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author:("Li, xiaoming")
1.  Elevated Expression of AKR1C3 Increases Resistance of Cancer Cells to Ionizing Radiation via Modulation of Oxidative Stress 
PLoS ONE  2014;9(11):e111911.
With the aim to elucidate the etiology of radioresistance, we explored the genetic alterations in non-radioresistant vs. resistant esophageal cancer cells acquired by long-term fractionated radiation. We found AKR1C3, an aldo-keto reductase expressed seldom in most human tissues, expressed higher in radioresistance-acquired cells. Suppression of AKR1C3 via RNAi or its chemical inhibitors restored the sensitivity of the acquired tumor cells and xenograft BALB/c nude mice to ionizing radiation (IR). Cellular monitoring of the oxidative stress in the AKR1C3-elevated cells indicated that IR-induced ROS accumulation and the concomitant DNA damage was significantly alleviated, and such protective consequence disappeared upon AKR1C3 knockdown. These findings uncover the potential involvement of AKR1C3 in removal of cellular ROS and explain, at least partially, the acquired radioresistance by AKR1C3 overexpression. A retrospective analysis of esophageal carcinomas also indicated a significant expression of AKR1C3 in radio-resistant but not radio-sensitive surgical samples. Our study may provide a potential biomarker for predicting prognosis of radiotherapy and even direct a targeted therapy for esophageal cancer and other tumors.
doi:10.1371/journal.pone.0111911
PMCID: PMC4242615  PMID: 25419901
2.  Development and Validation of a Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Three Papaya Viruses 
Viruses  2014;6(10):3893-3906.
Papaya ringspot virus (PRSV), Papaya leaf distortion mosaic virus (PLDMV), and Papaya mosaic virus (PapMV) produce similar symptoms in papaya. Each threatens commercial production of papaya on Hainan Island, China. In this study, a multiplex reverse transcription PCR assay was developed to detect simultaneously these three viruses by screening combinations of mixed primer pairs and optimizing the multiplex RT-PCR reaction conditions. A mixture of three specific primer pairs was used to amplify three distinct fragments of 613 bp from the P3 gene of PRSV, 355 bp from the CP gene of PLDMV, and 205 bp from the CP gene of PapMV, demonstrating the assay’s specificity. The sensitivity of the multiplex RT-PCR was evaluated by showing plasmids containing each of the viral target genes with 1.44 × 103, 1.79 × 103, and 1.91 × 102 copies for the three viruses could be detected successfully. The multiplex RT-PCR was applied successfully for detection of three viruses from 341 field samples collected from 18 counties of Hainan Island, China. Rates of single infections were 186/341 (54.5%), 93/341 (27.3%), and 3/341 (0.9%), for PRSV, PLDMV, and PapMV, respectively; 59/341 (17.3%) of the samples were co-infected with PRSV and PLDMV, which is the first time being reported in Hainan Island. This multiplex RT-PCR assay is a simple, rapid, sensitive, and cost-effective method for detecting multiple viruses in papaya and can be used for routine molecular diagnosis and epidemiological studies in papaya.
doi:10.3390/v6103893
PMCID: PMC4213569  PMID: 25337891
PRSV; PLDMV; PapMV; multiplex RT-PCR; detection; papaya
3.  Association between Serum Uric Acid Level and Metabolic Syndrome and Its Sex Difference in a Chinese Community Elderly Population 
Objective. This study aimed to evaluate the association between serum uric acid (SUA) levels within a normal to high range and the risk of metabolic syndrome (MetS) among community elderly and explore the sex difference. Design and Methods. A cross-sectional study was conducted in a representative urban area of Beijing between 2009 and 2010. A two-stage stratified clustering sampling method was used and 2102 elderly participants were included. Results. The prevalence of hyperuricemia and MetS was 16.7% and 59.1%, respectively. There was a strong association between hyperuricemia and four components of MetS in women and three components in men. Multiple logistic regression analysis showed ORs of hyperuricemia for MetS were 1.67 (95% CI: 1.11–2.50) in men and 2.73 (95% CI: 1.81–4.11) in women. Even in the normal range, the ORs for MetS increased gradually according to SUA levels. MetS component number also showed an increasing trend across SUA quartile in both sexes (P for trend < 0.01). Conclusion. This study suggests that higher SUA levels, even in the normal range, are positively associated with MetS among Chinese community elderly, and the association is stronger in women than men. Physicians should recognize MetS as a frequent comorbidity of hyperuricemia and take early action to prevent subsequent disease burden.
doi:10.1155/2014/754678
PMCID: PMC4129973  PMID: 25136366
4.  Occurrence of mycotoxins in feed ingredients and complete feeds obtained from the Beijing region of China 
Background
The current study was carried out to provide a reference for the control of mycotoxin contamination in feed ingredients and complete feeds for swine.
Methods
A total of 55 feed ingredients, including 14 corn, 13 wheat bran, 11 soybean meal and 17 dried distillers grains with solubles (DDGS) as well as 76 complete swine feeds including 7 creep feeds, 14 starter feeds, 14 grower feeds, 18 grower-finisher feeds, 10 gestating sow feeds, and 13 lactating sow feeds were randomly collected from 15 swine farms located in the Beijing region of China from July to August 2011. Immunoaffinity clean-up, using High-Performance Liquid Chromatography (HPLC) in combination with UV or Fluorescence Detection, was used for quantitative analysis of aflatoxin B1 (AFB1), deoxynivalenol (DON), zearalenone (ZEA) and ochratoxin A (OTA) in the ingredients and complete feeds.
