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1.  Protein AND Enzyme Gated Supramolecular Disassembly 
An amphiphilic nanoassembly was designed to respond to the concurrent presence of a protein and an enzyme. We present herein a system, where in the presence of these two stimuli causes a supramolecular disassembly and molecular release. This molecular release arises in the form a fluorescence response that has been shown to be specific. We also show that this system can be modified to respond only if light stimulus is also present in addition to the protein and the enzyme. Demonstration of such supramolecular disassembly principles could have wide implications in a variety of biological applications.
doi:10.1021/ja4108676
PMCID: PMC3985736  PMID: 24447098
2.  Brazilin inhibits amyloid β-protein fibrillogenesis, remodels amyloid fibrils and reduces amyloid cytotoxicity 
Scientific Reports  2015;5:7992.
Soluble amyloid β-protein (Aβ) oligomers, the main neurotoxic species, are predominantly formed from monomers through a fibril-catalyzed secondary nucleation. Herein, we virtually screened an in-house library of natural compounds and discovered brazilin as a dual functional compound in both Aβ42 fibrillogenesis inhibition and mature fibril remodeling, leading to significant reduction in Aβ42 cytotoxicity. The potent inhibitory effect of brazilin was proven by an IC50 of 1.5 ± 0.3 μM, which was smaller than that of (−)-epigallocatechin gallate in Phase III clinical trials and about one order of magnitude smaller than those of curcumin and resveratrol. Most importantly, it was found that brazilin redirected Aβ42 monomers and its mature fibrils into unstructured Aβ aggregates with some β-sheet structures, which could prevent both the primary nucleation and the fibril-catalyzed secondary nucleation. Molecular simulations demonstrated that brazilin inhibited Aβ42 fibrillogenesis by directly binding to Aβ42 species via hydrophobic interactions and hydrogen bonding and remodeled mature fibrils by disrupting the intermolecular salt bridge Asp23-Lys28 via hydrogen bonding. Both experimental and computational studies revealed a different working mechanism of brazilin from that of known inhibitors. These findings indicate that brazilin is of great potential as a neuroprotective and therapeutic agent for Alzheimer's disease.
doi:10.1038/srep07992
PMCID: PMC4303869  PMID: 25613018
3.  Protein AND Enzyme Gated Supramolecular Disassembly 
An amphiphilic nanoassembly was designed to respond to the concurrent presence of a protein and an enzyme. We present herein a system, where in the presence of these two stimuli supramolecular disassembly and molecular release occur. This molecular release arises in the form a fluorescence response that has been shown to be specific. We also show that this system can be modified to respond only if light stimulus is also present in addition to the protein and the enzyme. Demonstration of such supramolecular disassembly principles could have broad implications in a variety of biological applications.
doi:10.1021/ja4108676
PMCID: PMC3985736  PMID: 24447098
4.  Autoantibodies in Chinese patients with chronic hepatitis B: Prevalence and clinical associations 
AIM: To investigate the prevalence of autoantibodies and their associations with clinical features in Chinese patients with chronic hepatitis B (CHB).
METHODS: A total of 325 Chinese patients with CHB were enrolled in this retrospective, hospital-based study. Patients with chronic hepatitis C (CHC), autoimmune hepatitis (AIH), or primary biliary cirrhosis (PBC) were included, with healthy donors acting as controls. A panel of autoantibodies that serologically define AIH and PBC was tested by indirect immunofluorescence assay and line immunoassay. The AIH-related autoantibody profile included homogeneous anti-nuclear antibodies (ANA-H), smooth-muscle antibodies, anti-liver kidney microsome type 1, anti-liver cytosolic antigen type 1, and anti-soluble liver antigen/liver pancreas; the PBC-related antibodies were characterized by ANA-nuclear dots/membranous rim-like, anti-mitochondrial antibodies-M2 (AMA-M2), anti-BPO (recombinant antigen targeted by AMA-M2), anti-Sp100, anti-promyelocytic leukemia protein (anti-PML), and anti-gp210. The dichotomization of clustering was used to unequivocally designate the AIH or PBC profiles for each case. Anti-Ro52 antibodies were also tested.
RESULTS: The prevalence of any autoantibody in CHB amounted to 58.2%, which was similar to the 66.2% prevalence in CHC, significantly higher than the 6.7% in the healthy controls (P < 0.001), and lower than the 100% found in AIH and PBC (P = 0.004 and P < 0.001, respectively). There were more anti-PML and anti-gp210 antibodies among the CHB patients than the CHC patients (11.1% vs 0%, P = 0.003; 12.6% vs 0%, P < 0.001, respectively). The prevalence and titer of AMA, anti-BPO, anti-PML, and anti-gp210 were higher in PBC than in those with CHB. Among the CHB patients, the prevalence of ANA, especially ANA-H, was significantly lower in patients with compensated and decompensated cirrhosis compared with patients without cirrhosis. Thirty-eight cases of hepatocellular carcinoma (HCC) in CHB showed a significant difference compared with non-HCC patients in the prevalence of anti-PML (0% vs 12.5%, P = 0.013). Dichotomization of the autoantibodies revealed that the PBC profile was more prevalent in patients with CHB than in those with CHC, and that it was strongly correlated with both compensated and decompensated cirrhosis. In contrast, the prevalence of the AIH profile was significantly higher in non-cirrhosis patients with CHB than in those with compensated cirrhosis (18.5% vs 8.2%, P = 0.039). Moreover, the AIH profile was also closely associated with hepatitis B e-antigen positivity.
