Disruption of ubiquitin specific protease 26 gene causes male subfertility associated with spermatogenesis defects in mice†.

Ubiquitin-specific protease 26 (USP26) is an X-linked gene exclusively expressed in the testis and codes for the USP26, a peptidase enzyme that belongs to the deubiquitinating enzyme family. Recent studies have indicated that mutations in USP26 affect spermatogenesis and are associated with male infertility in humans and mice. However, the exact role of USP26 in spermatogenesis and how it affects male reproduction remains unknown.

Associations between occupation exposure to Formaldehyde and semen quality, a primary study.

Formaldehyde (FA), a ubiquitous environmental pollutant, has long been suspected of having male reproductive toxicity. However, FA male reproductive toxicity was inconclusive due to dearth of human studies. Therefore, we sought to investigate whether occupational exposure to FA affects semen quality.

[The change of alpha cell mass and its mechanism with diabetic progress].

Objective: To explore the change of a cell mass and its mechanism with diabetic progress.

Methods: Diabetic mice were killed by exsanguinations after 4, 12 and 20 weeks of diabetes, respectively. Indirect double immunofluorescences for Insulin/Ki67, or BrdU, Cleaved-Caspase 3, TUNEL were used to evaluate pancreatic alpha cell mass, regeneration and apoptosis of a cells.

[Diabetes induced by multiple low doses of STZ can be spontaneously recovered in adult mice].

In a previous study, we observed that multiple low doses of streptozotocin (STZ) can induce spontaneous restoration of normoglycemia in adult mouse diabetes models. In the present study, we sought to identify when spontaneous recovery from diabetes occurs and to disclose the changes in the diabetic data of diabetic mice induced by multiple low doses of streptozotocin (MLD-STZ mice). After inducing type 1 diabetes mellitus, radioimmunoassay, indirect immunofluorescence and hematoxylin-eosin staining were used to determine the diabetic data of MLD-STZ mice.

Correlation of Sperm Associated Antigen 11 (SPAG11) and its Isoforms with Varicocele in Rats.

Background: We undertook this study to investigate the variation relationship of sperm associated antigen 11 (Spag11) mRNA expression and SPAG11E protein in the epididymis and spermatozoa of experimental left varicocele (ELV) rats. These findings could contribute to the understanding of the role of epididymal proteins in sperm functions and the mechanism of male infertility induced by varicocele.

Methods: The ELV model was established in adolescent male Sprague-Dawley rats.

[Polymorphism of Usp26 correlates with idiopathic male infertility].

Objective: To study the correlation of nucleotide polymorphism in the ubiquitin-specific protease 26 (Usp26) gene with idiopathic male infertility and its action mechanism in spermatogenesis.

Methods: Based on the WHO criteria (4th ed.), we selected 41 patients with idiopathic infertility from 150 infertile males, and enlisted 50 normal fertile men as controls.

Vasoactive intestinal peptide protects against ischemic brain damage induced by focal cerebral ischemia in rats.

Vasoactive intestinal peptide (VIP) exerts neuroprotective effects under various neurotoxic conditions in vitro. In the present study, we investigated the effects of VIP on transient ischemic brain damage. Focal cerebral ischemia was induced using middle cerebral artery occlusion (MCAO) for 120 min in the adult rat brain.

[Effects of progesterone on intracellular free Ca2+ concentration in the spermatozoa of fertile men and patients with unexplained infertility].

Objective: To investigate the difference in the responsiveness of intracellular free Ca2+ concentration ([Ca2+]i) to progesterone in the spermatozoa of normal fertile men and patients with unexplained infertility.

Methods: Nine normal fertile men and 10 patients with unexplained infertility were selected in this study. After swim-up separation of the motile fraction and 2-hour in vitro capacitation, the spermatozoa were loaded with the fluorescent calcium indicator Fluo-3/AM (8.

[Recombinant lentivirus-mediated gene transfer of NT4-p53(N15)-Ant inhibits the growth of hepatocellular carcinoma cells in vitro].

Objective: To construct a recombinant lentivirus vector containing fusion gene NT4-p53(N15)-Ant and transfer it into HepG2 cancer cells for gene therapy.

