Researchers commonly use cyclization recombination enzyme/locus of X-over P1(Cre/loxP)technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and L...Researchers commonly use cyclization recombination enzyme/locus of X-over P1(Cre/loxP)technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis.However,the shortcomings of these genetic tools include high costs,lengthy experimental periods,and limited accessibility for researchers.Therefore,exploring alternative gene silencing techniques is of great practical value.In this study,we employed adeno-associated virus(AAV)as a vector for gene silencing in Sertoli and Leydig cells.Our findings demonstrated that AAV serotypes 1,8,and 9 exhibited high infection efficiency in both types of testis cells.Importantly,we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection.We achieved cell-specific knockouts of the steroidogenic acute regulatory(Star)and luteinizing hormone/human chorionic gonadotropin receptor(Lhcgr)genes in Leydig cells,but not in Sertoli cells,using AAV9-single guide RNA(sgRNA)-mediated gene editing in Rosa26-LSL-Cas9mice.Knockdown of androgen receptor(Ar)gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpinRNA(shRNA)-mediated targeting.Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.展开更多
Central precocious puberty secondary to Leydig cell tumors is rare in children. We retrospectively analyzed the mid- to long-term follow-up data of patients with Leydig cell tumors. The clinical data of 12 consecutive...Central precocious puberty secondary to Leydig cell tumors is rare in children. We retrospectively analyzed the mid- to long-term follow-up data of patients with Leydig cell tumors. The clinical data of 12 consecutive patients who were treated at Beijing Children’s Hospital, Capital Medical University (Beijing, China), between January 2016 and October 2023 were retrospectively reviewed. Clinical evaluations, including physical examination, hormone examination, serum tumor marker analysis, abdominal and scrotal ultrasound, chest X-ray, and bone age measurement, were conducted before surgery and at follow-up time points. Surgical approaches were selected according to the individual conditions. Patients with an abnormal hormonal status and suspected of having central precocious puberty were referred to endocrinologists to confirm the diagnosis. Subsequently, gonadotropin-releasing hormone analog therapy was proposed. The mean patient age was 81.3 (range: 40–140) months at the time of the operation. Ten patients had peripheral precocious puberty at admission. All patients had elevated preoperative testosterone levels, whereas tumor marker levels were normal. Testis-sparing surgery was performed in eleven patients, and radical orchiectomy was performed in one patient. The follow-up duration (mean ± standard deviation) was 36.2 ± 25.3 months. Five patients had central precocious puberty, with a mean duration of 3.4 (range: 1–6) months postoperatively. Three patients were receiving gonadotropin-releasing hormone analog therapy, and good suppression of puberty was observed. No risk factors were found for secondary central precocious puberty. There was a high prevalence of central precocious puberty secondary to Leydig cell tumors in our study. Gonadotropin-releasing hormone analog therapy has satisfactory treatment effects. Larger sample sizes and long-term follow-up are needed in future studies.展开更多
目的：探讨邻苯二甲酸二乙基己酯（DEHP）对新生小鼠睾丸及Leydig细胞形态结构及功能的影响。方法：DEHP分别以低、中、高3组剂量[100、200、500mg／（kg·d）]灌胃作用于怀孕12d到产后3d（GD12～PND3）的KM母鼠，观察DEHP对新生雄...目的：探讨邻苯二甲酸二乙基己酯（DEHP）对新生小鼠睾丸及Leydig细胞形态结构及功能的影响。方法：DEHP分别以低、中、高3组剂量[100、200、500mg／（kg·d）]灌胃作用于怀孕12d到产后3d（GD12～PND3）的KM母鼠，观察DEHP对新生雄性仔鼠体重、睾丸重量、Leydig细胞形态结构和3β-羟基类固醇脱氢酶（3β—HSD）活性、酶反应面积的影响。结果：DEHP作用于母鼠后，其雄性子代幼鼠体重和睾丸重量减轻，睾丸Leydig细胞形态、超微结构发生改变；高剂量组Leydig细胞数量明显增多；低、中剂量组睾酮合成关键酶3β-HSD酶活性下降，酶反应面积减小，但高剂量组在仔鼠出生后15d时酶活性降低[（吸光度值（0．154±0.011）1）8空白对照组（0．222±0.013），P〈0．01]，而酶反应面积增大[（6303．0±745．6）μm^2 vs 空白对照组（5091．4±214．4）μm^2，P〈0．01）]。结论：DEHP能影响新生雄性小鼠体重、睾丸重量、Leydig细胞的形态结构和3B—HSD活性，具有抗雄激素效应。展开更多
基金supported by the National Natural Science Foundation of China(No.82070872 and No.82370854 to JXL)Innovative and Entrepreneurial Team of Jiangsu Province(No.JSSCTD2021 to JXL)+1 种基金China Postdoctoral Science Foundation(2023M741790 to JP)Jiangsu Funding Program for Excellent Postdoctoral Talent(2023ZB558 to JP).