CONCLUSION: ANA-H could be an indicator of early-stage CHB. Dichotomizing the autoantibody profiles revealed that the PBC profile is strongly associated with cirrhosis in CHB.
doi:10.3748/wjg.v21.i1.283
PMCID: PMC4284347  PMID: 25574103
Autoantibodies; Chronic hepatitis B; Autoimmune hepatitis; Primary biliary cirrhosis; Cirrhosis; Hepatocellular carcinoma
5.  A Comparison between the Sixth and Seventh Editions of the UICC/AJCC Staging System for Nasopharyngeal Carcinoma in a Chinese Cohort 
PLoS ONE  2014;9(12):e116261.
Background
The International Union Against Cancer/American Joint Committee on Cancer (UICC/AJCC) TNM staging system of nasopharyngeal carcinoma (NPC) is the most important system for survival prediction. The TNM 7th edition UICC/AJCC TNM staging system for NPC was adopted in January 2009, and is now internationally recommended. In comparison with the TNM 6th edition, there were several revisions in the new edition staging system. This study aims to evaluate the prognostic value of the TNM 7th edition for NPC patients in comparison with the TNM 6th edition.
Method
Clinical data of 2,629 NPC patients from the Sun Yat-sen University Cancer Center between January 2006 and December 2010 were retrospectively collected and all the patients were restaged according to the criteria of the TNM 6th edition and TNM 7th edition UICC/AJCC staging manual. Univariate and multivariate COX proportional hazards analyses were applied to evaluate the prognostic values between adjacent stage categories of the TNM 6th edition and TNM 7th edition.
Results
In comparison with the TNM 6th edition, a significant alteration of the distribution of N categories was observed when the TNM 7th edition was applied (χ2 = 20.589, P<0.001), with 119 (119/670, 17.8%) patients up-staging from N0 to N1. With regard to T and overall stage, 37 (37/561, 6.6%) patients were down-staged from T2a with the TNM 6th edition to T1 with the TNM 7th edition, and finally two patients were up-staged to overall stage II (2/118, 1.7%). Moreover, the survival curves were significantly segregated (P<0.05) between T1 and T2 as well as N1 and N2 with the TNM 7th edition.
Conclusions
The TNM 7th edition led to a significant alteration in the distribution of N categories and it is superior to the TNM 6th edition in predicting the frequency of overall survival and distant metastasis-free survival.
doi:10.1371/journal.pone.0116261
PMCID: PMC4275293  PMID: 25536307
6.  Robust antiferromagnetism preventing superconductivity in pressurized (Ba0.61K0.39)Mn2Bi2 
Scientific Reports  2014;4:7342.
BaMn2Bi2 possesses an iso-structure of iron pnictide superconductors and similar antiferromagnetic (AFM) ground state to that of cuprates, therefore, it receives much more attention on its properties and is expected to be the parent compound of a new family of superconductors. When doped with potassium (K), BaMn2Bi2 undergoes a transition from an AFM insulator to an AFM metal. Consequently, it is of great interest to suppress the AFM order in the K-doped BaMn2Bi2 with the aim of exploring the potential superconductivity. Here, we report that external pressure up to 35.6 GPa cannot suppress the AFM order in the K-doped BaMn2Bi2 to develop superconductivity in the temperature range of 300 K–1.5 K, but induces a tetragonal (T) to an orthorhombic (OR) phase transition at ~20 GPa. Theoretical calculations for the T and OR phases, on basis of our high-pressure XRD data, indicate that the AFM order is robust in the pressurized Ba0.61K0.39Mn2Bi2. Both of our experimental and theoretical results suggest that the robust AFM order essentially prevents the emergence of superconductivity.
doi:10.1038/srep07342
PMCID: PMC4256658  PMID: 25475224
7.  Genetics, Receptor Binding Property, and Transmissibility in Mammals of Naturally Isolated H9N2 Avian Influenza Viruses 
PLoS Pathogens  2014;10(11):e1004508.
H9N2 subtype influenza viruses have been detected in different species of wild birds and domestic poultry in many countries for several decades. Because these viruses are of low pathogenicity in poultry, their eradication is not a priority for animal disease control in many countries, which has allowed them to continue to evolve and spread. Here, we characterized the genetic variation, receptor-binding specificity, replication capability, and transmission in mammals of a series of H9N2 influenza viruses that were detected in live poultry markets in southern China between 2009 and 2013. Thirty-five viruses represented 17 genotypes on the basis of genomic diversity, and one specific “internal-gene-combination” predominated among the H9N2 viruses. This gene combination was also present in the H7N9 and H10N8 viruses that have infected humans in China. All of the 35 viruses preferentially bound to the human-like receptor, although two also retained the ability to bind to the avian-like receptor. Six of nine viruses tested were transmissible in ferrets by respiratory droplet; two were highly transmissible. Some H9N2 viruses readily acquired the 627K or 701N mutation in their PB2 gene upon infection of ferrets, further enhancing their virulence and transmission in mammals. Our study indicates that the widespread dissemination of H9N2 viruses poses a threat to human health not only because of the potential of these viruses to cause an influenza pandemic, but also because they can function as “vehicles” to deliver different subtypes of influenza viruses from avian species to humans.
