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1.  Characterization of one sheep border disease virus in China 
Virology Journal  2015;12:15.
Border disease virus (BDV) causes border disease (BD) affecting mainly sheep and goats worldwide. BDV in goat herds suffering diarrhea was recently reported in China, however, infection in sheep was undetermined. Here, BDV infections of sheep herds in Jiangsu, China were screened; a BDV strain was isolated and identified from the sheep flocks in China. The genomic characteristics and pathogenesis of this new isolate were studied.
In 2012, samples from 160 animals in 5 regions of Jiangsu province of China were screened for the presence of BDV genomic RNA and antibody by RT-PCR and ELISA, respectively. 44.4% of the sera were detected positively, and one slowly grown sheep was analyzed to be pestivirus RNA positive and antibody-negative. The sheep kept virus positive and antibody negative in the next 6 months of whole fattening period, and was defined as persistent infection (PI). The virus was isolated in MDBK cells without cytopathic effect (CPE) and named as JSLS12-01. Near-full-length genome sequenced was 12,227 nucleotides (nt). Phylogenetic analysis based on 5'-UTR and Npro fragments showed that the strain belonged to genotype 3, and shared varied homology with the other 3 BDV strains previously isolated from Chinese goats. The genome sequence of JSLS12-01 also had the highest homology with genotype BDV-3 (the strain Gifhorn). Experimental infections of sheep had mild clinical signs as depression and short-period mild fever (5 days). Viremia was detected in 1–7 days post-infection (dpi), and seroconversion began after 14 dpi.
This study reported the genomic and pathogenesis characterizations of one sheep BDV strain, which confirmed the occurrence of BDV infection in Chinese sheep. This sheep derived BDV strain was classified as BDV-3, together with the goat derived strains in China. These results might be helpful for further understanding of BDV infection in China and useful for prevention and control of BDV infections in the future.
PMCID: PMC4329205
BDV; Complete genome sequence; Phylogenetic analysis; Experimental infection
2.  Crude extract of maggots: Antibacterial effects against Escherichia coli, underlying mechanisms, separation and purification 
AIM: To investigate the antibacterial effects of a crude extract of maggots against Escherichia coli (E. coli) and the underlying mechanisms, and to separate and purify the crude extract of maggots to assess the antibacterial effects of the active ingredients in the crude extract.
METHODS: Different concentrations of the crude extract of maggots were incubated with E. coli (O157:H7) and cultured. The optical density (OD) was measured at different time points to plot the OD-T curve. The effects of different concentrations of the crude extract on bacterial membrane permeability were determined by fluorescence probe technique. The effects of different concentrations of the crude extract on plasmid DNA replication were determined by agarose gel electrophoresis. DEAE-Sepharose ion exchange chromatography and Sephacryls-200HR gel filtration chromatography were used to separate and purify the crude extract of maggots. The molecular weight of proteins in the purified crude extract was determined by SDS-PAGD electrophoresis, and its antibacterial effects were determined by turbidimetric method.
RESULTS: The antibacterial effects of the crude extract of maggots at concentrations > 0.5 mg/mL were significant. The antibacterial effects of the crude extract at concentrations of 1.0, 1.5 and 2.0 mg/mL did not differ significantly. Fluorescence probe analysis showed that the rate of membrane permeability change was 1223.1% in bacteria incubated with 2 mg/mL of the crude extract, and 1300.0% in those incubated with 80 mg/mL of the crude extract. Plasmid DNA was undetectable in E. coli incubated with 2 and 80 mg/mL of the crude extract. A low molecular weight protein band (about 15 kDa) was detected in the crude extract of maggots and eluent, but not in eluant, from DEAE-Sepharose ion exchange chromatography. The antibacterial effects of the crude extract of maggots and eluent were superior to those of eluant, with the antibacterial effects of eluents being better than those of the crude extract of maggots. Of 24 tubes of filtrates, the antibacterial effects of filtrates in tubes 4, 5 and 11 were significantly higher than those of the control. The molecular weight of the protein in filtrates in tubes 4, 5 and 11 was about 15 kDa.
CONCLUSION: The crude extract of maggots exhibits obvious, dose-dependent antibacterial effects. The crude extract exerts antibacterial effects by changing the bacterial membrane permeability and inhibiting plasmid DNA replication. The protein that has antibacterial effects in the crude extract of maggots has a molecular weight of about 15 kDa.
