摘要: Temperature-dependent sex determination (TSD) is a type of environmental sex determination in which the sex of the embryos depends on the ambient temperature; however, the molecular mechanisms governing this process remain unknown. Aromatase, encoded by the cyp19a1 gene, which converts androgens into estrogens in animals, was considered to be the key gene for TSD. In this study, the 5'-flanking region of the cyp19a1 gene in Reeves' turtle (Mauremys reevesii) was cloned, and the promoter region was identified using the luciferase reporter assay. Then the eggs of Reeves' turtle were incubated at different temperatures (26 degrees C: male-biased temperature; 29 degrees C: non-sex-biased temperature and 32 degrees C: female-biased temperature). During the thermosensitive period, the adrenal kidney gonad complexes (AKG) were sampled. DNA methylation analysis of the AKG samples showed that the promoter region of the cyp19a1 gene was significantly de-methylated in the female-biased temperature regime (P<0.01). Quantitative analysis of the cyp19a1 gene and estrogen by qPCR and Elisa assay showed that the expression level of the cyp19a1 gene and estrogen content were both upregulated significantly at the female-biased temperature (P<0.01). These results indicated that the de-methylation response of the cyp19a1 gene to incubation temperature, especially at the female-biased temperature, could lead to temperature-specific expression of aromatase and increased estrogen levels, which may further determine gonadal development in Reeves' turtle. These findings provide insights into the genetic mechanisms underlying TSD.

摘要: Objective: Zinc oxide (ZnO) nanoparticles can exhibit toxicity towards organisms and oxidative stress is often hypothesized to be one of the most important factors. Nevertheless, the detailed mechanism of toxicity-induced by ZnO nanoparticles has not been completely addressed. The present study aimed to investigate the toxic effects of ZnO nanoparticles on the expression and activity of Na+/K+-ATPase and on potassium channel block. Materials and methods: In the present study, we explored the cytotoxic effect of ZnO nanoparticles on murine photoreceptor cells using lactate dehydrogenase (LDH) release assay, reactive oxygen species (ROS) determination, mitochondrial membrane potential (Delta phi m) measurement, delayed rectifier potassium current recordings and Na+/K+-ATPase expression and activity monitoring. Results: The results indicated that ZnO nanoparticles could increase the LDH release in medium, aggravate the ROS level within cells, collapse the Delta phi m, block the delayed rectifier potassium current, and attenuate the expressions of Na+/K+-ATPase at both mRNA and protein levels and its activity, and thus exert cytotoxic effects on murine photoreceptor cells, finally damaging target cells. Conclusion: Our findings will facilitate the understanding of the mechanism involved in ZnO nanoparticle-induced cytotoxicity in murine photoreceptor cells via potassium channel block and Na+/K+-ATPase inhibition.

摘要: Colon cancer-associated transcript 2 (CCAT2) was originally identified as an oncogenic long non-coding RNA in colorectal cancer. Since its discovery, the oncogenic role of CCAT2 has been increasingly demonstrated in human cancers. In this connection, CCAT2 upregulation is frequently reported and very often associated with tumour progression and poor clinical outcomes. Functionally, knockdown of CCAT2 could induce cancer cell apoptosis and suppress cell proliferation and invasiveness, suggesting that CCAT2 might be a therapeutic target. The present review summarized current literature concerning the expression and functional role of CCAT2 in human malignancies.

摘要: ObjectivesMicroRNAs (miRNAs) are considered as the cellular regulators which post-transcriptionally modulate gene expression in diverse biological processes including cell development and immunity. In this study, we investigated functions of miR-181d in dendritic cells (DCs) maturation, and the underlying mechanisms were also explored. Materials and methodsHere we did the miRNA screening in human DCs in response to lipopolysaccharides (LPS) by quantitative real-time PCR (qRT-PCR). The expressions of DCs maturation markers were measured after miRNA mimics transfections. The pharmacological inhibitors of signalling pathways were applied to examine miR-181d effect on DCs maturation by Western blot. Luciferase assay and mixed lymphocyte reaction (MLR) were also performed to reveal the target gene of miR-181d and test the viability of T cells treated with miR-181d transfected DCs. ResultsOverexpression of miR-181d per se is sufficient to promote DCs maturation, and up-regulate CD80 and CD83 expressions without LPS. Besides, we showed that miR-181d activated NF-B pathway and also promoted the expression of pro-inflammatory cytokine IL12 and TNF-. Inhibition of NF-B pathway suppressed DCs maturation. Luciferase reporter assay and target gene knockdown assay indicated that miR-181d targets regulator cylindromatosis (CYLD), a primary negative regulator of NF-B pathway. MLR assay showed that miR-181d-transfected DCs could promote T-cell proliferation than iDCs in vitro. ConclusionOur study demonstrates that miR-181d is required for DCs maturation through the activation of NF-B pathway by targeting CYLD.

摘要: Objectives: Present evidence has suggested that large tumour suppressor 2 (LATS2) is abnormally expressed in most human cancer. However, the clinical and prognostic value in hepatocellular carcinoma (HCC) is still unknown. Materials and methods: Large tumour suppressor 2 mRNA and protein expression levels in HCC tissues and cell lines were detected by qRT-PCR, immunohistochemistry or Western blot. The correlation between LATS2 expression and clinicopathological factors was analysed through immunohistochemistry. The function of LATS2 on HCC cell growth and mobility was explored through MTT, colony formation, Transwell migration and invasion assays. The molecular mechanism of LATS2 was screened and confirmed by qRT-PCR and Western blot. Results and conclusion: In this study, LATS2 mRNA and protein expressions were decreased in HCC tissues and cell lines compared with normal hepatic tissues and hepatic cell line. Low LATS2 expression was oppositely corrected with tumour stage, vascular invasion and metastasis. The univariate and multivariate analyses suggested that low LATS2 expression was an independent poor prognostic factor for HCC patients. The in vitro experiments showed that LATS2 regulated HCC cells migration and invasion, but had no effect on HCC cells proliferation. Meanwhile, LATS2 modulated metastasis-associated genes expression including E-cadherin, vimentin, snail, slug, MMP2 and MMP9. In conclusion, LATS2 is a prognostic biomarker and a tumour metastasis suppressor in HCC.