Methods: The gene of p53(N15)-Ant was obtained by T-vector method. After restriction enzyme digestion, the interest gene of p53(N15)-Ant was inserted in pBV220/NT4 vector and fusion gene of NT4-p53(N15)-Ant was subcloned into the plasmid of lentivirus and cotransferred into HEK-293 cells with helper plasmid.

NT4(Si)-p53(N15)-antennapedia induces cell death in a human hepatocellular carcinoma cell line.

Aim: To construct the recombinant lentivirus expression plasmid, pLenti6/V5-NT4 p53(N15)-antennapedia (Ant), and study its effect on HepG2 cells.

Methods: Plasmid pLenti6/V5-NT4 p53(N15)-Ant was constructed incorporating the following functional regions, including signal peptide sequence and pro-region of neurotrophin 4, N-terminal residues 12-26 of p53 and 17 amino acid drosophila carrier protein, Ant. Hepatocellular carcinoma (HepG2) cells were used for transfection.

[Experimental varicocele affects VEGF and Flt-1 protein expressions in the testis and epididymis of adolescent rats].

Objective: To investigate the effects of experimental left varicocele (ELV) on the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) proteins in the testis and epididymis of adolescent rats, and to find out the correlation of the two proteins with varicocele-induced male infertility.

Methods: We established the ELV model in adolescent male SD rats, and detected the expressions of VEGF and Flt-1 proteins in the testis and epididymis by immunohistochemistry at 2 and 4 weeks after surgery.

Results: Cell- and region-specific expressions of VEGF and Flt-1 were observed in the testis and epididymis of the ELV and control groups.

The expression of Usp26 gene in mouse testis and brain.

Deubiquitinating enzymes (DUBs) play an important role in ubiquitin-dependent processes as negative regulators of protein ubiquitination. Ubiquitin-specific protease 26 (USP26) is a member of this family. The expression of Usp26 in mammalian testis and in other tissues has yet to be fully elucidated.

[Effects of prepubertal exposure to estradiol benzoate on testicular development and function of Sprague-Dawley rats].

Objective: To investigate the effects of prepubertal exposure to estradiol benzoate (EB) on the testicular development and function of SD rats.

Methods: Fifty four 21-day-old male SD rats were randomly and equally divided into 2 experimental groups (AEa and AEb, n = 18 each) and one control group (AC, n = 18). The rats in the experimental groups were injected (s.

[Expressions of VEGF and Flt-1 in the testis, epididymis and epididymal sperm of adolescent rats].

Objective: To study the expressions of the vascular endothelial growth factor (VEGF) and its receptor fms-like tyrosine kinase-1 (Flt-1) in the testis, epididymis and epididymal sperm of adolescent rats and explore the functions of both the proteins in the male reproductive system.

Methods: The expressions of VEGF and Flt-1 were detected in 20 adolescent SD rats, immunohistochemical staining used for both the testis and the epididymis and immunofluorescent staining for sperm.

Results: VEGF and Flt-1 proteins were specifically present in the testis, epididymis and sperm.

Androgen receptor: a new player associated with apoptosis and proliferation of pancreatic beta-cell in type 1 diabetes mellitus.

Androgen receptor (AR) mediates a wide range of cellular processes, such as proliferation, differentiation and apoptosis. Here we sought to identify whether AR was located in pancreatic beta-cells and investigate its functions in type 1 diabetes induced by multiple low doses of streptozotocin. Double/triple immunofluorescence, Western blot and semi-quantitative RT-PCR were carried out to determine variances of AR expression in beta-cells and correlation between AR and apoptosis/proliferation of beta-cells with progress of diabetes.

[Effects of experimental left varicocele on SPAG11 mRNA and SPAG11E in the testis and epididymis of adolescent rats].

Objective: To investigate the effects of experimental left varicocele (ELV) on the expression of sperm associated antigen 11 (SPAG11) mRNA and its protein isomer SPAG11E in the testis and epididymis of adolescent rats, and to explore the mechanism of infertility caused by varicocele.

Methods: The experimental left varicocele model was established in the adolescent male Sprague-Dawley rats. Two and 4 weeks after the operation, the changes of SPAG11 mRNA and SPAG11E expression in the testis and epididymis were detected using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR).

[Effects of prepubertal exposure to diethylstilbestrol on testicular development and function of SD rats].