PMCID: PMC4316093  PMID: 25663770
Maggots; Antibacterial peptide; Antibacterial mechanism; Escherichia coli; Colorectal
3.  Mycobacterium tuberculosis 10-kDa co-chaperonin regulates the expression levels of receptor activator of nuclear factor-κB ligand and osteoprotegerin in human osteoblasts 
The aim of the present study was to investigate the effect of recombinant Mycobacterium tuberculosis (r-Mt) 10-kDa co-chaperonin (cpn10) on the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in third-generation cultured osteoblasts. The osteoblast-like cultures were isolated from bone fragments taken from patients undergoing surgery. Prior to stimulation with r-Mt cpn10, cells were incubated in serum-free medium for 24 h. r-Mt cpn10 was added into fresh serum-free medium, reaching final concentrations of 0.01–10 μg/ml. The levels of OPG were determined using enzyme-linked immunosorbent assay. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis was performed to determine the levels of RANKL and OPG mRNA. For measurement of the protein levels of OPG and RANKL, a western blotting assay was performed. r-Mt cpn10 downregulated the protein levels of OPG in the third generation cultured osteoblasts at a dose of 10 μg/ml. RT-qPCR revealed that the OPG mRNA level was decreased by 73% after 4 h and by 85.5% after 8 h following incubation with r-Mt cpn10 (10 μg/ml). Western blot analysis demonstrated similar results for the OPG protein level. In the third-generation cultured osteoblasts, the levels of RANKL mRNA and protein were increased by 2.6- and 1-fold, respectively, following incubation with r-Mt cpn10 (10 μg/ml). Furthermore, the RANKL/OPG ratio was markedly increased by r-Mt cpn10 (10 μg/ml) treatment. In conclusion, the results of the current study demonstrated that r-Mt cpn10 decreased the levels of OPG and increased the levels of RANKL in a dose- and time-dependent manner. Notably, the present study indicated that r-Mt cpn10 exerts its effect on osteoblastic cells by increasing the RANKL/OPG ratio.
PMCID: PMC4316961  PMID: 25667654
recombinant Mycobacterium tuberculosis 10-kDa co-chaperonin; osteoprotegerin; bone resorption; nuclear factor-κB ligand; osteoblast; bone tuberculosis
4.  Impacts of infection with different toxigenic Clostridium difficile strains on faecal microbiota in children 
Scientific Reports  2014;4:7485.
Increasing evidence suggests that altered intestinal microbial composition and function result in an increased risk of Clostridium difficile-associated diarrhoea (CDAD); however, the specific changes of intestinal microbiota in children suffering from CDAD and their associations with C. difficile strain toxigenicity are poorly understood. High-throughput pyrosequencing showed that reduced faecal bacterial diversity and dramatic shifts of microbial composition were found in children with CDAD. The Firmicutes/Bacteroidetes ratio was increased significantly in patients with CDAD, which indicated that dysbiosis of faecal microbiota was closely associated with CDAD. C. difficile infection resulted in an increase in lactate-producing phylotypes, with a corresponding decrease in butyrate-producing bacteria. The decrease in butyrate and lactate buildup impaired intestinal colonisation resistance, which increased the susceptibility to C. difficile colonisation. Strains of C. difficile which were positive for both toxin A and toxin B reduced faecal bacterial diversity to a greater degree than strains that were only toxin B-positive, and were associated with unusually abundant Enterococcus, which implies that the C. difficile toxins have different impacts on the faecal microbiota of children. Greater understanding of the relationships between disruption of the normal faecal microbiota and colonisation with C. difficile that produces different toxins might lead to improved treatment.
PMCID: PMC4265774  PMID: 25501371
5.  Vitamin A Supplementation in Early Life Enhances the Intestinal Immune Response of Rats with Gestational Vitamin A Deficiency by Increasing the Number of Immune Cells 
PLoS ONE  2014;9(12):e114934.
Vitamin A is a critical micronutrient for regulating immunity in many organisms. Our previous study demonstrated that gestational or early-life vitamin A deficiency decreases the number of immune cells in offspring. The present study aims to test whether vitamin A supplementation can restore lymphocyte pools in vitamin A-deficient rats and thereby improve the function of their intestinal mucosa; furthermore, the study aimed to identify the best time frame for vitamin A supplementation. Vitamin A-deficient pregnant rats or their offspring were administered a low-dose of vitamin A daily for 7 days starting on gestational day 14 or postnatal day 1, day 14 or day 28. Serum retinol concentrations increased significantly in all four groups that received vitamin A supplementation, as determined by high-performance liquid chromatography. The intestinal levels of secretory immunoglobulin A and polymeric immunoglobulin receptor increased significantly with lipopolysaccharide challenge in the rats that received vitamin A supplementation starting on postnatal day 1. The rats in this group had higher numbers of CD8+ intestinal intraepithelial lymphocytes, CD11C+ dendritic cells in the Peyer's patches and CD4+CD25+ T cells in the spleen compared with the vitamin A-deficient rats; flow cytometric analysis also demonstrated that vitamin A supplementation decreased the number of B cells in the mesenteric lymph nodes. Additionally, vitamin A supplementation during late gestation increased the numbers of CD8+ intestinal intraepithelial lymphocytes and decreased the numbers of B lymphocytes in the mesenteric lymph nodes. However, no significant differences in lymphocyte levels were found between the rats in the other two vitamin A supplement groups and the vitamin A-deficient group. In conclusion, the best recovery of a subset of lymphocytes in the offspring of gestational vitamin A-deficient rats and the greatest improvement in the intestinal mucosal immune response are achieved when vitamin A supplementation occurs during the early postnatal period.
