Rumen-protected glucose stimulates the secretion of reproductive hormones and the mTOR/AKT signaling pathway in the ovaries of early postpartum.

This study was conducted to determine the response of the reproductive hormones and the mTOR/AKT/PI3K pathway in the ovaries of postpartum dairy cows with dietary rumen-protected glucose (RPG). Twelve Holstein cows were randomly assigned to two groups (n = 6/group): the control group (CT) and the RPG group. Blood samples were collected on d 1, 7, and 14 after calving for the gonadal hormone assay. The expression of the gonadal hormones receptors and PI3K/mTOR/AKT pathways were detected using RT-PCR and Western blot. The RPG addition increased the plasma LH, E2, and P4 concentrations on d 14 after calving and upregulated the mRNA and protein expressions of the ERα, ERß, 17ß-HSD, FSHR, LHR, and CYP17A1 but downregulated StAR expression. Immunohistochemical analysis identified higher expressions of the FSHR and LHR in the ovaries of RPG-fed cows compared to CT cows. Furthermore, the protein expressions of p-AKT/AKT and p-mTOR/mTOR were significantly increased in the ovaries of RPG-fed cows compared to the CT group, but the addition of RPG did not alter the protein expression of p-PI3K/PI3K. In conclusion, the current results indicated that dietary RPG supplementation regulated gonadotropin secretion and stimulated expression of hormone receptors and the mTOR/AKT pathway in the ovaries of early postpartum dairy cows. RPG may be beneficial for the recovery of ovarian activity in post-calving dairy cows.

[An experiment study of in vitro induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells to kill SO-RB(50)].

OBJECTIVE: To investigate induced antitumor immune responses by vaccination of tumor lysate-pulsed dendritic cells (DC) to kill retinoblastoma cells SO-RB(50). We hope to offer new approach for the treatment of patients with retinoblastoma. METHODS: DC was pulsed with RB tumor lysates in vitro and incubated with autologous lymphocytes to induce antigen specific CTL (cytotoxic T lymphocyte, CTL). SO-RB(50) cells were used as target cells and Raji cells were used as control target cells. Cytotoxicity of CTL was evaluated by MTT method (methyl thiazolyl letrazolium). The specific cytotoxicity of CTL to SO-RB(50) and Raji cells was compared. The cytotoxicity of CTL from RB and normal subjects was compared between these two groups. RESULTS: Antigen specific CTL showed greater cytotoxicity to SO-RB(50) than Raji cells, the difference was statistically significant, P < 0.01. The cytotoxicity was dose-dependent to the ratio of CTL/target cell. The nonspecific cytotoxicity to Raji cells was the same in CTL from RB patients and normal subjects, P > 0.05. The specific cytotoxicity of CTL from RB patients to SO-RB(50) was weaker than that from the healthy subjects, P < 0.01. CONCLUSION: DC pulsed with RB tumor lysate in vitro can induce antigen specific CTL which can kill the SO-RB(50) target cells specifically. This method may have potential value of therapy for the RB patients.

Pluripotent embryonic stem cells developed into medulloepithelioma in nude mice eyes.

PURPOSE: The pluripotent embryonic stem cells can differentiate into various kinds of normal tissues. There is no previous report on the differentiation of embryonic stem cell in the intraocular environment. In this paper, the authors tried to investigate the intraocular growth character of mice embryonic stem cells in nude mice. METHODS: Murine embryonic stem cells were cultured and maintained in an undifferentiated state in vitro. They were transplanted into the right eyes of 20 nude mice by microinjection under operating microscope. Animal eye observation, light microscope and immunohistochemical examinations were implemented. RESULTS: Two to three days after transplantation, small pieces of gray-white material could be viewed in the vitreous cavity. Between the 15th and 20th day, the gray-white mass grew into the anterior chamber in 4 nude mice eyes. Then, the mass at the anterior chamber extended extraocularly. On the 30th day, a remarkable proptosis was observed in two of the four nude mice. In 6 to 45 days, the mice were executed for morphological examination which showed the following typical structures: (1) Undifferentiated cells with prominent nucleolius. (2) Flexner-Wintersteiner-like rosettes. (3) Medulloepithelioma-like structure: the cells were arranged in sheets, cords, tubes, and cysts. (4) Large, spindle-or astrocyte-like cells. (5) Cartilage-like structure. Immunohistochemically, most of the cells were highly positive in NSE staining and a few cells were moderately positive in GFAP staining. CONCLUSIONS: Both animal eye findings and morphologic examinations certificated that the transplanted embryonic stem cells could grow in the eyes of nude mice and differentiate into intraocular medulloepithelioma.