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1.  Pirt contributes to uterine contraction-induced pain in mice 
Molecular Pain  2015;11:57.
Uterine contraction-induced pain (UCP) represents a common and severe form of visceral pain. Nerve fibers that innervate uterine tissue express the transient receptor potential vanilloid channel 1 (TRPV1), which has been shown to be involved in the perception of UCP. The phosphoinositide-interacting regulator of TRP (Pirt) may act as a regulatory subunit of TRPV1. The intraperitoneal injection of oxytocin into female mice after a 6-day priming treatment with estradiol benzoate induces writhing responses, which reflect the presence of UCP. Here, we first compared writhing response between Pirt+/+ and Pirt−/− mice. Second, we examined the innervation of Pirt-expressing nerves in the uterus of Pirt−/− mice by immunofluorescence and two-photon microscopy. Third, we identified the soma of dorsal root ganglion (DRG) neurons that innerve the uterus using retrograde tracing and further characterized the neurochemical properties of these DRG neurons. Finally, we compared the calcium response of capsaicin between DRG neurons from Pirt+/+ and Pirt−/− mice. We found that the writhing responses were less intensive in Pirt−/− mice than in Pirt+/+ mice. We also observed Pirt-expressing nerve fibers in the myometrium of the uterus, and that retrograde-labeled cells were small-diameter, unmyelinated, and Pirt-positive DRG neurons. Additionally, we found that the number of capsaicin-responding neurons and the magnitude of evoked calcium response were markedly reduced in DRG neurons from Pirt−/− mice. Taken together, we speculate that Pirt plays an important role in mice uterine contraction-induced pain.
PMCID: PMC4574137  PMID: 26376721
Pirt; Uterine contraction-induced pain; Dorsal root ganglion; TRPV1
2.  Three-pathway combination for glutathione biosynthesis in Saccharomyces cerevisiae 
Glutathione (GSH), a pivotal non-protein thiol, can be biosynthesized through three pathways in different organisms: (1) two consecutive enzymatic reactions catalyzed by γ-glutamylcysteine synthetase (Gsh1 or GshA) and glutathione synthetase (Gsh2 or GshB); (2) a bifunctional γ-glutamylcysteine synthetase/glutathione synthetase (GshF); (3) an alternative condensation of γ-glutamyl phosphate synthesized by γ-glutamyl kinase (Pro1 or ProB) with cysteine to form γ-glutamylcysteine which was further conjugated to glycine by glutathione synthetase. The Gsh1 and Gsh2 of conventional GSH biosynthetic pathway or the bifunctional GshF reported previously have been independently modulated for GSH production. This study developed a novel three-pathway combination method to improve GSH production in Saccharomyces cerevisiae.
A bifunctional enzyme GshF of Actinobacillus pleuropneumoniae was functionally expressed in S. cerevisiae and Pro1 in proline biosynthetic pathway was exploited for improving GSH yield. Moreover, two fusion proteins Gsh2-Gsh1 and Pro1-GshB were constructed to increase the two-step coupling efficiency of GSH synthesis by mimicking the native domain fusion of GshF. The engineered strain W303-1b/FGP with three biosynthetic pathways presented the highest GSH concentration (216.50 mg/L) and GSH production of W303-1b/FGP was further improved by 61.37 % when amino acid precursors (5 mM glutamic acid, 5 mM cysteine and 5 mM glycine) were fed in shake flask cultures. In batch culture process, the recombinant strain W303-1b/FGP also kept high efficiency in GSH production and reached an intracellular GSH content of 2.27 % after 24-h fermentation.
The engineered strains harbouring three GSH pathways displayed higher GSH producing capacity than those with individually modulated pathways. Three-pathway combinatorial biosynthesis of GSH promises more effective industrial production of GSH using S. cerevisiae.
Electronic supplementary material
The online version of this article (doi:10.1186/s12934-015-0327-0) contains supplementary material, which is available to authorized users.
PMCID: PMC4574134  PMID: 26377681
Glutathione; Three-pathway combination; Combinatorial biosynthesis; Saccharomyces cerevisiae
3.  The Canadian Assessment of Physical Literacy: methods for children in grades 4 to 6 (8 to 12 years) 
BMC Public Health  2015;15:767.
Physical literacy is described as the motivation, confidence, physical competence, knowledge and understanding to value and engage in a physically active lifestyle. As such, it is expected that those who have greater physical literacy would be more likely to obtain the health benefits offered by habitual physical activity. A theoretical model and assessment battery, the Canadian Assessment of Physical Literacy (CAPL), for the assessment of childhood physical literacy had been proposed in theory but validity data were lacking. The purpose of this study was to explore validity evidence for the CAPL among children in grades 4 to 6.
CAPL validity was evaluated through three analyses that utilized cross-sectional data obtained through local schools in Eastern Ontario, Canada. A confirmatory factor analysis compared the data to the theoretical model. Patterns of association between self-reported age and gender and the CAPL total and domain scores were examined using regression models. Teacher ratings of participants’ knowledge, attitude and physical activity competence were compared to assessment results.