PMCID: PMC4263713  PMID: 25503794
6.  Clinical significance of chromosome 1p/19q loss of heterozygosity and Sox17 expression in oligodendrogliomas 
Objective: To study chromosome 1p/19q loss of heterozygosity (LOH) and Sox17 protein expression in oligodendrogliomas and correlate this loss with clinicopathological features. Methods: This study included 100 cases of oligodendrogliomas at the First Affiliated Hospital of Xinjiang Medical University from 2003 to 2014. The cases included paraffin-embedded tissues from 50 low-grade oligodendrogliomas and 50 anaplastic oligodendrogliomas. Chromosome 1p/19q LOH was detected by fluorescence in situ hybridization (FISH) and Sox17 protein expression was analyzed by immunohistochemistry. Clinicopathological characteristics of the oligodendrogliomas were compared and prognosis analyzed using Cox regression and Kaplan-Meier analyses. Results: The LOH positivity rate of 1p/19q was 52% in 50 cases of low-grade oligodendrogliomas and 68% in 50 cases of anaplastic oligodendrogliomas (P = 0.102). The rates of Sox17 expression were significantly different in oligodendrogliomas (82%) and anaplastic oligodendrogliomas (62%, P = 0.026). Single factor analysis determined that 1p/19q LOH (P = 0.000), Sox17 protein expression (P = 0.000), location (P = 0.001), chemotherapy (P = 0.000), and radiation therapy (P = 0.001) were associated with oligodendroglioma patient prognosis. Cox multiple factors regression analysis determined that 1p/19q LOH and Sox17 expression were independent prognostic factors of oligodendrogliomas. Conclusion: In this study, oligodendroglioma patients with 1p/19q LOH and Sox17 protein expression had a better prognosis. Thus, analysis of 1p/19q LOH and Sox17 protein expression could significantly enhance diagnostic accuracy, guide treatment, and improve the prognosis.
PMCID: PMC4313992
Oligodendroglioma; 1p/19q LOH; Sox17; prognosis; clinical features
7.  Chronic intake of high fish oil diet induces myeloid-derived suppressor cells to promote tumor growth 
Omega-3 polyunsaturated fatty acids enriched fish oil exerts beneficial anti-inflammatory effects in animal models with acute and chronic inflammatory diseases. Myeloid-derived suppressor cells (MDSCs), comprised of myeloid progenitors and precursors of myeloid cells, play vital roles in cancer. How fish oil affects the generation of MDSCs and the tumor development remains largely unexplored. Here, we show that dietary intake of high fish oil diet suppresses CD8+ T cells activation and proliferation in vivo via elevated levels of MDSCs. Mechanistically, high fish oil diet induces the expression of immunosuppressive cytokine IL-10 and promotes myelopoiesis in the spleen as well as other peripheral tissues. The immature myeloid cells in the spleen exhibit morphological and functional characteristics of MDSCs with the capability to downregulate CD8+ T cells activation. Depletion of MDSCs using anti-Gr-1 antibody decreases the growth of subcutaneously transferred B16 melanoma in mice on high fish oil diet. Interestingly, diet-induced production of MDSCs is not solely dependent of the spleen, as splenectomy has no effect on the tumor progress. Our data show that the liver functions as an alternative extramedullary hematopoiesis organ to support MDSCs differentiation and maintain tumor growth. Taken together, our study provides a novel insight into the physiological effects of fish oil and points to MDSCs as a possible mediator linking dietary fish oil intake and immunosuppression in cancer immunosurveillance.
PMCID: PMC4246012  PMID: 24691944
Polyunsaturated fatty acids; Fish oil; MDSCs; CD8+ T cell; Liver; Cancer
8.  Molecular Characterization and Expression Analysis of Triticum aestivum Squamosa-Promoter Binding Protein-Box Genes Involved in Ear Development 
Wheat (Triticum aestivum L.) is one of the most important crops in the world. Squamosa-promoter binding protein (SBP)-box genes play a critical role in regulating flower and fruit development. In this study, 10 novel SBP-box genes (TaSPL genes) were isolated from wheat ((Triticum aestivum L.) cultivar Yanzhan 4110). Phylogenetic analysis classified the TaSPL genes into five groups (G1–G5). The motif combinations and expression patterns of the TaSPL genes varied among the five groups with each having own distinctive characteristics: TaSPL20/21 in G1 and TaSPL17 in G2 mainly expressed in the shoot apical meristem and the young ear, and their expression levels responded to development of the ear; TaSPL6/15 belonging to G3 were upregulated and TaSPL1/23 in G4 were downregulated during grain development; the gene in G5 (TaSPL3) expressed constitutively. Thus, the consistency of the phylogenetic analysis, motif compositions, and expression patterns of the TaSPL genes revealed specific gene structures and functions. On the other hand, the diverse gene structures and different expression patterns suggested that wheat SBP-box genes have a wide range of functions. The results also suggest a potential role for wheat SBP-box genes in ear development. This study provides a significant beginning of functional analysis of SBP-box genes in wheat.