Results
DON and ZEA were the most prevalent mycotoxins found. DON was detected at percentages of 93, 92, 54, 100 and 97% with a mean level of 1.01, 0.44, 0.05, 1.36 and 0.65 ppm in the samples of corn, wheat bran, soybean meal, DDGS and complete feeds, respectively. The detected percentages of ZEA were 100, 100, 54, 100 and 100 with mean levels of 109.1, 14.9, 9.2, 882.7 and 58.9 ppb in the same samples. In the wheat bran and soybean meal samples, the content of all four mycotoxins were below the maximum limits set by Chinese regulations while the percentage of samples that exceeded regulatory limits were 7, 57 and 7% for corn, and 7, 14 and 3% for the complete feeds for AFB1, DON and OTA respectively. DDGS showed the most serious mycotoxin contamination and the percentage of samples that exceeded regulatory limits were 6, 88 and 41%, for AFB1, DON and ZEA, respectively.
Conclusions
This paper is the first to present data on the natural occurrence of AFB1, DON, ZEA and OTA in ingredients and complete feeds obtained from swine farms in China’s Beijing region. The data shows that feed ingredients and complete swine feeds obtained from these farms are most often contaminated with DON followed by contamination with AFB1 and ZEA.
doi:10.1186/2049-1891-5-37
PMCID: PMC4123309  PMID: 25101169
AflatoxinB1; Beijing region; Deoxynivalenol; Ochratoxin A; Swine feeds; Zearalenone
5.  Absence of 19 known hotspot oncogenic mutations in soft tissue clear cell sarcoma: two cases report with review of the literature 
Clear cell sarcoma (CCS) of the tendons and aponeuroses is a rare soft tissue sarcoma that morphologically resembles cutaneous malignant melanoma but exhibits a distinct molecular profile. Gastrointestinal (GI) CCS is extremely rare. In this study, two cases of CCS were presented: (1) left thumb and (2) jejunum. Case 1 manifested the characteristic CCS morphology. Case 2 was morphologically unusual and difficult to diagnose. Immunohistochemically, the two cases of tumor cells were diffusely positive for S100, vimentin, NSE protein, focal expression of CgA, and CAM2.5 protein. In case 1, the tumor cells were diffusely positive for HMB45, focal expression of CD56, and melan A antigen. Reverse transcriptase-polymerase chain reaction (RT-PCR) results confirmed the presence of the EWS/ATF1 translocation (type 1) in the two cases. Then, we detected 19 hotspot oncogenes in the two cases. To the best of our knowledge, this study is the first to apply a high-throughput OncoCarta panel 1.0 and MassARRAY system to detect 238 known mutations in 19 hotspot oncogenes in soft tissue clear cell sarcoma. In this study, no mutations were observed in these hotspot oncogenes in the two cases.
PMCID: PMC4152094  PMID: 25197404
Clear cell sarcoma; mutations; fuse gene
6.  Periostin promotes liver steatosis and hypertriglyceridemia through downregulation of PPARα 
The Journal of Clinical Investigation  2014;124(8):3501-3513.
Hepatosteatosis is characterized by an aberrant accumulation of triglycerides in the liver; however, the factors that drive obesity-induced fatty liver remain largely unknown. Here, we demonstrated that the secreted cell adhesion protein periostin is markedly upregulated in livers of obese rodents and humans. Notably, overexpression of periostin in the livers of WT mice promoted hepatic steatosis and hypertriglyceridemia. Conversely, both genetic ablation of periostin and administration of a periostin-neutralizing antibody dramatically improved hepatosteatosis and hypertriglyceridemia in obese mice. Overexpression of periostin resulted in reduced expression of peroxisome proliferator–activated receptor α (PPARα), a master regulator of fatty acid oxidation, and activation of the JNK signaling pathway. In mouse primary hepatocytes, inhibition of α6β4 integrin prevented activation of JNK and suppression of PPARα in response to periostin. Periostin-dependent activation of JNK resulted in activation of c-Jun, which prevented RORα binding and transactional activation at the Ppara promoter. Together, these results identify a periostin-dependent pathway that mediates obesity-induced hepatosteatosis.
doi:10.1172/JCI74438
PMCID: PMC4109546  PMID: 25003192
7.  Genetic Association Study with Metabolic Syndrome and Metabolic-Related Traits in a Cross-Sectional Sample and a 10-Year Longitudinal Sample of Chinese Elderly Population 
PLoS ONE  2014;9(6):e100548.
Background
The metabolic syndrome (MetS) has been known as partly heritable, while the number of genetic studies on MetS and metabolic-related traits among Chinese elderly was limited.
