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1.  Fasting Plasma Glucose at 24–28 Weeks to Screen for Gestational Diabetes Mellitus 
Diabetes Care  2013;36(7):2038-2040.
To evaluate the usefulness of a fasting plasma glucose (FPG) at 24–28 weeks’ gestation to screen for gestational diabetes mellitus (GDM).
The medical records and results of a 75-g 2-h oral glucose tolerance test (OGTT) of 24,854 pregnant women without known pre-GDM attending prenatal clinics in 15 hospitals in China were examined.
FPG cutoff value of 5.1 mmol/L identified 3,149 (12.1%) pregnant women with GDM. FPG cutoff value of 4.4 mmol/L ruled out GDM in 15,369 (38.2%) women. With use of this cutoff point, 12.2% of patients with mild GDM will be missed. The positive predictive value is 0.322, and the negative predictive value is 0.928.
FPG at 24–28 weeks’ gestation could be used as a screening test to identify GDM patients in low-resource regions. Women with an FPG between ≥4.4 and ≤5.0 mmol/L would require a 75-g OGTT to diagnose GDM. This would help to avoid approximately one-half (50.3%) of the formal 75-g OGTTs in China.
PMCID: PMC3687275  PMID: 23536582
2.  Preparation and functional studies of hydroxyethyl chitosan nanoparticles loaded with anti-human death receptor 5 single-chain antibody 
OncoTargets and therapy  2014;7:779-787.
To prepare hydroxyethyl chitosan nanoparticles loaded with anti-human death receptor 5 single-chain antibody, and study their characteristics, functions, and mechanisms of action.
Materials and methods
The anti-human death receptor 5 single-chain antibody was constructed and expressed. Protein-loaded hydroxyethyl chitosan nanoparticles were prepared, and their size, morphology, particle-size distribution and surface zeta potential were measured by scanning electron microscopy and laser particle-size analysis. Mouse H22 hepatocellular carcinoma cells were cultured, and growth inhibition was examined using the CellTiter-Blue cell-viability assay. Flow cytometry and Hoechst 33342 were employed to measure cell apoptosis. Kunming mice with H22 tumor models were treated with protein-loaded hydroxyethyl chitosan nanoparticles, and their body weight and tumor size were measured, while hematoxylin and eosin staining was used to detect antitumor effects in vivo and side effects from tumors.
The protein-loaded hydroxyethyl chitosan nanoparticles had good stability; the zeta potential was −24.2±0.205, and the dispersion index was 0.203. The inhibition of the protein-loaded hydroxyethyl chitosan nanoparticles on H22 growth was both time- and dose-dependent. Increased expressions of active caspase 8, active caspase 3, and BAX were detected following treatment. The average weight gain, tumor weight, and mean tumor volume of the protein and protein-loaded hydroxyethyl chitosan nanoparticle groups were significantly different (P<0.05) compared with the phosphate-buffered saline group.
The protein-loaded hydroxyethyl chitosan nanoparticles effectively suppressed tumor growth, indicating that nanotechnology has the potential for broad application in cancer therapy.
PMCID: PMC4039402  PMID: 24899816
anticancer effect; DR5; GCS-aDR5ScFv; H22
3.  Identification and Characterization of ABA Receptors in Oryza sativa 
PLoS ONE  2014;9(4):e95246.
Abscisic acid (ABA) is an essential phytohormone that regulates plant stress responses. ABA receptors in Arabidopsis thaliana (AtPYLs) have been extensively investigated by structural, biochemical, and in vivo studies. In contrast, relatively little is known about the ABA signal transduction cascade in rice. Besides, the diversities of AtPYLs manifest that the information accumulated in Arabidopsis cannot be simply adapted to rice. Thus, studies on rice ABA receptors are compulsory. By taking a bioinformatic approach, we identified twelve ABA receptor orthologs in Oryza sativa (japonica cultivar-group) (OsPYLs), named OsPYL1–12. We have successfully expressed and purified OsPYL1–3, 6 and 10–12 to homogeneity, tested the inhibitory effects on PP2C in Oryza sativa (OsPP2C), and measured their oligomerization states. OsPYL1–3 mainly exhibit as dimers and require ABA to inhibit PP2C’s activity. On the contrary, OsPYL6 retains in the monomer-dimer equilibrium state and OsPYL10–11 largely exist as monomers, and they all display an ABA-independent phosphatase inhibition manner. Interestingly, although OsPYL12 seems to be a dimer, it abrogates the phosphatase activity of PP2Cs in the absence of ABA. Toward a further understanding of OsPYLs on the ABA binding and PP2C inhibition, we determined the crystal structure of ABA-OsPYL2-OsPP2C06 complex. The bioinformatic, biochemical and structural analysis of ABA receptors in rice provide important foundations for designing rational ABA-analogues and breeding the stress-resistant rice for commercial agriculture.
