BACKGROUND Lymphadenectomy of the infrapyloric region remains technically demanding in laparoscopic radical gastrectomy.Traditional vessel-guided approaches often result in incomplete dissection and higher complicatio...BACKGROUND Lymphadenectomy of the infrapyloric region remains technically demanding in laparoscopic radical gastrectomy.Traditional vessel-guided approaches often result in incomplete dissection and higher complication rates,especially at station No.6.AIM To propose a mesentery-based strategy for infrapyloric lymphadenectomy and evaluate its safety,feasibility,and efficacy.METHODS By identifying key anatomical landmarks and defining the inferior mesenteric boundary of the pyloric region(right gastro-omental mesentery),this approach enables full exposure and en bloc resection of anterior and posterior mesenteric planes,with proximal ligation at the root of feeding vessels.A retrospective cohort study was conducted on 330 gastric cancer patients who underwent D2 lymphadenectomy(D2)from January 2020 to December 2021.Outcomes were compared between 165 patients treated with D2 plus complete mesogastric excision(D2+CME)and 165 matched controls receiving conventional D2.RESULTS The D2+CME group demonstrated significantly improved surgical outcomes,including shorter total operative time(279.19±45.50 minutes vs 301.25±52.30 minutes,P<0.001),reduced infrapyloric dissection time(22.24±3.80 minutes vs 27.58±4.20 minutes,P<0.001),and lower blood loss(4.71±1.12 mL vs 24.83±6.35 mL,P<0.001).More lymph nodes were retrieved overall(43.80±10.05 vs 37.25±8.80,P<0.001),particularly at station No.6(5.26±0.87 vs 4.14±0.41,P<0.001).Postoperative recovery indicators and hospital stay were comparable between groups,while the complication rate was significantly lower in the D2+CME group(20%vs 30.3%,P=0.042).CONCLUSION The mesentery-based approach enables safe pyloric lymphadenectomy.Systematic mesogastric excision improves operative efficiency and lymph node yield,especially at station No.6,offering potential oncological benefits in gastric cancer surgery.展开更多
Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collec...Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.展开更多
Androgen and androgen receptor （AR） play important roles in male spermatogenesis and fertility, yet detailed androgen/AR signals in Sertoli cells remain unclear. To identify AR target genes in Sertoli cells, we anal...Androgen and androgen receptor （AR） play important roles in male spermatogenesis and fertility, yet detailed androgen/AR signals in Sertoli cells remain unclear. To identify AR target genes in Sertoli cells, we analyzed the gene expression profiles of testis between mice lacking AR in Sertoli cells （S-AR-/y） and their littermate wild-type （WT） mice. Digital gene expression analysis identified 2276 genes downregulated and 2865 genes upregulated in the S-AR-/y mice testis compared to WT ones. To further nail down the difference within Sertoli cells, we first constructed Sertoli cell line TM4 with stably transfected AR （named as TM4/AR） and found androgens failed to transactivate AR in Sertoli TM4 and TM4/AR cells. Interestingly, additional transient transfection of AR-cDNA resulted in significant androgen responsiveness with TM4/AR cells showing 10 times more androgen sensitivity than TM4 cells. In the condition where maximal androgen response was demonstrated, we then analyzed gene expression and found the expression levels of 2313 genes were changed more than twofold by transient transfection of AR-cDNA in the presence of testosterone. Among these genes, 603 androgen-/ AR-regulated genes, including 164 upregulated and 439 downregulated, were found in both S-AR-/y mice testis and TM4/AR cells. Using informatics analysis, the gene ontology was applied to analyze these androgen-/AR-regulated genes to predict the potential roles of androgen/AR in the process of spermatogenesis. Together, using gene analysis in both S-AR-/y mice testis and TM4/AR cells may help us to better understand the androeen/AR signals in Sertoli cells and their influences in spermatogenesis.展开更多