The CAPL was completed by 963 children (55 % female) in grades 4, 5 and 6. Children were 8 to 12 years of age (mean 10.1 years), with 85 % of children approached agreeing to participate. A confirmatory factor analysis using data from 489 children with complete raw scores supported a model with four domains: engagement in physical activity (active and sedentary), physical competence (fitness and motor skill), motivation and confidence, and knowledge and understanding. Raw domain scores followed expected patterns for age and gender, providing evidence for their validity. Interpretive categories, developed from age and gender adjusted normative data, were not associated with age indicating that the CAPL is suitable for use across this age range. Children’s gender was associated with the physical competence, motivation and engagement in physical activity domain scores, indicating that further research is required regarding the gender adjustment of the raw CAPL scores. CAPL domain and total scores were statistically significantly associated with teacher ratings of the child’s motivation, attitudes, fitness, skill and overall physical activity.
CAPL offers a comprehensive assessment of engagement in physical activity, physical competence, motivation and confidence, and knowledge and understanding as components of childhood (grades 4 to 6, 8 to 12 years) physical literacy. Monitoring of these measures enhances our understanding of children’s physical literacy, and assists with the identification of areas where additional supports are required.
PMCID: PMC4532252  PMID: 26260572
4.  Smoking-related thoughts and microbehaviours and their predictive power for quitting: Findings from the International Tobacco Control (ITC) China Survey 
Tobacco control  2014;24(4):354-361.
Negative attitudes to smoking are well-established predictors of intentions to quit and quit behaviours, but less attention has been given to whether quitting is influenced by smoking-related thoughts and microbehaviours that reflect a concern about smoking.
This paper aimed to describe the occurrence of smoking-related thoughts and microbehaviours among Chinese smokers and to examine their predictive power for making quit attempts and sustained abstinence.
The data came from the first three waves of the International Tobacco Control China Survey. Four measures of recent thoughts about smoking and two microbehaviour measures (collectively referred to as micro indicators) were examined.
Most smokers (around three quarters) reported thinking about harms of smoking to themselves or to others at least occasionally, and an increasing minority reported the two microbehaviours of prematurely butting out cigarettes and forgoing them. All micro indicators were positively related to subsequent quit attempts in individual predictor analyses, but only serious thoughts about quitting and -butting out cigarettes had independent relationships. Overall, there was no clear relationship between these micro indicators and sustained abstinence.
There was a moderately high level of occurrence of recent smoking-related thoughts and microbehaviours among the Chinese adult smokers in the six cities studied. Like in the west, micro indicators of concern about smoking were positively associated with subsequent quit attempts, but unlike in the west, they were largely unrelated to sustained abstinence.
PMCID: PMC4532545  PMID: 24570098
health concern about smoking; micro-behaviour; smoking cessation; longitudinal research; China
5.  Keap1 modulates the redox cycle and hepatocyte cell cycle in regenerating liver 
Cell Cycle  2014;13(15):2349-2358.
Keap1 negatively controls the activity of transcription factor Nrf2. This Keap1/Nrf2 pathway plays a critical role in combating oxidative stress. We aimed at determining whether and how Keap1 modulates the cell cycle of replicating hepatocytes during liver regeneration. Two-thirds partial hepatectomy (PH) was performed on wild-type mice and Keap1+/− (Keap1 knockdown) mice. We found that, following PH, Keap1 knockdown resulted in a delay in S-phase entry, disruption of S-phase progression, and loss of mitotic rhythm of replicating hepatocytes. These events are associated with dysregulation of c-Met, EGFR, Akt1, p70S6K, Cyclin A2, and Cyclin B1 in regenerating livers. Astonishingly, normal regenerating livers exhibited the redox fluctuation coupled with hepatocyte cell cycle progression, while keeping Nrf2 quiescent. Keap1 knockdown caused severe disruption in both the redox cycle and the cell cycle of replicating hepatocytes. Thus, we demonstrate that Keap1 is a potent regulator of hepatic redox cycle and hepatocyte cell cycle during liver regeneration.
PMCID: PMC4128880  PMID: 25483186
Keap1; Nrf2; the redox cycle; the cell cycle; hepatocyte proliferation
6.  Bevacizumab promotes venous thromboembolism through the induction of PAI-1 in a mouse xenograft model of human lung carcinoma 
Molecular Cancer  2015;14:140.
An increased incidence of venous thromboembolism (VTE) is associated with anti-vascular endothelial growth factor (VEGF) treatment in cancer. However, the mechanism underlying this effect remains elusive. In this study, we examined the effect of bevacizumab, a humanized monoclonal antibody against VEGF-A, on VTE in a murine xenograft A549 cell tumor model.
Inferior vena cava stenosis model and FeCl3-induced saphenous vein thrombosis model were performed in a mouse xenograft models of human lung adenocarcinoma.
We found that treatment with bevacizumab significantly increased the thrombotic response to inferior vena cava obstruction and femoral vein injury. Plasminogen activator inhibitor (PAI-1) expression in tumors, plasma, and thrombi was significantly increased by bevacizumab. However, bevacizumab did not enhance VTE in PAI-1-deficient mice, suggesting that PAI-1 is a major mediator of bevacizumab’s prothrombotic effect. VEGF inhibited expression of PAI-1 by A549 cells, and this effect was neutralized by bevacizumab, suggesting that bevacizumab increases PAI-1 expression in vivo by blocking the inhibitory effect of VEGF on PAI-1 expression by tumor cells. Pharmacological inhibition of PAI-1 with PAI-039 blocked bevacizumab-induced venous thrombosis.