PMCID: PMC3990689  PMID: 24743650
4.  Aberrant SATB1 expression is associated with Epstein-Barr virus infection, metastasis and survival in human nasopharyngeal cells and endemic nasopharyngeal carcinoma 
Special AT-rich sequence-binding protein 1 (SATB1) has been identified as a key factor in the progression of some cancers, functioning as a global genome organizer and chromatin regulator. We examined the levels of SATB1 mRNA expression in NPC cell lines 5-8F (high metastasis) and 6-10B (low metastasis) and immortalized human nasopharyngeal epithelial cells NP69-SV40T by quantitative real-time PCR. We also examined the protein expression levels of SATB1 in 72 cases of nasopharyngeal carcinoma (NPC) tissues and 30 cases of normal nasopharyngeal (NNP) tissues by immunohistochemistry, and then assessed the correlations between SATB1 expression and clinicopathological factors. The expression level of SATB1 mRNA in 5-8F was much higher than those in 6-10B and NP69-SV40T (P < 0.05). The expression level of SATB1 mRNA in 6-10B was higher than in NP69-SV40T, but the difference was not statistically significant (P > 0.05). The positive expression rates of SATB1 protein in NPC (38/72, 52.8%) were significantly higher than in NNP (4/30, 13.3%) (P < 0.05). SATB1 protein levels in NPC were not associated with gender, age, and T stage (P > 0.05), but positively correlated with the titers of EBVCA-IgA, metastasis (N and M stage), recurrence, and survival (P < 0.05). Multivariate analysis showed that the overexpression of SATB1 protein is an independent prognostic factor for NPC. The expression levels of SATB1 were obviously upregulated in primary NPC tissues and human NPC cell lines. Therefore, SATB1 may be a valuable predictor in assessing the metastasis, recurrence, and prognosis of NPC.
PMCID: PMC4069958  PMID: 24966956
Nasopharyngeal carcinoma; SATB1; immunohistochemistry; quantitative real-time PCR; prognosis
5.  Phenethyl Isothiocyanate inhibits STAT3 activation in prostate cancer cells 
This study was undertaken to investigate the mechanism by which phenethyl isothiocyanate (PEITC), a natural compound from cruciferous vegetables, exhibits antitumor effect on prostate cancer cells. Cell proliferation, cell cycle, western blot, gene transfer and reporter assays were used to test the effects of PEITC on the growth and IL6/JAK/STAT3 pathway in prostate cancer. The result showed that PEITC significantly inhibited DU145 cell proliferation in a dose-dependent manner and induced the cell arrest at G2-M phase. PEITC inhibited both constitutive and interleukin 6 (IL-6)-induced STAT3 activity in DU145 cells. IL-6-stimulated phosphorylation of JAK2, an STAT3 upstream kinase, was also attenuated by PEITC. Moreover, an antioxidant reagent, N-acetyl-l-cysteine (NAC) which suppresses reactive oxygen species (ROS) generation, reversed the early inhibitory effects of PEITC on cell proliferation, constitutive or IL-6-mediated JAK-STAT3 phosphorylation in PCa cells. Taken together, our data demonstrated that PEITC can inhibit the activation of the JAK-STAT3 signal-cascade in prostate cancer cells and the underlying mechanism may be partially involved with blocking cellular ROS production during the early stage of the signaling activation by IL-6.
PMCID: PMC3964815  PMID: 19437484
Phenethyl isothiocyanate; IL-6; STAT3; Androgen independent growth
6.  Radiation Dose in the Thyroid and the Thyroid Cancer Risk Attributable to CT Scans for Pediatric Patients in One General Hospital of China 
Objective: To quantify the radiation dose in the thyroid attributable to different CT scans and to estimate the thyroid cancer risk in pediatric patients. Methods: The information about pediatric patients who underwent CT scans was abstracted from the radiology information system in one general hospital between 1 January 2012 and 31 December 2012. The radiation doses were calculated using the ImPACT Patient Dosimetry Calculator and the lifetime attributable risk (LAR) of thyroid cancer incidence was estimated based on the National Academies Biologic Effects of Ionizing Radiation VII model. Results: The subjects comprised 922 children, 68% were males, and received 971 CT scans. The range of typical radiation dose to the thyroid was estimated to be 0.61–0.92 mGy for paranasal sinus CT scans, 1.10–2.45 mGy for head CT scans, and 2.63–5.76 mGy for chest CT scans. The LAR of thyroid cancer were as follows: for head CT, 1.1 per 100,000 for boys and 8.7 per 100,000 for girls; for paranasal sinus CT scans, 0.4 per 100,000 for boys and 2.7 per 100,000 for girls; for chest CT scans, 2.1 per 100,000 for boys and 14.1 per 100,000 for girls. The risk of thyroid cancer was substantially higher for girls than for the boys, and from chest CT scans was higher than that from head or paransal sinus CT scans. Conclusions: Chest CT scans caused higher thyroid dose and the LAR of thyroid cancer incidence, compared with paransal sinus or head CT scans. Therefore, physicians should pay more attention to protect the thyroid when children underwent CT scans, especially chest CT scans.
PMCID: PMC3987004  PMID: 24608902
cancer risk; pediatric CT; radiation dose
7.  Association between Soy Isoflavone Intake and Breast Cancer Risk for Pre- and Post-Menopausal Women: A Meta-Analysis of Epidemiological Studies 
PLoS ONE  2014;9(2):e89288.