Methods
A cross-sectional analysis was performed among 2 014 aged participants from September 2009 to June 2010 in Beijing, China. An additional longitudinal study was carried out among the same study population from 2001 to 2010. Biochemical profile and anthropometric parameters of all the participants were measured. The associations of 23 SNPs located within 17 candidate genes (MTHFR, PPARγ, LPL, INSIG, TCF7L2, FTO, KCNJ11, JAZF1, CDKN2A/B, ADIPOQ, WFS1, CDKAL1, IGF2BP2, KCNQ1, MTNR1B, IRS1, ACE) with overweight and obesity, diabetes, metabolic phenotypes, and MetS were examined in both studies.
Results
In this Chinese elderly population, prevalence of overweight, central obesity, diabetes, dyslipidemia, hypertension, and MetS were 48.3%, 71.0%, 32.4%, 75.7%, 68.3% and 54.5%, respectively. In the cross-sectional analyses, no SNP was found to be associated with MetS. Genotype TT of SNP rs4402960 within the gene IGF2BP2 was associated with overweight (odds ratio (OR)  = 0.479, 95% confidence interval (CI): 0.316-0.724, p = 0.001) and genotype CA of SNP rs1801131 within the gene MTHFR was associated with hypertension (OR = 1.560, 95% CI: 1.194–2.240, p = 0.001). However, these associations were not observed in the longitudinal analyses.
Conclusions
The associations of SNP rs4402960 with overweight as well as the association of SNP rs1801131 with hypertension were found to be statistically significant. No SNP was identified to be associated with MetS in our study with statistical significance.
doi:10.1371/journal.pone.0100548
PMCID: PMC4069025  PMID: 24959828
8.  Significant Coronary Stenosis in Asymptomatic Chinese With Different Glycemic Status 
Diabetes Care  2013;36(6):1687-1694.
OBJECTIVE
To evaluate coronary artery stenosis in early diabetes or prediabetes asymptomatic of myocardial ischemia in community-dwelling Chinese adults.
RESEARCH DESIGN AND METHODS
Age- and sex-matched participants with normal glucose regulation (NGR), prediabetes, or diabetes diagnosed within 5 years, asymptomatic of coronary artery disease (CAD), were randomly selected from a community-dwelling Chinese population aged 40–60 years. Dual-source computed tomography coronary angiography was used to evaluate the existence and extent of coronary stenosis, which was considered significant if >50% narrowing of vessel lumen was detected.
RESULTS
After excluding uninterpretable segments attributable to motion artifacts, a total of 135 participants with NGR, 132 with prediabetes, and 134 with diabetes participated in data analysis. Significant coronary stenosis was detected in 10 (7.4%), 10 (7.6%), and 22 (16.4%) individuals with NGR, prediabetes, and diabetes, respectively (P for trend = 0.029). Diabetes, rather than prediabetes, was associated with a significant 2.34-fold elevated risk [odds ratio (OR) 2.34 (95% CI 1.01–5.43); P = 0.047] of significant coronary stenosis as compared with that associated with NGR. Levels of glucose evaluation were independently and significantly associated with risks of significant coronary stenosis in diabetes. Each 1-SD increase in fasting plasma glucose, 2-h postload plasma glucose, and HbA1c conveyed 2.11-fold, 1.73-fold, and 1.81-fold higher risks of significant coronary stenosis, respectively, after adjustment for other conventional cardiovascular risk factors.
CONCLUSIONS
Using a noninvasive CAD diagnostic modality such as dual-source computed tomography coronary angiography, we detected a markedly elevated risk of significant coronary stenosis with early diabetes in asymptomatic Chinese adults.
doi:10.2337/dc12-0977
PMCID: PMC3661849  PMID: 23462666
9.  Association of Serum Irisin with Metabolic Syndrome in Obese Chinese Adults 
PLoS ONE  2014;9(4):e94235.
Irisin, a recently identified novel myokine, drives brown-fat-like conversion of white adipose tissues and has been proposed to mediate beneficial effects of exercise on metabolism. Circulating irisin was significantly reduced in type 2 diabetes patients; however, no evidence is available about its association with metabolic syndrome (MetS) and effects of adiposity and muscle mass on circulating irisin have been controversial. Cross-sectional data on socio-demographic, lifestyle, clinical characteristics and serum irisin were collected for 1,115 community-living Chinese adults with central obesity. Associations of serum irisin with MetS (central obesity plus any two of the following four factors (raised blood pressure (BP), raised fasting plasma glucose (FPG), raised triglyceride (TG), and reduced HDL cholesterol) and each component of MetS were analyzed using multivariable logistic regression. Among the 1,115 obese Chinese adults with a mean age of 53.2(±7.2) years, serum irisin levels (log-transformed) were significantly reduced in subjects with MetS and raised FPG than their control groups (p = 0.034 and 0.041, respectively). After adjustment for potential confounders, serum irisin was significantly associated with reduced risks of MetS and raised FPG, with odds ratios (ORs) (95% CI) per standard deviation of log-transformed irisin of 0.796 (0.505–0.959, p = 0.027) and 0.873 (0.764–0.998, p = 0.046), respectively. Associations of irisin with raised BP, raised TG and reduced HDL were not statistically significant ((ORs) (95% CI): 0.733(0.454–1.182, p = 0.202), 0.954(0.838–1.086, p = 0.478) and 1.130(0.980–1.302, p = 0.092), respectively). Stepwise multivariable linear regression analysis showed that fasting insulin, HbA1c and albumin/globulin ratio were negatively associated with serum irisin level with statistical significance (all p-values <0.05) and waist circumference was negatively associated with serum risin with marginally statistical significance (p = 0.055). These results imply that irisin may play an important role in insulin resistance and MetS and should be confirmed in future prospective studies.
doi:10.1371/journal.pone.0094235
PMCID: PMC3978033  PMID: 24709991
10.  Yin Yang 1 Promotes Hepatic Gluconeogenesis Through Upregulation of Glucocorticoid Receptor 
Diabetes  2013;62(4):1064-1073.