Objective: To investigate the effects of prepubertal exposure to diethylstilbestrol (DES) on the testicular development and function of Sprague-Dawley (SD) rats.

Methods: Ninety 21-day-old male SD rats were randomly and equally divided into 4 experimental groups (Da, Db, Dc and Dd), which were injected with DES dissolved in corn oil at the dose of 0.01, 0.

[Effect of Astragalus polysaccharide on pancreatic cell mass in type 1 diabetic mice].

Objective: To study the effect of Astragalus polysaccharide (APS) on pancreatic beta cell mass in type 1 diabetic mice.

Method: Diabetic mice induced by multiple low dose streptozotocin (MLD-STZ) were administered either APS (100, 200, 400 mg x kg(-1) body weight) or saline intraperitoneally daily, and sacrificed after 15 or 30 days of treatment. Streptavidin-peroxidase immunohistochemical method with counterstain was performed to determine the effect of APS on insulitis.

[Construction and antitumor effect of recombinant adenovirus vector containing heterofusion gene NT4p53(N15)Ant].

Background & Objective: Peptide p53(N15)Ant from the amino-terminal of p53 fused with cell penetrating peptide antennapedia (Ant) can induce rapid cell death resembling necrosis in breast cancer and pancreatic cancer, whereas it is low cytotoxic to normal cells. This study was to construct an recombinant adenovirus vector containing NT4p53(N15)Ant gene and investigate its antitumor effects.

Methods: Adenovirus packaging system was used to construct Ad-NT4p53(N15)Ant by recombinant technique in vitro.

[Immunomodulatory effects of Astragalus polysaccharide in diabetic mice].

Objective: To study the immunomodulatory effects of Astragalus polysaccharide (APS) in type 1 diabetic mice.

Methods: A mouse model of type 1 diabetes mellitus was established by intraperitoneal injection of multiple low dose streptozotocin (MLD-STZ). The diabetic mice were intraperitoneally administered 100, 200, 400 mg/kg APS or 1 ml normal saline (NS) every day respectively, then the diabetic mice were sacrificed after 15 or 30 days of treatment.

Secretory expression of p53(N15)-Ant following lentivirus-mediated gene transfer induces cell death in human cancer cells.

p53 (N15)-Ant 32-peptide has been considered a novel and effective peptide for cancer therapy. To further enhance its anticancer effect and overcome the limitation of peptide therapy, a recombinant lentivirus was constructed in this study with the following strategies: the secretory expression of therapeutic peptide and lentivirus gene transfer system. The results demonstrated that LV-NT4(Si)-p53 (N15)-Ant could significantly suppress cell growth and induce rapid cell death in MCF-7 (overexpressed wild-type p53), HepG2 (wide-type p53), OVCAR-3 (mutant type p53) and H1299 (null p53) cells in time-dependent manners through successful gene transfer and secretory expression of therapeutic peptide at 48 h post-infection.

Novel mutations in ubiquitin-specific protease 26 gene might cause spermatogenesis impairment and male infertility.

Aim: To study the incidence of single nucleotide polymorphisms in ubiquitin-specific protease 26 (USP26) gene and its involvement in idiopathic male infertility in China.

Methods: Routine semen analysis was performed. Infertility factors such as immunological, infectious and biochemical disorders were examined to select patients with idiopathic infertility.

[Adenovirus-mediated killing of hepatocellular carcinoma HepG2 cells by heterogeneous fusion gene NT4p53(N15)Ant].

Unlabelled: OBJECTIVE To construct a recombinant adenovirus Ad.NT4p53(N15)Ant and explore its cytotoxic effect against hepatocellular carcinoma HepG2 cells in vitro.

Methods: The recombinant adenovirus containing the fusion gene of neurotrophin 4 (NT4)signal peptide, N-terminal residues (12-26) of p53 and 17 amino acid Drosophila homeobox protein Antennapedia (Ant) was constructed by gene cloning protocol.

[Radioligand binding assay of progesterone receptors on normal fertile human sperm membrane].

Objective: To investigate the progesterone-binding site on the normal fertile human sperm membrane after 2 hours of in vitro capacitation.

Methods: Viable spermatozoa were selected by a swim-up method. After 2 hours of in vitro capacitation, multipoint saturation binding experiments were performed.