Conclusions drawn from meta-analyses on the association between soy isoflavone intake and breast cancer risk for pre- and post-menopausal women are not fully consistent. These meta-analyses did not explore the influence of different study designs on the pooled results on the basis of distinguishing between pre- and post-menopausal women.
Methodology and Principal Findings
We performed a meta-analysis of 35 studies which reported results of association between soy isoflavone intake and breast cancer risk for pre- and/or post-menopausal women, calculated pooled odds ratios and their 95% confidence intervals of pre- and post-menopausal women respectively, and further explored soy isoflavone-breast cancer association on the basis of considering different study regions and designs. Summary results suggested that soy isoflavone intake has a protective effect against breast cancer for both pre- and post-menopausal women. However, they are influenced by study design and region. Pooled ORs of studies carried out in Asian countries suggested that soy isoflavone’s protective effect exist in both pre- and post-menopausal women (OR = 0.59, 95%CI: 0.48–0.69 for premenopausal women; OR = 0.59, 95%CI: 0.44–0.74 for postmenopausal women). However, there are some differences between the results pooled from different study designs for women in Asian countries (test for consistency, P = 0.04). Pooled OR of studies on postmenopausal women in Western countries suggested that soy isoflavone intake has a marginally significant protective effect (OR = 0.92; 95%CI: 0.83∼1.00), but further analyses stratifying by study design found no statistically significant association.
We meta-analyzed more and newer research results, and separated women according to menopausal status to explore soy isoflavone-breast cancer association. We founded that soy isoflavone intake could lower the risk of breast cancer for both pre- and post-menopausal women in Asian countries. However, for women in Western countries, pre- or post-menopausal, there is no evidence to suggest an association between intake of soy isoflavone and breast cancer.
PMCID: PMC3930722  PMID: 24586662
8.  Decreased Galectin-9 and Increased Tim-3 Expression Are Related to Poor Prognosis in Gastric Cancer 
PLoS ONE  2013;8(12):e81799.
Galectin-9 (Gal-9) induces adhesion and aggregation of certain cell types and inhibits the metastasis of tumor cells. T-cell immunoglobulin–and mucin domain-3–containing molecule 3 (TIM-3) plays a pivotal role in immune regulation. The aim of this study is to investigate Gal-9 and TIM-3 alterations in gastric cancer and their prognostic values.
Gal-9 and Tim-3 expression was evaluated using a tissue microarray immunohistochemistry method in 305 gastric cancers, of which 84 had paired adjacent normal samples. Cell lines SGC-7901, BGC-823, MGC-803, MKN45 and GES-1 were also stained. Correlations were analyzed between expression levels of Gal-9 and Tim-3 protein and tumor parameters or clinical outcomes.
Gal-9 and Tim-3 stained positive on tumor cells in 86.2% (263/305), and 60.0% (183/305) patients with gastric cancer, respectively. Gal-9 expression was significantly higher in cancer than in normal mucosa (P<0.001). Reduced Gal-9 expression was associated with lymph-vascular invasion, lymph node metastasis, distant metastasis and worse TNM staging (P = 0.034, P = 0.009, P = 0.002 and P = 0.043, respectively). In contrast, Tim-3 expression was significantly lower in cancer than in control mucosa (P<0.001). Patients with lymph-vascular invasion had higher expression levels of Tim-3 (P<0.001). Moreover, multivariate analysis shows that both high Gal-9 expression and low Tim-3 expression were significantly associated with long overall survival (P = 0.002, P = 0.010, respectively); the combination of Gal-9 and Tim-3 expression was an independent prognostic predictor for patients with gastric cancer (RR: 0.43; 95%CI: 0.20–0.93). H.pylori infection status was not associated with Gal-9 and Tim-3 expression (P = 0.102, P = 0.565).
The results suggest that expression of Gal-9 and Tim-3 in tumor cells may be a potential, independent prognostic factor for patients with gastric cancer. Gal-9 and TIM-3 may play an important part in the gastric carcinogenesis.
PMCID: PMC3858245  PMID: 24339967
9.  Recognition of methylated DNA by TAL effectors 
Cell Research  2012;22(10):1502-1504.
PMCID: PMC3463267  PMID: 22945353
10.  Peptidomic Analyses of Mouse Astrocytic Cell Lines and Rat Primary Cultured Astrocytes 
Journal of proteome research  2012;11(8):3965-3973.
Astrocytes play an active role in the modulation of synaptic transmission by releasing cell-cell signaling molecules in response to various stimuli that evoke a Ca2+ increase. We expand on recent studies of astrocyte intracellular and secreted proteins by examining the astrocyte peptidome in mouse astrocytic cell lines and rat primary cultured astrocytes, as well as those peptides secreted from mouse astrocytic cell lines in response to Ca2+-dependent stimulations. We identified 57 peptides derived from 24 proteins with LC–MS/MS and CE–MS/MS in the astrocytes. Among the secreted peptides, four peptides derived from elongation factor 1, macrophage migration inhibitory factor, peroxiredoxin-5, and galectin-1, were putatively identified by mass-matching to peptides confirmed to be found in astrocytes. Other peptides in the secretion study were mass-matched to those found in prior peptidomics analyses on mouse brain tissue. Complex peptide profiles were observed after stimulation, suggesting that astrocytes are actively involved in peptide secretion. Twenty-six peptides were observed in multiple stimulation experiments but not in controls and thus appear to be released in a Ca2+-dependent manner. These results can be used in future investigations to better understand stimulus-dependent mechanisms of astrocyte peptide secretion.