Collectively, these findings indicate that PAI-1 plays a role in VTE associated with antiangiogenic therapy and the inhibition of PAI-1 shows efficacy as a therapeutic strategy for the prevention of bevacizumab-associated VTE.
PMCID: PMC4517418  PMID: 26215730
Bevacizumab; Cancer; Plasminogen activator inhibitor 1; VEGF-A; Venous thromboembolism
7.  Reutericyclin producing Lactobacillus reuteri modulates development of fecal microbiota in weanling pigs 
Lactobacillus reuteri is used as probiotic culture in food and feed applications; however, strain specific properties of L. reuteri that mediate probiotic activity remain unknown. This study aimed to determine effects of feed fermentation with exopolysaccharide and reutericyclin producing L. reuteri on the transition of the gut microbiome of piglets after weaning. The reutericyclin and reuteran producing L. reuteri TMW1.656 was compared to the reutericyclin negative and levan producing L. reuteri LTH5794 and unfermented controls. Both strains were fermented at conditions supporting exopolysaccharide formation, or at conditions not supporting exopolysaccharide formation. Fecal microbiota were characterized by partial sequencing of 16S rRNA genes, and by quantitative PCR targeting clostridial toxins. The transition to solid food resulted in a transient increase of Proteobacteria to 12% of total bacteria, and increased bacterial diversity by increasing the abundance of anaerobic fiber fermenting Firmicutes. Three weeks after weaning, Prevotella and Lactobacillus were among the dominant bacterial genera. Feed fermentation with L. reuteri affected the abundance of few bacterial taxa and particularly reduced the abundance of Enterobacteriaceae (P < 0.05) when compared to unfermented controls. Reutericyclin producing L. reuteri increased the abundance of Dialister spp. and Mitsuokella spp. (P < 0.05) but did not influence the abundance of clostridial toxins in the feces. In conclusion, data on the contribution of specific metabolic activities of L. reuteri to probiotic activity will facilitate the strain selection for probiotic applications in food and feed.
PMCID: PMC4516970  PMID: 26284047
enterterotoxigenic Escherichia coli; ETEC; pigs; feed fermentation; reutericyclin; exopolysaccharides; Lactobacillus reuteri; probiotic
8.  Anesthesia management of surgery for sigmoid perforation and acute peritonitis patient following heart transplantation: case report 
Here we described a case in which a patient underwent emergency laparotomy for acute peritonitis and sigmoid perforation under general anesthesia with a history of heart transplantation. A good knowledge in the physiology of the transplanted heart is critical for effective and safe general anesthesia. We chose etomidate that have a weaker impact on cardiovascular function plus propofol for induction, and propofol plus cisatracurium for maintenance with intermittently analgesics and vasoactive drugs to facilitate the anesthesia. In addition, fluid input, electrolyte and acid-base balance were well adjusted during the whole procedure. The patient was in good condition after the surgery. In this case report we are aiming to provide some guidance for those scheduled for non-cardiac surgery after heart transplant.
PMCID: PMC4565380  PMID: 26379997
Anesthesia management; heart transplantation; sigmoid perforation; acute peritonitis patient
9.  The effects and mechanisms of SLC34A2 in tumorigenesis and progression of human non-small cell lung cancer 
SLC34A2 with highest expressions in lung, small intestine and kidney encoded a type 2b sodium-dependent phosphate transporter (NaPi-IIb). In lung, SLC34A2 only expressed in the apical membrane of type II alveolar epithelium cells (ATII cells) and played a pivotal role during the fetal lung development and embryonic development. ATII cells acting as multifunctional stem cells might transform into NSCLC after undergoing exogenous or endogenous factors. Increasing evidences showed that the genes performing critical roles during embryogenesis were also expressed during the development of cancer. In addition, recent research found the expression of SLC34A2 had a significant difference between the surgical samples of NSCLC and normal tissues, and SLC34A2 was down-regulated in lung adenocarcinoma cell line A549 and up-regulation expression of SLC34A2 could significantly inhibit cell viability and invasion of A549 in vitro. These results suggested SLC34A2 might play an important role in the development of NSCLC. However, the role of SLC34A2 in tumorigenesis and progression of NSCLC remains unknown.
Our study found that SLC34A2 was also significantly down-regulated in 14/15 of examined NSCLC tissues. Moreover, we found that expressions of SLC34A2 were reduced in six NSCLC cell lines for the first time. Our result also revealed a dramatic inhibitory effects of SLC34A2 on cell growth, migration and invasion of several NSCLC cell lines. SLC34A2 also strongly inhibited tumor growth and metastasis ability in A549 subcutaneous tumor model and lung metastasis model, respectively. Further studies found that the suppressive effects of SLC34A2 on tumorigenesis and progression might be associated with the down-regulation of related protein in PI3K/Akt and Ras/Raf/MEK signal pathway.