PMCID: PMC4239008  PMID: 24386921
Expression profile; grain yield; squamosa-promoter binding protein-box genes; Triticum aestivum
9.  TrkC expression predicts favorable clinical outcome in invasive ductal carcinoma of breast independent of NT-3 expression 
Background: TrkC, a member of neurotrophin receptor family, functions not only as an oncogene, but also act as a tumor suppressor via a manner of dependence receptor in human malignant tumors. Little is known on the action of TrkC for the clinical prognosis and the progression of breast cancer according to the availability of its ligand NT-3. We sought to investigate the prognostic relevance of NT-3-TrkC axis in breast cancer and estimate its role during the process of breast cancer progression. Methods: 236 cases of invasive ductal carcinoma (IDC), 60 pure ductal carcinoma in situ (DCIS) and 30 normal breast tissue (NBT) between 2004 and 2005 were included in the study. Spearman’s rank correlation test was used to analyze the association of NT-3-TrkC expression and breast cancer progression. The Kaplan-Meier method and Cox proportional hazards model were performed to identify the relevant prognostic factors. Results: 50.4% IDC tumors displayed absent or low TrkC expression, while 49.6% was high TrkC expression. TrkC expression was negatively associated with lymph node metastasis (P = 0.029) and tumor proliferation (P = 0.015). Patients with lower TrkC expressing tumors had a higher risk of recurrence (odds ratio, 0.401; 95% confidence interval, 0.207-0.778; P = 0.007). The layered analysis indicated that patients with high TrkC expression tumors had a favor disease-free survival whether NT-3 and TrkC were co-expressed or solitarily expressed in the tumor (P = 0.000). NT-3 was demonstrated to be not a predictor of IDC patients’ prognosis. But NT-3 expression was inversely correlated with the progression of breast cancer (r = -0.341, P = 0.000), since more IDC tumors showed high NT-3 expression than DCIS tumors (51.7% vs. 25.9%), while no NBT showed high NT-3 expression, as well. Conclusion: The study indicates TrkC expression reduces tumor relapse independent of NT-3 availability in the IDC. Elevated NT-3 expression contributes to the progression of breast cancer.
PMCID: PMC4266714  PMID: 25520870
TrkC; NT-3; invasive ductal carcinoma (IDC); breast; prognosis; progression; dependence receptor (DR)
10.  Effect of Continuous Cropping Generations on Each Component Biomass of Poplar Seedlings during Different Growth Periods 
The Scientific World Journal  2014;2014:618421.
In order to investigate the change rules and response characteristics of growth status on each component of poplar seedling followed by continuous cropping generations and growth period, we clear the biomass distribution pattern of poplar seedling, adapt continuous cropping, and provide theoretical foundation and technical reference on cultivation management of poplar seedling, the first generation, second generation, and third generation continuous cropping poplar seedlings were taken as study objects, and the whole poplar seedling was harvested to measure and analyze the change of each component biomass on different growth period poplar leaves, newly emerging branches, trunks and root system, and so forth. The results showed that the whole biomass of poplar seedling decreased significantly with the leaf area and its ratio increased, and the growth was inhibited obviously. The biomass aboveground was more than that underground. The ratios of leaf biomass and newly emerging branches biomass of first continuous cropping poplar seedling were relatively high. With the continuous cropping generations and growth cycle increasing, poplar seedling had a growth strategy to improve the ratio of root-shoot and root-leaf to adapt the limited soil nutrient of continuous cropping.
PMCID: PMC4225849  PMID: 25401150
11.  Prevalence of overweight and obesity and their associations with blood pressure among children and adolescents in Shandong, China 
BMC Public Health  2014;14(1):1080.
Obesity and high blood pressure (BP) are public health problems all over the world. Some studies have reported a positive association between them in children and adolescents. The purpose of this study was to assess the prevalence of overweight and obesity and their associations with BP among school children and adolescents in Shandong, an important province in eastern China.
In 2011, we conducted a cross-sectional population-representative survey in Shandong, China. A total of 4 898 children and adolescents aged 6–17 years were randomly selected from 140 counties/districts using a multistage random cluster sampling. Weight, height and BP were measured by a trained physician or pediatrician, and information about age, gender and place of residence was obtained using questionnaires. Obesity and high BP were defined according to age- and gender-specific Chinese reference data for children.
A total of 4 898 (100%) children and adolescents provided complete information. The prevalence of overweight, obesity and overweight plus obesity were 10.9%, 8.7% and 19.6%, respectively. Boys were more likely to be overweight or obese than girls (P < 0.05 for overweight; P < 0.001 for obesity). The prevalence of overweight plus obesity was highest among children aged 6–11 years (22.3%). BP and the prevalence of high BP increased with increasing body mass index (BMI). With age and sex adjusted, odds ratios (ORs) for high BP were [OR 2.2;95% CI 1.7–2.8) in overweight and [OR 3.6;95% CI 2.6–4.9] in obese children.
The representative survey confirms high prevalence of overweight and obesity among children and adolescents in Shandong. Childhood obesity is a strong risk factor for high BP. Intervention programs should be implemented to combat the growing obesity epidemic.