Author Summary
Avian influenza viruses continue to present challenges to human health. Recently the H7N9 and H10N8 viruses that are of low pathogenicity for poultry have caused human infections and deaths in China. H9N2 influenza virus have been isolated worldwide from wild and domestic avian species for several decades, and their low pathogenic nature to poultry made them a low priority for animal disease control, which has allowed them to continue to evolve and spread. Here, we investigated a series of H9N2 influenza viruses that were detected in live poultry markets in southern China. We found that these viruses are able to preferentially bind to the human-type receptor, and some of them can cause disease and transmit between ferrets by respiratory droplet. All the transmissible H9N2 viruses have a similar internal gene constellation, which was also present in the H7N9 and H10N8 viruses. Our study indicates that the widespread dissemination of H9N2 viruses poses a threat to human health not only because of the potential of these viruses to cause an influenza pandemic, but also because they can function as “vehicles” to deliver different subtypes of influenza viruses from avian species to humans.
doi:10.1371/journal.ppat.1004508
PMCID: PMC4239090  PMID: 25411973
8.  Repeated measures study of weekly and daily cytomegalovirus shedding patterns in saliva and urine of healthy cytomegalovirus-seropositive children 
BMC Infectious Diseases  2014;14(1):569.
Background
To better understand potential transmission risks from contact with the body fluids of children, we monitored the presence and amount of CMV shedding over time in healthy CMV-seropositive children.
Methods
Through screening we identified 36 children from the Atlanta, Georgia area who were CMV-seropositive, including 23 who were shedding CMV at the time of screening. Each child received 12 weekly in-home visits at which field workers collected saliva and urine. During the final two weeks, parents also collected saliva and urine daily.
Results
Prevalence of shedding was highly correlated with initial shedding status: children shedding at the screening visit had CMV DNA in 84% of follow-up saliva specimens (455/543) and 28% of follow-up urine specimens (151/539); those not shedding at the screening visit had CMV DNA in 16% of follow-up saliva specimens (47/303) and 5% of follow-up urine specimens (16/305). Among positive specimens we found median viral loads of 82,900 copies/mL in saliva and 34,730 copies/mL in urine (P = 0.01), while the viral load for the 75th percentile was nearly 1.5 million copies/mL for saliva compared to 86,800 copies/mL for urine. Younger age was significantly associated with higher viral loads, especially for saliva (P < 0.001). Shedding prevalence and viral loads were relatively stable over time. All children who were shedding at the screening visit were still shedding at least some days during weeks 11 and 12, and median and mean viral loads did not change substantially over time.
Conclusions
Healthy CMV-seropositive children can shed CMV for months at high, relatively stable levels. These data suggest that behavioral prevention messages need to address transmission via both saliva and urine, but also need to be informed by the potentially higher risks posed by saliva and by exposures to younger children.
doi:10.1186/s12879-014-0569-1
PMCID: PMC4240830  PMID: 25391640
Cytomegalovirus; Congenital; Awareness; Behavior; Pregnancy
9.  Role of the upper and lowest instrumented vertebrae in predicting the postoperative coronal balance in Lenke 5C patients after selective posterior fusion 
European Spine Journal  2013;22(11):2392-2398.
Purpose
To investigate which pre- and postoperative radiographic parameters are significantly correlated with the immediate postoperative coronal balance (CB) in Lenke 5C AIS patients, and to identify any radiographic parameter that is correlated with the ultimate CB at a minimum follow-up of 2 years.
Methods
Forty Lenke 5C AIS patients were recruited in the current study. Preoperative, immediate postoperative, and the latest follow-up radiographs were reviewed measuring various radiographic parameters related to UIV and LIV. The correlations between different parameters and CB were then studied.
Results
The average follow-up time was 35.2 months. Correlation analysis showed that the following radiographic parameters significantly associated with the immediate postoperative CB in Lenke 5C patients: preoperative CB (r = 0.66, p < 0.01), preoperative UIV translation (r = 0.61, p < 0.01), preoperative LIV tilt (r = 0.61, p < 0.01), postoperative UIV translation (r = 0.51, p < 0.05), and postoperative LIV tilt (r = 0.50, p < 0.05). At the last follow-up, only the final UIV tilt was inversely correlated with the ultimate CB (r = −0.58, p < 0.05). Seven patients presented with coronal imbalance immediately after surgery. However, only one of the seven patients presented with coronal imbalance at the last follow-up, and the other six achieved C7PL–CSVL distance within 10 mm.
Conclusions
In Lenke 5C patients, preoperative UIV translation and LIV tilt are two important parameters that can predict the immediate postoperative CB. During the postoperative follow-up, UIV tilt may play a very important role in compensating for postoperative coronal imbalance.
doi:10.1007/s00586-013-2808-3
PMCID: PMC3886519  PMID: 23708938
Idiopathic scoliosis; Lenke type 5 curve; Coronal balance; Lowest instrumented vertebra; Upper instrumented vertebra
10.  High-Resolution Mechanical Imaging of Glioblastoma by Multifrequency Magnetic Resonance Elastography 
PLoS ONE  2014;9(10):e110588.
Objective
To generate high-resolution maps of the viscoelastic properties of human brain parenchyma for presurgical quantitative assessment in glioblastoma (GB).