For the first time, our data indicated that SLC34A2 could exert significantly suppressive effects on tumorigenesis and progression of NSCLC. SLC34A2 might provide new insights for further understanding the early pathogenesis of human NSCLC.
PMCID: PMC4497375  PMID: 26156586
SLC34A2; ATII cells; NSCLC; Tumorigenesis and progression; Mechanism
10.  Hypoglycemic Effect of Ethanol and Ethyl Acetate Extract of Phellinus baumii Fruiting Body in Streptozotocin-Induced Diabetic Mice 
We investigated hypoglycemic effect of ethanol (EtOH) and ethyl acetate extract acetate (AcOEt) extracts in streptozotocin- (STZ-) induced diabetic mice. Our data showed the maximum inhibitory effect on the fasting plasma glucose (FPG) level was detected in STZ-induced diabetic mice administered with 400 mg/kg AcOEt extract of P. baumii. A lower glycated albumin (GA) level and a higher insulin level were observed in 400 mg/kg AcOEt and EtOH extract groups. Moreover, 400 mg/kg AcOEt and EtOH extract exhibited a stronger effect on increasing size and cell number of islets. The insulin expression level of β-cells and integrated optical density (IOD) value were significantly increased by the administration of 400 mg/kg AcOEt and EtOH extracts. Taken together, AcOEt and EtOH extracts of P. baumii fruiting body exhibited considerable hypoglycemic effect on STZ-induced diabetic mice.
PMCID: PMC4499395  PMID: 26221177
11.  Correction: The Interaction of the Atm Genotype with Inflammation and Oxidative Stress 
PLoS ONE  2015;10(6):e0130645.
PMCID: PMC4471270  PMID: 26083028
12.  Identification of a Novel Mutation in the COL2A1 Gene in a Chinese Family with Spondyloepiphyseal Dysplasia Congenita 
PLoS ONE  2015;10(6):e0127529.
Spondyloepiphyseal dysplasia congenita (SEDC) is an autosomal dominant chondrodysplasia characterized by disproportionate short-trunk dwarfism, skeletal and vertebral deformities. Exome sequencing and Sanger sequencing were performed in a Chinese Han family with typical SEDC, and a novel mutation, c.620G>A (p.Gly207Glu), in the collagen type II alpha-1 gene (COL2A1) was identified. The mutation may impair protein stability, and lead to dysfunction of type II collagen. Family-based study suggested that the mutation is a de novo mutation. Our study extends the mutation spectrum of SEDC and confirms genotype-phenotype relationship between mutations at glycine in the triple helix of the alpha-1(II) chains of the COL2A1 and clinical findings of SEDC, which may be helpful in the genetic counseling of patients with SEDC.
PMCID: PMC4452087  PMID: 26030151
13.  A Split-and-Merge-Based Uterine Fibroid Ultrasound Image Segmentation Method in HIFU Therapy 
PLoS ONE  2015;10(5):e0125738.
High-intensity focused ultrasound (HIFU) therapy has been used to treat uterine fibroids widely and successfully. Uterine fibroid segmentation plays an important role in positioning the target region for HIFU therapy. Presently, it is completed by physicians manually, reducing the efficiency of therapy. Thus, computer-aided segmentation of uterine fibroids benefits the improvement of therapy efficiency. Recently, most computer-aided ultrasound segmentation methods have been based on the framework of contour evolution, such as snakes and level sets. These methods can achieve good performance, although they need an initial contour that influences segmentation results. It is difficult to obtain the initial contour automatically; thus, the initial contour is always obtained manually in many segmentation methods. A split-and-merge-based uterine fibroid segmentation method, which needs no initial contour to ensure less manual intervention, is proposed in this paper. The method first splits the image into many small homogeneous regions called superpixels. A new feature representation method based on texture histogram is employed to characterize each superpixel. Next, the superpixels are merged according to their similarities, which are measured by integrating their Quadratic-Chi texture histogram distances with their space adjacency. Multi-way Ncut is used as the merging criterion, and an adaptive scheme is incorporated to decrease manual intervention further. The method is implemented using Matlab on a personal computer (PC) platform with Intel Pentium Dual-Core CPU E5700. The method is validated on forty-two ultrasound images acquired from HIFU therapy. The average running time is 9.54 s. Statistical results showed that SI reaches a value as high as 87.58%, and normHD is 5.18% on average. It has been demonstrated that the proposed method is appropriate for segmentation of uterine fibroids in HIFU pre-treatment imaging and planning.
PMCID: PMC4431844  PMID: 25973906
14.  Protective effect of high concentration of BN52021 on retinal contusion in cat eyes 
BMC Ophthalmology  2015;15:50.
Blunt injuries/contusion on eyes might cause retina blunt trauma. This study is to evaluate the protective function of BN52021 against retinal trauma.
A total of 70 cats, 6 months old, were divided into six groups: Group A to E (n = 12) and normal control (N) group (n = 10). The right eyes in Group A to E were contused. All experiments were performed under general anesthetization. Retrobulbar injections of medication in right eyes were performed. Cats were administrated with 0.5 mL of normal saline (NS), dimethyl sulphoxide, 0.2 g/L BN52021, 1 g/L BN52021 and 5 g/L BN52021, respectively. Cats in Group N were administrated with 0.5 mL of NS. Intraocular pressure (IOP), flash electroretinogram (ERG), and retinal nerve fiber layer (RNFL) thickness were measured. Hematoxylin and eosin (HE) staining and transmission electron microscope (TEM) were detected.
