sex determination.
文摘Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding of eukaryotic transcriptomes to a new level, because it can resolve both gene expression level and alternative splicing events simultaneously. To gain a better understanding of cellular differentiation in gonads, we analyzed mRNA profiles from Drosophila testes and ovaries using RNA-seq. We identified a set of genes that have sex-specific isoforms in wild-type （WT） gonads, including several transcription factors. We found that differentiation of sperms from undifferentiated germ cells induced a dramatic downregulation of RNA splicing factors. Our data confirmed that RNA splicing events are significantly more frequent in the undifferentiated cell-enriched bag of marbles （barn） mutant testis, but downregulated upon differentiation in WT testis. Consistent with this, we showed that genes required for meiosis and terminal differentiation in WT testis were mainly regulated at the transcriptional level, but not by alternative splicing. Unexpectedly, we observed an increase in expression of all families of chromatin remodeling factors and histone modifying enzymes in the undifferentiated cell-enriched bam testis. More interestingly, chromatin regulators and histone modifying enzymes with opposite enzymatic activities are coenriched in undifferentiated cells in testis, suggesting that these cells may possess dynamic chromatin architecture. Finally, our data revealed many new features of the Drosophila gonadal transcriptomes, and will lead to a more comprehensive understanding of how differential gene expression and splicing regulate gametogenesis in Drosophila. Our data provided a foundation for the systematic study of gene expression and alternative splicing in many interesting areas of germ cell biology in Droso- phila, such as the molecular basis for sexual dimorphism and the regulation of the proliferation vs terminal differentiation programs in germline stem cell lineages. The GEO accession number for the raw and analyzed RNA-seq data is GSE16960.
基金The work was supported by the National Natural Science Foundation of China, and the National Key Basic Research project （2004CB117400）the Program for New Century Excellent Talents in University and the Key Project of Chinese Ministry of Education （No. 2004.28）.
文摘Recent advances in the evolutionary genetics of sex determination indicate that the only molecular similarity in sex determination found so far among phyla is between the fly doublesex, worm mab-3 and vertebrate DMRTI（dsx- and mab3-related transcription factor 1）/DMY genes. Each of these factors encodes a zinc-finger-like DNA-binding motif, DM domain. Insights into the evolution and functions of human DMRT1 gene could reveal evolutionary mechanisms of sexual development. Here we report the identification and characterization of multiple isoforms of human DMRT1 in the testis. These transcripts encode predicted proteins with 373,275 and 175 amino acids and they were generated by alternative splicing at 3＇ region. Expression level of DMRTla is higher than those of both DMRTlb and c, and the DMR Tlc expression was the lowest in testis, based on comparisons of mean values from real-time fluorescent quantitative RT-PCR analysis. Both DMRTlb and c result from exonization of intronic sequences, including the exonization of an Alu element. A further search for Alu elements within the DMRT1 gene demonstrated that all 99 Alu elements are non-randomly distributed among the non-coding regions on both directions. These new characteristics of DMRT1 would have an important impact on the evolution of sexual development mechanisms.
基金Project (No. 2003B045) supported by the Health Bureau of ZhejiangProvince, China
文摘Objective： To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. Method： Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppier＇s method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein （CRP） was assayed by nepheloturbidity. Tumor necrosis factor-α（TNF-α）, lnterleukin-6 （IL-6）, lnterleukin-8 （IL-8）, lnterleukin-10 （IL-10）, Interleukin-18 （IL-18）, soluble intercellular adhesion molecule-1 （sICAM-1） and soluble vascular cell adhesion molecule-1 （sVCAM-1） were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. Results： Free testosterone was （6.337±3.371） pg/L in case group and （11.375±4.733）pg/L in control group, P〈0.01. No differences were found in total testosterone and estradiol. CRP was （27.294±10.238）mg/L in case group and （12.843±6.318） mg/L in control group, P〈0.01. IL-6 was （41.700±31.385）pg/L in case group and （25.396±20.772）pg/L in control group, P〈0.05. IL-8 was （89.249±58.357）pg/L in case group and （67.873±31.227）pg/L in control group, P〈0.05. sICAM-1 was （470.491±134.078）pg/L in case group and （368.487±97.183）pg/L in control group, P〈0.01. sVCAM-1 was （537.808±213.172）pg/L in case group and （457.275±157.273）pg/L in control group, P〈0.05. There were no differences in TNF-α, IL-10 and IL-18. Correlation analysis showed that FT （free testosterone） had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was -0.678. Conclusion： Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low tree testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male.
文摘Gonad development requires a coordinated soma-germline interaction that ensures renewal and differentiation of germline and somatic stem cells to ultimately produce mature gametes. The Drosophila tumour suppressor gene discs large （dig） encodes a septate junction protein functioning during epithelial polarization, asymmetric neuroblast division, and formation of neuromuscular junctions. Here, we report the role of dig in testis development and its critical function in somatic cyst cells （SCCs）. In these cells dig is primarily required for their survival and expansion, and contributes to spermatocyte cyst differentiation. Cell death primarily occurred in SCCs at the end of spermatogonial amplification at a time when Dig becomes restricted in wild-type （wt） testes to the distal somatic cells capping the growing spermatocyte cysts. RNAi depletion of dig transcripts in early SCCs fully prevented testis development, whereas depletion in late SCCs resulted in a breakdown of spermatocyte cyst structure and germ cell individualization. Specific dig expression in SCCs resulted in developmental rescue of dig mutant testes, whereas its expression in germ cells exerted no such effect, dig overexpression in wt testes led to spermatocyte cyst expansion at the expense of spermatogonial cysts. Our data demonstrate that dig is essentially required in SCCs for their survival, expansion, and differentiation, and for the encapsulation of the germline cells.