Methods
Twenty-two GB patients underwent routine presurgical work-up supplemented by additional multifrequency magnetic resonance elastography. Two three-dimensional viscoelastic parameter maps, magnitude |G*|, and phase angle φ of the complex shear modulus were reconstructed by inversion of full wave field data in 2-mm isotropic resolution at seven harmonic drive frequencies ranging from 30 to 60 Hz.
Results
Mechanical brain maps confirmed that GB are composed of stiff and soft compartments, resulting in high intratumor heterogeneity. GB could be easily differentiated from healthy reference tissue by their reduced viscous behavior quantified by φ (0.37±0.08 vs. 0.58±0.07). |G*|, which in solids more relates to the material's stiffness, was significantly reduced in GB with a mean value of 1.32±0.26 kPa compared to 1.54±0.27 kPa in healthy tissue (P = 0.001). However, some GB (5 of 22) showed increased stiffness.
Conclusion
GB are generally less viscous and softer than healthy brain parenchyma. Unrelated to the morphology-based contrast of standard magnetic resonance imaging, elastography provides an entirely new neuroradiological marker and contrast related to the biomechanical properties of tumors.
doi:10.1371/journal.pone.0110588
PMCID: PMC4206430  PMID: 25338072
11.  Experiences of Stigma and Discrimination among Caregivers of Persons with Schizophrenia in China: A Field Survey 
PLoS ONE  2014;9(9):e108527.
In China, caregivers for family members with schizophrenia play an important role in treatment and recovery but may experience stigma and discrimination simply because of their family relationship. The object of this study was to measure the degrees and correlates of stigma and discrimination experiences among this group. Four hundred twenty-seven caregivers participated in this hospital-based and cross-sectional study in Ningbo and Guangzhou, China. Data were collected by trained interviewers using fixed questionnaires. Stigma and discrimination experiences were measured by the Modified Consumer Experiences of Stigma Questionnaire (MCESQ). Caregivers’ social support was measured by the Social Support Rating Scale. Parametric analysis, nonparametric analysis and multivariate linear regression were used. The mean (SD) score of MCESQ was 2.44(0.45), 2.91(0.71) for stigma experiences and 1.97(0.37) for discrimination experiences on a five-point score (“1 = never” and “5 = very often”). Approximately 65% of caregivers reported that they tried to conceal their family members’ illness, and 71% lacked the support of friends. The experience of stigma was significantly negatively associated with the perceived social support of caregivers (standard β = −0.2,p<0.001). Caregivers who were children of the patients experienced fewer stigmas than other (standard β = −0.18, p<0.001). Urban residence (standard β = −0.12, p<0.01) and patients did not complete primary school education (standard β = −0.13, p<0.01) were negatively related with stigmas. In addition, stigma and discrimination was more experienced in Zhejiang than in Guangdong (p<0.05). In conclusion, this study performed that caregivers of people with schizophrenia in China experienced general stigmas and rare discrimination and found the relations with social support, kinship, patient’s educational level and regional differences. More interventions and supports should been given to caregivers who are lack of social support, who live in rural area and who are the patients’ parents, spouses or siblings.
doi:10.1371/journal.pone.0108527
PMCID: PMC4178170  PMID: 25259732
12.  Functional gene arrays-based analysis of fecal microbiomes in patients with liver cirrhosis 
BMC Genomics  2014;15(1):753.
Background
Human gut microbiota plays an important role in the pathogenesis of cirrhosis complications. Although the phylogenetic diversity of intestinal microbiota in patients with liver cirrhosis has been examined in several studies, little is known about their functional composition and structure.
Results
To characterize the functional gene diversity of the gut microbiome in cirrhotic patients, we recruited a total of 42 individuals, 12 alcoholic cirrhosis patients, 18 hepatitis B virus (HBV)-related cirrhosis patients, and 12 normal controls. We determined the functional structure of these samples using a specific functional gene array, which is a combination of GeoChip for monitoring biogeochemical processes and HuMiChip specifically designed for analyzing human microbiomes. Our experimental data showed that the microbial community functional composition and structure were dramatically distinctive in the alcoholic cirrhosis. Various microbial functional genes involved in organic remediation, stress response, antibiotic resistance, metal resistance, and virulence were highly enriched in the alcoholic cirrhosis group compared to the control group and HBV-related cirrhosis group. Cirrhosis may have distinct influences on metabolic potential of fecal microbial communities. The abundance of functional genes relevant to nutrient metabolism, including amino acid metabolism, lipid metabolism, nucleotide metabolism, and isoprenoid biosynthesis, were significantly decreased in both alcoholic cirrhosis group and HBV-related cirrhosis group. Significant correlations were observed between functional gene abundances and Child-Pugh scores, such as those encoding aspartate-ammonia ligase, transaldolase, adenylosuccinate synthetase and IMP dehydrogenase.
Conclusions
Functional gene array was utilized to study the gut microbiome in alcoholic and HBV-related cirrhosis patients and controls in this study. Our array data indicated that the functional composition of fecal microbiomes was heavily influenced by cirrhosis, especially by alcoholic cirrhosis. This study provides new insights into the functional potentials and activity of gut microbiota in cirrhotic patients with different etiologies.
