The lncRNA Gm8097 is associated with hypospermatogenesis.

Objective: To investigate whether long non-coding RNA (lncRNA) Gm8097 (LncGm8097) is associated with male infertility. Methods: The expression and bilogical role of LncGm8097 were investigated. Results: LncGm8097 expression was down-regulated in the testis tissues with moderate and severe hypospermatogenesis compared with those with normal spermatogenesis and mild hypospermatogenesis (p<0.05). LncGm8097 down-regulation significantly promoted apoptosis and inhibited proliferation in GC1 and GC2 cells. In addition, LncGm8097 was significantly down-regulated in mouse model of hypospermatogenesis and correlated with cell apoptosis and proliferation. LncGm8097 was located immediately upstream of PRPS2, and correlated with Bcl-2/P53/caspase 6/caspase 9 signal pathway. Conclusion: LncGm8097 down-regulation correlates with hypospermatogenesis, which may offer new insights into the pathogenesis of male infertility.

[Dynamic changes of the PGAM1 expression in the mouse testis exposed to single heat stress].

OBJECTIVE: To investigate the expression of phosphoglycerate mutase 1 (PGAM1) in the mouse testis after exposure to single heat stress (SHS). METHODS: We randomly assigned 32 C57 male mice to an SHS (n = 16) and a control group (n = 16), the former bathed in water at 43 ℃ and the latter at 25 ℃ for 15 minutes. At 1 and 7 days after exposure, we harvested the testicular tissue for observation of the morphological changes of testicular cells by HE staining and determination of the location and expression of the PGAM1 protein by immunohistochemistry and Western blot. RESULTS: The testis volume of the mice were reduced significantly, the spermatogenic tubules were disorganized, and the cells were reduced in number after heat stress and basically disappeared after 7 days. Immunohistochemistry showed extensive expression of the PGAM1 protein in the testicular spermatogenic tubules of the SHS-exposed mice, significantly higher than in the control group at 1 day after exposure, which was down-regulated in the testis tissue at 7 days, but still markedly higher than that in the control. Western blot exhibited significantly up-regulated expression of the PGAM1 protein after heat stress compared with that in the control group. CONCLUSIONS: The expression of the PGAM1 protein undergoes dynamic changes in the mouse testis after exposed to single heat stress, which is related to heat stress-induced proliferation and division of testicular spermatogenic cells.

[Expression of phosphoribosyl pyrophosphate synthase 2 in human testis tissue].

OBJECTIVE: To investigate the expression of phosphoribosyl pyrophosphate synthase 2 (PRPS2) in the human testis and its clinical significance. METHODS: Using quantitative real-time PCR (qRT-PCR) and immunohistochemistry, we detected the expression of PRPS2 mRNA in the testis tissue of the men with normal spermatogenesis or mile, moderate or severe hypospermatogenesis (HS) and that of the PRPS2 protein in the testicular biopsy tissue of 67 adult males. Then, we analyzed the relationship of the PRPS2 expressions with the testicular histological types and clinical parameters of the subjects. RESULTS: The expression of PRPS2 mRNA in the testis tissue was significantly higher in the normal spermatogenesis group than in the moderate and severe HS groups (P < 0.01). The positive expression of the PRPS2 protein was 70.0% in the normal spermatogenesis group, 66.7% in the mild HS group, 50.0% in the moderate HS group and 23.8% in the severe HS group, significantly higher in the normal spermatogenesis and mild HS groups than in the moderate and severe HS groups (P < 0.01). No significant correlation, however, was observed between the PRPS2 expression and clinical parameters of the subjects (P > 0.05). CONCLUSIONS: PRPS2 is lowly expressed in the testis tissue of the men with hypospermatogenesis and its expression level may help the diagnosis of male infertility and the prediction of the spermatogenic function of the testis.

Discovery and Evaluation of Pyrazolo[3,4-d]pyridazinone as a Potent and Orally Active Irreversible BTK Inhibitor.

The identification and lead optimization of a series of pyrazolo[3,4-d]pyridazinone derivatives are described as a novel class of potent irreversible BTK inhibitors, resulting in the discovery of compound 8. Compound 8 exhibited high potency against BTK kinase and acceptable PK profile. Furthermore, compound 8 demonstrated significant in vivo efficacy in a mouse-collagen-induced arthritis (CIA) model.

UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis.