PMCID: PMC3434970  PMID: 22742998
glia; astrocytes; secreted peptides; peptidomics; LC–MS; CE–MS; Ca2+-dependent stimulation
11.  High Duty Cycle to Low Duty Cycle: Echolocation Behaviour of the Hipposiderid Bat Coelops frithii 
PLoS ONE  2013;8(5):e62938.
Laryngeally echolocating bats avoid self-deafening (forward masking) by separating pulse and echo either in time using low duty cycle (LDC) echolocation, or in frequency using high duty cycle (HDC) echolocation. HDC echolocators are specialized to detect fluttering targets in cluttered environments. HDC echolocation is found only in the families Rhinolophidae and Hipposideridae in the Old World and in the New World mormoopid, Pteronotus parnellii. Here we report that the hipposiderid Coelops frithii, ostensibly an HDC bat, consistently uses an LDC echolocation strategy whether roosting, flying, or approaching a fluttering target rotating at 50 to 80 Hz. We recorded the echolocation calls of free-flying C. frithii in the field in various situations, including presenting bats with a mechanical fluttering target. The echolocation calls of C. frithii consisted of an initial narrowband component (0.5±0.3 ms, 90.6±2.0 kHz) followed immediately by a frequency modulated (FM) sweep (194 to 113 kHz). This species emitted echolocation calls at duty cycles averaging 7.7±2.8% (n = 87 sequences). Coelops frithii approached fluttering targets more frequently than did LDC bats (C.frithii, approach frequency  = 40.4%, n = 80; Myotis spp., approach frequency  = 0%, n = 13), and at the same frequency as sympatrically feeding HDC species (Hipposideros armiger, approach rate  = 53.3%, n = 15; Rhinolophus monoceros, approach rate  = 56.7%, n = 97). We propose that the LDC echolocation strategy used by C. frithii is derived from HDC ancestors, that this species adjusts the harmonic contents of its echolocation calls, and that it may use both the narrowband component and the FM sweep of echolocations calls to detect fluttering targets.
PMCID: PMC3663840  PMID: 23717396
12.  Increasing trends in central obesity among Chinese adults with normal body mass index, 1993–2009 
BMC Public Health  2013;13:327.
Central obesity is thought to be more pathogenic than overall obesity and studies have shown that the association between waist circumference (WC) and mortality was strongest in those with a normal body mass index (BMI). The objective of our study was to determine secular trends in the prevalence of central obesity (WC ≥ 90 cm for men and ≥ 80 cm for women) among Chinese adults with normal BMI from 1993 to 2009 and to examine the impact of performance of combined BMI and WC on the prevalence of obesity in Chinese adults.
We used data from the China Health and Nutrition Survey (CHNS) conducted from 1993 to 2009. From which we included a total of 52023 participants aged ≥ 18 years.
The age-standardized prevalence of central obesity among Chinese adults with BMI < 25 kg/m2 increased from 11.9% in 1993 to 21.1% in 2009 (P for linear trend <0.001). The upward trends were noted in both genders, all ages, rural/urban settings, and education groups (all P for linear trend <0.001), with greater increments in men, participants aged 18–64 years, and rural residents (P for interaction terms survey × sex, survey × age, and survey × rural/urban settings were 0.042, 0.003, and < 0.001, respectively). Trends in the prevalence of central obesity were similar when a more stringent BMI < 23 kg/m2 cut point (Asian cut point) was applied. Central obesity is associated with a higher risk of incident hypertension within normal BMI category. More than 65% individuals with obesity would be missed if solely BMI was measured.
We observed an upward trend in the prevalence of central obesity among participants with normal BMI irrespective of sex, age, rural/urban settings, and education level. Central obesity is associated with a higher risk of incident hypertension within normal BMI category. Approximately two thirds of the individuals with obesity would be missed if WC was not measured. It is, therefore, urgent to emphasize the importance of WC as a measure to monitor the prevalence of obesity.
PMCID: PMC3626835  PMID: 23575244
Body mass index; Waist circumference; Central obesity; General obesity; CHNS
13.  A case report of hydatid cysts containing aspergillus 
Journal of Thoracic Disease  2013;5(2):E25-E27.
PMCID: PMC3621937  PMID: 23585951
14.  Prevalence of Hypertension in Chinese Cities: A Meta-Analysis of Published Studies 
PLoS ONE  2013;8(3):e58302.
Hypertension has been recognized as a health concern for developing countries. However, there are no current nationwide surveys on the prevalence of hypertension in China (the latest nationwide survey was ten years ago). The goal of this study was to estimate the pooled prevalence of hypertension in Chinese cities.