Gluconeogenesis is critical in maintaining blood glucose levels in a normal range during fasting. In this study, we investigated the role of Yin Yang 1 (YY1), a key transcription factor involved in cell proliferation and differentiation, in the regulation of hepatic gluconeogenesis. Our data showed that hepatic YY1 expression levels were induced in mice during fasting conditions and in a state of insulin resistance. Overexpression of YY1 in livers augmented gluconeogenesis, raising fasting blood glucose levels in C57BL/6 mice, whereas liver-specific ablation of YY1 using adenoviral shRNA ameliorated hyperglycemia in wild-type and diabetic db/db mice. At the molecular level, we further demonstrated that the major mechanism of YY1 in the regulation of hepatic glucose production is to modulate the expression of glucocorticoid receptor. Therefore, our study uncovered for the first time that YY1 participates in the regulation of hepatic gluconeogenesis, which implies that YY1 might serve as a potential therapeutic target for hyperglycemia in diabetes.
doi:10.2337/db12-0744
PMCID: PMC3609554  PMID: 23193188
11.  cAMP Signaling Prevents Podocyte Apoptosis via Activation of Protein Kinase A and Mitochondrial Fusion 
PLoS ONE  2014;9(3):e92003.
Our previous in vitro studies suggested that cyclic AMP (cAMP) signaling prevents adriamycin (ADR) and puromycin aminonucleoside (PAN)-induced apoptosis in podocytes. As cAMP is an important second messenger and plays a key role in cell proliferation, differentiation and cytoskeleton formation via protein kinase A (PKA) or exchange protein directly activated by cAMP (Epac) pathways, we sought to determine the role of PKA or Epac signaling in cAMP-mediated protection of podocytes. In the ADR nephrosis model, we found that forskolin, a selective activator of adenylate cyclase, attenuated albuminuria and improved the expression of podocyte marker WT-1. When podocytes were treated with pCPT-cAMP (a selective cAMP/PKA activator), PKA activation was increased in a time-dependent manner and prevented PAN-induced podocyte loss and caspase 3 activation, as well as a reduction in mitochondrial membrane potential. We found that PAN and ADR resulted in a decrease in Mfn1 expression and mitochondrial fission in podocytes. pCPT-cAMP restored Mfn1 expression in puromycin or ADR-treated podocytes and induced Drp1 phosphorylation, as well as mitochondrial fusion. Treating podocytes with arachidonic acid resulted in mitochondrial fission, podocyte loss and cleaved caspase 3 production. Arachidonic acid abolished the protective effects of pCPT-cAMP on PAN-treated podocytes. Mdivi, a mitochondrial division inhibitor, prevented PAN-induced cleaved caspase 3 production in podocytes. We conclude that activation of cAMP alleviated murine podocyte caused by ADR. PKA signaling resulted in mitochondrial fusion in podocytes, which at least partially mediated the effects of cAMP.
doi:10.1371/journal.pone.0092003
PMCID: PMC3958405  PMID: 24642777
12.  Involvement of Multiple Types of Dehydrins in the Freezing Response in Loquat (Eriobotrya japonica) 
PLoS ONE  2014;9(1):e87575.
Dehydrins (DHNs) are a family of plant proteins typically induced in response to stress conditions that cause cellular dehydration, such as low temperatures, high salinity, and drought. Loquat (Eriobotrya japonica) is a perennial fruit crop that blossoms during winter. Loquat fruitlets are frequently injured by freezing. To evaluate the role of the EjDHNs in freezing resistance in loquat fruitlets, two cultivars of loquat, the freezing-sensitive ‘Ninghaibai’ (FS-NHB) and the freezing-tolerant ‘Jiajiao’ (FT-JJ), were analyzed under induced freezing stress. Freezing stress led to obvious accumulation of reactive oxygen species and considerable lipid peroxidation in membranes during the treatment period. Both these phenomena were more pronounced in ‘FS-NHB’ than in ‘FS-JJ.’ Immunogold labeling of dehydrin protein was performed. DHN proteins were found to be concentrated mainly in the vicinity of the plasma membrane, and the density of the immunogold labeling was significantly higher after freezing treatment, especially in the more freezing-tolerant cultivar ‘FT-JJ.’ Seven DHNs, showing four different structure types, were obtained from loquat fruitlets and used to study the characteristics of different EjDHN proteins. These DHN proteins are all highly hydrophilic, but they differ significantly in size, ranging from 188 to 475 amino acids, and in biochemical properties, such as theoretical pI, aliphatic index, and instability index. Freezing treatment resulted in up-regulation of the expression levels of all seven EjDHNs, regardless of structure type. The accumulation of the transcripts of these EjDHN genes was much more pronounced in ‘FT-JJ’ than in ‘FS-NHB.’ Altogether, this study provides evidence that EjDHNs are involved in the cryoprotection of the plasma membrane during freeze-induced dehydration in loquat fruitlets.
doi:10.1371/journal.pone.0087575
PMCID: PMC3909202  PMID: 24498141
13.  Expression, refolding and preliminary X-ray crystallographic analysis of equine MHC class I molecule complexed with an EIAV-Env CTL epitope 
The equine MHC class I molecule was crystallized in complex with β2-microglobulin and a CTL epitope and X-ray diffraction data were collected to 2.3 Å resolution.
