An enzymatic activity regulation-based clusterzyme sensor array for high-throughput identification of heavy metal ions.

The accurate identification and sensitive quantification of heavy metal ions are of great significance, considering that pose a serious threat to environment and human health. Most array-based sensing platforms, to date, utilize nanozymes as sensing elements, but few studies have explored the application of the peroxidase-like activity of clusterzymes in identification of multiple analytes. Herein, for the first time, we developed a clusterzyme sensor array utilizing gold nanoclusters (AuNCs) as sensing elements for five heavy metal ions identification including Hg2+, Pb2+, Cu2+, Cd2+ and Co2+. The heavy metal ions can differentially regulate the peroxidase-like activity of AuNCs, and that can be converted into colorimetric signals with 3,3',5,5'-tetramethylbenzidine (TMB) as the chromogenic substrate. Subsequently, the generated composite responses can be interpreted by combining pattern recognition algorithms. The developed clusterzyme sensor array can identify five heavy metal ions at concentrations as low as 0.5 µM and their multi-component mixtures. Especially, we demonstrated the successful identification of multiple heavy metal ions in tap water and traditional Chinese medicine, with an accuracy of 100% in blind test. This study provided a simple and effective method for identification and quantification of heavy metal ions, rendering a promising technique for environmental monitoring and drug safety assurance.

[Effect of traditional Chinese medicine in attenuating chronic kidney disease and its complications by regulating gut microbiota-derived metabolite trimethylamine N-oxide: a review].

Trimethylamine N-oxide(TMAO), a metabolite of gut microbiota, is closely associated with chronic kidney disease(CKD). It can aggravate the kidney injury and promote the occurrence of complications of CKD mainly by inducing renal fibroblast activation, vascular endothelial inflammation, macrophage foaming, platelet hyperreactivity, and inhibition of reverse cholesterol transport. Thus it is of great significance for clinical treatment of CKD to regulate circulating TMAO and alleviate its induced body damage. Currently, therapeutic strategies for TMAO regulation include dietary structure adjustment, lifestyle intervention, intestinal microflora regulation, and inhibition of intestinal trimethylamine synthesis and liver trimethylamine oxidation. Chinese medicinal herbs have the clinical advantage of multi-component and multi-target effects, and application of traditional Chinese medicine(TCM) to synergistically regulating TMAO and improving CKD via multiple pathways has broad prospects. This study systematically reviewed the clinical relevance and mechanism of TMAO in aggravating CKD renal function deterioration and complication progression. In addition, the effect and mechanism of TCM in improving TMAO-induced kidney injury, cardiovascular disease, hyperlipidemia, thrombosis and osteoporosis were summarized. The results provided a theoretical basis for TCM in attenuating gut microbiota-derived metabolite TMAO and improving CKD, as well as a basis and direction for in-depth clinical development and mechanism research in the future.

Mechanism for the restoration of degraded typical steppe by nitrogen and phosphorus co-addition.

The reduction of soil nutrient content is one of the major reasons caused grassland degradation in China. Nutrient addition is thus considered as an effective measure for the restoration of degraded grasslands. However, over-fertilization can lead to decrease in plant diversity. To clarify the appropriate amount of nutrient addition and the underlying mechanism that promotes grassland restoration, we set up a nitrogen and phosphorus co-addition experiment in a degraded typical steppe of Inner Mongolia, and examined the responses at community, functional group and species levels to nutrient addition. The results showed that nutrient addition enhanced biomass while did not reduce species richness at the community level. The biomass showed a saturation response with the increases of nutrient addition, which approached saturation under the 12.0 g N·m-2, 3.8 g P·m-2 treatment. Species richness increased significantly under the lower nutrient treatments (N <9.6 g·m-2, P < 3.0 g·m-2) compared with the control, while the two high nutrient treatments did not alter species richness. At the functional group level, biomass and abundance of perennial rhizome grasses increased significantly with the increases of nutrient addition levels. Biomass and density of annuals increased significantly under high nutrient addition levels. However, the abundance and biomass of perennial bunchgrasses and perennial forbs were rarely affected. At the species level, six target species responded differently to nutrient addition. Biomass of Leymus chinensis was significantly increased due to the increase of population density and individual biomass. Biomass of Stipa grandis, Agropyron cristatum and Cleistogenes squarrosa change little. Biomass of Potentilla acaulis and Carex korshinskyi were reduced due to the decreases in individual biomass and population density, respectively. As a measure of restoring degraded grassland, nutrient addition could significantly increase biomass and species diversity, decrease biomass of the degradation indicator species, and increase biomass of perennial rhizomes grasses.