No significant difference was observed in IOP levels among groups. Comparing with cats in Group N, those in Group A to E showed significant lower amplitudes of rod a- and b-waves (P < 0.05). Amplitudes of rod a- and b-waves were increased by administration of high concentration of BN52021 (≥1 g/L). Moreover, high concentration of BN52021 decreased the RNFL thickness increased by contusion. Axons in RNFL in Group E arranged neatly at 7 days after modeling.
The degenerated axons caused by contusion were repaired by BN52021. The administration of high concentration of (≥1 g/L) BN52021 could partially repair retinal function in contused cat eyes.
PMCID: PMC4440277  PMID: 25956877
Retina trauma; BN52021; Electroretinogram; Intraocular pressure; Retinal nerve fiber layer
15.  Effect of Glutamate on the Vestibulo-Solitary Projection after Sodium Nitroprusside-Induced Hypotension in Conscious Rats 
Orthostatic hypotension is most common in elderly people, and its prevalence increases with age. Attenuation of the vestibulo-sympathetic reflex (VSR) is commonly associated with orthostatic hypotension. In this study, we investigated the role of glutamate on the vestibulo-solitary projection of the VSR pathway to clarify the pathophysiology of orthostatic hypotension. Blood pressure and expression of both pERK and c-Fos protein were evaluated in the nucleus tractus solitarius (NTS) after microinjection of glutamate into the medial vestibular nucleus (MVN) in conscious rats with sodium nitroprusside (SNP)-induced hypotension that received baroreceptor unloading via sinoaortic denervation (SAD). SNP-induced hypotension increased the expression of both pERK and c-Fos protein in the NTS, which was abolished by pretreatment with glutamate receptor antagonists (MK801 or CNQX) in the MVN. Microinjection of glutamate receptor agonists (NMDA or AMPA) into the MVN increased the expression of both pERK and c-Fos protein in the NTS without causing changes in blood pressure. These results indicate that both NMDA and AMPA receptors play a significant role in the vestibulo-solitary projection of the VSR pathway for maintaining blood pressure, and that glutamatergic transmission in this projection might play a key role in the pathophysiology of orthostatic hypotension.
PMCID: PMC4422969  PMID: 25954134
c-Fos protein; Glutamate; Nucleus tractus solitarius; Orthostatic hypotension; Vestibulosympathetic reflex
16.  Remodeling of Kv1.5 Channel in Right Atria from Han Chinese Patients with Atrial Fibrillation 
The incidence of atrial fibrillation (AF) in rheumatic heart diseases (RHD) is very high and increases with age. Occurrence and maintenance of AF are very complicated process accompanied by many different mechanisms. Ion-channel remodeling, including the voltage-gated potassium channel Kv1.5, plays an important role in the pathophysiology of AF. However, the changes of Kv1.5 channel expression in Han Chinese patients with RHD and AF remain poorly understood. The aim of the present study was to investigate whether the Kv1.5 channels of the right atria may be altered with RHD, age, and sex to contribute to AF.
Right atrial appendages were obtained from 20 patients with normal cardiac functions who had undergone surgery, and 26 patients with AF. Subjects were picked from 4 groups: adult and aged patients in normal sinus rhythm (SR) and AF. Patients were divided into non-RHD and RHD groups or men and women groups in normal SR and AF, respectively. The expression of Kv1.5 protein and messenger RNA (mRNA) were measured using Western blotting and polymerase chain reaction (PCR) method, respectively.
Compared with the SR group, the expression of Kv1.5 protein decreased significantly in the AF group. However, neither Kv1.5 protein nor KCNA5 mRNA had significant differences in adult and aged groups, non-RHD and RHD group, and men and women group of AF.
The expression of Kv1.5 channel protein changes with AF but not with age, RHD, and sex in AF.
PMCID: PMC4424910  PMID: 25918274
Aging; Atrial Fibrillation; Kv1.5 Potassium Channel; Rheumatic Heart Disease; Sex
17.  Amino acid substitutions in the neuraminidase protein of an H9N2 avian influenza virus affect its airborne transmission in chickens 
Veterinary Research  2015;46(1):44.
Cases of H9N2 avian influenza virus (AIV) in poultry are increasing throughout many Eurasian countries, and co-infections with other pathogens have resulted in high morbidity and mortality in poultry. Few studies have investigated the genetic factors of virus airborne transmission which determine the scope of this epidemic. In this study, we used specific-pathogen-free chickens housed in isolators to investigate the airborne transmissibility of five recombinant H9N2 AIV rescued by reverse genetic technology. The results show that airborne transmission of A/Chicken/Shandong/01/2008 (SD01) virus was related to the neuraminidase (NA) gene, and four amino acid mutations (D368E, S370L, E313K and G381D) within the head region of the SD01 NA, reduced virus replication in the respiratory tract of chickens, reduced virus NA activity, and resulted in a loss of airborne transmission ability in chickens. Similarly, reverse mutations of these four amino acids in the NA protein of r01/NASS virus, conferred an airborne transmission ability to the recombinant virus. We conclude that these four NA residues may be significant genetic markers for evaluating potential disease outbreak of H9N2 AIV, and propose that immediate attention should be paid to the airborne transmission of this virus.
