DNA methylation promotes the expression of PPARγ transcript 1 at least in part by preventing NRF1 binding to the promoter P1 of chicken PPARγ gene.

Peroxisome proliferator-activated receptor gamma (PPARγ) is a master regulator of adipogenesis. Our previous study revealed that chicken PPARγ has 3 alternative promoters named as P1, P2, and P3, and the DNA methylation of promoter P3 was negatively associated with PPARγ mRNA expression in abdominal adipose tissue (AAT). However, the methylation status of promoters P1 and P2 is unclear. Here we assessed promoter P1 methylation status in AAT of Northeast Agricultural University broiler lines divergently selected for abdominal fat content (NEAUHLF). The results showed that promoter P1 methylation differed in AAT between the lean and fat lines of NEAUHLF at 7 wk of age (p < 0.05), and AAT expression of PPARγ transcript 1 (PPARγ1), which was derived from the promoter P1, was greatly higher in fat line than in lean line at 2 and 7 wk of age. The results of the correlation analysis showed that P1 methylation was positively correlated with PPARγ1 expression at 7 wk of age (Pearson's r = 0.356, p = 0.0242), suggesting P1 methylation promotes PPARγ1 expression. To explore the underlying molecular mechanism of P1 methylation on PPARγ1 expression, bioinformatics analysis, dual-luciferase reporter assay, pyrosequencing, and electrophoresis mobility shift assay (EMSA) were performed. The results showed that transcription factor NRF1 repressed the promoter activity of the unmethylated P1, but not the methylated P1. Of all the 4 CpGs (CpG48, CpG49, CpG50, and CpG51), which reside within or nearby the NRF1 binding sites of the P1, only CpG49 methylation in AAT was remarkably higher in the fat line than in lean line at 7 wk of age (3.18 to 0.57, p < 0.05), and CpG49 methylation was positively correlated with PPARγ1 expression (Pearson's r = 0.3716, p = 0.0432). Furthermore, EMSA showed that CpG49 methylation reduced the binding of NRF1 to the P1. Taken together, our findings illustrate that P1 methylation promotes PPARγ1 expression at least in part by preventing NRF1 from binding to the promoter P1.

Low-dose vitamin A therapy on T lymphocyte function in neonatal pneumonia.

OBJECTIVE: We aimed at studying the effect of adjuvant therapy with low-dose vitamin A on the function of T lymphocytes in neonatal pneumonia. PATIENTS AND METHODS: We recruited 60 cases of neonatal pneumonia which were randomly divided in two equal groups. The control group was treated with conventional anti-inflammatory therapy and aerosol inhalation. The experimental group received oral vitamin A soft capsules for 7 days. RESULTS: Pre-treatment levels vitamin A level and vitamin A deficiency disorders (VADD) percentage revealed no differences between the two groups. The treatment course for the experimental group was shorter than the control group. Serum IgM, IgG, and glutathione peroxidase (GSH-Px) levels were increased, whereas the levels of malondialdehyde were decreased in the experimental group after treatment. The control group showed no changes in these factors. After treatment, both groups showed increased percentages of CD4+ and CD8+ T cells, but the experimental group showed a larger increase. CONCLUSIONS: Neonatal pneumonia is often accompanied by a low level of vitamin A, and adjuvant therapy can shorten its disease course, improve IgM and IgG levels, and improve anti-oxidative and cellar immune function.

A dual-label time-resolved fluorescence immunoassay for the simultaneous determination of cystatin C and ß2-microglobulin in urine.

BACKGROUND: Contrast media is widely used in clinical diagnostic and interventional procedures, but may cause damage to the kidney, that is, contrast-induced nephropathy. This study was to establish a dual-label time-resolved fluorescence immunoassay (TRFIA) for the simultaneous determination of renal function markers cystatin-C (Cys-C) and ß2-microglobulin (ß2-MG) for the early diagnosis and follow-up surveillance of contrast-induced nephropathy. METHODS: A sandwich immunoassay was used to detect the concentration of Cys-C, and the competitive immunoassay was used to detect the concentration of ß2-MG in 50 samples of urine. The performance of this dual-label TRFIA was evaluated and compared with commercial assays. RESULTS: The sensitivity for Cys-C detection was 1.26 ng/ml, the average recovery was 99.36%; The sensitivity for ß2-MG detection was 2.13 ng/ml, the average recovery was 100.18%. Bland-Altman analysis showed that the dual-label TRFIA method and the commercial kits had a good agreement, suggesting they can be used interchangeably in clinical urine analysis. CONCLUSION: The present dual-label TRFIA has high sensitivity, specificity and accuracy in clinical sample analysis. This method can be used for the early diagnosis and follow-up surveillance of the contrast-induced nephropathy.

[Evaluate the prevalence of trachoma in high risk area of China].

