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author:("Wu, jiangyou")
1.  Evaluation of the 3D Finite Element Method Using a Tantalum Rod for Osteonecrosis of the Femoral Head 
Background
The aim of this study was to contrast the collapse values of the postoperative weight-bearing areas of different tantalum rod implant positions, fibula implantation, and core decompression model and to investigate the advantages and disadvantages of tantalum rod implantation in different ranges of osteonecrosis in comparison with other methods.
Material/Methods
The 3D finite element method was used to establish the 3D finite element model of normal upper femur, 3D finite element model after tantalum rod implantation into different positions of the upper femur in different osteonecrosis ranges, and other 3D finite element models for simulating fibula implant and core decompression.
Results
The collapse values in the weight-bearing area of the femoral head of the tantalum rod implant model inside the osteonecrosis area, implant model in the middle of the osteonecrosis area, fibula implant model, and shortening implant model exhibited no statistically significant differences (p>0.05) when the osteonecrosis range was small (60°). The stress values on the artificial bone surface for the tantalum rod implant model inside the osteonecrosis area and the shortening implant model exhibited statistical significance (p<0.01).
Conclusions
Tantalum rod implantation into the osteonecrosis area can reduce the collapse values in the weight-bearing area when osteonecrosis of the femoral head (ONFH) was in a certain range, thereby obtaining better clinical effects. When ONFH was in a large range (120°), the tantalum rod implantation inside the osteonecrosis area, shortening implant or fibula implant can reduce the collapse values of the femoral head, as assessed by other methods.
doi:10.12659/MSM.890920
PMCID: PMC4266390  PMID: 25479830
Femoral Neoplasms; Finite Element Analysis; Tantalum
2.  Effect of the knee position during wound closure after total knee arthroplasty on early knee function recovery 
Objective
This study investigated the effect of the knee position during wound closure on early knee function recovery after total knee arthroplasty (TKA).
Methods
This study included 80 primary total knee arthroplasties due to osteoarthritis. The patients were randomized according to the type of wound closure: extension group for full extension and flexion group for 90° flexion. The incision of articular capsule was marked for precise wound alignment. In the flexion group, the knee was kept in high flexion for 1 to 2 min after wound closure. The two groups were treated with the same postoperative rehabilitation exercises. The range of motion (ROM), visual analogue scale (VAS) score of anterior knee pain, Knee Society Score (KSS) and postoperative complications were assessed at 6 weeks, 3 months and 6 months, postoperatively.
Results
At 6 weeks and 3 months postoperatively, the ROM in flexion group was 98.95 ± 10.33° and 110.05 ± 4.93° respectively, with 87.62 ± 8.92° and 95.62 ± 6.51° in extension group, respectively; The VAS score of anterior knee pain in flexion group was 2.02 ± 1.38 and 2.21 ± 0.87, respectively, with 2.57 ± 1.07 and 2.87 ± 0.83 in extension group, respectively. The ROM and VAS pain score of the two groups were significantly different at these two time points, with no significant difference at 6 months postoperatively. The two groups were not significantly different in KSS, and no apparent complication was observed at three time points.
Conclusion
Marking the articular capsule incision, wound closure in flexion and high flexion after wound closure can effectively decrease anterior knee pain after TKA and promote the early recovery of ROM.
doi:10.1186/s13018-014-0079-2
PMCID: PMC4237872  PMID: 25149657
Anterior knee pain; Range of motion; Wound closure; Total knee arthroplasty
3.  Three-dimensional virtual reality simulation of periarticular tumors using Dextroscope reconstruction and simulated surgery: A preliminary 10-case study 
Background
Dextroscope® three-dimensional (3D) imaging has been extensively used to generate virtual reality (VR) workspaces for neurosurgery and laparoscopy, but few applications have been reported for orthopedic surgery. Here, we investigated orthopedic periarticular tumor surgery planning and anatomical characteristics using a Dextroscope.
Material/Methods
Patients undergoing surgery for periarticular tumors (n=10) between October 2008 and June 2010 were enrolled and presurgically subjected to computed tomography (CT), magnetic resonance imaging (MRI), and MRI angiography (MRI-A). Imaging data were transferred and integrated in a Dextroscope to produce a VR simulation. The presurgical 3D anatomical reconstructions and intraoperative anatomical characteristics (virtual vs. actual data) and surgical approach (virtual vs. actual situation) measurement and subjective appearance were compared.
Results
Anatomical characteristics in the area of interest and tumor diameters in all 3 planes (superior-inferior, medial-lateral, and anteroposterior) were consistent between virtual and actual data (3.92±1.22, 1.96±0.53, and 1.73±0.44 vs. 3.92±1.13, 1.91±0.44, and 1.81±0.41; P=0.99, 0.24, and 0.09, respectively). However, the virtual surgical situations were inconsistent with the actual intraoperative situation in many cases, leading to complications. The resolutions of the original CT, MRI, and MRI-A images directly correlated with 3D simulation quality, with soft tissues most poorly represented. Tumor tissue imaging quality in 3D varied extensively by tumor type.