PMCID: PMC4216350  PMID: 25326029
Prevalence; Overweight; Obesity; Adolescents; Blood pressure
12.  Altered Fecal Microbiota Composition Associated with Food Allergy in Infants 
Increasing evidence suggests that perturbations in the intestinal microbiota composition of infants are implicated in the pathogenesis of food allergy (FA), while the actual structure and composition of the intestinal microbiota in human beings with FA remain unclear. Microbial diversity and composition were analyzed with parallel barcoded 454 pyrosequencing targeting the 16S rRNA gene hypervariable V1-V3 regions in the feces of 34 infants with FA (17 IgE mediated and 17 non-IgE mediated) and 45 healthy controls. Here, we showed that several key FA-associated bacterial phylotypes, but not the overall microbiota diversity, significantly changed in infancy fecal microbiota with FA and were associated with the development of FA. The proportion of abundant Bacteroidetes, Proteobacteria, and Actinobacteria phyla were significantly reduced, while the Firmicutes phylum was highly enriched in the FA group (P < 0.05). Abundant Clostridiaceae 1 organisms were prevalent in infants with FA at the family level (P = 0.016). FA-enriched phylotypes negatively correlated with interleukin-10, for example, the genera Enterococcus and Staphylococcus. Despite profound interindividual variability, levels of 20 predominant genera were significantly different between the FA and healthy control groups (P < 0.05). Infants with IgE-mediated FA had increased levels of Clostridium sensu stricto and Anaerobacter and decreased levels of Bacteroides and Clostridium XVIII (P < 0.05). A positive correlation was observed between Clostridium sensu stricto and serum-specific IgE (R = 0.655, P < 0.001). The specific microbiota signature could distinguish infants with IgE-mediated FA from non-IgE-mediated ones. Detailed microbiota analysis of a well-characterized cohort of infants with FA showed that dysbiosis of fecal microbiota with several FA-associated key phylotypes may play a pathogenic role in FA.
PMCID: PMC3993190  PMID: 24532064
13.  Effect of r-Mt-Cpn10 on human osteoblast cells 
Objective: To observe the effect of recombinant mycobacterium tuberculosis heat shock protein 10 (r-Mt-Cpn10) on human osteoblast proliferation, cell cycle, alkaline phosphatase, calcium nodules and the expression of Receptor Activator of Nuclear Factor KB Ligand (RANKL) and Osteoprotegerin (OPG). Methods: Osteoblasts were cultured in the medium with different concentration of r-Mt-Cpn10. No drug was added to the medium in the control group. The effect of r-Mt-Cpn10 on osteoblast proliferation was detected by MTT. The 3rd generation of osteoblasts was taken and detected the effect on the activity of osteoblasts secreted alkaline phosphatase on 1, 3, 5, 7 and 9 d of cell culture. The effects of different concentrations of r-Mt-Cpn10 on the expression of RANKL and OPG were detected. Results: The r-Mt-Cpn10 blocked osteoblasts in the G2/M phase and G1 to S phase. Compared with the control group, the r-Mt-Cpn10 with different concentrations inhibited the proliferation and alkaline phosphatase activity of osteoblast (P<0.05), the number of calcium nodules formation was significantly reduced. The r-Mt-Cpn10 increased the expression of RANKL in a dose-dependent manner and reduced the expression of OPG (P<0.01). Conclusion: The inhibition of r-Mt-Cpn10 on the osteoblast proliferation and alkaline phosphatase activity was achieved by osteoblasts arrest in G2/M phase and G1 to S phase, it can also regulate the expression of RANKL and OPG which affecting local bone metabolic balance.
PMCID: PMC4211789  PMID: 25356139
Human osteoblasts; proliferation; RANKL; OPG; alkaline phosphatase activity
14.  miR-148b functions as a tumor suppressor in non-small cell lung cancer by targeting carcinoembryonic antigen (CEA) 
Aberrant expression of miR-148b has been found in several types of cancer, but its expression and potential biologic role in non-small cell lung cancer (NSCLC) are still largely unknown. Here, we found that miR-148b was commonly under-expressed in human non-small celllung cancer (NSCLC) specimens and cell lines. The overexpression of miR-148b dramatically suppressed NSCLC cell proliferation and migration. Furthermore, miR-148b could regulate carcinoembryonic antigen (CEA) expression by luciferase reporter assay. On the other hand, CEA was widely up-regulated in NSCLC specimens, and its mRNA levels were inversely correlated with miR-148b expression. These suggest that CEA expression may be regulated by miR-148b. Collectively, our findings indicate miR-148b is low expression in NSCLC cells, which results in CEA overexpression and disease progression in NSCLC patients.
PMCID: PMC4161539  PMID: 25232379
NSCLC; CEA; miR-148b; migration; tumor suppressors
15.  Estimating daily salt intake based on 24 h urinary sodium excretion in adults aged 18–69 years in Shandong, China 
BMJ Open  2014;4(7):e005089.
24 h urinary sodium extretion was used to estimate the daily salt intake of shandong residents aged from 18 to 69 years in China.
20 selected counties/districts in Shandong stratified by geographic region (Eastern, Central Southern and North Western) and residence type (urban vs rural).
Among 2184 randomly selected adults, 2061 provided usable 24 h urine samples. Urine volume <500 mL or male creatinine <3.81 (female creatinine <4.57) are not included in the analysis.
The mean sodium level excreted over 24 h was 237.61 mmol (95% CI 224.77 to 250.44) mmol. Overall, the estimated mean salt intake was 13.90 g/day (95% CI 13.15 to 14.65). The mean salt intake among rural residents was higher than that among urban residents (14.00 vs 13.68 g; p<0.01). Salt intake in men was higher than that in women (14.40 vs 13.37 g; p<0.01). Approximately 96% of the survey participants had a dietary salt intake of ≥6 g/day.