In order to clarify the structure and the peptide-presentation characteristics of the equine major histocompatibility complex (MHC) class I molecule, a complex of equine MHC class I molecule (ELA-A1 haplotype, 7-6 allele) with mouse β2-­microglobulin and the cytotoxic T lymphocyte (CTL) epitope Env-RW12 (RVEDVTNTAEYW) derived from equine infectious anaemia virus (EIAV) envelope protein (residues 195–206) was refolded and crystallized. The crystal, which belonged to space group P21, diffracted to 2.3 Å resolution and had unit-cell parameters a = 82.5, b = 71.4, c = 99.8 Å, β = 102.9°. The crystal structure contained two molecules in the asymmetric unit. These results should help to determine the first equine MHC class I molecule structure presenting an EIAV CTL epitope.
doi:10.1107/S1744309111038139
PMCID: PMC3253827  PMID: 22232164
equine MHC class I molecule
14.  Increasing Prevalence of Metabolic Syndrome in a Chinese Elderly Population: 2001–2010 
PLoS ONE  2013;8(6):e66233.
Objective
The information on the changes of prevalence of MetS in China is limited. Our objective was to assess a 10-year’s change of the prevalence of MetS in a Chinese elderly population between 2001 and 2010.
Methods
We conducted two cross-sectional surveys in a representative sample of elderly population aged 60 to 95 years in Beijing in 2001 and 2010 respectively. MetS was defined according to the 2009 harmonizing definition.
Results
A total of 2,334 participants (943 male, 1,391 female) in 2001 and 2,102 participants (848 male, 1,254 female) in 2010 completed the survey. The prevalence of MetS was 50.4% (95%CI: 48.4%–52.4%) in 2001 and 58.1% (95%CI: 56.0%–60.2%) in 2010. The absolute change of prevalence of MetS was 7.7% over the 10-year’s period (p<0.001). The syndrome was more common in female than male in both survey years. Among the five components, hypertriglyceridemia and low HDL-C had increased most, with an increase of 14.8% (from 29.4% to 44.2%) and 9.9% (from 28.3% to 38.2%) respectively. The adjusted ORs of MetS for CHD, stroke and CVD were 1.67(95%CI: 1.39–1.99), 1.50(95%CI: 1.19–1.88) and 1.70(95%CI: 1.43–2.01) respectively in 2001, and were 1.74(95%CI: 1.40–2.17), 1.25(95%CI: 0.95–1.63) and 1.52(95%CI: 1.25–1.86) respectively in 2010.
Conclusion
The prevalence of MetS is high and increasing rapidly in this Chinese elderly population. Participants with Mets and its individual components are at significantly elevated ORs for CVD. Urgent public health actions are needed to control MetS and its components, especially for dislipidemia.
doi:10.1371/journal.pone.0066233
PMCID: PMC3688874  PMID: 23824753
15.  Orthostatic Hypotension and the Risk of Congestive Heart Failure: A Meta-Analysis of Prospective Cohort Studies 
PLoS ONE  2013;8(5):e63169.
Background
Orthostatic hypotension (OH) has been related to the increased risk of future congestive heart failure (CHF) events. However, the overall quantitative estimate of predictive ability of OH for CHF has not been determined. We therefore performed a meta-analysis to investigate the association between OH and incident CHF.
Methods
Prospective cohort studies relevant to the aim of the study were identified by searching of Medline and Embase databases up to December 25, 2012 without restrictions and by reviewing the reference lists from retrieved articles.
Results
A total of 51270 subjects and 3603 incident CHF cases from 4 prospective cohorts were included in the meta-analysis. Using random effect model, the pooled result indicated that presence of OH at baseline was significantly associated with an increased risk for future CHF outcomes (adjusted hazard ratio: 1.30, 95% confidence interval 1.09–1.55; p = 0.004). Results of stratified analysis suggested that the association between OH and CHF incidence seemed to be significant in middle-age subjects, or the individuals with hypertension and diabetes at baseline, but did not significant in the elderly subjects or those without hypertension or diabetes.