We systematically reviewed published epidemiologic studies on the prevalence of hypertension in Chinese cities through meta-analysis. We searched for studies published between January 2002 and June 2012 using PubMed and two Chinese electronic publication libraries. The keywords ‘hypertension’ and ‘prevalence’ were used. Before pooling prevalence of hypertension, all raw prevalence data was age adjusted to the 2010 China standard population. Prevalence estimates were stratified by sex and geographic area.
27 studies were identified with of a total of 195,027 study participants. The overall pooled prevalence of hypertension was 21.5% (19.4%, 23.6%). Subgroup analyses showed the following results north 25.8% (21.6%, 30.0%), south 20.4% (18.6%, 22.2%); male 22.2% (19.3%, 25.1%), female 19.9% (17.6%, 22.1%); large cities 18.9% (15.7%, 22.1%), medium-sized cities 24.6% (19.9%, 29.4%), small cities 20.6% (17.5%, 23.7%); study years in 2002–2006, 21.9% (18.9%, 24.8%), and study year in 2007–2011, 20.6% (17.3%, 23.9%).
Comparing data from several previous national hypertension surveys, the prevalence of hypertension is higher in cities than the Chinese national average. Subgroup studies also found a higher prevalence of hypertension in northern cities and among males. Also, the prevalence of hypertension in medium-sized and small cities is likely to increase faster than in large cities.
PMCID: PMC3590128  PMID: 23484011
15.  Neuropeptidomics: Mass spectrometry-based qualitative and quantitative analysis 
Neuropeptidomics refers to a global characterization approach for the investigation of neuropeptides, often under specific physiological conditions. Neuropeptides comprise a complex set of signaling molecules that are involved in regulatory functions and behavioral control in the nervous system. Neuropeptidomics is inherently challenging because neuropeptides are spatially, temporally and chemically heterogeneous, making them difficult to predict in silico from genomic information. Mature neuropeptides are produced from intricate enzymatic processing of precursor proteins/prohormones via a range of post-translational modifications, resulting in multiple final peptide products from each prohormone gene. Although there are several methods for targeted peptide studies, mass spectrometry (MS), with its qualitative and quantitative capabilities, is ideally suited to the task. MS provides fast, sensitive, accurate, and high-throughput peptidomic analyses of neuropeptides without requiring prior knowledge of the peptide sequences. Aided by liquid chromatography (LC) separations and bioinformatics, MS is quickly becoming a leading technique in neuropeptidomics. This chapter describes several LC-MS analytical methods to identify, characterize and quantify neuropeptides, while emphasizing the sample preparation steps so integral to experimental success.
PMCID: PMC3587730  PMID: 21922411
sample preparation; liquid chromatography (LC); mass spectrometry (MS); peptide identification; stable isotope labeling; quantitation
16.  Increased expression of tyrosine phosphatase SHP-2 in Helicobacter pylori-infected gastric cancer 
AIM: To explore the alteration of tyrosine phosphatase SHP-2 protein expression in gastric cancer and to assess its prognostic values.
METHODS: Three hundred and five consecutive cases of gastric cancer were enrolled into this study. SHP-2 expression was carried out in 305 gastric cancer specimens, of which 83 were paired adjacent normal gastric mucus samples, using a tissue microarray immunohistochemical method. Correlations were analyzed between expression levels of SHP-2 protein and tumor parameters or clinical outcomes. Serum anti-Helicobacter pylori (H. pylori) immunoglobulin G was detected with enzyme-linked immunosorbent assay. Cox proportional hazards model was used to evaluate prognostic values by compassion of the expression levels of SHP-2 and disease-specific survivals in patients.
RESULTS: SHP-2 staining was found diffuse mainly in the cytoplasm and the weak staining was also observed in the nucleus in gastric mucosa cells. Thirty-two point five percent of normal epithelial specimen and 62.6% of gastric cancer specimen were identified to stain with SHP-2 antibody positively (P < 0.001). Though SHP-2 staining intensities were stronger in the H. pylori (+) group than in the H. pylori (-) group, no statistically significant difference was found in the expression levels of SHP-2 between H. pylori (+) and H. pylori (-) gastric cancer (P = 0.40). The SHP-2 expression in gastric cancer was not significantly associated with cancer stages, lymph node metastases, and distant metastasis of the tumors (P = 0.34, P = 0.17, P = 0.52). Multivariate analysis demonstrated no correlation between SHP-2 expression and disease-free survival (P = 0.86).
CONCLUSION: Increased expression of SHP-2 protein in gastric cancer specimen suggesting the aberrant up-regulation of SHP-2 protein might play an important role in the gastric carcinogenesis.
PMCID: PMC3558584  PMID: 23382639
Gastric cancer; SH2-containing protein tyrosine phosphatase 2; Expression; Helicobacter pylori
17.  The Association between Cultural Orientation and Drinking Behaviors among University Students in Wuhan, China 
PLoS ONE  2013;8(1):e54796.