The salt intake in Shandong is alarmingly higher than the current recommended amount (6 g/day). Thus, effective interventions to reduce salt intake levels to combat the increasing burden of non-communicable diseases need to be developed and implemented.
PMCID: PMC4120319  PMID: 25037642
16.  Randomized double-blind trial of prophylactic topical Evozac® Calming Skin Spray for gefitinib-associated acne-like eruption 
OncoTargets and therapy  2014;7:1261-1266.
“Gefitinib” is a first-generation epidermal growth factor receptor tyrosine-kinase inhibitor. More than half of patients receiving gefitinib develop acne-like eruption. Evozac® Calming Skin Spray (Evaux Laboratoires, Évaux-les-Bains, France) is made of Évaux thermal spring water and commonly used for the treatment of dermatological toxicities caused by anti-epidermal growth factor receptor therapy. The aim of the study reported here was to test the effect of Evozac Calming Skin Spray on the prevention of rash in patients receiving gefitinib.
Non-small-cell lung cancer patients preparing to initiate gefitinib therapy were randomly assigned to apply Evozac Calming Skin Spray or physiological saline to the face three times a day. The treatment was started on the same day as initiation of gefitinib therapy and continued for 4 weeks.
A total of 51 patients in the Evozac Calming Skin Spray group and 50 patients in the physiological saline group completed the study per the protocol. The number of facial lesions peaked at the end of 3 weeks in both groups. There were significantly fewer lesions in the Evozac Calming Skin Spray group than in the physiological saline group at the end of 1 week (0.25 versus [vs] 1.10, P=0.031) and 3 weeks (6.67 vs 12.26, P=0.022). Patients from the Evozac Calming Skin Spray group also developed fewer facial lesions at the end of 2 weeks and 4 weeks, however, the difference was not statistically significant. At the end of 4 weeks, fewer patients from the Evozac Calming Skin Spray group developed rash of grade 2 or greater severity (17.6% vs 36.0%, P=0.037), or experienced rash-associated symptoms (13.7% vs 34.0%, P=0.017).
Prophylactic treatment with Evozac Calming Skin Spray appears to decrease the number of facial lesions at the peak of the rash, reduce the incidence of grade 2 or more severe rash and relieve rash-associated symptoms.
PMCID: PMC4096493  PMID: 25031540
dermatological toxicities; facial rash lesions; rash severity; rash-associated symptoms
17.  Glutamate and Lipid Metabolic Perturbation in the Hippocampi of Asymptomatic Borna Disease Virus-Infected Horses 
PLoS ONE  2014;9(6):e99752.
Borna disease virus (BDV) is a neurotropic, enveloped, non-segmented, negative-stranded RNA virus that infects a wide variety of vertebrate species from birds to humans across a broad global geographic distribution. Animal symptomatology range from asymptomatic infection to behavioral abnormalities to acute meningoencephalitis. Asymptomatic BDV infection has been shown to be more frequent than conventionally estimated. However, the molecular mechanism(s) underyling asymptomatic BDV infection remain largely unknown. Here, based on real-time quantitative PCR and Western blotting, a total of 18 horse hippocampi were divided into BDV-infected (n = 8) and non-infected control (n = 10) groups. A gas chromatography coupled with mass spectrometry (GC-MS) metabolomic approach, in conjunction with multivariate statistical analysis, was used to characterize the hippocampal metabolic changes associated with asymptomatic BDV infection. Multivariate statistical analysis showed a significant discrimination between the BDV-infected and control groups. BDV-infected hippocampi were characterized by lower levels of D-myo-inositol-1-phosphate, glutamate, phosphoethanolamine, heptadecanoic acid, and linoleic acid in combination with a higher level of ammonia. These differential metabolites are primarily involved in glutamate and lipid metabolism. These finding provide an improved understanding of hippocampal changes associated with asymptomatic BDV infection.
PMCID: PMC4067290  PMID: 24956478
18.  Small-molecule TrkB receptor agonists improve motor function and extend survival in a mouse model of Huntington's disease 
Human Molecular Genetics  2013;22(12):2462-2470.
Huntington's disease (HD) is a fatal neurodegenerative disease characterized by abnormal motor coordination, cognitive decline and psychiatric disorders. This disease is caused by an expanded CAG trinucleotide repeat in the gene encoding the protein huntingtin. Reduced levels of brain-derived neurotrophic factor (BDNF) in the brain, which results from transcriptional inhibition and axonal transport deficits mediated by mutant huntingtin, have been suggested as critical factors underlying selective neurodegeneration in both HD patients and HD mouse models. BDNF activates its high-affinity receptor TrkB and promotes neuronal survival; restoring BDNF signaling is thus of particular therapeutic interest. In the present study, we evaluated the ability of a small-molecule TrkB agonist 7,8-dihydroxyflavone (7,8-DHF) and its synthetic derivative 4′-dimethylamino-7,8- dihydroxyflavone (4′-DMA-7,8-DHF) to protect neurons in the well-characterized N171-82Q HD mouse model. We found that chronic administration of 7, 8-DHF (5 mg/kg) or 4′-DMA-7,8-DHF (1 mg/kg) significantly improved motor deficits, ameliorated brain atrophy and extended survival in these N171-82Q HD mice. Moreover, 4′-DMA-7,8-DHF preserved DARPP32 levels in the striatum and rescued mutant huntingtin-induced impairment of neurogenesis in the N171-82Q HD mice. These data highlight consideration of TrkB as a therapeutic target in HD and suggest that small-molecule TrkB agonists that penetrate the brain have high potential to be further tested in clinical trials of HD.