Effect of traditional Chinese medicine in attenuating chronic kidney disease and its complications by regulating gut microbiota-derived metabolite trimethylamine N-oxide: a review / 中国中药杂志

Trimethylamine N-oxide(TMAO), a metabolite of gut microbiota, is closely associated with chronic kidney disease(CKD). It can aggravate the kidney injury and promote the occurrence of complications of CKD mainly by inducing renal fibroblast activation, vascular endothelial inflammation, macrophage foaming, platelet hyperreactivity, and inhibition of reverse cholesterol transport. Thus it is of great significance for clinical treatment of CKD to regulate circulating TMAO and alleviate its induced body damage. Currently, therapeutic strategies for TMAO regulation include dietary structure adjustment, lifestyle intervention, intestinal microflora regulation, and inhibition of intestinal trimethylamine synthesis and liver trimethylamine oxidation. Chinese medicinal herbs have the clinical advantage of multi-component and multi-target effects, and application of traditional Chinese medicine(TCM) to synergistically regulating TMAO and improving CKD via multiple pathways has broad prospects. This study systematically reviewed the clinical relevance and mechanism of TMAO in aggravating CKD renal function deterioration and complication progression. In addition, the effect and mechanism of TCM in improving TMAO-induced kidney injury, cardiovascular disease, hyperlipidemia, thrombosis and osteoporosis were summarized. The results provided a theoretical basis for TCM in attenuating gut microbiota-derived metabolite TMAO and improving CKD, as well as a basis and direction for in-depth clinical development and mechanism research in the future.

The Gut Microbiota and Traditional Chinese Medicine: A New Clinical Frontier on Cancer.

Gut microbiota is a complex microecosystem, which is called the second genome of the human body. Herbal medicine can balance tumor-suppressing bacteria and tumor-promoting bacteria and exert its anti-cancer effect by regulating gut microbiota. Traditional Chinese medicine (TCM) has a history of thousands of years in prevention and treatment of diseases in China. In recent decades, TCM has been shown to have an obvious advantage in prolonging the survival time and improving the living quality of patients with cancer. Notably, gut microbiota has become a new pathway to understanding TCM. In this review, we will focus on gut microbiota and tumor progression, especially the diversity, functionality and metabolites of gut microbiota affected by TCM in various cancer. We will also discuss the potential mechanism of gut microbiota for exploring TCM in anti-cancer effect. This article aims to comprehensively review the anti-cancer research of TCM by regulating gut microbiota, and address future perspectives and challenges of gut microbiota in TCM intervention for cancer.

Corrigendum to "Yiqi Huayu Jiedu Decoction Inhibits the Invasion and Metastasis of Gastric Cancer Cells through TGF-ß/Smad Pathway".

[This corrects the article DOI: 10.1155/2017/1871298.].