Conclusions
Our meta-analysis confirmed that presence of OH is related to a significant increased risk for development of CHF in the future. Studies are needed to explore the potential mechanisms underlying this association. More importantly, screen for OH may be of great clinical significance for the early identification of subjects at higher risk for development of CHF.
doi:10.1371/journal.pone.0063169
PMCID: PMC3652866  PMID: 23675460
16.  Chebulagic Acid, a Hydrolyzable Tannin, Exhibited Antiviral Activity in Vitro and in Vivo against Human Enterovirus 71 
Human enterovirus 71 is one of the major causative agents of hand, foot and mouth disease in children under six years of age. Presently, no vaccines or antiviral drugs have been clinically available to employ against EV71. In this study, we demonstrate that treatment with chebulagic acid reduced the viral cytopathic effect on rhabdomyosarcoma cells with an IC50 of 12.5 μg/mL. The utilization of the chebulagic acid treatment on mice challenged with a lethal dose of enterovirus 71 was able to efficiently reduce mortality and relieve clinical symptoms through the inhibition of viral replication. Chebulagic acid may represent a potential therapeutic agent to control infections to enterovirus 71.
doi:10.3390/ijms14059618
PMCID: PMC3676802  PMID: 23644889
chebulagic acid; enterovirus 71; hydrolyzable tannin; antiviral activity
17.  Loss of Steroid Receptor Coactivator-3 attenuates carbon tetrachloride-induced murine hepatic injury and fibrosis 
Hepatic fibrosis, a disease characterized by altered accumulation of extracellular matrix, can cause cirrhosis and liver failure. There is growing interest in the impact of coactivators on hepatic fibrogenesis. Here, we provided genetic evidence that mice lacking steroid receptor coactivator-3 (SRC-3) were protected against carbon tetrachloride (CCl4) induced acute liver necrosis and chronic hepatic fibrosis. After acute CCl4 treatment, SRC-3−/− mice showed attenuated profibrotic response and hepatocyte apoptosis, while hepatocyte proliferation was elevated in SRC-3−/− mice versus SRC-3+/+ mice. Similarly, chronically CCl4-treated SRC-3−/− mice showed significant weakening of inflammatory infiltrates, hepatic stellate cell activation and collagen accumulation in the liver compared to SRC-3+/+ mice. Further investigation revealed that TGFβ1/Smad signaling pathway was impaired in the absence of SRC-3. Moreover, the expression levels of SRC-3, as assessed in human tissue microarray of liver diseases, correlated positively with degrees of fibrosis. These data revealed that SRC-3−/− mice were resistant to CCl4 induced acute and chronic hepatic damage and TGFβ1/Smad signaling was suppressed in the lack of SRC-3. Our results established an essential involvement of SRC-3 in liver fibrogenesis, which might provide new clues to the future treatment of hepatic fibrosis.
doi:10.1038/labinvest.2009.51
PMCID: PMC3620314  PMID: 19488034
Chronic hepatic fibrosis; Smad2/3; Steroid receptor coactivator-3; Tissue microarray; Transforming growth factor β1
18.  Maskless and low-destructive nanofabrication on quartz by friction-induced selective etching 
Nanoscale Research Letters  2013;8(1):140.
A low-destructive friction-induced nanofabrication method is proposed to produce three-dimensional nanostructures on a quartz surface. Without any template, nanofabrication can be achieved by low-destructive scanning on a target area and post-etching in a KOH solution. Various nanostructures, such as slopes, hierarchical stages and chessboard-like patterns, can be fabricated on the quartz surface. Although the rise of etching temperature can improve fabrication efficiency, fabrication depth is dependent only upon contact pressure and scanning cycles. With the increase of contact pressure during scanning, selective etching thickness of the scanned area increases from 0 to 2.9 nm before the yield of the quartz surface and then tends to stabilise after the appearance of a wear. Refabrication on existing nanostructures can be realised to produce deeper structures on the quartz surface. Based on Arrhenius fitting of the etching rate and transmission electron microscopy characterization of the nanostructure, fabrication mechanism could be attributed to the selective etching of the friction-induced amorphous layer on the quartz surface. As a maskless and low-destructive technique, the proposed friction-induced method will open up new possibilities for further nanofabrication.
doi:10.1186/1556-276X-8-140
PMCID: PMC3620578  PMID: 23531381
Maskless; Nanofabrication; Friction-induced selective etching; Quartz; 81.16.Rf; 68.37.Ps
19.  Active Smoking, Passive Smoking, and Risk of Nonalcoholic Fatty Liver Disease (NAFLD): A Population-Based Study in China 
Journal of Epidemiology  2013;23(2):115-121.
Background
The effect of active smoking on development of nonalcoholic fatty liver disease (NAFLD) is controversial, and there are limited clinical data on the relationship between passive smoking and NAFLD. We investigated whether active and passive smoking are associated with NAFLD.
Methods
A total of 8580 subjects (2691 men) aged 40 years or older participated in a community-based survey in Shanghai, China. Information on active and passive smoking was collected using a validated questionnaire. NAFLD was diagnosed by abdominal B-mode ultrasound testing and serum liver enzymes.
Results
NAFLD prevalence was 29.4% in never smokers, 34.2% in former smokers, 27.8% in light smokers (<20 cigarettes/day), 30.8% in moderate smokers (20–39 cigarettes/day), and 43.5% in heavy smokers (≥40 cigarettes/day). Fully adjusted logistic regression analyses revealed that, as compared with never smoking, former and heavy smoking were associated with increased risk of prevalent NAFLD, with odds ratios of 1.45 (95% CI 1.05–2.00) and 2.29 (95% CI 1.30–4.03), respectively. Active smoking and body mass index (BMI) had a synergistic effect on the risk of prevalent NAFLD; the combination of these risk factors was associated with the highest observed odds ratio for NAFLD: 8.58. In never-smoking women, passive smoking during both childhood and adulthood was associated with a 25% increase in the risk of prevalent NAFLD (OR = 1.25, 95% CI 1.05–1.50) as compared with no passive smoking.
