Differential analysis of proteomic profiles between cryptorchid and normal mouse testes / 生理学报

To screen factors related to spermatogonial stem cell (SSC) proliferation, and to investigate the mechanism of infertility caused by cryptorchidism, ten-day-old Kunming (KM) mice were used and experimental cryptorchidism was conducted. On the 35th day after cryptorchid operation, the left testes were fixed in Bouin's fluid and used for histological analysis. The testes of 45-day-old mice were subjected to the same histological analysis, and it was found that they contained germ cells at every stage of development, from SSCs to sperm, indicating that the animals were fully sexually mature at this age. While in experimental cryptorchid mice, the spermatogenesis was arrested at the stage of spermatocytes, and only spermatogonia and primary spermatocytes were present in cryptorchid testes. The proportion of spermatogonia to other types of germ cells was much higher than that in sexually mature mice. On the other hand, the right testes were used for proteomic analysis. The total protein in testes was extracted on the 35th day after cryptorchid operation. The differentially expressed proteins in cryptorchid mice and sexually mature mice were screened and compared by the proteomic techniques. Through the separation of two-dimensional gel electrophoresis (2-DE), 20 differential protein spots were found, and 9 of them were digested and identified by the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrum. In cryptorchid mice, 6 out of 9 proteins were down-regulated, and 3 were up-regulated. Among these proteins, 4 proteins were identified, and they were Stathmin, phosphatidylethanolamine-binding protein1 (PEBP1), HES-related basic helix-loop-helix protein (HERP), and one unnamed protein (we temporarily named it Px). More Stathmin, PEBP1 and Px were expressed in sexually mature mice than in experimental cryptorchid mice. But HERP1 was the other way round. In the present study, we have screened 4 proteins related to cryptorchidism. It is helpful to study the mechanism of SSC proliferation and infertility caused by cryptorchidism.

Relationship between the adnormal expression of KAI 1 gene and carcinogenesis and development of thyroid papillary carcinoma / 肿瘤

Objective: To investigate the relationship between the expression of KAI 1 gene and the carcinogenesis and development of thyroid papillary carcinoma (PTC). Methods: The expressions of KAI1 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical method (Envision™) in 59 cases of malignant and benign thyroid tissues, respectively. Their correlation with clinical pathological parameters were analyzed. Results: The level of KAI1 mRNA expression was 3.59 ± 1.57 in PTC, and the expression of KAI1 protein was positive in 24 cases (66.67%). Both the mRNA and the protein expressions of KAI1 were significantly higher than those of follicular adenoma (P <0.01, P <0.05), multinodular goiters (P < 0.01, P < 0.01) and normal thyroid tissues (P < 0.01, P < 0.01). The expressions of KAI1 mRNA and protein had significant correlation with the lymph node metastasis and the tumor size of patients with thyroid papillary carcinoma (P < 0.05). The correlation between KAI1 mRNA and protein are significant positive (r = 0.486, P < 0.01). Conclusion: The abnormal expression of KAI1 mRNA and protein are related with the carcinogenesis and development of thyroid papillary carcinoma. It provides the basis for evaluating the lymph node metastasis and prognosis of thyroid papillary carcinoma.

17beta-estradiol stimulates proliferation of spermatogonia in experimental cryptorchid mice / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate whether estrogen stimulates the proliferation of spermatogonia or induces spermatogenesis in cryptorchid mice.</p><p><b>METHODS</b>Mice were surgically rendered cryptorchid, then treated with different doses of 17beta-estradiol (E2) s.c. once a day. Mice were killed at sexual maturity (45 days of age), and histological analysis and immunofluorescence were performed. Serum follicle stimulating hormone (FSH), estradiol, testosterone and luteinizing hormone (LH) were measured.</p><p><b>RESULTS</b>Low doses of E2 had no notable effect on spermatogonia, but at higher doses, E2 stimulated the proliferation of spermatogonia.</p><p><b>CONCLUSION</b>E2 has a dose-related mitogenic effect on spermatogonia.</p>

Cloning of Bmi1 cDNA from mouse testis and its expression in E. coli BL21 / 中华男科学杂志

<p><b>OBJECTIVE</b>To produce BMI1 polyclonal antibody, mouse Bmi1 cDNA was cloned from mouse testis and expressed in E. coli BL21.</p><p><b>METHODS</b>Bmi1 gene was amplified from mouse testis by RT-PCR and inserted into the prokaryotic expression vector pET-28c(+). Subsequently the recombined vector was transformed and expressed in E. coli BL21 (DE3) and the immunogenicity of recombined protein BMI1 (rBMI1) was tested by Western blot.</p><p><b>RESULTS</b>Mouse Bmi1 cDNA of 975 bp was successfully cloned and recombined. E. coli BL21 strains expressed rBMI1 were screened. The expression protein amounted to 12% of the total bacterial protein after induced with IPTG, which included inclusion body and soluble protein. Inclusion body was the major pattern of the expression that amounted to 71% of the insoluble protein. Western blot analysis showed that rBMI1 could be specially recognized by mouse monoclonal IgG1 anti-BMI1 and His-tag antibody.</p><p><b>CONCLUSION</b>There was expression of Bmi1 gene in mouse testis. Mouse Bmi1 cDNA was successfully cloned and expressed prokaryoticly.</p>