文摘Researchers commonly use cyclization recombination enzyme/locus of X-over P1(Cre/loxP)technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis.However,the shortcomings of these genetic tools include high costs,lengthy experimental periods,and limited accessibility for researchers.Therefore,exploring alternative gene silencing techniques is of great practical value.In this study,we employed adeno-associated virus(AAV)as a vector for gene silencing in Sertoli and Leydig cells.Our findings demonstrated that AAV serotypes 1,8,and 9 exhibited high infection efficiency in both types of testis cells.Importantly,we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection.We achieved cell-specific knockouts of the steroidogenic acute regulatory(Star)and luteinizing hormone/human chorionic gonadotropin receptor(Lhcgr)genes in Leydig cells,but not in Sertoli cells,using AAV9-single guide RNA(sgRNA)-mediated gene editing in Rosa26-LSL-Cas9mice.Knockdown of androgen receptor(Ar)gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpinRNA(shRNA)-mediated targeting.Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
文摘Central precocious puberty secondary to Leydig cell tumors is rare in children. We retrospectively analyzed the mid- to long-term follow-up data of patients with Leydig cell tumors. The clinical data of 12 consecutive patients who were treated at Beijing Children’s Hospital, Capital Medical University (Beijing, China), between January 2016 and October 2023 were retrospectively reviewed. Clinical evaluations, including physical examination, hormone examination, serum tumor marker analysis, abdominal and scrotal ultrasound, chest X-ray, and bone age measurement, were conducted before surgery and at follow-up time points. Surgical approaches were selected according to the individual conditions. Patients with an abnormal hormonal status and suspected of having central precocious puberty were referred to endocrinologists to confirm the diagnosis. Subsequently, gonadotropin-releasing hormone analog therapy was proposed. The mean patient age was 81.3 (range: 40–140) months at the time of the operation. Ten patients had peripheral precocious puberty at admission. All patients had elevated preoperative testosterone levels, whereas tumor marker levels were normal. Testis-sparing surgery was performed in eleven patients, and radical orchiectomy was performed in one patient. The follow-up duration (mean ± standard deviation) was 36.2 ± 25.3 months. Five patients had central precocious puberty, with a mean duration of 3.4 (range: 1–6) months postoperatively. Three patients were receiving gonadotropin-releasing hormone analog therapy, and good suppression of puberty was observed. No risk factors were found for secondary central precocious puberty. There was a high prevalence of central precocious puberty secondary to Leydig cell tumors in our study. Gonadotropin-releasing hormone analog therapy has satisfactory treatment effects. Larger sample sizes and long-term follow-up are needed in future studies.
文摘目的：探讨邻苯二甲酸二乙基己酯（DEHP）对新生小鼠睾丸及Leydig细胞形态结构及功能的影响。方法：DEHP分别以低、中、高3组剂量[100、200、500mg／（kg·d）]灌胃作用于怀孕12d到产后3d（GD12～PND3）的KM母鼠，观察DEHP对新生雄性仔鼠体重、睾丸重量、Leydig细胞形态结构和3β-羟基类固醇脱氢酶（3β—HSD）活性、酶反应面积的影响。结果：DEHP作用于母鼠后，其雄性子代幼鼠体重和睾丸重量减轻，睾丸Leydig细胞形态、超微结构发生改变；高剂量组Leydig细胞数量明显增多；低、中剂量组睾酮合成关键酶3β-HSD酶活性下降，酶反应面积减小，但高剂量组在仔鼠出生后15d时酶活性降低[（吸光度值（0．154±0.011）1）8空白对照组（0．222±0.013），P〈0．01]，而酶反应面积增大[（6303．0±745．6）μm^2 vs 空白对照组（5091．4±214．4）μm^2，P〈0．01）]。结论：DEHP能影响新生雄性小鼠体重、睾丸重量、Leydig细胞的形态结构和3B—HSD活性，具有抗雄激素效应。