Ubiquitin conjugating enzyme (E2) is crucial for mediating N-terminal ubiquitination. Recent study reports that UBE2W is involved in male infertility. However, the correlation between UBE2W expression and hypospermatogenesis is unclear. The present study is to explore the biological role of UBE2W and its association with hypospermatogenesis. Results showed that the sexpression levels of UBE2W in mouse testes were gradually elevated from 2 to 10 weeks, while were significantly deceased in the testes with hypospermatogenesis. When UBE2W expression was successfully down-regulated in spermatogenic cells, the rate of apoptosis was significantly increased and the P53/Bcl-2/caspase 6/caspase 9 signal pathways were activated. Thus, these data indicate that UBE2W down-regulation promotes cell apoptosis and correlates with hypospermatogenesis, which may be helpful for the diagnosis of male infertility.

Phosphoribosyl-pyrophosphate synthetase 2 (PRPS2) depletion regulates spermatogenic cell apoptosis and is correlated with hypospermatogenesis.

Phosphoribosyl-pyrophosphate synthetase 2 (PRPS2) is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis. Recent studies report that PRPS2 is involved in male infertility. However, the role of PRPS2 in hypospermatogenesis is unknown. In this study, the relationship of PRPS2 with hypospermatogenesis and spermatogenic cell apoptosis was investigated. The results showed that PRPS2 depletion increased the number of apoptotic spermatogenic cells in vitro. PRPS2 was downregulated in a mouse model of hypospermatogenesis. When PRPS2 expression was knocked down in mouse testes, hypospermatogenesis and accelerated apoptosis of spermatogenic cells were noted. E2F transcription factor 1 (E2F1) was confirmed as the target gene of PRPS2 and played a key role in cell apoptosis by regulating the P53/Bcl-xl/Bcl-2/Caspase 6/Caspase 9 apoptosis pathway. Therefore, these data indicate that PRPS2 depletion contributes to the apoptosis of spermatogenic cells and is associated with hypospermatogenesis, which may be helpful for the diagnosis of male infertility.

PGAM1 knockdown is associated with busulfan­induced hypospermatogenesis and spermatogenic cell apoptosis.

Phosphoglycerate mutase 1 (PGAM1) is reported to be involved in spermatogenic dysfunction. However, the association between PGAM1 and busulfan­induced hypospermatogenesis and spermatogenic cell apoptosis remains unclear. The aim of the current study was to investigate the association between PGAM1 expression and busulfan­induced hypospermatogenesis, and the effect of PGAM1 expression on spermatogenic cell apoptosis. PGAM1 expression was detected in mouse models of busulfan­induced hypospermatogenesis by western blotting, reverse transcription­quantitative polymerase chain reaction and immunohistochemistry. Then, spermatogenic cell apoptosis in mouse models of busulfan­induced hypospermatogenesis was assessed by TUNEL assay. The effect and potential mechanism of PGAM1 downregulation on spermatogenic cells were further investigated. The results indicated that PGAM1 expression was significantly downregulated in the mouse models of busulfan­induced hypospermatogenesis, compared with those with normal spermatogenesis (P<0.05). Furthermore, the TUNEL assay revealed that the apoptosis of spermatogenic cells was accelerated in the mouse model of busulfan­induced hypospermatogenesis. In addition, PGAM1 knockdown promoted the apoptosis of spermatogenic cells in vitro, which was associated with the P53/Caspase 3/Caspase 6/Caspase 9 signaling pathway. In conclusion, these data indicate that PGAM1 knockdown is associated with busulfan­induced hypospermatogenesis and contributes to spermatogenic cell apoptosis by regulating the P53/Caspase 3/Caspase 6/Caspase 9 signaling pathway.

Downregulation of lncRNA PVT1 expression inhibits proliferation and migration by regulating p38 expression in prostate cancer.

Long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) has been reported to be overexpressed in prostate cancer cells and associated with tumorigenesis in various types of cancer. However, the biological function of lncRNA PVT1 remains largely unknown. The aim of the present study was to investigate the effect of lncRNA PVT1 expression on the proliferation and migration of prostate cancer cells. Stably transfected prostate cancer cells with downregulated expression of lncRNA PVT1 were constructed by an efficient siRNA fragment, followed by confirmation by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Proliferation was assessed using CCK-8, colony formation and xenograft assays, and cell migration was evaluated using a wound healing assay. The PathScan® Intracellular Signaling Array kit was utilized to explore the underlying molecular mechanisms of lncRNA PVT1 expression in prostate cancer cells. RT-qPCR results confirmed that the lncRNA PVT1 expression level was successfully knocked down in prostate cancer cells. When lncRNA PVT1 expression was downregulated in prostate cancer cells, proliferation and migration were significantly inhibited, compared with the control lncRNA PVT1 group. Furthermore, PVT1 knockdown decreased the phosphorylation of p38 in DU145 cells. Therefore, the present study demonstrated that lncRNA PVT1 downregulation inhibits the proliferation and migration of prostate cancer cells, and is associated with p38 phosphorylation.