This study examines the association between cultural orientation and drinking behaviors among university students. Cultural orientation is the measure of how the cultural values of individuals living in their own society are influenced by cultural values introduced from the outside.
In 2011, a cross-sectional survey collected data from 1279 university students from six universities in central China. Participants used a likert scale to rank a series of statements reflecting cultural values from the previously validated Chinese Cultural Orientation Scale and answered questions about their drinking behaviors and socio-demographic characteristics.
Statistically significant differences in cultural orientation were observed for gender, hometown and type of university attendance. Traditional-oriented students were more likely to be occasional drinkers or nondrinkers, while marginal-oriented students, bicultural-oriented students and western-oriented students were more likely to be regular drinkers. Bicultural orientation (OR = 1.80, P<0.05) and marginal orientation (OR = 1.64, P<0.05) increased the likelihood of the student being regular drinking, compared to students with traditional orientations. Males (OR = 4.40, P<0.05) had a higher likelihood of regular drinking than females, graduate students (OR = 2.59, P<0.05) had a higher likelihood of regular drinking than undergraduates, students from urban areas (OR = 1.79, P<0.05) had a higher likelihood of regular drinking than those from towns/rural areas, and students attending key universities (OR = 0.48, P<0.05) had a lower likelihood of regular drinking than those attending general universities.
Cultural orientation influences drinking behaviors. Traditional cultural orientation was associated with less drinking while western cultural orientation, marginal cultural orientation and bicultural orientation were associated with more drinking. The role of gender, hometown and university attendance is partially moderated through the influence of cultural orientation. The relationship between a traditional cultural orientation and alcohol drinking suggests that traditional Chinese cultural values should be examined for their role in possibly reducing alcohol-related risks through education and policy initiatives.
PMCID: PMC3554628  PMID: 23359611
18.  A Survey of Congenital Heart Disease and Other Organic Malformations Associated with Different Types of Orofacial Clefts in Eastern China 
PLoS ONE  2013;8(1):e54726.
A high incidence of orofacial clefts is reported in China, but no data has shown the relation between cleft types and the incidence of other defects so far. The aim of this study is to assess the incidence of congenital heart diseases and other organic defects associated with different types of orofacial clefts.
Methodology and Principal Findings
All children with orofacial clefts, which were sought out from the Health Information System of Shanghai Ninth People's Hospital between 1st Jan 2009 and 30th Dec 2011, were enrolled in this study. All subjects underwent a thorough examination and grouped by the cleft phenotype. The numbers and types of other organic defects were recorded and analyzed statistically using SPSS 17.0. Of 2180 cases reported as having orofacial clefts, 657 (30.1%) had other congenital abnormalities, which were significantly more common in cleft palate (47.9% (329/687)) than that in cleft lip (10.6% (80/755)) or cleft lip and palate (33.6% (248/738)) (P<0.01). In subgroups, unilateral cleft lip and palate had a statistically higher incidence of associated abnormalities than bilateral cleft lip and palate (P<0.01). The most common malformation was congenital heart disease, which counted 45.1% (296/657) of all malformations. Disorders of the central nervous system (14.3%(94/657)) and Skeletal anomalies (13.1%(86/657)) were also frequently associated. Additionally, the most common defect in heart was atrial septal defect, which was 39.7% (118/296) of all congenital heart diseases.
Conclusions and Significance
As the high incidence of heart defects and other organic abnormalities in the children with cleft palate in Eastern China, special attention should be paid to them and echocardiography should be a proposed examination in the evaluation of children with cleft palate before any surgical correction being executed.
PMCID: PMC3549991  PMID: 23349958
19.  Development and Validation of a Tuberculosis Medication Adherence Scale 
PLoS ONE  2012;7(12):e50328.
Medication adherence is critical in Tuberculosis (TB) treatment success, but existing tools are inadequate in identifying non-adherents, reasons for non-adherence or interventions to improve adherence. This study intended to fill the gap by developing and validating a TB medication adherence scale (TBMAS).
An initial 41-item TBMAS was designed through review of literature, consultation from an 8-member clinical expert panel and a 15-patient focus group, and pilot-testing in 25 TB patients. The questionnaire was validated in 438 patients who visited 23 community health centers for TB treatment in Wuhan from September 1, 2010, to August 31, 2011, using pharmacy refill records in a 15-week period as external criteria for medication adherence. After removing redundant and cross-loading items, the internal consistency, reliability and validity of TBMAS in identifying non-adherents were examined.
The final TBMAS included 30 items scored on a 5-point Likert scale, and these items were loaded in nine distinct factors that explained 65% of cumulative variance among respondents. Cronbach's alpha, test-retest reliability and split-half reliability were 0.87, 0.83, and 0.85, respectively. Convergent validity was supported by statistically significant associations between TBMAS scores and adherence measured by pharmacy refill records. Receiver Operating Characteristics curve analysis suggested a cut-off point at 113, with which TBMAS showed a positive predictive value of 65.5% and sensitivity of 82.9% in identifying non-adherents.
TBMAS demonstrated satisfactory internal consistency, reliability and validity in identifying TB patients with poor adherence and potential causes for non-adherence.