The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent ...The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent is certain to be of value. The aim of this study was to achieve targeted inactivation of viral E7 gene in keratinocytes using the reprogrammed clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas） 9 system. To accomplish this, a universal CRISPR-Cas9 system for targeting both HPV6/11 E7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E7-transfromed keratinocytes, the expression level of E7 protein was measured using western-blot analysis and the sequence of the E7 gene was determined using Sanger sequencing. Cell proliferation was analyzed by CCK-8 assay, and cell apoptosis was evaluated by Hoechst 33258 staining, flow cytometry analysis and ELISA assay. The results indicated that both HPV6/11 E7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E7 led to inhibition of cell proliferation and induction of apoptosis in E7-transfromed keratinocytes but not in normal keratinocytes. Our data suggested that the reprogrammed CRISPR-Cas9 system has the potential for the development of an adjuvant therapy for genital warts.展开更多
This study assesses the long-term outcomes in Han Chinese patients with clinical stage I non-seminomatous germ cell testicular cancer （CSI NSGCT） treated with surveillance, retroperitoneal lymph node dissection （RP...This study assesses the long-term outcomes in Han Chinese patients with clinical stage I non-seminomatous germ cell testicular cancer （CSI NSGCT） treated with surveillance, retroperitoneal lymph node dissection （RPLND） and adjuvant chemotherapy. We retrospectively evaluated 89 patients with a mean age of 26.5 years. After orchiectomy, 37 patients were treated with surveillance, 34 underwent RPLND and 18 were managed with chemotherapy. The overall survival rate, the recurrence-free survival rate and the risk factors were evaluated. The median follow-up length was 92 months （range： 6-149 months）. Thirteen of the 89 patients （14.6%） had relapses, and one died by the evaluation date. The overall survival rate was 98.9%. The cumulative 4-year recurrence-free rates were 80.2%, 92.0% and 100% for the surveillance, RPLND and chemotherapy groups, respectively. The disease-free period tended to be briefer in patients with a history of cryptorchidism and those with stage Is. Therefore, surveillance, RPLND and adjuvant chemotherapy might be reliable strategies in compliant patients with CSI NSGCT. Surveillance should be recommended for patients with the lowest recurrence rate, especially those without lymphovascular invasion. This study might aid the establishment of a standard therapy for CSI NSGCT in China.展开更多
Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months...Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months） were performed with mouse whole genome Affymetrix chip. The characteristics of the selected gene were analyzed by various bioinformatic tools. The expression profile of the selected gene was identified by RT-PCR. Results By analyzing the hybridization signals, a gene with a differential expression in the developmental stages of testis was identified. This gene was designated as TSF22. The full length cDNA of 1 597 bp contained an open reading frame of 570 bp which encoded a putative protein of 190 amino acids and a molecular weight of 22.106 kD. RT-PCR analysis revealed that TSF22 mRNA was exclusively expressed in mice testis. Conclusions TSF22, functions as a testis-specific transcription factor, may play important roles during spermatogenesis.展开更多
Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene seri...Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene series in workshop was determined by gas chromatography. As an internal exposure dose of benzene, the concentration of trans, trans-muconic acid （ttMA） was determined by high performance liquid chromatography. DNA was detected by modified single cell gel electrophoresis （SCGE）. Results The air concentrations of benzene, toluene and xylene at the workplace were 86.49±2.83 mg/m^3, 97.20±3.52 mg/m^3 and 97.45± 2.10 mg/m^3, respectively. Urinary ttMA in exposed group （1.040 ± 0.617 mg/L） was significantly higher than that of control group （0.819 ± 0.157 mg/L）. The percentage of head DNA, determined by modified SCGE method, significantly decreased in the exposed group （n=13, 70.18% ± 7.36%） compared with the control （n=16, 90.62% ± 2.94%）（P〈0.001）. Conclusion The modified SCGE method can be used to investigate the damage of sperm DNA. As genotoxin and reprotoxins, benzene had direct effect on the germ cells during the spermatogenesiss.展开更多