Electronic supplementary material
The online version of this article (doi:10.1186/1471-2164-15-753) contains supplementary material, which is available to authorized users.
doi:10.1186/1471-2164-15-753
PMCID: PMC4171554  PMID: 25179593
End-stage liver disease; Intestines; Microbial communities; Alcohol; Microarray
13.  Prediction and functional analysis of the sweet orange protein-protein interaction network 
BMC Plant Biology  2014;14:213.
Background
Sweet orange (Citrus sinensis) is one of the most important fruits world-wide. Because it is a woody plant with a long growth cycle, genetic studies of sweet orange are lagging behind those of other species.
Results
In this analysis, we employed ortholog identification and domain combination methods to predict the protein-protein interaction (PPI) network for sweet orange. The K-nearest neighbors (KNN) classification method was used to verify and filter the network. The final predicted PPI network, CitrusNet, contained 8,195 proteins with 124,491 interactions. The quality of CitrusNet was evaluated using gene ontology (GO) and Mapman annotations, which confirmed the reliability of the network. In addition, we calculated the expression difference of interacting genes (EDI) in CitrusNet using RNA-seq data from four sweet orange tissues, and also analyzed the EDI distribution and variation in different sub-networks.
Conclusions
Gene expression in CitrusNet has significant modular features. Target of rapamycin (TOR) protein served as the central node of the hormone-signaling sub-network. All evidence supported the idea that TOR can integrate various hormone signals and affect plant growth. CitrusNet provides valuable resources for the study of biological functions in sweet orange.
doi:10.1186/s12870-014-0213-7
PMCID: PMC4236729  PMID: 25091279
Protein-protein interaction; Ortholog; Domain; Modular; Plant hormone
14.  Sanmiao formula inhibits chondrocyte apoptosis and cartilage matrix degradation in a rat model of osteoarthritis 
Sanmiao formula (SM) is a basic prescription for the treatment of gouty and rheumatoid arthritis that has been used in China over a long period of history. However, there is no evidence associating SM with the treatment of osteoarthritis (OA). In this study, a characterization of the anti-OA effect of SM was conducted using an in vivo rat model induced by anterior cruciate ligament transection and medial meniscus resection (ACLT plus MMx), together with in vitro studies using chondrocytes for further molecular characterization. Rats subjected to ACLT plus MMx were treated with SM at doses of 0.63, 1.25 and 2.5 g/kg per day for three or six weeks. SM treatment significantly inhibited the histopathological changes of articular cartilage damage and synovial inflammation in the rats following ACLT plus MMx. SM (2.5 g/kg) clearly inhibited chondrocyte apoptosis and prevented cartilage matrix degradation, which was indicated by the increased proteoglycan and collagen content, particularly with regard to type II collagen expression in articular cartilage. Furthermore, SM (2.5 g/kg) markedly inhibited the release of interleukin (IL)-1β, tumor necrosis factor-α and nitric oxide in serum, while simultaneously increasing the levels of bone morphogenetic protein-2 and transforming growth factor-β in the circulation. Notably, SM (2.5 g/kg) clearly attenuated the OA-augmented expression of matrix metalloproteinase (MMP)-13 and augmented the OA-reduced expression of tissue inhibitor of metalloproteinase (TIMP)-1 in the knee joints. In addition, SM significantly reduced the proportion of early and late apoptotic and sub-G1 phase cells, and clearly decreased the expression of MMP-13 and increased that of TIMP-1 at the mRNA and protein levels in IL-1β-induced chondrocytes. These findings provide the first evidence that SM effectively treats OA by inhibiting chondrocyte apoptosis, cartilage matrix degradation and the inflammatory response.
doi:10.3892/etm.2014.1862
PMCID: PMC4151698  PMID: 25187798
Sanmiao formula; chondrocytes; proteoglycan; collagen; inflammatory cytokines; metalloproteinases; tissue inhibitors of metalloproteinase
15.  MicroRNA-132 silencing decreases the spontaneous recurrent seizures 
Objective: This study aimed to investigate the role of microRNA-132 in the epileptogenesis. Methods: Antagomir-132 (Ant-132) was used to silence the expression of miR-132 and non-targeting scrambled sequence (Scr) as a control. Rats were randomly divided into ant-132 group and Scr group in which rats were pretreated with An-132 and Scr, respectively, and then induced temporal lobe epilepsy (TLE) by Li-Pilo. Behavioral observation was done, and results showed the changes in spontaneous recurrent seizures in the chronic phase between two groups. Bax and Bcl-2 were detected aiming to evaluate the neuronal apoptosis. NPY staining was done to investigate the mossy fiber sprouting (MFS). Golgi staining was used to assess the changes in the dendritic morphology. Results: Our study showed that ant-132 induced miR-132 silencing in rats could increase the on-set epilepsy threshold and suppress the numbers of spontaneous recurrent seizures. The number of apoptotic neurons and MFS reduced after miR-132 silencing. In addition, the dendrites of neurons were highly suppressed in the CA3 region of the hippocampus. Conclusions: miR-132 silencing suppresses the spontaneous seizures. The better outcome may result from the neuroprotective effect and the inhibition of MFs-CA3 pathway following miR-132 silencing. Thus, miR-132 may serve as a potential target for the development of anti-epileptic drugs.