The effect of Jianpi Yangzheng Xiaozheng Decoction and its components on gastric cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Jianpi Yangzheng Xiaozheng Decoction (JPYZXZ) is an empirical compound prescription based on the theory of traditional Chinese medicine. JPYZXZ, which is "Qi-invigorating, spleen-strengthening and stasis-removing," can improve the quality of life of gastric cancer patients and prolong their survival; however, the exact mechanism underlying the antitumor effects of this compound is still not clear. AIM OF THE STUDY: The aim of this study is to clearly define the effect of JPYZXZ and its components, Jianpi Yangzheng Decoction (JPYZ) and Xiao Zheng San Jie Decoction (XZSJ), on inhibiting the progression of gastric cancer. MATERIALS AND METHODS: The effect of JPYZXZ and its components on the motility of gastric cancer MGC-803 cells was measured by MTT, adhesion, transwell assays and wound-healing assays. JPYZXZ, JPYZ and XZSJ were administered to 615 mice with gastric cancer xenografts, and their effect on the inhibition of subcutaneous transplantation was analyzed. THP-1 monocyte cells were used to establish tumor-associated macrophage (TAM) models. The polarized state of the TAMs was detected by Flow Cytometry, ELISA and Immunohistochemistry. The mRNA and protein expression of tumor epithelial-mesenchymal transition (EMT) and TAM-related genes was determined by Real-time PCR and Western Blot, respectively. RESULTS: We determined that both JPYZXZ and its components inhibited the progress of gastric cancer in vitro, and JPYZXZ was clearly more effective than JPYZ or XZSJ. The in vivo results demonstrated that the JPYZXZ and XZSJ group exhibited a significant decrease in the tumor weight compared to the control group. Further analysis indicated that JPYZXZ was more active than JPYZ or XZSJ in inhibiting the gastric cancer EMT transformation both in vivo and in vitro. However, JPYZ was more effective compared with JPYZXZ for inducing the phenotypic change in macrophages from M2 to M1. CONCLUSIONS: Our results demonstrate that both JPYZXZ and its components prevent the progress of gastric cancer. JPYZXZ inhibits the gastric cancer EMT more effectively than JPYZ and XZSJ, but JPYZ primarily works to regulate the phenotypic change in macrophages from M2 to M1.

Effect and Mechanism of Dendrobii Officinalis Caulis in Intervening and Preventing Myocardial Fibrosis in Rats with Heart Failure and Heart Qi Deficiency Syndrome After Ischemia-reperfusion / 中国实验方剂学杂志

Objective:To investigate the expressions of cardiac cycle, myocardial pathology, galectin-3 (Gal-3),transforming growth factor-β (TGF-β),Smad homologue 3 recombinant protein (Smad3) in rats with heart failure and heart failure after ischemia-reperfusion, and the intervention effect of Dendrobii Officinalis Caulis (DOC) myocardial fibrosis in model rats. Method:A rat model of heart failure and Qi deficiency was established through ligation of the left anterior descending coronary artery. The rats were divided into blank group, model group, valsartan group (9.43 mg·kg-1) and DOC group (10 mg·kg-1), with 10 in each group. The blank group and the model group were given an equal volume of physiological saline. The changes in left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic dimension(LVESD), left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS) of the cardiac cycle of rats in each group were recorded by high-resolution ultrasound system. The carboxyterrninal propeptide of type I procollagen (PICP), and carboxyterrninal propeptide of type Ⅲ procollagen (PⅢNP)were detected by enzyme-linked immunosorbent assay (ELISA) kit. The morphological changes of myocardial cells were observed by hematoxylin-eosin (HE) staining. The changes of myocardial fiber tissue and collagen were observed by Masson staining. Western blot was used to detect the protein expressions of Gal-3, TGF-β, Smad3. Result:Compared with the blank group, the levels of LVEDD, LVEF, and LVFS were lower in the model group (PPβ, and Smad3 were decreased (PPPPβ and Smad3 were lower than those in the model group (PPConclusion:DOC can effectively inhibit myocardial fibrosis in rats with heart failure and heart Qi deficiency syndrome after ischemia-reperfusion. The mechanism may be correlated with the reduction of the expressions of Gal-3, TGF-β and Smad3.

Yiqi Huayu Jiedu Decoction Inhibits the Invasion and Metastasis of Gastric Cancer Cells through TGF-ß/Smad Pathway.