Conclusions
Anatomical structures of periarticular tumors can be reconstructed using the Dextroscope system with good accuracy in the case of simple fenestration, increasing treatment individualization, surgical competence level, and potentially reducing intraoperative complications. However, further specialization of VR tools for use in orthopedic applications that involve specialized tools and procedures, such as drilling and implant placement, are urgently need.
doi:10.12659/MSM.8897770
PMCID: PMC4076173  PMID: 24961404
Reconstruction; Virtual Surgery; Virtual Reality Exposure Therapy; Computer Simulation
4.  Viral Etiologies of Hospitalized Acute Lower Respiratory Infection Patients in China, 2009-2013 
PLoS ONE  2014;9(6):e99419.
Background
Acute lower respiratory infections (ALRIs) are an important cause of acute illnesses and mortality worldwide and in China. However, a large-scale study on the prevalence of viral infections across multiple provinces and seasons has not been previously reported from China. Here, we aimed to identify the viral etiologies associated with ALRIs from 22 Chinese provinces.
Methods and Findings
Active surveillance for hospitalized ALRI patients in 108 sentinel hospitals in 24 provinces of China was conducted from January 2009-September 2013. We enrolled hospitalized all-age patients with ALRI, and collected respiratory specimens, blood or serum collected for diagnostic testing for respiratory syncytial virus (RSV), human influenza virus, adenoviruses (ADV), human parainfluenza virus (PIV), human metapneumovirus (hMPV), human coronavirus (hCoV) and human bocavirus (hBoV).
We included 28,369 ALRI patients from 81 (of the 108) sentinel hospitals in 22 (of the 24) provinces, and 10,387 (36.6%) were positive for at least one etiology. The most frequently detected virus was RSV (9.9%), followed by influenza (6.6%), PIV (4.8%), ADV (3.4%), hBoV (1.9), hMPV (1.5%) and hCoV (1.4%). Co-detections were found in 7.2% of patients. RSV was the most common etiology (17.0%) in young children aged <2 years. Influenza viruses were the main cause of the ALRIs in adults and elderly. PIV, hBoV, hMPV and ADV infections were more frequent in children, while hCoV infection was distributed evenly in all-age. There were clear seasonal peaks for RSV, influenza, PIV, hBoV and hMPV infections.
Conclusions
Our findings could serve as robust evidence for public health authorities in drawing up further plans to prevent and control ALRIs associated with viral pathogens. RSV is common in young children and prevention measures could have large public health impact. Influenza was most common in adults and influenza vaccination should be implemented on a wider scale in China.
doi:10.1371/journal.pone.0099419
PMCID: PMC4063718  PMID: 24945280
5.  Lipopolysaccharide Induces Immune Activation and SIV Replication in Rhesus Macaques of Chinese Origin 
PLoS ONE  2014;9(6):e98636.
Background
Chronic immune activation is a hallmark of progressive HIV infection and a key determinant of immunodeficiency in HIV-infected individuals. Bacterial lipopolysaccharide (LPS) in the circulation has been implicated as a key factor in HIV infection-related systemic immune activation. We thus investigate the impact of LPS on systemic immune activation in simian immunodeficiency virus (SIV)-infected rhesus macaques of Chinese origin.
Methods
The animals were inoculated intravenously with SIVmac239. The levels of plasma viral load and host inflammatory cytokines in PBMC were measured by real-time RT-PCR. CD4/CD8 ratio and systemic immune activation markers were examined by flow cytometric analysis of PBMCs. White blood cell and neutrophil counts and C Reactive Protein levels were determined using biochemistry analyzer. The plasma levels of LPS were determined by Tachypleus Amebocyte Lysate (TAL) test.
Results
The animals inoculated with SIVmac239 became infected as evidenced by the increased plasma levels of SIV RNA and decreased CD4/CD8 ratio. LPS administration of SIV-infected animals induced a transient increase of plasma SIV RNA and immune activation, which was indicated by the elevated expression of the inflammatory cytokines and CD4+HLA-DR+ T cells in PBMCs.
Conclusions
These data support the concept that LPS is a driving factor in systemic immune activation of HIV disease.
doi:10.1371/journal.pone.0098636
PMCID: PMC4053387  PMID: 24918575
6.  Inhibition of hepatitis C virus by an M1GS ribozyme derived from the catalytic RNA subunit of Escherichia coli RNase P 
Virology Journal  2014;11:86.
Background
Hepatitis C virus (HCV) is a human pathogen causing chronic liver disease in about 200 million people worldwide. However, HCV resistance to interferon treatment is one of the important clinical implications, suggesting the necessity to seek new therapies. It has already been shown that some forms of the catalytic RNA moiety from E. coli RNase P, M1 RNA, can be introduced into the cytoplasm of mammalian cells for the purpose of carrying out targeted cleavage of mRNA molecules. Our study is to use an engineering M1 RNA (i.e. M1GS) for inhibiting HCV replication and demonstrates the utility of this ribozyme for antiviral applications.