PMCID: PMC4132124  PMID: 25126160
Temporal lobe epilepsy; microRNA-132; antagomirs; mossy fber sprouting
16.  Syndrome Differentiation of Diabetes by the Traditional Chinese Medicine according to Evidence-Based Medicine and Expert Consensus Opinion 
In Chinese medicine, diabetes belongs to the category of “Xiaoke disease (disease with symptoms of frequent drinking and urination)”; in the traditional sense, its pathogenesis is “Yin deficiency and dryness-heat.” However, over time, changes in the social environment and lifestyle have also changed the use of traditional Chinese medicine (TCM) in diabetes. In this study, we performed diabetes syndrome differentiation using TCM according to evidence-based medicine and expert consensus opinion.
doi:10.1155/2014/492193
PMCID: PMC4123514  PMID: 25132859
17.  Naringin administration inhibits platelet aggregation and release by reducing blood cholesterol levels and the cytosolic free calcium concentration in hyperlipidemic rabbits 
This study investigated the effects of naringin on platelet aggregation and release in hyperlipidemic rabbits, and the underlying mechanisms. The safety of naringin was also investigated. The rabbits were orally administered 60, 30 or 15 mg/kg of naringin once a day for 14 days after being fed a high fat/cholesterol diet for four weeks. Following the two weeks of drug administration, the degree of platelet aggregation induced by arachidonic acid, adenosine diphosphate and collagen was significantly reduced by naringin at certain doses compared with those in the rabbits of the model group (P<0.01). The levels of P-selectin and platelet factor 4 (PF4) also decreased following treatment with naringin compared with those of the model group. Certain doses of naringin significantly reduced the total cholesterol (TC) levels and elevated the ratio of high-density lipoprotein cholesterol to TC compared with those in the model group, and significantly decreased the cytosolic free calcium concentration ([Ca2+]i). No significant difference in the coagulation function was observed between the control and drug-treatment groups. These results indicate that naringin improved platelet aggregation and inhibited the excessive release of P-selectin and PF4 in hyperlipidemic rabbits. This study suggests that the antiplatelet effect of naringin may be due to its ability to regulate the levels of blood cholesterol and [Ca2+]i in platelets. Naringin also did not cause bleeding in the hyperlipidemic rabbits.
doi:10.3892/etm.2014.1794
PMCID: PMC4113534  PMID: 25120631
naringin; hyperlipidemic; aggregation; cytosolic free calcium concentration; P-selectin; platelet factor 4
18.  The Routine Utilization of Dental Care during Pregnancy in Eastern China and the Key Underlying Factors: A Hangzhou City Study 
PLoS ONE  2014;9(6):e98780.
Objectives
Oral diseases are associated with adverse pregnancy outcomes. The routine utilization of dental care (RUDC) during pregnancy is an effective way to improve pregnant women’s oral health, and thus safeguard the health of their babies. As China has one fifth of the world’s population, it is especially meaningful to encourage RUDC there. However, the status of RUDC in China and the key underlying factors are largely unknown.
Methods
This cross-sectional survey investigated the current status of RUDC during pregnancy and the key underlying factors in Hangzhou City, Zhejiang Province, eastern China. We collected participants’ demographics, individual oral-hygiene behaviors, individual lifestyle, oral-health conditions and attitudes, and also their RUDC during pregnancy. Binary Logistic Regression Analysis was used to analyze the key underlying factors.
Results
Only 16.70% of the participants reported RUDC during pregnancy. The percentage of RUDC was significantly lower among pregnant women with the following characteristics: aged 30 or less, an annual household income under $8,000, brushing once a day or less, never flossing or rinsing the mouth, paying no attention to pregnancy-related oral-health knowledge, and being dissatisfied with one’s individual dental hygiene behavior.
Conclusions
RUDC during pregnancy is very low in eastern China and is greatly influenced not only by a woman’s age, annual income, individual hygiene behavior, but also by her attention and attitudes to oral health. To improve this population’s access to and use of dental care during pregnancy, appropriate programs and policies are urgently needed.
doi:10.1371/journal.pone.0098780
PMCID: PMC4046992  PMID: 24901520
19.  BMP-Functionalised Coatings to Promote Osteogenesis for Orthopaedic Implants 
The loss of bone integrity can significantly compromise the aesthetics and mobility of patients and can be treated using orthopaedic implants. Over the past decades; various orthopaedic implants; such as allografts; xenografts and synthetic materials; have been developed and widely used in clinical practice. However; most of these materials lack intrinsic osteoinductivity and thus cannot induce bone formation. Consequently; osteoinductive functionalisation of orthopaedic implants is needed to promote local osteogenesis and implant osteointegration. For this purpose; bone morphogenetic protein (BMP)-functionalised coatings have proven to be a simple and effective strategy. In this review; we summarise the current knowledge and recent advances regardingBMP-functionalised coatings for orthopaedic implants.
doi:10.3390/ijms150610150
PMCID: PMC4100145  PMID: 24914764
bone morphogenetic protein; coating; orthopaedic implant; osteogenesis
20.  Expression of p-CREB and activity-dependent miR-132 in temporal lobe epilepsy 
Objective: The mechanisms underlying the pathogenesis of intractable epilepsy (IE) are still unclear. This study aimed to investigate the role of phosphorylation of Cyclase Response Element Binding Protein (p-CREB)/microRNA-132 (miR-132) signal pathway in the epileptogenesis. Methods: Temporal lobe epilepsy (TLE) was induced in rodents, and the brain tissues were collected. P-CREB expression was detected by Western blot assay and immunochemistry at 6 h, 24 h, 3 d, 7 d, 14 d, 30 d and 60 d after induced status epilepsy (SE), respectively, and in patients with TLE. The expression of miR-132 was detected by RT-PCR. Results: The p-CREB and miR-132 were highly expressed in both rats and patients with TLE as compared to controls. Conclusions: The expression of p-CREB and miR-132 increases in epilepsy rats and patients, suggesting a possible pathogenetic role of p-CREB and miR-132 in TLE via modulating the dendritic plasticity.