PMCID: PMC3520953  PMID: 23251363
Current biology : CB  2011;21(18):1515-1524.
In Drosophila the bHLH protein DIMM coordinates the molecular and cellular properties of all major neuroendocrine cells, irrespective of the secretory peptides they produce. When expressed by non-neuroendocrine neurons, DIMM confers the major properties of the Regulated Secretory Pathway and converts such cells away from fast neurotransmission and towards a neuroendocrine state.
We first identified 134 transcripts upregulated by DIMM in embryos, then evaluated them systematically using diverse assays (including embryo in situ hybridization, in vivo ChIP, and cell-based transactivation assays). We conclude that of 11 strong candidates, six are strongly and directly controlled by DIMM in vivo. The six targets include several large dense-core vesicle (LDCV) proteins, but also proteins in non-LDCV compartments such as the RNA-associated protein MAELSTROM. In addition, a functional in vivo assay, combining transgenic RNAi with MS-based peptidomics, revealed that three DIMM targets are especially critical for its action: These include two well-established LDCV proteins, the amidation enzyme PHM and the ascorbate-regenerating electron transporter Cytochrome-b561-1. The third key DIMM target, CAT-4 (CG13248), has not previously been associated with peptide neurosecretion – it encodes a putative cationic amino acid transporter, closely related to the SLIMFAST Arginine transporter. Finally, we compared transcripts upregulated by DIMM with those normally enriched in DIMM neurons of the adult brain and found an intersection of 18 DIMM-regulated genes, which included all six direct DIMM targets.
The results provide a rigorous molecular framework with which to describe the fundamental regulatory organization of diverse neuroendocrine cells.
PMCID: PMC3184372  PMID: 21885285
Dimmed; peptidergic neuron; bHLH; Drosophila; CAT-4; Phm; Cytochrome b561
21.  Genome-Wide Association Study Identified CNP12587 Region Underlying Height Variation in Chinese Females 
PLoS ONE  2012;7(9):e44292.
Human height is a highly heritable trait considered as an important factor for health. There has been limited success in identifying the genetic factors underlying height variation. We aim to identify sequence variants associated with adult height by a genome-wide association study of copy number variants (CNVs) in Chinese.
Genome-wide CNV association analyses were conducted in 1,625 unrelated Chinese adults and sex specific subgroup for height variation, respectively. Height was measured with a stadiometer. Affymetrix SNP6.0 genotyping platform was used to identify copy number polymorphisms (CNPs). We constructed a genomic map containing 1,009 CNPs in Chinese individuals and performed a genome-wide association study of CNPs with height.
We detected 10 significant association signals for height (p<0.05) in the whole population, 9 and 11 association signals for Chinese female and male population, respectively. A copy number polymorphism (CNP12587, chr18:54081842-54086942, p = 2.41×10−4) was found to be significantly associated with height variation in Chinese females even after strict Bonferroni correction (p = 0.048). Confirmatory real time PCR experiments lent further support for CNV validation. Compared to female subjects with two copies of the CNP, carriers of three copies had an average of 8.1% decrease in height. An important candidate gene, ubiquitin-protein ligase NEDD4-like (NEDD4L), was detected at this region, which plays important roles in bone metabolism by binding to bone formation regulators.
Our findings suggest the important genetic variants underlying height variation in Chinese.
PMCID: PMC3434125  PMID: 22957059
22.  Early growth response-2 expression in uterine leiomyoma cells: regulation and function 
Fertility and sterility  2011;96(2):439-444.
To investigate the regulation of Egr-2 by TGF-β3 and its functions in cultured human uterine leiomyoma smooth muscle (LSM) cells.
Laboratory research.
Academic medical center.
Primary leiomyoma cells from patients with symptomatic leiomyomata.
Tissue culture followed by RNA and protein analysis.
Main outcome Measure(s)
Cell proliferation, alteration in ECM component expression.
In vivo mRNA levels of Egr-2 were significantly higher in leiomyoma tissues compared with matched myometrial tissues and correlated significantly with TGF-β3 mRNA levels in leiomyoma tissues. In primary LSM cells, TGF-β3 significantly induced Egr-2 gene expression in a dose- and time- dependent manner. Small interfering RNA knockdown of Egr-2 markedly increased level of the proliferation marker proliferating cell nuclear antigen and the expression of proto-oncogene c-myc. On the other hand, ablation of Egr-2 stimulated collagen 1A1 and 3A1 transcription and inhibited dematopontin gene expression. However, the mRNA levels of α-smooth muscle actin and fibronectin were not affected by Egr-2 knockdown.
We demonstrated that TGF-β3 regulated Egr-2 gene expression and presented evidence that Egr-2 decreases collagen production and stimulates dermatopontin gene expression.
PMCID: PMC3143242  PMID: 21703609
TGF-β3; Egr-2; collagen; dermatopontin; c-myc; PCNA; leiomyomata
23.  Association of polymorphism of PTPN 11 encoding SHP-2 with gastric atrophy but not gastric cancer in Helicobacter pylori seropositive Chinese population 
BMC Gastroenterology  2012;12:89.