Results
By analyzing the sequence and structure of the 5′ untranslated region of HCV RNA, a putative cleavage site (C67-G68) was selected for ribozyme designing. Based on the flanking sequence of this site, a targeting M1GS ribozyme (M1GS-HCV/C67) was constructed by linking a custom guide sequence (GS) to the 3′ termini of catalytic RNA subunit (M1 RNA) of RNase P from Escherichia coli through an 88 nt-long bridge sequence. In vitro cleavage assays confirmed that the engineered M1GS ribozyme cleaved the targeted RNA specifically. Moreover, ~85% reduction in the expression levels of HCV proteins and >1000-fold reduction in viral growth were observed in supernatant of cultured cells that transfected the functional ribozyme. In contrast, the HCV core expression and viral growth were not significantly affected by a “disabled” ribozyme (i.e. M1GS-HCV/C67*). Moreover, cholesterol-conjugated M1GS ribozyme (i.e. Chol-M1GS-HCV/C67) showed almost the same bioactivities with M1GS-HCV/C67, demonstrating the potential to improve in vivo pharmacokinetic properties of M1GS-based RNA therapeutics.
Conclusion
Our results provide direct evidence that the M1GS ribozyme can function as an antiviral agent and effectively inhibit gene expression and multiplication of HCV.
doi:10.1186/1743-422X-11-86
PMCID: PMC4038377  PMID: 24885776
Ribozyme; RNase P; Hepatitis C virus; 5′ UTR; Antiviral
7.  Transcriptome profiling confirmed correlations between symptoms and transcriptional changes in RDV infected rice and revealed nucleolus as a possible target of RDV manipulation 
Virology Journal  2014;11:81.
Background
Rice dwarf virus (RDV) is the causal agent of rice dwarf disease, which limits rice production in many areas of south East Asia. Transcriptional changes of rice in response to RDV infection have been characterized by Shimizu et al. and Satoh et al.. Both studies found induction of defense related genes and correlations between transcriptional changes and symptom development in RDV-infected rice. However, the same rice cultivar, namely Nipponbare belonging to the Japonic subspecies of rice was used in both studies.
Methods
Gene expression changes of the indica subspecies of rice, namely Oryza sativa L. ssp. indica cv Yixiang2292 that show moderate resistance to RDV, in response to RDV infection were characterized using an Affymetrix Rice Genome Array. Differentially expressed genes (DEGs) were classified according to their Gene Ontology (GO) annotation. The effects of transient expression of Pns11 in Nicotiana benthaminana on the expression of nucleolar genes were studied using real-time PCR (RT-PCR).
Results
856 genes involved in defense or other physiological processes were identified to be DEGs, most of which showed up-regulation. Ribosome- and nucleolus related genes were significantly enriched in the DEGs. Representative genes related to nucleolar function exhibited altered expression in N. benthaminana plants transiently expressing Pns11 of RDV.
Conclusions
Induction of defense related genes is common for rice infected with RDV. There is a co-relation between symptom severity and transcriptional alteration in RDV infected rice. Besides ribosome, RDV may also target nucleolus to manipulate the translation machinery of rice. Given the tight links between nucleolus and ribosome, it is intriguing to speculate that RDV may enhance expression of ribosomal genes by targeting nucleolus through Pns11.
doi:10.1186/1743-422X-11-81
PMCID: PMC4032362  PMID: 24885215
RDV; Transcriptome profiling; Pns11; Nucleolus
8.  Co-Circulation and Genomic Recombination of Coxsackievirus A16 and Enterovirus 71 during a Large Outbreak of Hand, Foot, and Mouth Disease in Central China 
PLoS ONE  2014;9(4):e96051.
A total of 1844 patients with hand, foot, and mouth disease (HFMD), most of them were children of age 1–3-year-old, in Central China were hospitalized from 2011 to 2012. Among them, 422 were infected with coxsackievirus A16 (CVA16), 334 were infected with enterovirus 71 (EV71), 38 were co-infected with EV71 and CVA16, and 35 were infected with other enteroviruses. Molecular epidemiology analysis revealed that EV71 and CVA16 were detected year-round, but EV71 circulated mainly in July and CVA16 circulated predominantly in November, and incidence of HFMD was reduced in January and February and increased in March. Clinical data showed that hyperglycemia and neurologic complications were significantly higher in EV71-infected patients, while upper respiratory tract infection and C-reactive protein were significantly higher in CVA16-associated patients. 124 EV71 and 80 CVA16 strains were isolated, among them 56 and 68 EV71 strains were C4a and C4b, while 25 and 55 CVA16 strains were B1a and B1b, respectively. Similarity plots and bootscan analyses based on entire genomic sequences revealed that the three C4a sub-genotype EV71 strains were recombinant with C4b sub-genotype EV71 in 2B–2C region, and the three CVA16 strains were recombinant with EV71 in 2A–2B region. Thus, CVA16 and EV71 were the major causative agents in a large HFMD outbreak in Central China. HFMD incidence was high for children among household contact and was detected year-round, but outbreak was seasonal dependent. CVA16 B1b and EV71 C4b reemerged and caused a large epidemic in China after a quiet period of many years. Moreover, EV71 and CVA16 were co-circulated during the outbreak, which may have contributed to the genomic recombination between the pathogens. It should gain more attention as there may be an upward trend in co-circulation of the two pathogens globally and the new role recombination plays in the emergence of new enterovirus variants.
doi:10.1371/journal.pone.0096051
PMCID: PMC4002479  PMID: 24776922
9.  Rescue of Wild-type Mumps Virus from a Strain Associated with Recent Outbreaks Helps to Define the Role of the SH ORF in the Pathogenesis of Mumps Virus 
Virology  2011;417(1):126-136.