PMCID: PMC3658168  PMID: 23446639
19.  Zinc dependent endopeptidases of matrix metalloproteinases-9 expressions is associated with tumor metastases of oral squamous cell carcinoma in Chinese population: a meta-analysis 
Matrix metalloproteinase-9 (MMP-9) is an important member of zinc dependent endopeptidases family and is considered to be involved in the invasion and metastasis of cancer cells. Many studies were published to assess the prognostic role of MMP-9 expression in patients with oral squamous cell Carcinoma, but the findings from those studies were inconsistent in Chinese population. We searched eligible studies in Pubmed, Embase, and Web of Science databases. Six studies with a total of 556 patients were finally included into the meta-analysis. The pooled odds ratios (OR) with the corresponding 95% confidence interval (95% CIs) for positive rate of MMP-9 were calculated by using meta-analysis. Overall, MMP-9 positive expression was associated with tumor metastases in patients with oral squamous cell Carcinoma (fixed-effects OR 4.24, 95% CI 2.25-7.99, P < 0.001; random-effects OR 4.35, 95% CI 2.31-8.21, P < 0.001). Our results indicated that MMP-9 expression is associated with tumor metastases in patients with oral squamous cell carcinoma, and patients with higher MMP-9 expression have less tumor metastases.
PMCID: PMC4100962  PMID: 25035776
MMP-9; tumor metastases; oral squamous cell carcinoma
20.  Acupuncture at the Taixi (KI3) acupoint activates cerebral neurons in elderly patients with mild cognitive impairment 
Neural Regeneration Research  2014;9(11):1163-1168.
Our previous findings have demonstrated that acupuncture at the Taixi (KI3) acupoint in healthy youths can activate neurons in cognitive-related cerebral cortex. Here, we investigated whether acupuncture at this acupoint in elderly patients with mild cognitive impairment can also activate neurons in these regions. Resting state and task-related functional magnetic resonance imaging showed that the pinprick senstation of acupuncture at the Taixi acupoint differed significantly between elderly patients with mild cognitive impairment and healthy elderly controls. Results showed that 20 brain regions were activated in both groups of participants, including the bilateral anterior cingulate gyrus (Brodmann areas [BA] 32, 24), left medial frontal cortex (BA 9, 10, 11), left cuneus (BA 19), left middle frontal gyrus (BA 11), left lingual gyrus (BA 18), right medial frontal gyrus (BA 11), bilateral inferior frontal gyrus (BA 47), left superior frontal gyrus (BA11), right cuneus (BA 19, 18), right superior temporal gyrus (BA 38), left subcallosal gyrus (BA 47), bilateral precuneus (BA 19), right medial frontal gyrus (BA 10), right superior frontal (BA 11), left cingulate gyrus (BA 32), left precentral gyrus (BA 6), and right fusiform gyrus (BA 19). These results suggest that acupuncture at the Taixi acupoint in elderly patients with mild cognitive impairment can also activate some brain regions.
PMCID: PMC4146092  PMID: 25206776
nerve regeneration; acupuncture; acupoint; Taixi (KI3); acupoint specificity; mild cognitive impairment; functional MRI; resting state; cognitive function; brain function; NSFC; neural regeneration
21.  The Association Between Early Menarche and Offspring’s Obesity Risk in Early Childhood was Modified by Gestational Weight Gain 
Obesity (Silver Spring, Md.)  2013;22(1):19-23.
Study effect of maternal gestational weight gain (GWG) on the relationship between maternal menarcheal age (MMA) and child growth and overweight risk and examine socio-demographics characteristics of excessive GWG.
Design and Methods
The relationships between GWG and MMA in 54,184 women and their children’s growth trajectories during first 5 years of life (2000–2005) in south China were tested using longitudinal data analysis with mixed models and logistic regression.
Average MMA was 14.8 (1.3) years; 36.3% of the women had excessive GWG. Excessive GWG interacted with adverse effects of early MMA (if ≤ 13 years), leading to the most rapid growth in offspring and highest risk of overweight at age 4–5 (OR = 5.2 [2.0–13.5]) than others. Women with early menarche, high-education, urban residence, and a routine job were more likely to have excessive GWG than the others.
GWG modify the association between early MMA and offspring’s growth and overweight. Controlling for GWG may reduce the adverse influence of early MMA and its own adverse influence on childhood health.
PMCID: PMC4036118  PMID: 23836480
22.  Stem Cell-Like Circulating Tumor Cells Indicate Poor Prognosis in Gastric Cancer 
BioMed Research International  2014;2014:981261.