The interaction between Src homology 2 domain-containing protein tyrosine phosphatase (SHP-2) of gastric epithelial cells and cagA from H. pylori plays a crucial role in developments of gastric atrophy and gastric cancer. This study aimed to investigate the association of haplotype tagging SNPs (htSNPs) in the PTPN11 gene encoding SHP-2 with gastric atrophy and gastric cancer in Chinese population.
The subjects comprised 414 patients with gastric cancer, 109 individuals with gastric atrophy and 923 healthy controls. Blood was collected from October 2008 to October 2010. Five htSNPs rs2301756, rs12423190, rs12229892, rs7958372 and rs4767860 from the PTPN11 gene were selected and genotyped by Taqman assay. Serum Ig G antibodies to H. pylori were detected by ELISA. Gastric atrophy was screened by the levels of serum pepsinogenIandII, and confirmed by endoscopy and histopatholgical examinations. Odds ratio (ORs) and 95% confidence intervals (CIs) were calculated by a multivariate logistic regression.
Among H. pylori seropositive subjects, age and gender-adjusted OR of gastric atrophy was 2.47 (95%CI 1.13-4.55, P = 0.02) for CC genotype compared with CT/TT genotypes, suggesting a recessive model of genetic risk for rs12423190. The prevalence of H. pylori seropositivity were significantly higher in groups of gastric cancer and gastric atrophy compared to the control group (70.3% vs. 75.2% vs. 49.7%, P <0.001). However, the distributions of genotypes and haplotypes in patients with gastric cancer were not significantly different from healthy controls.
Our study provides the first evidence that rs12423190 polymorphism of the PTPN11 gene is significantly associated with an increased risk of gastric atrophy in H. pylori infected Chinese Han population, suggesting that rs12423190 polymorphism could be used as a useful marker of genetic susceptibility to gastric atrophy among H. pylori infected subjects. The biological roles of this polymorphism require a further investigation.
PMCID: PMC3509400  PMID: 22788847
24.  Significance of decoy receptor 3 (Dcr3) and external-signal regulated kinase 1/2 (Erk1/2) in gastric cancer 
BMC Immunology  2012;13:28.
Decoy receptor 3 (DcR3), a member of the tumor necrosis factor receptor (TNFR) superfamily, is associated with anti-tumor immunity suppression. It is highly expressed in many tumors, and its expression can be regulated by the MAPK/MEK/ERK signaling pathway. The MAPK/MEK/ERK pathway has been reported to be a regulator in tumor occurrence, development and clonal expansion. External-signal regulated kinase (ERK) is a vital member of this pathway.
The expression of DcR3 and ERK1/2 in tumor tissues of gastric cancer patients was significantly higher than the non-cancerous group (P < 0.05). There was no statistical difference among tumor tissues from patients with different ages or gender, and even of different differentiation (P > 0.05). However, in patients with stage I gastric cancer, the DcR3 and ERK1/2 levels were significantly lower than patients with more advanced stages.
DcR3 and ERK1/2 play a vital role in the development of gastric cancer, and they may be new markers for indicating the efficiency of gastric cancer treatment in the future.
PMCID: PMC3459731  PMID: 22672288
25.  Evaluation of malignancy using Ki-67, p53, EGFR and COX-2 expressions in gastrointestinal stromal tumors 
AIM: To investigate the role of expressions of Ki-67, p53, epidermal growth factor receptor (EGFR) and cyclooxygenase-2 (COX-2) in gastrointestinal stromal tumor (GIST) grading and prognosis.
METHODS: Tumor tissue was collected retrospectively from 96 patients with GIST. Antibodies against Ki-67, p53, EGFR and COX-2 were used for immunohistochemical staining. Tumor grading was designated according to a consensus system and the staining was quantified in 3 categories for each antibody in the statistical analysis.
RESULTS: The Ki-67 expression in GISTs was significantly associated with the size of the tumors, mitotic rate and the risk of malignancy (χ2 = 15.51, P = 0.02; χ2 = 22.27, P < 0.001; χ2 = 20.05; P < 0.001). The p53 expression was also significantly correlated with mitotic rate and the risk of malignancy (χ2 = 9.92, P = 0.04; χ2 = 9.97; P = 0.04). Over-expression of Ki-67 was strongly correlated with poor survival (χ2 = 10.44, P = 0.006), but no correlation was found between the expression of p53, EGFR or COX-2 and survival. Multivariate analysis further demonstrated that Ki-67 expression (relative risk = 15.78, 95% CI: 4.25-59.37) could be used as an independent prognostic value for GIST patients. Adjuvant imatinib therapy could improve clinical outcomes in the patients with high risk and intermediate risk of recurrence after complete tumor resections (median survival time: 52 mo vs 37 mo, χ2 = 7.618, P = 0.006).
CONCLUSION: Our results indicated that the expression of Ki-67 could be used as an independent prognostic factor for GIST patients.
PMCID: PMC3360457  PMID: 22654456
Gastrointestinal stromal tumor; Prognosis; Ki-67 alteration; p53; Epidermal growth factor receptor

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