Mumps virus (MuV) causes acute infections in humans. In recent years, MuV has caused epidemics among highly vaccinated populations. The largest outbreak in the U.S. in the past 20 years occurred in 2005–2006 with over reported 5,000 cases which the majority of the cases was in vaccinated young adults. We sequenced the complete genome of a representative strain from the epidemic (MuV-IA). MuV-IA is a member of genotype G, the same genotype of MuV that was associated with the outbreak in the UK in 2004–2005. We constructed a reverse genetics system for MuV-IA (rMuV-IA), and rescued a virus lacking the open reading frame (ORF) of the SH gene (rMuVΔSH). rMuVΔSH infection in L929 cells induced increased NF-κB activation, TNF-α production and apoptosis compared to rMuV-IA. rMuVΔSH was attenuated in an animal model. These results indicated that the SH ORF of MuV plays a significant role in interfering with TNF-α signaling and viral pathogenesis during virus infection.
doi:10.1016/j.virol.2011.05.003
PMCID: PMC3959920  PMID: 21676427
10.  Tetherin has negligible activity in restricting hepatitis C virus in hepatocytes 
Innate immunity  2011;18(3):398-405.
We investigated the ability of tetherin, a recently identified antiviral factor, in restricting hepatitis C virus (HCV) in the Japanese fulminant hepatitis-1 (JFH-1) infectious cell culture system. Human hepatocytes (Huh7, Huh7.5.1) expressed low levels of endogenous tetherin, which could be induced by IFN-α. However, tetherin contributes little to IFN-α-mediated anti-HCV JFH-1 activity. Although tetherin could inhibit Vpu-deleted HIV-1 release, it had negligible activity in restricting HCV JFH-1 release from hepatocytes, which was evidenced by unaffected levels of intracellular/extracellular HCV RNA and infectious virus. The failure of tetherin’s anti-HCV activity could not be related to the counteraction of HCV, as HCV infection of hepatocytes affected neither tetherin expression nor anti-HIV function of tetherin. These observations imply that tetherin has negligible activity in the restriction of HCV JFH-1 in human hepatocytes.
doi:10.1177/1753425911412984
PMCID: PMC3937259  PMID: 21940748
Hepatitis C virus; innate immunity; interferon; tetherin; virus release
11.  PolyC-Binding Protein 1 Interacts with 5′-Untranslated Region of Enterovirus 71 RNA in Membrane-Associated Complex to Facilitate Viral Replication 
PLoS ONE  2014;9(1):e87491.
Enterovirus 71 (EV71) is one causative agent of hand, foot, and mouth disease (HFMD), which may lead to severe neurological disorders and mortality in children. EV71 genome is a positive single-stranded RNA containing a single open reading frame (ORF) flanked by 5′-untranslated region (5′UTR) and 3′UTR. The 5′UTR is fundamentally important for virus replication by interacting with cellular proteins. Here, we revealed that poly(C)-binding protein 1 (PCBP1) specifically binds to the 5′UTR of EV71. Detailed studies indicated that the RNA-binding K-homologous 1 (KH1) domain of PCBP1 is responsible for its binding to the stem-loop I and IV of EV71 5′UTR. Interestingly, we revealed that PCBP1 is distributed in the nucleus and cytoplasm of uninfected cells, but mainly localized in the cytoplasm of EV71-infected cells due to interaction and co-localization with the viral RNA. Furthermore, sub-cellular distribution analysis showed that PCBP1 is located in ER-derived membrane, in where virus replication occurred in the cytoplasm of EV71-infected cells, suggesting PCBP1 is recruited in a membrane-associated replication complex. In addition, we found that the binding of PCBP1 to 5′UTR resulted in enhancing EV71 viral protein expression and virus production so as to facilitate viral replication. Thus, we revealed a novel mechanism in which PCBP1 as a positive regulator involved in regulation of EV71 replication in the host specialized membrane-associated replication complex, which provides an insight into cellular factors involved in EV71 replication.
doi:10.1371/journal.pone.0087491
PMCID: PMC3906175  PMID: 24489926
12.  Downregulation of RelA (p65) by Rapamycin Inhibits Murine Adipocyte Differentiation and Reduces Fat Mass of C57BL/6J Mice despite High Fat Diet 
ISRN Obesity  2014;2014:540582.
