Hepatitis B virus X protein promotes the development of liver fibrosis and hepatoma through downregulation of miR-30e targeting P4HA2 mRNA.

Hepatitis B virus (HBV)-induced liver necrosis takes great part in liver cirrhosis progression. However, less is known about whether hepatitis B virus X protein (HBx) has effect on liver fibrosis. Here, we report that HBV leads to liver fibrosis and hepatocarcinogenesis through miR-30e targeting P4HA2. HBV transgenic mouse was treated by CCl4 to generate a model of liver fibrosis. A crucial enzyme catalyzing collagen formation, prolyl 4-hydroxylase subunit α2 (P4HA2) was evaluated by immunohistochemistry, western blotting or quantitative reverse transcription-PCR analysis. The function of HBV-modulated P4HA2 in hepatoma cell growth in vitro and in vivo was analyzed by EdU, MTT, colony-forming assay and animal transplantation assay. HBV transgenic mice exhibited more collagen deposition in liver after intraperitoneal injection of CCl4. P4HA2 was dramatically augmented in liver samples of HBV transgenic mice, clinical liver cirrhosis and liver cancer patients. Mechanistically, HBx was capable of inducing P4HA2 through suppressing miR-30e, in which miR-30e could target P4HA2 mRNA 3' untranslated region in liver cancer cells. HBx inhibited the miR-30e expression through increasing methylation of CpG islands in its promoter mediated by EZH2-formed complexes. Functionally, HBx-elevated P4HA2 enhanced the collagen deposition in the liver in vivo and in vitro, leading to liver fibrosis and liver cancer progression. In conclusion, HBx promotes the development of liver fibrosis and hepatocellular carcinoma through miR-30e targeting P4HA2 mRNA. We provide novel perspective on how HBx induces liver fibrosis.

[Effect of blastocyst quality on the strategy of single blastocyst transfer in frozen-thawed cycles].

OBJECTIVE: To investigate the effect of blastocyst quality on the strategy of single blastocyst transfer in frozen-thawed cycles. METHODS: A retrospective analysis was performed in Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region on clinical data of single frozen-thawed blastocyst transfer cycles from January 2008 to December 2013. All cycles were divided into four groups (AA, AB/BA, BB, BC/CB) according to the blastocyst score, then the clinical outcomes were compared between groups. And on this basis, the clinical outcomes were further explored when the group of outcomes with single blastocyst transfer wasn't ideal, which would diverted to transfer two blastocyst. RESULTS: In single frozen blastocyst transfer cycles, the clinical pregnancy rate of each group with the blastocyst scored AA, AB/BA, BB, BC/CB were 61.4% (470/765), 51.2% (330/645), 40.5% (407/1 005), 22.9% (60/262), live births rate in each group were 52.2% (399/765), 41.2% (266/645), 30.4% (306/1 005), 13.7% (36/262), and the abortion rate were 13.6% (64/470), 16.7% (55/330), 21.4% (87/407), 35.0%(21/60), separately. This showed that the clinical pregnancy rate and live births rate decreased significantly with the decline of blastocyst quality (P<0.01), but the abortion rate showed significant upward trend (P<0.01). When single blastocyst scored ≥BB grade transferred, an acceptable clinical pregnancy rate (>40%) and live births rate (>30%) could be obtained, however, the clinical pregnancy rate of 22.9% and live births rate of 13.7% could only be acquired when blastocyst scored BC/CB only transferred one embryo, which significant lower than those of each group scored ≥BB grade (P<0.01). So, after that, the blastocyst scored BC/CB were further divided into two groups (single blastocyst transferred versus two blastocyst transferred) to investigate, then the result showed that the clinical pregnancy rate [22.9% versus 38.5%(67/174),P<0.01] and live births rate [13.7% versus 30.5%(16/67),P<0.01] were significantly increased in the group of two blastocyst transferred compared with the group of one blastocyst transferred, and the abortion rate was also significantly decreased from 35.0% to 17.9% (12/67;P<0.05). So when two blastocyst scored BC/CB were transferred, the clinical outcomes were similar to the group of one blastocyst scored BB transferred (P>0.05). CONCLUSIONS: Of single blastocyst transfer in frozen-thawed cycles, the clinical pregnancy rate and liver births rate showed significant upward trend, but the abortion rate showed significant downward trend, with the decline of blastocyst quality. When the blastocyst scored ≥BB grade, the single blastocyst transfer could be considered to be performed.

Variation of plasma levels of endothelin, calcitonin gene-related peptide, nitric oxide, and malondialdehyde in acute myocardial ischemia reperfusion injury in a rabbit model.

We examined the variation in plasma levels of endothelin (ET), calcitonin gene-related peptide (CGRP), nitric oxide (NO), and malondialdehyde (MDA), as well as superoxide dismutase (SOD) activity, in acute myocardial ischemia reperfusion injury in a rabbit model. Seventy rabbits were randomly assigned into 3 groups. Open-chest surgery (OCS) was performed for all rabbits. Group A (N = 20) received sham-surgery, group B (N = 25) was the reperfusion group, and group C (N = 25) was the infarction group. At 12 h after chest clo-sure, plasma ET levels in groups B and C were clearly increased, while CGRP levels were clearly decreased, particularly in group B. At 24 h after chest closure, ET levels were higher than before OCS, while there was no significant difference between groups B and C. ET in group B was decreased, while that in group C was increased at 12 h. No significant difference in CGRP was observed between 12 and 24 h after chest closure. NO levels in groups B and C at 12 h after chest closure were significantly decreased compared to those before OCS. NO levels in group B at 24, 48, and 72 h were significantly lower than those at 12 h, while those of group C were not significantly changed after 12 h. Dynamic monitoring and comparison of plasma levels of ET, CGRP, NO, and MDA as well as SOD activity revealed that appropriate intervention of these factors may reduce reperfusion injury.