Background. Yiqi Huayu Jiedu Decoction (YHJD) can obviously improve the quality of life of those patients with gastric cancer and prolong their survival. Methods. In vitro experiments, we observe YHJD's effect on the cells' proliferation by MTT assay. Cell adhesion assay, wound-healing assay, and Transwell invasion assay serve to detect its influence on cells' adhesion, migration, and invasion, respectively. Inhibitor (10 µM/L of SB431542) and activator (10 ng/mL of TGF-ß) of TGF-ß/Smad pathway were used to estimate whether YHJD's impact on the biological behavior of gastric cancer cells was related to TGF-ß/Smad pathway. In in vivo studies, YHJD was administered to the nude mice transplanted with gastric cancer to observe its effect on the tumor. Western blotting and immunohistochemical assay were used to test relevant cytokines of TGF-ß/Smad pathway and epithelial-mesenchymal transition (EMT) in MGC-803 cells and the tumor bearing nude mice. Results. YHJD inhibited proliferation, adhesion, migration, and invasion of MGC-803 gastric cancer cells in vitro. In in vivo studies, YHJD reduced the volume of the transplanted tumors. It also enhanced the expression of E-cadherin and decreased the levels of N-cadherin, TGF-ß, Snail, and Slug in both MGC-803 cells and the transplanted tumor by western blot assay. The immunohistochemical assay revealed that YHJD raised E-cadherin in the tumors of the mice; on the contrary, the expression of N-cadherin, Twist, vimentin, TGF-ßR I, p-Smad2, p-Smad3, Snail, and Slug reduced. Conclusion. YHJD can effectively inhibit the invasion and metastasis of gastric cancer cells. The mechanism may be related to TGF-ß/Smad pathway.

Role of TAZ in genistein induced osteoblastogenic differentiation of mouse bone marrow-derived mesenchymal stem cells / 中华内分泌代谢杂志

Objective To investigate the role of transcriptional-coactivator with PDZ-binding motif( TAZ) in genistein-induced osteoblastogenic differentiation of mouse bone marrow-derived mesenchymal stem cells ( BMSCs) .Methods Mouse BMSCs were cultured in phenol red-freeα-MEM containing osteogenic supplements for inducing osteogenic differentiation.BMSCs were transfected with siRNA-TAZ and treated with genistein.The temporal sequence of osteoblastic differentiation in BMSCs cultures was assayed by measuring alkaline phosphatase activity (ALP) and calcium deposition.The mRNA expression of bone sialoprotein ( BSP) and osteocalcin ( OC) were detected by reverse transcription-polymerase chain reaction(RT-PCR).The binding interaction between TAZ and cbfa1 was identified by co-immunoprecipitation.Results TAZ expression was detected during the induction of osteogenic differentiation, the ALP activity and calcium deposition were significantly decreased in BMSCs which were transfected with siRNA-TAZ.Genistein(0.01-1 μmol/L) exhibited a dose-dependent effect on TAZ expression in mouse BMSCs cultures.Treatment with genistein ( 1 μmol/L ) resulted in increased ALP avtivity and calcium deposition of BMSC cultures as function of time.Genistein(1μmol/L) also promoted the nuclear localization of TAZ and augmented the interaction between TAZ and cbfa1, and by which upregulated cbfa1-mediated gene expression such as BSP and OC.However, the ALP avtivity and calcium deposition, as well as the expression of BSP and OC were not promoted by genistein in BMSCs transfected with siRNA-TAZ.Conclusion These data suggest that the TAZ plays an important role in genistein-induced osteoblastic differentiation of mouse BMSCs cultures.

Design, synthesis, anti-TMV, fungicidal, and insecticidal activity evaluation of 1,2,3,4-tetrahydro-ß-carboline-3-carboxylic acid derivatives based on virus inhibitors of plant sources.

By drawing the creation ideas of botanical pesticides, a series of tetrahydro-ß-carboline-3-carboxylic acid derivatives were designed and synthesized, and first evaluated for their anti-TMV, fungicidal and insecticidal activities. Most of these derivatives exhibited good antiviral activity against TMV both in vitro and in vivo. Especially, the activities of compounds 8 and 15 in vivo were higher than that of ribavirin. The compound 8 exhibited more than 70% fungicidal activities against Cercospora arachidicola Hori, Alternaria solani, Bipolaris maydis, and Rhizoctonia solani at 50mg/kg, compounds 16 and 20 exhibited more than 60% insecticidal activities against Mythimna separate and Ostrinia nubilalis.

[Inhibitory effect of cinnamaldehyde on invasion capacities of human breast cancer cell line MDA-MB-435S and its relation with regulating the expression of miR-27a].