Electronic supplementary material
The online version of this article (doi:10.1186/s13567-014-0142-3) contains supplementary material, which is available to authorized users.
PMCID: PMC4404070  PMID: 25928577
18.  Assessment of visual outcomes of cataract surgery in Tujia nationality in Xianfeng County, China 
To evaluate the visual outcome and factors influencing visual outcome of manual small incision cataract surgery (MSICS) in the rural area in the Xianfeng County.
Eighty-two eyes of 82 patients who underwent cataract surgery performed by using MSICS technique were identified. Data collected included each patient's age, gender, the level of education. Uncorrected and corrected distance visual acuity (UDVA and CDVA) at presentation and at 1, 6, 8wk postoperatively, pre-existing eye disease, operative findings and complications, the risk factors were evaluated.
In 82 patients, the average age was 69.6±0.6y, illiterate were 52 (63.4%). Of 82 eyes, pseudophakia was present in 77 eyes (93.9%). At 1wk postoperatively, 47 eyes (57.3%) had the UDVA of ≥6/18, and 52 eyes (63.4%) had the CDVA of ≥6/18. At 6 to 8wk postoperatively, 50 eyes (61.0%) had UDVA of ≥6/18, and 57 eyes (69.5%) had the CDVA of ≥6/18. Postoperative visual status was significantly related to the co-morbidities, such as corneal pathology, glaucoma (P<0.001). Operative complications, such as posterior capsule opacity and cystoid macular edema were main operative cause for the poor visual outcome.
MSICS provides a good visual recovery in our study but the vision outcome did not fulfill the standards proposed by WHO, which highlights the need for an improvement in local socioeconomic understanding, population education and surgery quality.
PMCID: PMC4413593  PMID: 25938043
cataract surgery; manual small incision cataract surgery; visual outcome; co-morbidities; operative complications
19.  Ferulic Acid Attenuates TGF-β1-Induced Renal Cellular Fibrosis in NRK-52E Cells by Inhibiting Smad/ILK/Snail Pathway 
Renal fibrosis is a common cause of renal dysfunction with chronic kidney disease. Central to this process is epithelial-mesenchymal transformation (EMT) of proximal tubular epithelial cells driven by transforming growth factor-β1 (TGF-β1) signaling. The present study aimed to investigate the effect of Ferulic acid (FA) on EMT of renal proximal tubular epithelial cell line (NRK-52E) induced by TGF-β1 and to elucidate its underlying mechanism against EMT related to TGF-β1/Smads pathway. The NRK-52E cells were treated for 48 h with TGF-β1 (5 ng/mL) in different concentrations of FA (0 to 200 µM). Fibronectin, a mesenchymal marker, was assessed by western blotting. Western blotting was also used to examine the EMT markers (E-cadherin, and α-smooth muscle actin (α-SMA)), signal transducer (p-Smad2/3), and EMT initiator (Snail). ILK was also assayed by western blotting. The results showed that TGF-β1 induced spindle-like morphological transition in NRK-52E cells. Smad2/3 signaling pathway activation, increased fibronectin, α-SMA, ILK, and Snail expression, and decreased E-cadherin expression in TGF-β1-treated NRK-52E cells. FA efficiently blocked P-Smad2/3 activation and attenuated all these EMT changes induced by TGF-β1. These findings suggest that FA may serve as a potential fibrosis antagonist for renal proximal tubule cells by inhibiting EMT process.
PMCID: PMC4408646  PMID: 25949265
20.  Hepatitis C Virus (HCV) NS3 Sequence Diversity and Antiviral Resistance-Associated Variant Frequency in HCV/HIV Coinfection 
Antimicrobial Agents and Chemotherapy  2014;58(10):6079-6092.
HIV coinfection accelerates disease progression in chronic hepatitis C and reduces sustained antiviral responses (SVR) to interferon-based therapy. New direct-acting antivirals (DAAs) promise higher SVR rates, but the selection of preexisting resistance-associated variants (RAVs) may lead to virologic breakthrough or relapse. Thus, pretreatment frequencies of RAVs are likely determinants of treatment outcome but typically are below levels at which the viral sequence can be accurately resolved. Moreover, it is not known how HIV coinfection influences RAV frequency. We adopted an accurate high-throughput sequencing strategy to compare nucleotide diversity in HCV NS3 protease-coding sequences in 20 monoinfected and 20 coinfected subjects with well-controlled HIV infection. Differences in mean pairwise nucleotide diversity (π), Tajima's D statistic, and Shannon entropy index suggested that the genetic diversity of HCV is reduced in coinfection. Among coinfected subjects, diversity correlated positively with increases in CD4+ T cells on antiretroviral therapy, suggesting T cell responses are important determinants of diversity. At a median sequencing depth of 0.084%, preexisting RAVs were readily identified. Q80K, which negatively impacts clinical responses to simeprevir, was encoded by more than 99% of viral RNAs in 17 of the 40 subjects. RAVs other than Q80K were identified in 39 of 40 subjects, mostly at frequencies near 0.1%. RAV frequency did not differ significantly between monoinfected and coinfected subjects. We conclude that HCV genetic diversity is reduced in patients with well-controlled HIV infection, likely reflecting impaired T cell immunity. However, RAV frequency is not increased and should not adversely influence the outcome of DAA therapy.