PMCID: PMC4073747  PMID: 24995086
Temporal lobe epilepsy; phosphorylation of cyclase response element binding protein; microRNA-132; mossy fiber sprouting
21.  The value of Apolipoprotein B/Apolipoprotein A1 ratio for metabolic syndrome diagnosis in a Chinese population: a cross-sectional study 
Background
The apoB/apoA1 ratio has been reported to be associated with the metabolic syndrome (MetS), and it may be a more convenient biomarker in MetS predicting. However, whether apoB/apoA1 ratio is a better indicator of metabolic syndrome than other biomarkers and what is the optimal cut-off value of apoB/apoA1 ratio as an indicator of metabolic syndrome in Chinese population remain unknown. Thus, we carried out the current study to assess the predictive value of apoB/apoA1 ratio and determine the optimal cut-off value of apoB/apoA1 ratio for diagnosing MetS in a Chinese population.
Method
We selected 1,855 subjects with MetS and 6,265 individuals without MetS based on the inclusion and exclusion criteria from the China Health Nutrition Survey (CHNS) in 2009. MetS was identified based on the diagnostic criteria of International Diabetes Federation (2005). Logistic regression was used to estimate the association between the apoB/apoA1 ratio and risk of MetS, and receiver operating characteristics (ROC) curve analysis was performed to test the predictive value of apoB/apoA1 ratio and calculate the appropriate cut-off value.
Results
Compared with the lowest quartile of apoB/apoA1 ratio, subjects in the fourth quartile had a higher risk of MetS in both men [odds ratio (OR) = 2.64, 95% confidence interval (CI) =1.82-3.83] and women (OR = 5.18, 95% CI = 3.87-6.92) after adjustment for potential confounders. The optimal cut-off value of apoB/apoA1 ratio for MetS detection was 0.85 in men and 0.80 in women. Comparisons of ROC curves indicated that apoB/apoA1 ratio was better than traditional biomarkers in predicting MetS.
Conclusion
Our results suggest that, apoB/apoA1 ratio has a promising predictive effectiveness in detection of MetS. An apoB/apoA1 ratio higher than 0.85 in men and 0.80 in women may be a promising and convenient marker of MetS.
doi:10.1186/1476-511X-13-81
PMCID: PMC4041140  PMID: 24886173
Metabolic syndrome; Apolipoprotein ratio; ROC curve
22.  Knowledge of, attitudes towards, and practice relating to schistosomiasis in two subtypes of a mountainous region of the People’s Republic of China 
Background
Schistosomiasis japonica is still endemic in the People’s Republic of China (P.R. China) in five provinces of lake and marshland regions and in two provinces of mountainous regions. Studies elucidated that individual and community perception, attitudes towards schistosomiasis, and hygiene behaviors were crucial factors for preventing schistosomiasis. This study sought to assess the knowledge of, attitudes towards, and practices (KAP) relating to schistosomiasis in two subtypes of a mountainous region in Eryuan County, Yunnan Province, P.R. China. The study’s aim is to make suggestions for establishing more specific and effective control measures for disease transmission and interruption in two subtypes of a mountainous region with low-level infection rates.
Methods
A cross-sectional study of 3,000 inhabitants was carried out in the Yongle (plateau basin) and Xinzhuang (plateau canyon) communities of Eryuan County, Yunnan Province in November and December 2011. Stratified cluster random sampling was undertaken using a uniform set of quantitative questionnaires administered by trained assistants. This was further supported with qualitative data from in-depth interviews (IDIs) conducted with ten farmers and ten students. All participants were examined for schistosomiasis using both a serological test (indirect hemagglutination assay [IHA]) and a stool examination (Kato-Katz).
Results
The total schistosomiasis knowledge rate in Yongle (83.4%) was significantly lower than that in Xinzhuang (95.5%). In both communities, among the respondents aged 15 years or below, more than one third didn’t know the name, endemic areas, and animal reservoirs of schistosomiasis. The majority of respondents in Eryuan acquired their schistosomiasis knowledge from doctors, followed by handouts and hearing from others. The infection rate was once the highest in Yongle, but is now the highest in Xinzhuang, where there are more risk factors for schistosomiasis, such as frequently grazing cattle, digging vegetables or cutting grass in the field, as well as raising cattle by free grazing.
Conclusion
In short, Eryuan County’s overall knowledge rate of schistosomiasis was found to be high. Due to various dominating risk factors, different control strategies should be designed keeping in mind the two different subtypes of endemic areas for schistosomiasis in mountainous regions, namely plateau basins and plateau canyons.
doi:10.1186/2049-9957-3-16
PMCID: PMC4064289  PMID: 24955240
Schistosomiasis; Knowledge; Attitude; Practice; Eryuan; P.R. China
23.  Cell-to-cell transmission of pathogenic proteins in neurodegenerative diseases 
Nature medicine  2014;20(2):130-138.