Objective To analyze the expression of SPACA4 in human and mice. Methods Testes cRNA samples from Balb/c mice of different postnatal days were performed with mouse affymetrix chip to screen the expression of SPACA4 in...Objective To analyze the expression of SPACA4 in human and mice. Methods Testes cRNA samples from Balb/c mice of different postnatal days were performed with mouse affymetrix chip to screen the expression of SPACA4 in mice. Sub-quantitative RT-PCR and bioinformatic tools were used here to describe the expression profile of SPACA4 in mice and human. Results The results of gene chip analysis indicated that the expression of mSPACA4 began after d 35 of postnatal testis in mice. Sub-quantitative RT-PCR assay showed that SPACA4 gene expressed exclusively in mouse and human testis, and mouse mSPACA4 gene expressed after d 35 of postnatal testis that was consistency with the results of gene chip analysis. By bioinformatics analysis, mSPACA4 is located in cell membrane （34.8%） or plasma membrane （34.8%）, the signal peptide cleavage site between position 19 and 20 amino acids, transmembrane region between 2-20 and 101-126 amino acids, respectively, on mSPACA4 protein. Conclusion mSPACA4 and hSPACA4 were testis-specific genes, and the expression of mSPACA4 begins after d 35 of postnatal testis in mice. SPACA4 is a candidate for targeting in a sperm-based contraceptive vaccine.展开更多
Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hy...Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hybridized with mouse whole genome affymetrix chip to screen the spermatogenesisrelated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. The mRNA expression of Dkkll at different stages of testis development and different tissues in mouse were analyzed by RT-PCR. The protein localization of Dkkll in mouse testis was assesed by immunohistoehemistry. Results By analyzing the gene chip signals of mouse testis aged 4 d, 9 d, 18 d, 35 d, 54 d and 6 months, Dkkll was identified with a differential expression in the develop- mental stages of testis. RT-PCR analysis showed that the expression of Dkkll mRNA was firstly detected on 15 d testis tissue and gradually upregulated during the testis developing to the adult stage. The Dkkll protein was predominantly located in spermatocytes and round spermatids in mouse testis. Conclusion The expression of Dkkll is gradually upregulated during the development of mouse testis and corresponds to the mouse spermatogenesis. It may play a critical role in male mammalian spermatogenesis.展开更多
Objective To assess the effectiveness of testosterone undecanoate on sperm motility and pregnancy incidence in men with asthenospermia. Methods A clinical trial was performed. Fifty men with asthenospermia were includ...Objective To assess the effectiveness of testosterone undecanoate on sperm motility and pregnancy incidence in men with asthenospermia. Methods A clinical trial was performed. Fifty men with asthenospermia were included to receive placebo （control group, n=9） or T undecanoate 80 mg/d （study group, n=41）. Pregnancy incidence and sperm characteristics after 1, 2 and 3 months of medication and 3 months after the end of the trial were measured. Results Compared with the placebo, T undecanoate treatment produced a satisfactory improvement of seminal motility （F=55.904, P=0.000）. In study group, the incidence of pregnancy was 28.2% while the incidence of pregnancy in control group was 11.1%. In study group, 26patients took T undecanoate for more than 3 months. In the 3 months, semen volume showed no statistical difference （F=1.206, P=0.312） before and after treatment, sperm concentration （F=0.023, P=0.000） and motility a, b, a ＋b （P=0.000） showed statistical differences. Motility grade a showed significantly higher increment in 2 and 3 months after treatment than 1 month and there was no statistical difference between 2 and 3 months. So did grade b. Conclusion The results indicate that T undecanoate increases semjnal motility, leading to a higher incidence of pregnancy in couples with infertility related to asthenoaspermia.展开更多