Rapamycin (RAPA) is a clinical immunosuppressive agent first reported in the literature in 1975 after its discovery in a soil sample from the island of Rapa Nui. Aside from the well-documented effects of RAPA on cell division and immunologic response, the literature reveals it to have negative effects on adipocyte and osteocyte differentiation as well. Understanding of the molecular effects of RAPA on cell differentiation is fragmentary in regard to these cell lineages. In this paper, we examined a potential mechanism for RAPA's effects on adipocyte differentiation in vitro and in vivo. The data point to a unique role of Rel A (p65)—a component of the NF-κB system—in mediating this event. In murine adipose derived stem cell cultures (muADSCs) from C57BL/6J mice, RAPA was found to selectively downregulate RelA/p65, mammalian target of rapamycin (mTOR), and do so in a dose-dependent manner. This implies a novel role for RelA in adipocyte biology. Intracellular lipid accumulation—as subjectively observed—was also decreased in muADSCs treated with RAPA. Mice treated with RAPA had reduced overall body weight and reduced size of both intraabdominal and subcutaneous fat pads. When treated with RAPA, mice fed a high fat diet did not develop obesity and were not different from their regular diet controls in terms of body weight. These results suggested that RAPA inhibits adipogenesis and lipogenesis of muADSCs resulting in a prevention of obesity in C57BL/6J mice. This inhibition is strong enough to negate the effects of a high fat diet and seems to act by downregulating the RelA/p65 mTOR signaling pathway—a key component of the NF-κB family.
doi:10.1155/2014/540582
PMCID: PMC3920817  PMID: 24587943
13.  Infection of Mice, Ferrets, and Rhesus Macaques with a Clinical Mumps Virus Isolate 
Journal of Virology  2013;87(14):8158-8168.
In recent years, many mumps outbreaks have occurred in vaccinated populations worldwide. The reasons for these outbreaks are not clear. Animal models are needed to investigate the causes of outbreaks and to understand the pathogenesis of mumps virus (MuV). In this study, we have examined the infection of three animal models with an isolate of mumps virus from a recent outbreak (MuV-IA). We have found that while both ferrets and mice generated humoral and cellular immune responses to MuV-IA infection, no obvious signs of illness were observed in these animals; rhesus macaques were the most susceptible to MuV-IA infection. Infection of rhesus macaques via both intranasal and intratracheal routes with MuV-IA led to the typical clinical signs of mumps 2 weeks to 4 weeks postinfection. However, none of the infected macaques showed any fever or neurologic signs during the experimental period. Mumps viral antigen was detected in parotid glands by immunohistochemistry (IHC). Rhesus macaques represent the best animal model for the study of mumps virus pathogenesis.
doi:10.1128/JVI.01028-13
PMCID: PMC3700206  PMID: 23678169
14.  China’s Wetlands: Conservation Plans and Policy Impacts 
Ambio  2012;41(7):782-786.
doi:10.1007/s13280-012-0280-7
PMCID: PMC3472011  PMID: 22457078
15.  Contribution of Small RNA Pathway Components in Plant Immunity 
Small RNAs regulate a multitude of cellular processes, including development, stress responses, metabolism, and maintenance of genome integrity, in a sequence-specific manner. Accumulating evidence reveals that host endogenous small RNAs and small RNA pathway components play important roles in plant immune responses against various pathogens, including bacteria, fungi, oomycetes, and viruses. Small-RNA-mediated defense responses are regulated through diverse pathways and the components of these pathways, including Dicer-like proteins, RNA-dependent RNA polymerases, Argonaute proteins, and RNA polymerase IV and V, exhibit functional specificities as well as redundancy. In this review, we summarize the recent insights revealed mainly through the examination of two model plants, Arabidopsis and rice, with a primary focus on our emerging understanding of how these small RNA pathway components contribute to plant immunity.
doi:10.1094/MPMI-10-12-0255-IA
PMCID: PMC3752434  PMID: 23489060
16.  Engineered External Guide Sequences Are Highly Effective in Inhibiting Gene Expression and Replication of Hepatitis B Virus in Cultured Cells 
PLoS ONE  2013;8(6):e65268.
External guide sequences (EGSs) are RNA molecules that consist of a sequence complementary to a target mRNA and recruit intracellular ribonuclease P (RNase P), a tRNA processing enzyme, for specific degradation of the target mRNA. We have previously used an in vitro selection procedure to generate EGS variants that efficiently induce human RNase P to cleave a target mRNA in vitro. In this study, we constructed EGSs from a variant to target the overlapping region of the S mRNA, pre-S/L mRNA, and pregenomic RNA (pgRNA) of hepatitis B virus (HBV), which are essential for viral replication and infection. The EGS variant was about 50-fold more efficient in inducing human RNase P to cleave the mRNA in vitro than the EGS derived from a natural tRNA. Following Salmonella-mediated gene delivery, the EGSs were expressed in cultured HBV-carrying cells. A reduction of about 97% and 75% in the level of HBV RNAs and proteins and an inhibition of about 6,000- and 130-fold in the levels of capsid-associated HBV DNA were observed in cells treated with Salmonella vectors carrying the expression cassette for the variant and the tRNA-derived EGS, respectively. Our study provides direct evidence that the EGS variant is more effective in blocking HBV gene expression and DNA replication than the tRNA-derived EGS. Furthermore, these results demonstrate the feasibility of developing Salmonella-mediated gene delivery of highly active EGS RNA variants as a novel approach for gene-targeting applications such as anti-HBV therapy.
doi:10.1371/journal.pone.0065268
PMCID: PMC3680410  PMID: 23776459
17.  Hepatitis C Virus Activates Bcl-2 and MMP-2 Expression through Multiple Cellular Signaling Pathways 
Journal of Virology  2012;86(23):12531-12543.
