SNP-based susceptibility-resistance association and mRNA expression regulation analyses of tlr7 to grass carp Ctenopharyngodon idella reovirus.

Eleven single nucleotide polymorphisms (SNP) in Ctenopharyngodon idella toll-like receptor 7 (citlr7) gene, containing two in the 5'-flanking region, three within the single intron and six distributed in the coding sequence (CDS), were identified. A case-control study of 73 susceptible individuals and 67 resistant individuals was conducted to test the SNPs-based susceptibility-resistance association and mRNA expression of citlr7 to grass carp reovirus (GCRV), showing that both 820 A/G and 1726 A/G were significantly correlative sites in genotype (P < 0·05). Multifactor dimensionality reduction (MDR) analysis suggested the exertion of antiviral effects of 820 A/G might rely on SNPs interactions of citlr7 and C. idella toll-like receptor 8 (citlr8). Combining the mortality rate and citlr7 mRNA expression, it was suggested that 1726 GG-genotyped individuals might be more resistant than 1726 A/G genotyped individuals, indicating the selection on synonymous mutations in 1726 A/G might be susceptibility-resistance-type specific. In addition, haplotype analysis uncovered no significantly correlative haplotypes in citlr7. These findings may provide an in-depth insight for the further functional research of citlr7. The potential genetic markers identified may contribute to the molecular and transgenic breeding of C. idella.

[Expression of mRNA and protein of Klotho gene in placental tissue of macrosomia and its relationship with birth weight of neonates].

OBJECTIVE: To explore the the expression of Klotho mRNA and protein in placenta of macrosomia and its relationship with the birth weight of neonates. METHODS: The cases were from November 2014 to March 2015 in Shengjing Hospital of China Medical University, divided into 4 groups: the gestational diabetes with macrosomia group (GM), the gestational diabetes with normal birth weight group (GN), the normal pregnancy with macrosomia group (NM) and the normal pregnancy with normal birth weight group (NN). Klotho mRNA and protein expression in the placenta were detected by immunohistochemistry SP method, real-time fluorescent quantitative PCR and western blot, respectively, and were compared among the 4 groups. RESULTS: (1) Immunohistochemical detection showed the positive rate of Klotho protein was significantly higher in the placenta of GM (93%,28/30) than in the GN (73%,22/30; P<0.05). The positive rate was significantly higher in the placenta of NM (97%,29/30) than in the NN (80%,24/30; P<0.05). (2) Real-time fluorescent quantitative PCR showed the Klotho mRNA expression was significantly higher in the placenta of GM (4.3 ± 3.1) than in the GN (2.1 ± 2.4; P<0.05). The Klotho mRNA expression was also significantly higher in the placenta of NM (4.8± 3.4) than in the NN (2.6± 3.3; P<0.05). (3) Western blot showed the Klotho protein expression was significantly higher in the placenta of GM (1.27±0.90) than in the GN (0.64±0.24; P<0.05). It was also significantly higher in the placenta of NM (2.51±3.52) than in the NN (0.77±0.37; P<0.05). (4) There were no significant differences in the expression of Klotho mRNA and protein between GM and NM, GN and NN (P>0.05). CONCLUSIONS: The up-regulation of Klotho gene may be associated with macrosomia. The relationship is not affected by the complication of gestational diabetes.

Sorafenib combined with percutaneous radiofrequency ablation for the treatment of medium-sized hepatocellular carcinoma.

OBJECTIVE: Sorafenib, an oral multikinase inhibitor, is the proved therapy method for patients with advanced hepatocellular carcinoma (HCC). Based on heat delivery, Radiofrequency ablation (RFA) has been found to achieve complete neoplasm necrosis. It is the most widely performed percutaneous therapy for HCC. However, Study associated combined Sorafenib with RFA therapy for patients with advanced HCC has never been reported. The aim of present study is to explore the efficacy and safety of sorafenib combined with RFA therapy for the patients with medium-sized HCC. PATIENTS AND METHODS: A total of 62 patients diagnosed as HCC were involved in this study. All patients were randomly assigned to sorafenib and RFA (n=30) or RFA-alone (n=32) treatment groups. Treatment outcomes, including recurrence rates, time to progression (TTP) and adverse reactions induced by sorafenib were observed and recorded to assess the efficacy and safety of the combination method. RESULTS: During the overall follow-up period, the recurrence rate of the combination subgroup was 56.7% (17/30), and that of the RFA-alone subgroup was 87.5% (28/32) (p < 0.01). The median TTP was 17.0 months in the combination therapy vs. 6.1 months in the RFA-alone (p < 0.05). Hand-foot skin reactions were reported by 83.3% (25/30) of patients and 46.7% (14/30) reported diarrhea while the most adverse events (AEs) were mild to moderate in the combination subgroup. CONCLUSIONS: Sorafenib combined with RFA significantly decreased recurrence rates and prolonged the survival time of medium-sized HCC patients. The combination therapy is safer and more effective than the control without unexpected side effects. Furthermore, the earlier application, the better results were.

Identification and expression of the laboratory of genetics and physiology 2 gene in common carp Cyprinus carpio.

In this study, a laboratory of genetics and physiology 2 gene (lgp2) from common carp Cyprinus carpio was isolated and characterized. The full-length complementary (c)DNA of lgp2 was 3061 bp and encoded a polypeptide of 680 amino acids, with an estimated molecular mass of 77 341·2 Da and a predicted isoelectric point of 6·53. The predicted protein included four main overlapping structural domains: a conserved restriction domain of bacterial type III restriction enzyme, a DEAD-DEAH box helicase domain, a helicase super family C-terminal domain and a regulatory domain. Real-time quantitative polymerase chain reaction (PCR) showed widespread expression of lgp2, mitochondrial antiviral signalling protein (mavs) and interferon transcription factor 3 (irf3) in tissues of nine organs. lgp2, mavs and irf3 expression levels were significantly induced in all examined organs by infection with koi herpesvirus (KHV). lgp2, mavs and irf3 messenger (m)RNA levels were significantly up-regulated in vivo after KHV infection, and lgp2 transcripts were also significantly enhanced in vitro after stimulation with synthetic, double-stranded RNA polyinosinic polycytidylic [poly(I:C)]. These findings suggest that lgp2 is an inducible protein involved in the innate immune defence against KHV in C. carpio. These results provide the basis for further research into the role and mechanisms of lgp2 in fishes.