Aim:To investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes.Methods:Immunodefic...Aim:To investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes.Methods:Immunodeficient nude mice were used as hosts for allografting of neonatal mouse testes and xenografting of human fetal testicular tissues. Stepwise development and stage-specific gene expression of germ cells in allografts were systematically evaluated and parallel compared with those in intact mice by periodically monitoring the graft status with measurement of graft weight,histological analysis and determination of five stage-specific genes.Human testicular tissues from 20 and 26 weeks fetuses were used for the xenografting study.Histological analysis of xenografts was performed 116 and 135 d after the grafting procedure.Results:In the allografting study,progressive increase in tissue volume and weight as well as in tubule diameter in grafts was observed;the appearance time of various germ cells in seminiferous tubules,including spermatogonia,spermatocytes,round and elongate spermatids and sperm,was comparable with that in intact donors;the initiation of gene transcription in grafts showed a similar trend as in normal mice.Graft weight ceased to increase after 7-8 weeks and degradation of grafts was observed after 5 weeks with progressive damage to seminiferous epithelium.In the xenografting study using immature human testicular tissues,graft survival and development was indicated by increasing graft weight,Sertoli cells differentiation into advanced stage,germ cells migration and location to the basal lamina and formation of a niche-like structure.Conclusion:The developmental course and gene expression pattern of germ cells in allografts were similar to those in intact mice.The best time point for retrieval of mouse sperm from grafts was 5-7 weeks after grafting procedure.An accelerated development of immature human testicular cells could be achieved by ectopic xenografting of human testes.(Asian J Androl 2006 Jul;8:393-403)展开更多
Gravitational wave astronomy has opened a new window to the universe,allowing us to explore cosmic phenomena with unprecedented precision.Current detectors like LIGO[1],Virgo[1],KAGRA[1],LISA[2],Taiji[3],and Tian Qin[...Gravitational wave astronomy has opened a new window to the universe,allowing us to explore cosmic phenomena with unprecedented precision.Current detectors like LIGO[1],Virgo[1],KAGRA[1],LISA[2],Taiji[3],and Tian Qin[3]have made significant strides in gravitational wave detection.However,the demand for more sensitive and comprehensive detection capabilities continues to grow.展开更多
基金Supported by the Natural Science Foundation Program in Fujian Province of China,No.2023J011726.
文摘BACKGROUND Lymphadenectomy of the infrapyloric region remains technically demanding in laparoscopic radical gastrectomy.Traditional vessel-guided approaches often result in incomplete dissection and higher complication rates,especially at station No.6.AIM To propose a mesentery-based strategy for infrapyloric lymphadenectomy and evaluate its safety,feasibility,and efficacy.METHODS By identifying key anatomical landmarks and defining the inferior mesenteric boundary of the pyloric region(right gastro-omental mesentery),this approach enables full exposure and en bloc resection of anterior and posterior mesenteric planes,with proximal ligation at the root of feeding vessels.A retrospective cohort study was conducted on 330 gastric cancer patients who underwent D2 lymphadenectomy(D2)from January 2020 to December 2021.Outcomes were compared between 165 patients treated with D2 plus complete mesogastric excision(D2+CME)and 165 matched controls receiving conventional D2.RESULTS The D2+CME group demonstrated significantly improved surgical outcomes,including shorter total operative time(279.19±45.50 minutes vs 301.25±52.30 minutes,P<0.001),reduced infrapyloric dissection time(22.24±3.80 minutes vs 27.58±4.20 minutes,P<0.001),and lower blood loss(4.71±1.12 mL vs 24.83±6.35 mL,P<0.001).More lymph nodes were retrieved overall(43.80±10.05 vs 37.25±8.80,P<0.001),particularly at station No.6(5.26±0.87 vs 4.14±0.41,P<0.001).Postoperative recovery indicators and hospital stay were comparable between groups,while the complication rate was significantly lower in the D2+CME group(20%vs 30.3%,P=0.042).CONCLUSION The mesentery-based approach enables safe pyloric lymphadenectomy.Systematic mesogastric excision improves operative efficiency and lymph node yield,especially at station No.6,offering potential oncological benefits in gastric cancer surgery.
基金We would like to thank Mr Jian-Rong Zhang, Mr Li-Bing Zhang and Dr Zhen-Dong Yu for technical assistance. This work was supported by grants from the National Natural Science Foundation of China （No. 30500543）, Ministry of Education ＂985 project＂ （No. 985-2-054-29）, and Shenzhen Foundation of Science ＆ Technology （JH200505270413B）.
文摘Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.