Conclusions
Passive smoking and heavy active smoking are associated with prevalent NAFLD in middle-aged and elderly Chinese. Active smoking and BMI have a synergistic effect on prevalent NAFLD.
doi:10.2188/jea.JE20120067
PMCID: PMC3700247  PMID: 23399520
active tobacco smoking; passive tobacco smoking; fatty liver
20.  Impaired Lung Function Is Associated with Increased Carotid Intima-Media Thickness in Middle-Aged and Elderly Chinese 
PLoS ONE  2013;8(2):e53153.
Background
Impairment of lung function was reported to be associated with cardiovascular disease (CVD). The aim of the present study is to evaluate the relationship between lung function and carotid intima-media thickness (cIMT) in participants without chronic pulmonary disease.
Methodology and Principal Findings
A total of 6,423 participants aged 40 years and above were recruited from Jiading District, Shanghai, China. Lung function, evaluated by forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) was measured with standard spirometry. CIMT was measured with high-resolution ultrasonography by trained physicians. Mean values of FVC (% pred) and FEV1 (% pred) in participants with elevated cIMT were significantly lower than in those without (0.92±0.20 vs. 0.99±0.19, 0.83±0.24 vs. 0.90±0.22; both p-values < 0.0001). The levels of cIMT in the lowest quartile of FVC (% pred) and FEV1 (% pred) were markedly higher than in the second, third and fourth quartile, respectively (p < 0.0001 for all). The lowest quartile of FVC (% pred) and FEV1 (% pred) was associated with increased odds of elevated cIMT, with the fully adjusted odds ratio of 1.34 and 1.41 (95% confidence interval (CI) 1.09–1.65, p  =  0.006, 95% CI 1.15–1.72, p  =  0.0008), respectively.
Conclusions and Significance
Impaired lung function is associated with elevated cIMT in middle-aged and elderly Chinese. These findings suggest the need to screen impairment of lung function in people without respiratory disease for the presence of subclinical atherosclerosis in CVD prevention.
doi:10.1371/journal.pone.0053153
PMCID: PMC3574141  PMID: 23457443
21.  Assessing the value of bilateral inferior petrosal sinus sampling in the diagnosis and treatment of a complex case of Cushing's disease 
Summary
A 41-year-old female visited Ruijin Hospital because her face was swollen for more than 2 months. The patient was initially diagnosed with Cushing's disease (CD). Several examinations, including a dexamethasone suppression test (DST) at 2 mg and 8 mg, pituitary MRI, abdominal CT, punch biopsy of adrenal masses, and bilateral inferior petrosal sinus sampling (BIPSS), were performed, but the findings were not consistent with the clinical presentation. Ultimately, the patient underwent surgery and recovered. In this case, BIPSS was a useful way to diagnosis CD and suggested the exact location of a pituitary adenoma to Neurosurgery. BIPSS should be a required test for cases of CD that cannot be definitively diagnosed with just an MRI and 8 mg DST before surgery.
doi:10.5582/irdr.2013.v2.1.24
PMCID: PMC4204576  PMID: 25343097
Cushing's syndrome; Cushing's disease; bilateral inferior petrosal sinus sampling (BIPSS)
22.  Effect of Fish Oil Supplementation on Fasting Vascular Endothelial Function in Humans: A Meta-Analysis of Randomized Controlled Trials 
PLoS ONE  2012;7(9):e46028.
Background
Effect of fish oil supplementation on flow-mediated dilation, an index of endothelial function in humans, remains controversial. We performed a meta-analysis to determine whether fish oil supplementation could improve endothelial function.
Methods
Human intervention studies were identified by systematic searches of Medline, Embase, Cochrane's library and references of related reviews and studies. A random-effect model was applied to estimate the pooled results. Meta-regression and subgroup analyses were performed to evaluate the impact of study characteristics on the effect of fish oil supplementation on flow-mediated dilation.
Results
A total of sixteen records with 1,385 subjects were reviewed. The results of the pooled analysis showed that fish oil supplementation significantly improved flow-mediated dilation (weighed mean difference: 1.49%, 95% confidence interval 0.48% to 2.50%, p = 0.004). Meta-regression and subgroup analysis suggested that the quality of included studies were inversely related to the overall effect (regression coefficient  = −1.60, p = 0.04), and the significance of the effect was mainly driven by the studies with relatively poor quality. Sensitivity analysis including only double-blind, placebo-controlled studies indicated fish oil supplementation has no significant effect on endothelial function (weighed mean difference: 0.54%, 95% confidence interval −0.25% to 1.33%, p = 0.18). Besides, normoglycemic subjects or participants with lower diastolic blood pressure seemed to be associated with remarkable improvement of endothelial function after fish oil supplementation.
Conclusions
Although current evidence suggested a possible role of fish oil in improving endothelial function, large-scale and high-quality clinical trials are needed to evaluate these effects before we can come to a definite conclusion.
doi:10.1371/journal.pone.0046028
PMCID: PMC3448723  PMID: 23029372
23.  Effects of fish oil supplementation on inflammatory markers in chronic heart failure: a meta-analysis of randomized controlled trials 
Background
Effects of fish oil on systematic inflammation in chronic heart failure remain unclear. In this meta-analysis, we aimed to evaluate the influence of fish oil supplementation on circulating levels of inflammatory markers in patients with chronic heart failure.