PMCID: PMC4187920  PMID: 25092699
21.  The Efficacy of Traditional Chinese Medical Exercise for Parkinson’s Disease: A Systematic Review and Meta-analysis 
PLoS ONE  2015;10(4):e0122469.
Background and Objective
Several studies assessed the efficacy of traditional Chinese medical exercise in the management of Parkinson’s disease (PD), but its role remained controversial. Therefore, the purpose of this systematic review is to evaluate the evidence on the effect of traditional Chinese medical exercise for PD.
Seven English and Chinese electronic databases, up to October 2014, were searched to identify relevant studies. The PEDro scale was employed to assess the methodological quality of eligible studies. Meta-analysis was performed by RevMan 5.1 software.
Fifteen trials were included in the review. Tai Chi and Qigong were used as assisting pharmacological treatments of PD in the previous studies. Tai Chi plus medication showed greater improvements in motor function (standardized mean difference, SMD, -0.57; 95% confidence intervals, CI, -1.11 to -0.04), Berg balance scale (BBS, SMD, -1.22; 95% CI -1.65 to -0.80), and time up and go test (SMD, -1.06; 95% CI -1.44 to -0.68). Compared with other therapy plus medication, Tai Chi plus medication also showed greater gains in motor function (SMD, -0.78; 95% CI -1.46 to -0.10), BBS (SMD, -0.99; 95% CI -1.44 to -0.54), and functional reach test (SMD, -0.77; 95% CI -1.51 to -0.03). However, Tai Chi plus medication did not showed better improvements in gait or quality of life. There was not sufficient evidence to support or refute the effect of Qigong plus medication for PD.
In the previous studies, Tai Chi and Qigong were used as assisting pharmacological treatments of PD. The current systematic review showed positive evidence of Tai Chi plus medication for PD of mild-to-moderate severity. So Tai Chi plus medication should be recommended for PD management, especially in improving motor function and balance. Qigong plus medication also showed potential gains in the management of PD. However, more high quality studies with long follow-up are warrant to confirm the current findings.
PMCID: PMC4382160  PMID: 25830664
22.  Prevalence of Hepatitis B and Knowledge About Hepatitis B Among Migrant Workers in Shandong Province, China: A Cross-Sectional Study 
China is a country with a high prevalence of hepatitis B. As a special population, migrant workers are more vulnerable to hepatitis B.
The present study was conducted to gain insight into the prevalence of hepatitis B and knowledge about hepatitis B among migrant workers in Shandong Province, China, as well as to explore a series of strategies for preventing and controlling the spread and prevalence of hepatitis B.
Patients and Methods:
A cross-sectional study was conducted without age restriction, in Shandong Province in China. The sample-size was determined scientifically. The study population was selected using the random multistage cluster sampling. Personal information, including sex, age, ethnicity, marital status, education level, years of duration of stay in Jinan and health insurance were obtained from 2065 migrant workers. Moreover, blood samples were collected for hepatitis B infection screening. Knowledge about hepatitis B was assessed using a self-administered questionnaire. Correct response rates were calculated.
Of 2065 migrant workers tested for Hepatitis B Surface Antigen (HBsAg), 167 (126 men and 41 women) tested positive for HBsAg; the overall prevalence was 8.1%. The prevalence rates in men and women were 9.6% and 5.4%, respectively. The prevalence rates in subjects 18 - 30 years old and 30 years older were 3.5% and 9.3%, respectively. There were statistically significant differences between those groups (P < 0.05). Correct response rates for the transmission of hepatitis B questions were low, especially for a question regarding whether hepatitis B can be transferred by sexual contact (36.8%) and whether it can be transferred from mothers to infants (33.9%). A total of 80.9% of migrant workers correctly responded that vaccination is effective for hepatitis B. However, 68% of migrants also had the misconception that it is necessary to receive a booster vaccination.
The hepatitis B virus infection rate in migrant workers is higher than average infection rates in China, and these workers’ knowledge regarding hepatitis B is poor. It is urgent that an appropriate program be undertaken for the prevention and control of hepatitis B among migrant workers.
PMCID: PMC4443392  PMID: 26023348
Transients and Migrants; Hepatitis B; Knowledge; Cross-Sectional Studies; Questionnaires
23.  The PDGF-D/miR-106a/Twist1 pathway orchestrates epithelial-mesenchymal transition in gemcitabine resistance hepatoma cells 
Oncotarget  2015;6(9):7000-7010.
Emerging evidence demonstrates that platelet-derived growth factor-D (PDGF-D) plays a critical role in epithelial-mesenchymal transition (EMT) and drug resistance in hepatocellular carcinoma (HCC) cells. However, the underlying mechanism has not been fully elucidated. The objective is to explore the molecular mechanism of PDGF-D-mediated EMT in drug resistance HCC cells. To achieve our goal, we used multiple approaches including Western blotting, real-time RT-PCR, wound healing assay, invasion assay, luciferase activity assay, transfection, and immunohistochemistry. We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells. Moreover, PDGF-D markedly inhibited miR-106a expression and subsequently upregulated Twist1 expression. Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients. Our findings provide a possible molecular mechanism for understanding GR chemoresistance in HCC cells. Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.