基金This work was supported by the National Natural Science Foundation of China （no. 30971636）, and the George H. Whipple Professorship Endowment, and National Science Council, Talwan, China （96-2314-B-182A-023-MY2 and 97- 2314-B-182A-077-MY3）. Supplementary Information accompanies the paper on Asian lournal of Andrology website （http：Hwww.nature.com/aja）.
文摘Androgen and androgen receptor （AR） play important roles in male spermatogenesis and fertility, yet detailed androgen/AR signals in Sertoli cells remain unclear. To identify AR target genes in Sertoli cells, we analyzed the gene expression profiles of testis between mice lacking AR in Sertoli cells （S-AR-/y） and their littermate wild-type （WT） mice. Digital gene expression analysis identified 2276 genes downregulated and 2865 genes upregulated in the S-AR-/y mice testis compared to WT ones. To further nail down the difference within Sertoli cells, we first constructed Sertoli cell line TM4 with stably transfected AR （named as TM4/AR） and found androgens failed to transactivate AR in Sertoli TM4 and TM4/AR cells. Interestingly, additional transient transfection of AR-cDNA resulted in significant androgen responsiveness with TM4/AR cells showing 10 times more androgen sensitivity than TM4 cells. In the condition where maximal androgen response was demonstrated, we then analyzed gene expression and found the expression levels of 2313 genes were changed more than twofold by transient transfection of AR-cDNA in the presence of testosterone. Among these genes, 603 androgen-/ AR-regulated genes, including 164 upregulated and 439 downregulated, were found in both S-AR-/y mice testis and TM4/AR cells. Using informatics analysis, the gene ontology was applied to analyze these androgen-/AR-regulated genes to predict the potential roles of androgen/AR in the process of spermatogenesis. Together, using gene analysis in both S-AR-/y mice testis and TM4/AR cells may help us to better understand the androeen/AR signals in Sertoli cells and their influences in spermatogenesis.
文摘The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent is certain to be of value. The aim of this study was to achieve targeted inactivation of viral E7 gene in keratinocytes using the reprogrammed clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas） 9 system. To accomplish this, a universal CRISPR-Cas9 system for targeting both HPV6/11 E7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E7-transfromed keratinocytes, the expression level of E7 protein was measured using western-blot analysis and the sequence of the E7 gene was determined using Sanger sequencing. Cell proliferation was analyzed by CCK-8 assay, and cell apoptosis was evaluated by Hoechst 33258 staining, flow cytometry analysis and ELISA assay. The results indicated that both HPV6/11 E7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E7 led to inhibition of cell proliferation and induction of apoptosis in E7-transfromed keratinocytes but not in normal keratinocytes. Our data suggested that the reprogrammed CRISPR-Cas9 system has the potential for the development of an adjuvant therapy for genital warts.
文摘This study assesses the long-term outcomes in Han Chinese patients with clinical stage I non-seminomatous germ cell testicular cancer （CSI NSGCT） treated with surveillance, retroperitoneal lymph node dissection （RPLND） and adjuvant chemotherapy. We retrospectively evaluated 89 patients with a mean age of 26.5 years. After orchiectomy, 37 patients were treated with surveillance, 34 underwent RPLND and 18 were managed with chemotherapy. The overall survival rate, the recurrence-free survival rate and the risk factors were evaluated. The median follow-up length was 92 months （range： 6-149 months）. Thirteen of the 89 patients （14.6%） had relapses, and one died by the evaluation date. The overall survival rate was 98.9%. The cumulative 4-year recurrence-free rates were 80.2%, 92.0% and 100% for the surveillance, RPLND and chemotherapy groups, respectively. The disease-free period tended to be briefer in patients with a history of cryptorchidism and those with stage Is. Therefore, surveillance, RPLND and adjuvant chemotherapy might be reliable strategies in compliant patients with CSI NSGCT. Surveillance should be recommended for patients with the lowest recurrence rate, especially those without lymphovascular invasion. This study might aid the establishment of a standard therapy for CSI NSGCT in China.