OBJECTIVE: To determine the prevalence of trachoma in urban and rural area in China. To provide the data evidence for the strategies of eliminating trachoma in China. METHODS: Survey was conducted in 13 suspect trachoma high prevalence provinces according to the World Health Organization trachoma grading system and Trachoma Rapid Assessment(TRA). RESULTS: From 2004 to 2007, a total number of examined children which were younger than 10 years old was 59 630. The prevalence of TF was 0.94%. To sum up the data of 2004 and 2005, the prevalence of active trachoma was 1.71% for children.TT and CO was not reported. The results for subjects older than 50 years old showed that the prevalence of TT was 0.34%. We examined 26 857 adults in both 2004 and 2005.The prevalence of TF, TI, TS and CO was 0.03%,0.08%,0.88% and 0.05% respectively. CONCLUSIONS: The prevalence of active trachoma of younger children was under 5%. However,the prevalence of TT of the adults was not reached the target.

miR-196a-2 rs11614913 polymorphism is associated with vitiligo by affecting heterodimeric molecular complexes of Tyr and Tyrp1.

Tyrosinase and tyrosinase-related protein 1 (Tyr-Tyrp1) complex plays a critical role in the synthesis of melanin intermediates, which involves the production of reactive oxygen species (ROS) and contributes to the development of vitiligo. Based on our previous observation that rs11614913 single nucleotide polymorphism (SNP) in miR-196a-2 could affect the risk of vitiligo by influencing Tyrp1, we hypothesized that the same SNP could also regulate the level of Tyr in vitiligo. The aim of this study was to evaluate the potential association between rs11614913 SNP in miR-196a-2 and serum Tyr level in vitiligo and the regulatory role of miR-196a-2 in the expression of Tyr in melanocytes. The serum Tyr level was detected in 116 patients with vitiligo and 116 controls by ELISA plate assay. The expression level of Tyrp1 and Tyr in PIG1(normal melanocyte cell lines) cells was analyzed by western blotting. The ROS level and apoptosis rate in PIG1 cells transfected with si-Tyr or control siRNA were tested by flow cytometry. The results show that the individuals with TT+TC genotypes in miR-196a-2 and higher Tyr level in serum had an increased risk of vitiligo compared with those who had the CC genotype and lower Tyr level (P < 0.001). Furthermore, the rs11614913 C allele in miR-196a-2 enhanced its inhibitory regulation on the expression of Tyr, the down-regulation of which in melanocytes successfully reduced the intracellular ROS levels and the apoptosis rate. In conclusion, our findings suggest that miR-196a-2 polymorphisms can regulate the Tyr levels, which influences the susceptibility of vitiligo.

Complete genome sequence of a tobacco isolate of the tobacco vein banding mosaic virus strain prevailing in China.

Tobacco vein banding mosaic virus (TVBMV) is a species of the largest plant virus genus Potyvirus. Its incidence has been increasing in Chinese tobacco-growing area. TVBMV isolates can be clustered into three genetic groups that are corresponding with their geographical origin. We have reported the complete genomic sequence of TVBMV isolate YND with unique NIb/CP cleavage site. Here, we determined and analyzed the complete genomic sequence of isolate HN39, which was collected from tobacco in Henan Province and represented Chinese prevalent strain of TVBMV. HN39 has similar host range with YND, but induce mild vein banding symptom in Nicotiana tabacum cv. Samsun. The genome of TVBMV-HN39 is composed of 9,570 nucleotides, excluding the poly(A) tail. It contains a large ORF of 9,240 nucleotides and encode a polyprotein of 3,079 amino acids. The putative NIa-Pro cleavage site for NIb/CP is Q/G. The identities between the complete genomes of isolates HN39 and YND were 90.0% at nucleotide level and 95.4% at amino acid level. As for other potyviruses, HN39 shared the highest identity with wild tomato mosaic virus (WTMV) at complete genomic level, while different genes shared the highest identities with different potyviruses. This is the second complete genomic sequence of TVBMV reported.

Modelling and predicting the effect of temperature, water activity and pH on growth of Streptococcus iniae in Tilapia.

AIMS: To evaluate and model the growth of Streptococcus iniae affect by temperatures (10-45 degrees C), water activity (A(w); 0.995-0.957), and pH (5-8). METHODS AND RESULTS: Temperatures, A(w), and pH were adjusted. The behaviour of S. iniae was studied and modelled. Growth curves were fitted by using logistic, Gompertz, and Baranyi models. The maximum growth rates obtained from the primary model were then modelled as a function of temperature, A(w), and pH using the Belehradek-type models for secondary model. The optimum values for growth were found to be in the range of 35-40 degrees C, A(w) 0.995-1, and pH 6-7. The statistical characteristics of the models were validated by r(2), mean square error, bias, and accuracy factors. The results of validation indicated that Baranyi model performed the best. CONCLUSIONS: The effect of temperature, A(w)/NaCl, pH control of S. iniae in tilapia could be satisfactorily predicted under current experimental conditions, and the proposed models could serve as a tool for this purpose. SIGNIFICANCE AND IMPACT OF THE STUDY: The suggested predictive model can be used for risk assessment concerning S. iniae in tilapia.