Expression of PGAM1 in renal clear cell carcinoma and its clinical significance.

OBJECTIVE: This study is aimed to evaluate the expression of phosphoglycerate mutase 1 (PGAM1) in normal kidney and clear cell renal cell carcinoma (CCRCC), also to evaluate the correlation between PGAM1 expression and clinicopathological features in CCRCC. METHODS: PGAM1 expression was detected in 80 cases of normal kidney and 192 cases of CCRCC by immunohistochemistry (IHC). Meanwhile, PGAM1 expression measured in 8 cases of CCRCC and matched normal kidney tissues by Western blot. Then, the correlation between PGAM1 expression and clinicalpathological features was analyzed in CCRCC. RESULTS: IHC results exhibited that the high-expression rate of PGAM1 in CCRCC tissues was 45.8%, which was significantly higher than those in normal kidney tissues (32.5%, P=0.044). Meanwhile, PGAM1 expression in CCRCC was significantly greater compared with those in normal kidney by Western blot. Moreover, PGAM1 expression was significantly associated with age, tumor size and T stage in CCRCC. CONCLUSION: PGAM1 is highly expressed in CCRCC and correlated with clinicalpathological features, which may contribute to tumor formation and progression.

Biochemical hormone parameters in seminal and blood plasma samples correlate with histopathologic properties of testicular biopsy in azoospermic patients.

OBJECTIVE: To investigate the relationship between histopathologic patterns of testicular biopsy and biochemical semen and blood plasma parameters, including neutral a-glucosidase (NAG), fructose, follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and prolactin (PRL) in patients with azoospermia. MATERIALS AND METHODS: A total of 471 azoospermic patients with definitive pathologic diagnosis were enrolled in this study. Six biochemical parameters, including 2 seminal (NAG and fructose) and 4 blood (FSH, LH, testosterone, and PRL) plasma markers, were analyzed. RESULTS: NAG, fructose, FSH, and LH levels were significantly higher in patients with Sertoli-cell-only (SCO) syndrome and severe hypospermatogenesis than in those with normal spermatogenesis or mild hypospermatogenesis (P <.05). In addition, NAG levels positively correlated with fructose amounts in azoospermic patients (P <.05); a significant correlation between FSH and LH levels was also observed in azoospermic patients. Furthermore, PRL levels were higher in SCO syndrome patients compared with subjects showing normal spermatogenesis and the levels positively correlated with NAG, FSH, and LH amounts. However, testosterone levels in SCO syndrome patients were significantly reduced compared with individuals having normal spermatogenesis. CONCLUSION: The levels of biochemical parameters in seminal (NAG and fructose) and blood (FSH, LH, testosterone, and PRL) plasma samples correlate with the histologic diagnosis in azoospermic patients, providing potential benefits in predicting the pathologic diagnosis of male infertility.

PGAM1 is Involved in Spermatogenic Dysfunction and Affects Cell Proliferation, Apoptosis, and Migration.

OBJECTIVE: To investigate the correlation between PGAM1 and spermatogenic dysfunction and to evaluate the effect of expression of PGAM1 on the function of germ cells. METHODS: Expression of PGAM1 was detected in 40 cases of infertile males with definite pathological diagnosis and 12 cases of mouse models with spermatogenic dysfunction by immunohistochemistry. Then, cell proliferation, apoptosis, and migration were evaluated when expression of PGAM1 was knocked down by a specific small interfering RNA in GC1 and TM4 cells. RESULTS: The positive rates of PGAM1 in patients with normal spermatogenesis, mild hypospermatogenesis, severe hypospermatogenesis, and Sertoli cell-only syndrome were 90%, 80%, 10%, 100%, respectively, and the difference was significant (P < .001). Meanwhile, expression of PGAM1 was found to be significantly decreased in mouse models with spermatogenic dysfunction. Moreover, when expression of PGAM1 was knocked down in GC1 cells, the proliferation and migration were significantly inhibited, but the rate of apoptosis was significantly increased. Furthermore, PGAM1 downregulation in TM4 cells significantly inhibited proliferation and promoted apoptosis but didn't affect migration. CONCLUSION: PGAM1 correlates with spermatogenic distinction and affects the function of cell proliferation, apoptosis and migration.