基金This study was supported by grants from Chinese Natural Science Funds(No.30471728,No.30500543)Natural Science Funds of Guangdong Province(No.04007303)
文摘Objective To identify genes that involved in spermatogenesis. Methods In order to screen the testis-specific genes, testes cDNA samples from BALB/c mice of different postnatal days （days 4, 9, 18, 35, 54 and 6 months） were performed with mouse whole genome Affymetrix chip. The characteristics of the selected gene were analyzed by various bioinformatic tools. The expression profile of the selected gene was identified by RT-PCR. Results By analyzing the hybridization signals, a gene with a differential expression in the developmental stages of testis was identified. This gene was designated as TSF22. The full length cDNA of 1 597 bp contained an open reading frame of 570 bp which encoded a putative protein of 190 amino acids and a molecular weight of 22.106 kD. RT-PCR analysis revealed that TSF22 mRNA was exclusively expressed in mice testis. Conclusions TSF22, functions as a testis-specific transcription factor, may play important roles during spermatogenesis.
文摘Objective To assess the effect of benzene on sperm DNA damage ;Methods Twenty-seven benzene-exposed workers were selected as exposed group and 35 normal sperm donors as control group. Air concentration of benzene series in workshop was determined by gas chromatography. As an internal exposure dose of benzene, the concentration of trans, trans-muconic acid （ttMA） was determined by high performance liquid chromatography. DNA was detected by modified single cell gel electrophoresis （SCGE）. Results The air concentrations of benzene, toluene and xylene at the workplace were 86.49±2.83 mg/m^3, 97.20±3.52 mg/m^3 and 97.45± 2.10 mg/m^3, respectively. Urinary ttMA in exposed group （1.040 ± 0.617 mg/L） was significantly higher than that of control group （0.819 ± 0.157 mg/L）. The percentage of head DNA, determined by modified SCGE method, significantly decreased in the exposed group （n=13, 70.18% ± 7.36%） compared with the control （n=16, 90.62% ± 2.94%）（P〈0.001）. Conclusion The modified SCGE method can be used to investigate the damage of sperm DNA. As genotoxin and reprotoxins, benzene had direct effect on the germ cells during the spermatogenesiss.
基金This study was supported by grants from National Natural Science Foundation of China (No.30700824, No.30770810)973 Program(2008CB517412)Guangdong Natural Science Foundation (No.7008952)
文摘Objective To analyze the expression of SPACA4 in human and mice. Methods Testes cRNA samples from Balb/c mice of different postnatal days were performed with mouse affymetrix chip to screen the expression of SPACA4 in mice. Sub-quantitative RT-PCR and bioinformatic tools were used here to describe the expression profile of SPACA4 in mice and human. Results The results of gene chip analysis indicated that the expression of mSPACA4 began after d 35 of postnatal testis in mice. Sub-quantitative RT-PCR assay showed that SPACA4 gene expressed exclusively in mouse and human testis, and mouse mSPACA4 gene expressed after d 35 of postnatal testis that was consistency with the results of gene chip analysis. By bioinformatics analysis, mSPACA4 is located in cell membrane （34.8%） or plasma membrane （34.8%）, the signal peptide cleavage site between position 19 and 20 amino acids, transmembrane region between 2-20 and 101-126 amino acids, respectively, on mSPACA4 protein. Conclusion mSPACA4 and hSPACA4 were testis-specific genes, and the expression of mSPACA4 begins after d 35 of postnatal testis in mice. SPACA4 is a candidate for targeting in a sperm-based contraceptive vaccine.
