Male germline stem cells (mGSCs) are stem cells present in male testis responsible for spermatogenesis during their whole life. Studies have shown that mGSCs can be derived in vitro and resemble embryonic stem cells (ESCs) properties both in the mouse and humans. However, little is know about these cells in domestic animals. Here we report the first successful establishment of goat GSCs derived from 2-5-month fetal testis, and developmental potential assay of these cells both in vitro and in vivo. These cells express pluripotent markers such as Oct4, Sox2, C-myc, and Tert when cultured as human ESCs conditions. Embryoid bodies (EBs) formed by goat mGSCs were induced with 2 × 10(-6) M retinoic acid (RA). Immunofluorescence analysis showed that some cells inside of the EBs were positive for meiosis marker-SCP3, STRA8, and germ cell marker-VASA, and haploid marker-FE-J1, PRM1, indicating their germ cell lineage differentiation. Some cells become elongated sperm-like cells after induction. Approximately 34.88% (30/86) embryos showed cleavage and four embryos were cultured on murine fibroblast feeder and formed small embryonic stem like colonies. However, most stalled at four-cell stage after intracytoplasmic sperm injection (ICSI) of these cells. Transplantation of DAPI labeled mGSCs into the seminiferous tubules of busulfan-treated mice, and showed that mGSCs can colonize, self-renew, and differentiate into germ cells. Thus, we have established a goat GSC cell line and these cells could be differentiated into sperm-like cells in vivo and sperms in vitro, providing a promising platform for generation of transgenic goat for production of specific humanized proteins.

Boule is a conserved gene in meiosis, which encodes RNA binding protein required for spermatocyte meiosis. Deletion of Boule was found to block meiosis in spermatogenesis, which contributes to infertility. Up to date, the expression and function of Boule in the goat testis are not known. The objectives of this study were to investigate the expression pattern of Boule in dairy goat testis and their function in male germline stem cells (mGSCs). The results first revealed that the expression level of Boule in adult testes was significantly higher than younger and immature goats, and azoospermia and male intersex testis. Over-expression of Boule promoted the expression of meiosis-related genes in dairy goat mGSCs. The expression of Stra8 was up-regulated by over-expression of Boule analyzed by Western blotting and Luciferase reporter assay. While, Cdc25a, the downstream regulator of Boule, was found not to affect the expression of Stra8, and our data illustrated that Cdc25a did not regulate meiosis via Stra8. The expression of Stra8 and Boule was up-regulated by RA induction. Taken together, results suggest the Boule plays an important role in dairy goat spermatogenesis and that over-expression of Boule may promote spermatogenesis and meiosis in dairy goat.

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Objective: This study aimed to provide support for the extensive application of Distress Thermometer (DT) in advanced cancer inpatients with pain and explored factors associated with high DT scores among this population. Methods: Advanced cancer patients with pain were recruited from Department of Pain Relief in Tianjin Cancer Hospital and Institute, China. They completed the DT with problems list and HADS within 48 h after admission. The cutoff score of DT was evaluated against Hospital Anxiety and Depression Scale (HADS) for its sensitivity and specificity by using receiver operating characteristic (ROC) curves. Multiple logistic regression model analysis was performed to investigate correlates of DT scores.Results: Four hundred forty one inpatients with mixed diagnoses were recruited.Referring to the cutoff of 15 on HADS, DT cutoff score of 5 yielded AUC of 0.757, with an optimal sensitivity of 0.861 and specificity of 0.531. Using the cutoff scores of greater than or equal to 5, 70.5% of the patients were distressed. Logistic regression analysis of DT found that the breakthrough pain, poorer KPS, higher pain degree, and emotional problems were the predictive factor for current distress.Conclusion: DT is efficacious in screening for psychological distress in advanced cancer inpatients with pain. Psychological distress is prevalent with a cutoff score of greater than or equal to five. To better identify the distressed cancer patients with pain, pain degree, performance status, and emotional problems should be considered together.