Hepatitis C virus (HCV) infection is associated with numerous liver diseases and causes serious global health problems, but the mechanisms underlying the pathogenesis of HCV infections remain largely unknown. In this study, we demonstrate that signal transducer and activator of transcription 3 (STAT3), matrix metalloproteinase-2 (MMP-2), and B-cell lymphoma 2 (Bcl-2) are significantly stimulated in HCV-infected patients. We further show that HCV activates STAT3, MMP-2, Bcl-2, extracellular regulated protein kinase (ERK), and c-Jun N-terminal kinase (JNK) in infected Huh7.5.1 cells. Functional screening of HCV proteins revealed that nonstructural protein 4B (NS4B) is responsible for the activation of MMP-2 and Bcl-2 by stimulating STAT3 through repression of the suppressor of cytokine signaling 3 (SOCS3). Our results also demonstrate that multiple signaling cascades, including several members of the protein kinase C (PKC) family, JNK, ERK, and STAT3, play critical roles in the activation of MMP-2 and Bcl-2 mediated by NS4B. Further studies revealed that the C-terminal domain (CTD) of NS4B is sufficient for the activation of STAT3, JNK, ERK, MMP-2, and Bcl-2. We also show that amino acids 227 to 250 of NS4B are essential for regulation of STAT3, JNK, ERK, MMP-2, and Bcl-2, and among them, three residues (237L, 239S, and 245L) are crucial for this regulation. Thus, we reveal a novel mechanism underlying HCV pathogenesis in which multiple intracellular signaling cascades are cooperatively involved in the activation of two important cellular factors, MMP-2 and Bcl-2, in response to HCV infection.
doi:10.1128/JVI.01136-12
PMCID: PMC3497616  PMID: 22951829
18.  HCV-Induced miR-21 Contributes to Evasion of Host Immune System by Targeting MyD88 and IRAK1 
PLoS Pathogens  2013;9(4):e1003248.
Upon recognition of viral components by pattern recognition receptors, such as the toll-like receptors (TLRs) and retinoic acid-inducible gene I (RIG-I)-like helicases, cells are activated to produce type I interferon (IFN) and proinflammatory cytokines. These pathways are tightly regulated by the host to prevent an inappropriate cellular response, but viruses can modulate these pathways to proliferate and spread. In this study, we revealed a novel mechanism in which hepatitis C virus (HCV) evades the immune surveillance system to proliferate by activating microRNA-21 (miR-21). We demonstrated that HCV infection upregulates miR-21, which in turn suppresses HCV-triggered type I IFN production, thus promoting HCV replication. Furthermore, we demonstrated that miR-21 targets two important factors in the TLR signaling pathway, myeloid differentiation factor 88 (MyD88) and interleukin-1 receptor-associated kinase 1 (IRAK1), which are involved in HCV-induced type I IFN production. HCV-mediated activation of miR-21 expression requires viral proteins and several signaling components. Moreover, we identified a transcription factor, activating protein-1 (AP-1), which is partly responsible for miR-21 induction in response to HCV infection through PKCε/JNK/c-Jun and PKCα/ERK/c-Fos cascades. Taken together, our results indicate that miR-21 is upregulated during HCV infection and negatively regulates IFN-α signaling through MyD88 and IRAK1 and may be a potential therapeutic target for antiviral intervention.
Author Summary
Hepatitis C virus (HCV), a major cause of chronic hepatitis, end-stage cirrhosis, and hepatocellular carcinoma, has chronically infected 200 million people worldwide and 3–4 million more each year. When triggered by viral infection, host cells produce type I interferon (IFN) and proinflammatory cytokines to antagonize the virus. Despite extensive research, the mechanism underlying HCV immune system evasion remains elusive. Our results provided the first direct evidence that microRNA-21 (miR-21) feedback inhibits type I IFN signaling when cells are challenged with HCV, thus promoting the infection. MicroRNA is a kind of endogenous non-coding small RNA that regulates a wide range of biological processes and participate in innate and adaptive immune responses through complementarily pairing with target mRNA, which can regulate its expression or translation. Currently, miRNAs have intrigued many scientists as potent targets or therapeutic agents for diseases. In our study, the targets of miR-21, myeloid differentiation factor 88 (MyD88) and interleukin-1 receptor-associated kinase 1 (IRAK1), which are important for HCV-induced type I IFN production, have also been found. Moreover, we identified a transcription factor, AP-1, which is partly responsible for miR-21 induction in response to HCV infection. Taken together, our research has provided new insights into understanding the effects of miRNA on host-virus interactions, and revealed a potential therapeutic target for antiviral intervention.
doi:10.1371/journal.ppat.1003248
PMCID: PMC3635988  PMID: 23633945
19.  Use of extended curettage with osteotomy and fenestration followed by reconstruction with conservation of muscle insertion in the treatment of Enneking stage II locally aggressive bone tumor of the proximal extremities: resection and treatment of bone tumors 
Background
The purpose of this study was to investigate the clinical efficacy of extended resection with osteotomy, fenestration and conservation of muscle (tendon) insertion in the treatment of bone tumors.