A common feature of many neurodegenerative diseases is the deposition of β-sheet-rich amyloid aggregates formed by proteins specific to these diseases. These protein aggregates are thought to cause neuronal dysfunction, directly or indirectly. Recent studies have strongly implicated cell-to-cell transmission of misfolded proteins as a common mechanism for the onset and progression of various neurodegenerative disorders. Emerging evidence also suggests the presence of conformationally diverse ‘strains’ of each type of disease protein, which may be another shared feature of amyloid aggregates, accounting for the tremendous heterogeneity within each type of neurodegenerative disease. Although there are many more questions to be answered, these studies have opened up new avenues for therapeutic interventions in neurodegenerative disorders.
doi:10.1038/nm.3457
PMCID: PMC4011661  PMID: 24504409
24.  Public Health Workers and Vaccination Coverage in Eastern China: A Health Economic Analysis 
Background: Vaccine-preventable diseases cause more than one million deaths among children under 5 years of age every year. Public Health Workers (PHWs) are needed to provide immunization services, but the role of human resources for public health as a determinant of vaccination coverage at the population level has not been assessed in China. The objective of this study was to test whether PHW density was positively associated with childhood vaccination coverage in Zhejiang Province, East China. Methods: The vaccination coverage rates of Measles Containing Vaccine (MCV), Diphtheria, Tetanus and Pertussis combined vaccine (DTP), and Poliomyelitis Vaccine (PV) were chosen as the dependent variables. Vaccination coverage data of children aged 13–24 months for each county in Zhejiang Province were taken from the Zhejiang Immunization Information System (ZJIIS). Aggregate PHW density was an independent variable in one set of regressions, and Vaccine Personnel (VP) and other PHW densities were used separately in another set. Data on densities of PHW and VP were taken from a national investigation on EPI launched by Ministry of Health of China in 2013. We controlled other determinants that may influence the vaccination coverage like Gross Domestic Product (GDP) per person, proportion of migrant children aged <7 years, and land area. These data were taken from Zhejiang Provincial Bureau of Statistics and ZJIIS. Results: PHW density was significantly influence the coverage rates of MCV [Adjusted Odds Ratio(AOR) = 4.29], DTP3(AOR = 2.16), and PV3 (AOR = 3.30). However, when the effects of VPs and other PHWs were assessed separately, we found that VP density was significantly associated with coverage of all three vaccinations (MCV AOR = 7.05; DTP3 AOR = 1.82; PV3 AOR = 4.83), while other PHW density was not. Proportion of migrant children < 7 years and Land area were found as negative and significant determinants for vaccination coverage, while GDP per person had no effect on vaccination coverage. Conclusions: A higher density of PHWs (VP) would improve the availability of immunization services over time and space, which may increase the possibility of achieving a higher childhood vaccination coverage rate. It was indicated that the level of GDP per person had no association with the improved vaccination coverage after controlling for other potential factors. Our findings implicated that PHW density was a major constraint on immunization coverage in Zhejiang Province.
doi:10.3390/ijerph110505555
PMCID: PMC4053906  PMID: 24859680
public health worker; vaccination personnel; immunization; vaccination coverage; determinants
25.  Bisphenol AF-Induced Endogenous Transcription Is Mediated by ERα and ERK1/2 Activation in Human Breast Cancer Cells 
PLoS ONE  2014;9(4):e94725.
Bisphenol AF (BPAF)-induced transcriptional activity has been evaluated by luciferase reporter assay. However, the molecular mechanism of BPAF-induced endogenous transcription in human breast cancer cells has not been fully elucidated. In the present study, we investigated the effect and mechanism of BPAF-induced endogenous transcription detected by real-time PCR in human breast cancer cells. We found that BPAF stimulated transcription of estrogen responsive genes, such as trefoil factor 1 (TFF1), growth regulation by estrogen in breast cancer 1 (GREB1) and cathepsin D (CTSD), through dose-dependent and time-dependent manners in T47D and MCF7 cells. Gene-silencing of ERα, ERβ and G protein-coupled estrogen receptor 1 (GPER) by small interfering RNA revealed that BPAF-induced endogenous transcription was dependent on ERα and GPER, implying both genomic and nongenomic pathways might be involved in the endogenous transcription induced by BPAF. ERα-mediated gene transcription was further confirmed by inhibition of ER activity using ICI 182780 in ERα-positive T47D and MCF7 cells as well as overexpression of ERα in ERα-negative MDA-MB-231 breast cancer cells. Moreover, we utilized Src tyrosine kinase inhibitor PP2 and two MEK inhibitors PD98059 and U0126 to elucidate the rapid nongenomic activation of Src/MEK/ERK1/2 cascade on endogenous transcription. Our data showed that BPAF-induced transcription could be significantly blocked by PP2, PD98059 and U0126, suggesting activation of ERK1/2 was also required to regulate endogenous transcription. Taken together, these results indicate that BPAF-induced endogenous transcription of estrogen responsive genes is mediated through both genomic and nongenomic pathways involving the ERα and ERK1/2 activation in human breast cancer cells.
doi:10.1371/journal.pone.0094725
PMCID: PMC3984236  PMID: 24727858

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