Three new piscidins from orange-spotted grouper (Epinephelus coioides): Phylogeny, expression and functional characterization.

The present study reports the identification, and characterization of three new putative piscidin paralogues, ecPis-2, ecPis-3 and ecPis-4, from orange-spotted grouper (Epinephelus coioides). The cDNA of the three piscidins with the 207, 216, and 231 nt open reading frame encoded respectively a 68-, 71-, and 76-amino acid preprotein consisting of the predicted signal peptide, and putative mature peptide and prodomain. The phylogenetic analysis indicated that multiple piscidin paralogues in one fish species are highly diversified, the analysis suggested that the piscidins should be a family belonging to the superfamily of ancient cationic, linear, and amphipathic host defence peptides widespread across invertebrate and vertebrate taxa comprising insect cecropins and ceratotoxins, and the amphibian dermaseptins. The synthetic putative mature peptides, ecPis-2S, ecPis-3S and ecPis-4S, had strong activities against bacterial and fungal species. EcPis-3S exhibited powerful activity against the infective stage of Cryptocaryon irritans, theronts. The full length ecPis-2 and ecPis-4 by removal of signal peptide, ecPis-2L and ecPis-4L respectively, had potency against bacterial, fungal and parasitic species. The peptide ecPis-2S was proved to exist in spleen of orange-spotted grouper by HPLC followed by ESI-LCMS analysis. Basal transcriptions of ecPis-2, ecPis-3 and ecPis-4 were detected not only in the potential sites of pathogen entry such as gills, skin and intestine, but also in tissues such as head kidney, trunk kidney, blood cells, and spleen with highly abundant immune cells, however different paralogues expressed constitutively with different levels in the tissues. In addition, the expression of ecPis-2, ecPis-3 and ecPis-4 was upregulated in orange-spotted grouper challenged by Vibrio Parahaemolyticus, in different tissues at different time point after bacteria injection. These results support ecPis-2, ecPis-3 and ecPis-4 being the important immune-related genes in orange-spotted grouper innate immune system and playing multifunctional and complementary roles following their structural and functional diversification, and expression pattern difference. Finally, this study facilitates the evaluation of ecPis-2S, 2L, ecPis-3S, and ecPis-4S, -4L as potential templates of therapeutic agents against pathogens.

The expression of cytoglobin as a prognostic factor in gliomas: a retrospective analysis of 88 patients.

BACKGROUND: Evidence suggests that cytoglobin (Cygb) may function as a tumor suppressor gene. METHODS: We immunohistochemically evaluated the expression of Cygb, phosphatidylinositol-3 kinase (PI-3K), phosphorylated (p)-Akt, Interleukin-6 (IL-6), tumor necrosis factor-α (TNFα) and vascular endothelial growth factor (VEGF) in 88 patients with 41 high-grade gliomas and 47 low-grade gliomas. Intratumoral microvessel density (IMD) was also determined and associated with clinicopathological factors. RESULTS: Low expression of Cygb was significantly associated with the higher histological grading and tumor recurrence. A significant negative correlation emerged between Cygb expression and PI3K, p-Akt, IL-6, TNFα or VEGF expression. Cygb expression was negatively correlated with IMD. There was a positive correlation between PI3K, p-Akt, IL-6, TNFα and VEGF expression with IMD.High histologic grade, tumor recurrence, decreased Cygb expression, increased PI3K expression, increased p-Akt expression and increased VEGF expression correlated with patients' overall survival in univariate analysis. However, only histological grading and Cygb expression exhibited a relationship with survival of patients as independent prognostic factors of glioma by multivariate analysis. CONCLUSIONS: Cygb loss may contribute to tumor recurrence and a worse prognosis in gliomas. Cygb may serve as an independent predictive factor for prognosis of glioma patients.

Systematic comparison of cervical and extra-cervical surgical approaches for endoscopic thyroidectomy.

OBJECTIVE: To evaluate the clinical application of different surgical approaches for endoscopic thyroidectomy and provide more rational treatment criteria. METHODS: Collect all randomized controlled trials, multi-center studies, clinical controlled trials, clinical trials and other comparative studies of endoscopic thyroidectomy with a large sample size in different databases through an established search strategy, make a systematic analysis of all the included literature. RESULTS: This study selected 12 publications for analysis from more than 800 articles: these included six publications describing cervical thyroidectomy (A) and six publications describing extra-cervical thyroidectomy (B). Conversion to open surgery occurred in 29 patients in group A and only 4 in group B (p < 0.001). The patients in group A experienced shorter hospital stays than patients in group B (1.90 ± 0.80 vs. 4.03 ± 0.99 days, p < 0.001), and there was shorter operating time in group A (p < 0.001). Hemorrhage occurred in 3 cases in group A and 8 cases in group B (p = 0.04), Seroma occurred in 25 cases in group B but in no cases in group A (p < 0.001). Postoperative cosmetic results evaluated by verbal response scales (VRS) registered showed: group A (3.35 ± 0.60) and group B (3.74 ± 0.50; p < 0.001). Other complications such as recurrent laryngeal nerve injury and hypocalcemia showed no significant differences. CONCLUSIONS: Evaluation of the different surgical approaches for endoscopic thyroidectomy shows that the incidence of hemorrhage and seroma are higher in the extra-cervical group, but the rate of conversion to conventional open surgery is significantly higher in the cervical group. Furthermore, patients who undergo extra-cervical endoscopic thyroidectomy are associated with longer operating time and hospital stays; however, these studies suggest that the extra-cervical surgical approach for endoscopic thyroidectomy is preferable for dealing with more kinds of thyroid tumor and leaving no scars on neck.