Methods
Human randomized controlled trials, which compared the effects of fish oil supplementation with placebo in patients with chronic heart failure, were identified by systematic search of Medline, Embase, Cochrane’s library and references cited in related reviews and studies up to November 2011. Outcome measures comprised the changes of circulating inflammatory markers. Meta-analysis was performed with the fixed-effect model or random-effect model according to the heterogeneity.
Results
A total of seven trials with eight study arms were included. The pooled results indicated circulating levels of tumor necrosis factor α (SMD = -0.62, 95% CI -1.08 to -0.16, p = 0.009), interleukin 1 (SMD = -1.24, 95% CI -1.56 to -0.91, p < 0.001) and interleukin 6 (SMD = -0.81, 95% CI -1.48 to -0.14, p = 0.02) were significantly decreased after fish oil supplementation; however, high sensitivity C reactive protein, soluble intracellular adhesion molecular 1 and vascular cell adhesion molecular 1 were not significantly affected. Meta-regression and subgroup analysis results suggested the difference in dose of fish oil and follow-up duration might influence the effects of fish oil on tumor necrosis factor α and interleukin 6. Greater reduction of these two markers might be achieved in patients taking fish oil of a higher dose (over 1000 mg/day) or for a longer duration (over 4 months).
Conclusions
Limited evidence suggests anti-inflammation may be a potential mechanism underlying the beneficial effects of fish oil for chronic heart failure. Further large-scale and adequately powered clinical trials are needed to confirm these effects.
doi:10.1186/1471-2261-12-77
PMCID: PMC3507701  PMID: 22994912
Fish oil; Omega 3 polyunsaturated fatty acids; Heart failure; Inflammatory markers; Meta-analysis
24.  Structural Changes of Gut Microbiota during Berberine-Mediated Prevention of Obesity and Insulin Resistance in High-Fat Diet-Fed Rats 
PLoS ONE  2012;7(8):e42529.
Berberine, a major pharmacological component of the Chinese herb Coptis chinensis, which was originally used to treat bacterial diarrhea, has recently been demonstrated to be clinically effective in alleviating type 2 diabetes. In this study, we revealed that berberine effectively prevented the development of obesity and insulin resistance in high-fat diet (HFD)-fed rats, which showed decreased food intake. Increases in the levels of serum lipopolysaccharide-binding protein, monocyte chemoattractant protein-1, and leptin and decrease in the serum level of adiponectin corrected for body fat in HFD-fed rats were also significantly retarded by the co-administration of berberine at 100 mg/kg body weight. Bar-coded pyrosequencing of the V3 region of 16S rRNA genes revealed a significant reduction in the gut microbiota diversity of berberine-treated rats. UniFrac principal coordinates analysis revealed a marked shift of the gut microbiota structure in berberine-treated rats away from that of the controls. Redundancy analysis identified 268 berberine-responding operational taxonomic units (OTUs), most of which were essentially eliminated, whereas a few putative short-chain fatty acid (SCFA)-producing bacteria, including Blautia and Allobaculum, were selectively enriched, along with elevations of fecal SCFA concentrations. Partial least square regression models based on these 268 OTUs were established (Q2>0.6) for predicting the adiposity index, body weight, leptin and adiponectin corrected for body fat, indicating that these discrete phylotypes might have a close association with the host metabolic phenotypes. Taken together, our findings suggest that the prevention of obesity and insulin resistance by berberine in HFD-fed rats is at least partially mediated by structural modulation of the gut microbiota, which may help to alleviate inflammation by reducing the exogenous antigen load in the host and elevating SCFA levels in the intestine.
doi:10.1371/journal.pone.0042529
PMCID: PMC3411811  PMID: 22880019
25.  High-Throughput Construction of Intron-Containing Hairpin RNA Vectors for RNAi in Plants 
PLoS ONE  2012;7(5):e38186.
With the wide use of double-stranded RNA interference (RNAi) for the analysis of gene function in plants, a high-throughput system for making hairpin RNA (hpRNA) constructs is in great demand. Here, we describe a novel restriction-ligation approach that provides a simple but efficient construction of intron-containing hpRNA (ihpRNA) vectors. The system takes advantage of the type IIs restriction enzyme BsaI and our new plant RNAi vector pRNAi-GG based on the Golden Gate (GG) cloning. This method requires only a single PCR product of the gene of interest flanked with BsaI recognition sequence, which can then be cloned into pRNAi-GG at both sense and antisense orientations simultaneously to form ihpRNA construct. The process, completed in one tube with one restriction-ligation step, produced a recombinant ihpRNA with high efficiency and zero background. We demonstrate the utility of the ihpRNA constructs generated with pRNAi-GG vector for the effective silencing of various individual endogenous and exogenous marker genes as well as two genes simultaneously. This method provides a novel and high-throughput platform for large-scale analysis of plant functional genomics.
doi:10.1371/journal.pone.0038186
PMCID: PMC3364983  PMID: 22675447

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