Methods
A total of 15 patients with locally aggressive bone tumors (Enneking stage II) in the adjacent muscle (tendon) insertion of the proximal extremity were enrolled in the present study (mean age of 29 years). Extended curettage of lesions with osteotomy, fenestration and/or conservation of muscle (tendon) insertion and internal fixation with a bone graft or bone cement was performed at stage I. Postsurgical brace protection was used for 4 to 12 weeks and the patients were periodically followed-up by X-ray and functional assessment. Recurrence, postsurgical Enneking score and outcome rating were assessed.
Results
Treated cases included 15 patients aged 29 ±7.75 years (range, 18 to 42) with a male to female ratio of 8:7. Six had a femoral tumor and nine had a humeral tumor. These tumors comprised three chondroblastomas, five giant-cell tumors and seven aneurysmal bone cysts. Follow-up for 48 ±12.95 months (range, 25 to 72) revealed that 13 of 15 (87%) patients exhibited no recurrence. Local recurrence was observed in a patient with an aneurysmal bone cyst (nine months) and one with a giant-cell tumor (12 months). Mean Enneking scores were 27 ±4.07 (range, 18 to 29). Except for the patient with the recurrent giant-cell tumor, all patients reported good (13%, 2 out of 15) or very good (80%, 12 out of 15) outcomes. Very good outcomes were reported in 92% of patients (12 out of 13) without recurrence.
Conclusions
The procedures used in this study achieved high clinical efficacy, complete lesion removal, reduced recurrence and good restoration of joint function in patients with primary locally aggressive Enneking stage II bone tumors of the proximal extremities.
doi:10.1186/1477-7819-11-54
PMCID: PMC3618005  PMID: 23497479
Bone tumor; Conservation of muscle insertion; Extended curettage; Proximal extremity; Reconstruction; Resection
20.  Effective Inhibition of Human Immunodeficiency Virus 1 Replication by Engineered RNase P Ribozyme 
PLoS ONE  2012;7(12):e51855.
Using an in vitro selection procedure, we have previously isolated RNase P ribozyme variants that efficiently cleave an mRNA sequence in vitro. In this study, a variant was used to target the HIV RNA sequence in the tat region. The variant cleaved the tat RNA sequence in vitro about 20 times more efficiently than the wild type ribozyme. Our results provide the first direct evidence that combined mutations at nucleotide 83 and 340 of RNase P catalytic RNA from Escherichia coli (G83 -> U83 and G340 -> A340) increase the overall efficiency of the ribozyme in cleaving an HIV RNA sequence. Moreover, the variant is more effective in reducing HIV-1 p24 expression and intracellular viral RNA level in cells than the wild type ribozyme. A reduction of about 90% in viral RNA level and a reduction of 150 fold in viral growth were observed in cells that expressed the variant, while a reduction of less than 10% was observed in cells that either did not express the ribozyme or produced a catalytically inactive ribozyme mutant. Thus, engineered ribozyme variants are effective in inhibiting HIV infection. These results also demonstrate the potential of engineering RNase P ribozymes for anti-HIV application.
doi:10.1371/journal.pone.0051855
PMCID: PMC3530568  PMID: 23300569
21.  A Space-For-Time (SFT) Substitution Approach to Studying Historical Phenological Changes in Urban Environment 
PLoS ONE  2012;7(12):e51260.
Plant phenological records are crucial for predicting plant responses to global warming. However, many historical records are either short or replete with data gaps, which pose limitations and may lead to erroneous conclusions about the direction and magnitude of change. In addition to uninterrupted monitoring, missing observations may be substituted via modeling, experimentation, or gradient analysis. Here we have developed a space-for-time (SFT) substitution method that uses spatial phenology and temperature data to fill gaps in historical records. To do this, we combined historical data for several tree species from a single location with spatial data for the same species and used linear regression and Analysis of Covariance (ANCOVA) to build complementary spring phenology models and assess improvements achieved by the approach. SFT substitution allowed increasing the sample size and developing more robust phenology models for some of the species studied. Testing models with reduced historical data size revealed thresholds at which SFT improved historical trend estimation. We conclude that under certain circumstances both the robustness of models and accuracy of phenological trends can be enhanced although some limitations and assumptions still need to be resolved. There is considerable potential for exploring SFT analyses in phenology studies, especially those conducted in urban environments and those dealing with non-linearities in phenology modeling.
doi:10.1371/journal.pone.0051260
PMCID: PMC3517568  PMID: 23236460
22.  The Effects of Landscape Variables on the Species-Area Relationship during Late-Stage Habitat Fragmentation 
PLoS ONE  2012;7(8):e43894.