基金supported by the Medical Research Foundation of Guangdong Province(B2014426)the Qingyuan Foundation of Science&Technology(2012B011204127)+1 种基金the National Natural Science Foundation of China(No.81170613,No.81270740)Shenzhen Basic Research Funds for Distinguished Young Scientists(JC201005260216A)
文摘Objective To investigate the expression and the distribution of Dickkopf-likel （Dkkll） protein during the development of mouse testis. Methods Testes eDNA samples from BALB/c mice in different postnatal days were hybridized with mouse whole genome affymetrix chip to screen the spermatogenesisrelated genes. The characteristics of the selected genes were analyzed by various bioinformatics tools. The mRNA expression of Dkkll at different stages of testis development and different tissues in mouse were analyzed by RT-PCR. The protein localization of Dkkll in mouse testis was assesed by immunohistoehemistry. Results By analyzing the gene chip signals of mouse testis aged 4 d, 9 d, 18 d, 35 d, 54 d and 6 months, Dkkll was identified with a differential expression in the develop- mental stages of testis. RT-PCR analysis showed that the expression of Dkkll mRNA was firstly detected on 15 d testis tissue and gradually upregulated during the testis developing to the adult stage. The Dkkll protein was predominantly located in spermatocytes and round spermatids in mouse testis. Conclusion The expression of Dkkll is gradually upregulated during the development of mouse testis and corresponds to the mouse spermatogenesis. It may play a critical role in male mammalian spermatogenesis.
基金supported by Planned Science and Technology Project of Shenzhen (No.200902069)
文摘Objective To assess the effectiveness of testosterone undecanoate on sperm motility and pregnancy incidence in men with asthenospermia. Methods A clinical trial was performed. Fifty men with asthenospermia were included to receive placebo （control group, n=9） or T undecanoate 80 mg/d （study group, n=41）. Pregnancy incidence and sperm characteristics after 1, 2 and 3 months of medication and 3 months after the end of the trial were measured. Results Compared with the placebo, T undecanoate treatment produced a satisfactory improvement of seminal motility （F=55.904, P=0.000）. In study group, the incidence of pregnancy was 28.2% while the incidence of pregnancy in control group was 11.1%. In study group, 26patients took T undecanoate for more than 3 months. In the 3 months, semen volume showed no statistical difference （F=1.206, P=0.312） before and after treatment, sperm concentration （F=0.023, P=0.000） and motility a, b, a ＋b （P=0.000） showed statistical differences. Motility grade a showed significantly higher increment in 2 and 3 months after treatment than 1 month and there was no statistical difference between 2 and 3 months. So did grade b. Conclusion The results indicate that T undecanoate increases semjnal motility, leading to a higher incidence of pregnancy in couples with infertility related to asthenoaspermia.
文摘Aim:To investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes.Methods:Immunodeficient nude mice were used as hosts for allografting of neonatal mouse testes and xenografting of human fetal testicular tissues. Stepwise development and stage-specific gene expression of germ cells in allografts were systematically evaluated and parallel compared with those in intact mice by periodically monitoring the graft status with measurement of graft weight,histological analysis and determination of five stage-specific genes.Human testicular tissues from 20 and 26 weeks fetuses were used for the xenografting study.Histological analysis of xenografts was performed 116 and 135 d after the grafting procedure.Results:In the allografting study,progressive increase in tissue volume and weight as well as in tubule diameter in grafts was observed;the appearance time of various germ cells in seminiferous tubules,including spermatogonia,spermatocytes,round and elongate spermatids and sperm,was comparable with that in intact donors;the initiation of gene transcription in grafts showed a similar trend as in normal mice.Graft weight ceased to increase after 7-8 weeks and degradation of grafts was observed after 5 weeks with progressive damage to seminiferous epithelium.In the xenografting study using immature human testicular tissues,graft survival and development was indicated by increasing graft weight,Sertoli cells differentiation into advanced stage,germ cells migration and location to the basal lamina and formation of a niche-like structure.Conclusion:The developmental course and gene expression pattern of germ cells in allografts were similar to those in intact mice.The best time point for retrieval of mouse sperm from grafts was 5-7 weeks after grafting procedure.An accelerated development of immature human testicular cells could be achieved by ectopic xenografting of human testes.(Asian J Androl 2006 Jul;8:393-403)
文摘Gravitational wave astronomy has opened a new window to the universe,allowing us to explore cosmic phenomena with unprecedented precision.Current detectors like LIGO[1],Virgo[1],KAGRA[1],LISA[2],Taiji[3],and Tian Qin[3]have made significant strides in gravitational wave detection.However,the demand for more sensitive and comprehensive detection capabilities continues to grow.