OBJECTIVE: To explore the inhibitory effect of cinnamaldehyde on invasion capacities of human breast cancer cell line MDA-MB-435S and its relation with regulating the expression of miR-27a. METHODS: The effect of cinnamaldehyde on invasive capacities of MDA-MB-435S was measured by Transwell matrigel invasion assay. The effect of miR-27a expression on invasive capabilities of MDA-MB-435S, the intervention of cinnamaldehyde in the miR-27a expression, and its relation with its effect on invasive capabilities were defected with liposome 2000 transinfection miRNA27a mimics/inhibitors, real time-polymerase chain reaction (Real-time PCR), and Transwell chamber model. RESULTS: Compared with the control group, the number of cells passing through the transwell chamber was more significantly reduced after treated by cinnamaldehyde for 12 h (P < 0.05). The miR-27a expression was 962.07 times and 40% of that of the control group after transinfected by miR-27a mimics and miR-27a inhibitors. After transinfected by miR-27a inhibitors, the number of cells passing through the transwell chamber was more significantly reduced (P < 0.05). The miR-27a expression of MDA-MB-435S was down-regulated by 12-h treatment of cinnamaldehyde (2(-deltaCt) = 0.56, 0.18, 0.18, respectively). The number of miR-27a mimics transinfection pretreated MDA-MB-435S cells passing through the transwell chamber increased more obviously than the number of un-pretreated MDA-MB-435S cells in the control group (P < 0.05). CONCLUSIONS: Cinnamaldehyde could inhibit invasive capabilities of human breast cancer cell line MDA-MB-435S. The over-expression of miR-27a played an important role in the invasive capability of MDA-MB-435S. The inhibition of cinnamaldehyde on invasive capabilities of MDA-MB-435S cells was correlated with down-regulating the expression of miR-27a.

[Effect of Chinese herbal therapy on T-lymphocytes of IgA nephropathy patients: a clinical observation].

OBJECTIVE: To observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN). METHODS: Totally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry. RESULTS: Compared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05). CONCLUSION: Chinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.

Effect of Chinese herbal therapy on T-lymphocytes of IgA nephropathy patients: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN).</p><p><b>METHODS</b>Totally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry.</p><p><b>RESULTS</b>Compared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05).</p><p><b>CONCLUSION</b>Chinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.</p>

[Relations between attitude and practice of smoking and the training program regarding tobacco control among community medical staff members in Hangzhou, Zhejiang province].

OBJECTIVE: To investigate the relations between training and both the attitude and practice on smoking control among community medical staff members in Hangzhou, Zhejiang province. METHODS: Three representative districts including Xiacheng, Gongshu and Westlake were chosen from Hangzhou city. Questionnaire survey was applied to collect information from the related community medical staff members. The survey mainly contained three aspects: knowledge, attitude and practice regarding smoking control involved in the community medical activities. Availability and application of the resources on smoking cessation were also studied. Logistic regression analysis was applied to explore the factors associated with the smoking control training programs. Differences of rates between groups were assessed with chi-square statistics. Wilcoxon rank sum test was used to study the relationships among knowledge, attitude and practice related to smoking control programs, targeted to the community medical staff members. RESULTS: Eight hundred forty-six community medical workers were involved. Sixty-five percent of the community medical staff members had learned related knowledge on smoking control. Proportion of the community medical staff who had taken lessons on smoking control with 3-10 working years was 1.77 times more than the ones with experience less than two years (OR = 1.77, 95% CI: 1.25-2.51). Eighty-eight point seven percent of the medical staff who had received smoking control training programs were identified with the consciousness that they should advise the patients to quit smoking, comparing to the proportion 81.60% (Z=-2.87, P=0.00) in the control group. In terms of the practice regarding smoking control, data showed that 21.62% of the medical staff who had received smoking control training programs would provide 'how to quit smoking' to more than 90% of the smoking patients, while the proportion in the control group was 10.65% (Z = -5.68, P = 0.00). The use of drugs, traditional Chinese medicine therapy and the smoking cessation hotline rate were all less than 30%. CONCLUSION: The training programs being used on smoking control seemed useful in improving the consciousness and practice towards the smoking control programs during their medical activities among the community medical staff members.