PMCID: PMC4466665  PMID: 25760076
Hepatocellular carcinoma; EMT; PDGF-D; miR-106a; Twist
24.  Superparamagnetic core/shell GoldMag nanoparticles: size-, concentration- and time-dependent cellular nanotoxicity on human umbilical vein endothelial cells and the suitable conditions for magnetic resonance imaging 
GoldMag nanoparticles (GMNPs) possess the properties of colloid gold and superparamagnetic iron oxide nanoparticles, which make them useful for delivery, separation and molecular imaging. However, because of the nanometer effect, GMNPs are highly toxic. Thus, the biosafety of GMNPs should be fully studied prior to their use in biomedicine. The main purpose of this study was to evaluate the nanotoxicity of GMNPs on human umbilical vein endothelial cells (HUVECs) and determine a suitable size, concentration and time for magnetic resonance imaging (MRI).
Transmission electron microscopy showed that GMNPs had a typical shell/core structure, and the shell was confirmed to be gold using energy dispersive spectrometer analysis. The average sizes of the 30 and 50 nm GMNPs were 30.65 ± 3.15 and 49.23 ± 5.01 nm, respectively, and the shell thickness were 6.8 ± 0.65 and 8.5 ± 1.36 nm, respectively. Dynamic light scattering showed that the hydrodynamic diameter of the 30 and 50 nm GMNPs were 33.2 ± 2.68 and 53.12 ± 4.56 nm, respectively. The r2 relaxivity of the 50 nm GMNPs was 98.65 mM−1 s−1, whereas it was 80.18 mM−1 s−1 for the 30 nm GMNPs. The proliferation, cytoskeleton, migration, tube formation, apoptosis and ROS generation of labeled HUVECs depended on the size and concentration of GMNPs and the time of exposure. Because of the higher labeling rate, the 50 nm GMNPs exhibited a significant increase in nanotoxicity compared with the 30 nm GMNPs at the same concentration and time. At no more than 25 μg/mL and 12 hours, the 50 nm GMNPs exhibited no significant nanotoxicity in HUVECs, whereas no toxicity was observed at 50 μg/mL and 24 hours for the 30 nm GMNPs.
These results demonstrated that the nanotoxicity of GMNPs in HUVECs depended on size, concentration and time. Exposure to larger GMNPs with a higher concentration for a longer period of time resulted in a higher labeling rate and ROS level for HUVECs. Coupled with r2 relaxivity, it was suggested that the 50 nm GMNPs are more suitable for HUVEC labeling and MRI, and the suitable concentration and time were 25 μg/mL and 12 hours.
PMCID: PMC4387586  PMID: 25890315
Superparamagnetic; Core/shell GoldMag nanoparticles; Human umbilical vein endothelial cells; Cellular nanotoxicity; Reactive oxygen species; Labeling efficiency; Magnetic resonance imaging
25.  The effectiveness of Tai Chi for patients with Parkinson’s disease: study protocol for a randomized controlled trial 
Trials  2015;16:111.
Parkinson’s disease (PD) is a common degenerative neurological disorder that causes loss of independence and decreased quality of life. The prevalence of PD tends to increase with age. In China, the morbidity rate of PD among people aged more than 65 years old is 1.70%. As an important component of traditional Chinese Qigong exercises, Tai Chi is a popular and safe exercise, especially for older adults in China. And it may result in promising gains for PD patients. However, current evidence is insufficient to inform the use of Tai Chi in the management of PD. Therefore, the aim of this trial is to systematically evaluate the effect of Tai Chi on PD and determine whether Tai Chi is an eligible exercise program for Chinese PD patients.
A single-blind, parallel randomized controlled trial will be conducted. One hundred and forty-two patients with PD will be randomly assigned to a Tai Chi group (n = 71) or routine exercise group (n = 71). Subjects will participate in supervised study programs 3 times per week for 2 months and will be followed for an additional 6 months after formal training stops. The primary outcome measures include Berg Balance Scale, Timed Up and Go Test and Six-Minute Walk Test, which are known to be valid and reliable clinical instruments. The Unified Parkinson’s Disease Rating Scale Motor Section and Parkinson’s Disease Questionnaire-39 will be used as the secondary outcome measure. All outcomes will be measured at baseline, 2 and 8 months. The sample for this trial (N = 142) will provide relevant information to detect the improvement of balance, gait and quality of life in either of the 2 exercise groups.
Findings from this study will provide insights into the effects of Tai Chi in people with PD. The information gained from this project has the potential to influence the clinical decisions of Chinese doctors, and will provide clear evidence as to whether Tai Chi should be advocated in people with PD.
Trial registration
The trial was registered at (ChiCTR-TRC-14004549) on 22 April 2014.
PMCID: PMC4377011  PMID: 25873334
Parkinson’s disease; Tai Chi; Exercise; Balance; Gait; Quality of life

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