Circulating tumor cells (CTCs), which have stem cell-like characteristics, might play a crucial role in cancer metastasis. CD44 has been identified as gastric cancer (GC) stem cell (CSC) marker. Here, the prognostic significance of CD44-positive CTCs in GC patients was investigated. CTCs were detected in 27 of 45 GC patients. The presence of CTCs was significantly associated with lymph node metastasis, distant metastasis, and recurrence (P = 0.007, P = 0.035, and P = 0.035, resp.). Nineteen of the 27 CTC-positive patients had CD44-positive CTCs. These patients were more likely to develop metastasis and recurrence than patients with CD44-negative CTCs. CD44-positive CTC counts were higher in recurrent patients than in the nonrecurrent ones (means 4.8 and 1.9, resp.; P = 0.010). Furthermore, 13 of 19 patients with CD44-positive CTCs developed recurrent disease, and the mean time to recurrence was shorter than that in patients with CD44-negative CTCs (10.54 ± 5.55 and 19.13 ± 9.72 months, resp.; P = 0.04). COX proportional hazards model indicated that the presence of CD44-positive CTCs and TNM stage were independent predictors of recurrence for GC (P = 0.030 and 0.008). So identifying the stem cell-like CTC subset may provide more clinically useful prognostic information than only detecting CTCs.
PMCID: PMC4054962  PMID: 24963492
23.  Predictors of recurrence in breast cancer subtypes with negative lymph node in a Chinese population 
To establish a series of objective parameters to predict the risk of relapse from axillary lymph node-negative (ANN) breast cancer, and evaluate the patterns of recurrence according to molecular subtypes, we collected information on 2126 consecutive breast cancer patients operated between 2002 and 2006. In this case-control study, 212 patients experiencing recurrence or breast cancer related death were defined as ‘poor group’. Another 212 patients were selected from the remaining cases with stratified sampling method to comprise the ‘good group’. Significant differences were found in vascular invasion, grade and molecular subtype between the two groups. Expression of ER and PR in the ‘poor group’ was lower (P < 0.05). However, positive rates of Ki67, p53 and VEGF in the ‘poor group’ were higher (P < 0.05). Multivariate analysis revealed that molecular subtype, expression of VEGF, tumor grade, and vascular invasion were closely correlated with bad outcome. Analysis of the ‘poor group’ demonstrated that ‘HER2 positive’ and ‘triple negative’ subtypes more commonly suffered from distant metastases and death. No metastasis was found in patients with pure invasive papillary carcinoma, invasive cribriform carcinoma or adenoid cystic carcinoma, whereas the diagnoses of invasive micropapillary carcinoma, invasive apocrine carcinoma, invasive papillary carcinoma mixed with invasive ductal carcinoma, or metaplastic carcinoma were correlated with distant metastasis and death. In conclusion, molecular subtype and expression of VEGF are useful markers for predicting prognosis of ANN breast cancer patients. ‘Luminal A-like’ subtype has better outcome than others. Moreover, molecular subtypes have different recurrence patterns.
PMCID: PMC4097226  PMID: 25031741
Breast cancer; molecular subtype; p53; VEGF; recurrence
24.  Oncogenic roles of carbonic anhydrase IX in human nasopharyngeal carcinoma 
Carbonic anhydrase IX (CA IX), a hypoxia-inducible protein in tumors, has been shown to be valuable for the prognosis of nasopharyngeal carcinoma (NPC). However, the function and mechanism of CA IX has been not explored in NPC. Here, we found that CA IX was detected at higher levels in NPC cells and tissues than their corresponding partners. Furthermore, the cell growth, migration and invasion in vitro were altered with shRNA or overexpression of CA IX in NPC cells. More importantly, the metastatic ability of NPC cells stably expressing CA IX was significantly enhanced using the hepatic metastasis model of nude mice in vivo. Finally, the mTOR pathway was indicated to be involved in such effects of CA IX on NPC. This is the first evidence that CA IX may promote the NPC metastasis to potentially be a therapeutic target for NPC, and that the inhibitory molecules of CA IX and/or the mTOR pathway alone or combination with both may be worth to have a clinical trial for the patients with NPC.
PMCID: PMC4097237  PMID: 25031713
CA IX; NPC; growth; metastasis; mTOR
25.  Sterols and Terpenoids from Viburnum odoratissimum 
A new stigmasterol type natural product, viburodorol A (1), along with eleven known sterols and terpenoids (2–12), were isolated from the aerial parts of Viburnum odoratissimum. The structure of 1 was elucidated on the basis of comprehensive spectroscopic analysis. It’s noteworthy that compound 2, the major constituent of this plant, can significantly stimulate glucose absorption in insulin resistant HepG2 cells without affecting cell viability. Furthermore, this compound can also restore the glucose absorption in DXMS-induced insulin resistant 3T3-L1 cells.
Graphical Abstract
Electronic supplementary material
The online version of this article (doi:10.1007/s13659-014-0021-7) contains supplementary material, which is available to authorized users.
PMCID: PMC4050308  PMID: 24955299
Viburnum odoratissimum; Insulin sensitizing activity; 6α-Hydroxy-lup-20(29)-en-3-on-28-oic acid; Viburodorol A

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