Relations between attitude and practice of smoking and the training program regarding tobacco control among community medical staff members in Hangzhou, Zhejiang province / 中华流行病学杂志

Objective To investigate the relations between training and both the attitude and practice on smoking control among community medical staff members in Hangzhou,Zhejiang province.Methods Three representative districts including Xiacheng,Gongshu and Westlake were chosen from Hangzhou city.Questionnaire survey was applied to collect information from the related community medical staff members.The survey mainly contained three aspects:knowledge,attitude and practice regarding smoking control involved in the community medical activities.Availability and application of the resources on smoking cessation were also studied.Logistic regression analysis was applied to explore the factors associated with the smoking control training programs.Differences of rates between groups were assessed with chi-square statistics.Wilcoxon rank sum test was used to study the relationships among knowledge,attitude and practice related to smoking control programs,targeted to the community medical staff members.Results Eight hundred forty-six community medical workers were involved.Sixty-five percent of the community medical staff members had learned related knowledge on smoking control.Proportion of the community medical staff who had taken lessons on smoking control with 3-10 working years was 1.77 times more than the ones with experience less than two years (OR=1.77,95％CI:1.25-2.51).Eighty-eight point seven percent of the medical staff who had received smoking control training programs were identified with the consciousness that they should advise the patients to quit smoking,comparing to the proportion 81.60％ (Z=-2.87,P=0.00) in the control group.In terms of the practice regarding smoking control,data showed that 21.62％ of the medical staff who had received smoking control training programs would provide ‘how to quit smoking' to more than 90％ of the smoking patients,while the proportion in the control group was 10.65％ (Z=-5.68,P=0.00).The use of drugs,traditional Chinese medicine therapy and the smoking cessation hotline rate were all less than 30％.Conclusion The training programs being used on smoking control seemed useful in improving the consciousness and practice towards the smoking control programs during their medical activities among the community medical staff members.

[Study on knowledge, attitudes and use of the pre-packaged food nutrition labeling in Hangzhou supermarket consumers in 2010].

OBJECTIVE: To study the prevalence rates of knowledge, attitudes and use of the nutrition labeling and related influential factors in Hangzhou supermarket consumers. METHODS: Using a self-developed survey questionnaire, randomly selected customers were conducted a face-to-face interview program in a large supermarket of Hangzhou city. RESULTS: 586 people were interviewed, including 202 males and 384 females, with the mean age as 41.6 ± 17.23 years. The accuracy rates of 6 questions related to the 'ability of choosing healthier food by using the Nutrition Facts Labels' were as follows: dietary fiber (71.84%), fat (70.99%), calcium (60.75%), salt (58.36%), energy (50.85%) and sugar (39.42%). The support rates of the three attitudes indicators were "support marking the Nutrition Facts Label" (90.44%), "support marking the nutrition claims" (87.03%) and "want to know more relative knowledge" (77.13%). There were 58.36% and 80.03% first-time buyers who would read the Nutrition Facts Food Labels and the Nutrition Claims. Through logistic model analysis, male, youth, low-educated people were found to be the risk factors related to the nutrition label reading behavior. CONCLUSION: The capacity of the reading, understanding and correct application of nutrition labeling among consumers was not satisfactory. Improvement on the readability and the authority of nutrition labeling and the development of the national nutrition knowledge and health education programs were in urgent needs.

Inhibition of dengue virus by targeting viral NS4B protein.

We report a novel inhibitor that selectively suppresses dengue virus (DENV) by targeting viral NS4B protein. The inhibitor was identified by screening a 1.8-million-compound library using a luciferase replicon of DENV serotype 2 (DENV-2). The compound specifically inhibits all four serotypes of DENV (50% effective concentration [EC(50)], 1 to 4 µM; and 50% cytotoxic concentration [CC(50)], >40 µM), but it does not inhibit closely related flaviviruses (West Nile virus and yellow fever virus) or nonflaviviruses (Western equine encephalomyelitis virus, Chikungunya virus, and vesicular stomatitis virus). A mode-of-action study suggested that the compound inhibits viral RNA synthesis. Replicons resistant to the inhibitor were selected in cell culture. Sequencing of the resistant replicons revealed two mutations (P104L and A119T) in the viral NS4B protein. Genetic analysis, using DENV-2 replicon and recombinant viruses, demonstrated that each of the two NS4B mutations alone confers partial resistance and double mutations confer additive resistance to the inhibitor in mammalian cells. In addition, we found that a replication defect caused by a lethal NS4B mutation could be partially rescued through trans complementation. The ability to complement NS4B in trans affected drug sensitivity when a single cell was coinfected with drug-sensitive and drug-resistant viruses. Mechanistically, NS4B was previously shown to interact with the viral NS3 helicase domain; one of the two NS4B mutations recovered in our resistance analysis-P104L-abolished the NS3-NS4B interaction (I. Umareddy, A. Chao, A. Sampath, F. Gu, and S. G. Vasudevan, J. Gen. Virol. 87:2605-2614, 2006). Collectively, the results suggest that the identified inhibitor targets the DENV NS4B protein, leading to a defect in viral RNA synthesis.

Development and characterization of a stable luciferase dengue virus for high-throughput screening.

To facilitate dengue virus (DENV) drug discovery, we developed a stable luciferase reporter DENV-2. A renilla luciferase gene was engineered into the capsid-coding region of an infectious cDNA clone of DENV-2. Transfection of BHK-21 cells with the cDNA clone-derived RNA generated high titers (>10(6)PFU/ml) of luciferase reporter DENV-2. The reporter virus was infectious to a variety of cells, producing robust luciferase signals. Compared with wild-type virus, the reporter virus replicated slower in both mammalian Vero and mosquito C6/36 cells. To examine the stability of the reporter virus, we continuously passaged the virus on Vero cells for five rounds. All passaged viruses stably maintained the luciferase gene, demonstrating the stability of the reporter virus. Furthermore, we found that the passaged virus accumulated a mutation (T108M) in viral NS4B gene that could enhance viral RNA replication in a cell-type specific manner. Using the reporter virus, we developed a HTS assay in a 384-well format. The HTS assay was validated with known DENV inhibitors and showed a robust Z' factor of 0.79. The Luc-DENV-2 HTS assay allows screening for inhibitors of all steps of the viral life cycle. The reporter virus will also be a useful tool for studying DENV replication and pathogenesis.

Inhibition of dengue virus through suppression of host pyrimidine biosynthesis.

Viral replication relies on the host to supply nucleosides. Host enzymes involved in nucleoside biosynthesis are potential targets for antiviral development. Ribavirin (a known antiviral drug) is such an inhibitor that suppresses guanine biosynthesis; depletion of the intracellular GTP pool was shown to be the major mechanism to inhibit flavivirus. Along similar lines, inhibitors of the pyrimidine biosynthesis pathway could be targeted for potential antiviral development. Here we report on a novel antiviral compound (NITD-982) that inhibits host dihydroorotate dehydrogenase (DHODH), an enzyme required for pyrimidine biosynthesis. The inhibitor was identified through screening 1.8 million compounds using a dengue virus (DENV) infection assay. The compound contains an isoxazole-pyrazole core structure, and it inhibited DENV with a 50% effective concentration (EC(50)) of 2.4 nM and a 50% cytotoxic concentration (CC(50)) of >5 µM. NITD-982 has a broad antiviral spectrum, inhibiting both flaviviruses and nonflaviviruses with nanomolar EC(90)s. We also show that (i) the compound inhibited the enzymatic activity of recombinant DHODH, (ii) an NITD-982 analogue directly bound to the DHODH protein, (iii) supplementing the culture medium with uridine reversed the compound-mediated antiviral activity, and (iv) DENV type 2 (DENV-2) variants resistant to brequinar (a known DHODH inhibitor) were cross resistant to NITD-982. Collectively, the results demonstrate that the compound inhibits DENV through depleting the intracellular pyrimidine pool. In contrast to the in vitro potency, the compound did not show any efficacy in the DENV-AG129 mouse model. The lack of in vivo efficacy is likely due to the exogenous uptake of pyrimidine from the diet or to a high plasma protein-binding activity of the current compound.