Few studies have focused explicitly on the later stages of the fragmentation process, or “late-stage fragmentation”, during which habitat area and patch number decrease simultaneously. This lack of attention is despite the fact that many of the anthropogenically fragmented habitats around the world are, or soon will be, in late-stage fragmentation. Understanding the ecological processes and patterns that occur in late-stage fragmentation is critical to protect the species richness in these fragments. We investigated plant species composition on 152 islands in the Thousand Island Lake, China. A random sampling method was used to create simulated fragmented landscapes with different total habitat areas and numbers of patches mimicking the process of late-stage fragmentation. The response of the landscape-scale species-area relationship (LSAR) to fragmentation per se was investigated, and the contribution of inter-specific differences in the responses to late-stage fragmentation was tested. We found that the loss of species at small areas was compensated for by the effects of fragmentation per se, i.e., there were weak area effects on species richness in landscapes due to many patches with irregular shapes and high variation in size. The study also illustrated the importance of inter-specific differences for responses to fragmentation in that the LSARs of rare and common species were differently influenced by the effects of fragmentation per se. In conclusion, our analyses at the landscape scale demonstrate the significant influences of fragmentation per se on area effects and the importance of inter-specific differences for responses to fragmentation in late-stage fragmentation. These findings add to our understanding of the effects of habitat fragmentation on species diversity.
doi:10.1371/journal.pone.0043894
PMCID: PMC3427301  PMID: 22937120
23.  The V Protein of Mumps Virus Plays a Critical Role in Pathogenesis 
Journal of Virology  2012;86(3):1768-1776.
Mumps virus (MuV) causes an acute infection in humans characterized by a wide array of symptoms ranging from relatively mild manifestations, such as parotitis, to more-severe complications, such as meningitis and encephalitis. Widespread mumps vaccination has reduced mumps incidence dramatically; however, outbreaks still occur in vaccinated populations. The V protein of MuV, when expressed in cell culture, blocks interferon (IFN) expression and signaling and interleukin-6 (IL-6) signaling. In this work, we generated a recombinant MuV incapable of expressing the V protein (rMuVΔV). The rescued MuV was derived from a clinical wild-type isolate from a recent outbreak in the United States (MuVIowa/US/06, G genotype). Analysis of the virus confirmed the roles of V protein in blocking IFN expression and signaling and IL-6 signaling. We also found that the rMuVIowa/US/06ΔV virus induced high levels of IL-6 expression in vitro, suggesting that V plays a role in reducing IL-6 expression. In vivo, the rMuVIowa/US/06ΔV virus was highly attenuated, indicating that the V protein plays an essential role in viral virulence.
doi:10.1128/JVI.06019-11
PMCID: PMC3264346  PMID: 22090137
24.  Activation of the Ras/Raf/MEK Pathway Facilitates Hepatitis C Virus Replication via Attenuation of the Interferon-JAK-STAT Pathway 
Journal of Virology  2012;86(3):1544-1554.
Hepatitis C virus (HCV) is a major cause of chronic liver diseases worldwide, often leading to the development of hepatocellular carcinoma (HCC). Constitutive activation of the Ras/Raf/MEK pathway is responsible for approximately 30% of cancers. Here we attempted to address the correlation between activation of this pathway and HCV replication. We showed that knockdown of Raf1 inhibits HCV replication, while activation of the Ras/Raf/MEK pathway by V12, a constitutively active form of Ras, stimulates HCV replication. We further demonstrated that this effect is regulated through attenuation of the interferon (IFN)-JAK-STAT pathway. Activation of the Ras/Raf/MEK pathway downregulates the expression of IFN-stimulated genes (ISGs), attenuates the phosphorylation of STAT1/2, and inhibits the expression of interferon (alpha, beta, and omega) receptors 1 and 2 (IFNAR1/2). Furthermore, we observed that HCV infection activates the Ras/Raf/MEK pathway. Thus, we propose that during HCV infection, the Ras/Raf/MEK pathway is activated, which in turn attenuates the IFN-JAK-STAT pathway, resulting in stimulation of HCV replication.
doi:10.1128/JVI.00688-11
PMCID: PMC3264379  PMID: 22114332
25.  Radius neck-to-humerus trochlea transposition elbow reconstruction after proximal ulnar metastatic tumor resection: case and literature review 
Wide en bloc excision of proximal ulna sections is used to treat traumatic and pathological fractures of the ulna, though poor standardization of clinical treatment often results in long-term failure of such reconstructed biomechanical structures. In order to provide insight into effective ulnar reconstructive treatments, the case of an 80-year-old Chinese Han male presenting with pathological fracture caused by a proximal ulnar metastatic tumor concurrent with metastatic renal cancer complicated by occurrence in the brain and lungs is reported and contrasted with alternative treatment techniques. Wide resectioning of the proximal ulna and reconstruction with local radius neck-to-humerus trochlea transposition resulted in preservation of functionality, sensitivity, and biomechanical integrity after postsurgical immobilization, 6 weeks of passive- and active-assisted flexion, and extension with a hinged brace. The resultant Musculoskeletal Tumor Society rating score was 25 of 30 (83 %). Full sensitivity and mobility of the left hand and elbow (10° to 90° with minimally impaired supination and pronation) was restored with minimal discomfort. No evidence of local recurrence or other pathological complications were observed within a 1-year follow-up period. Efficient reconstruction of osseous and capsuloligamentous structures in the elbow is often accomplished by allografts, prosthesis, and soft tissue reconstruction, though wide variations in risk and prognosis associated with these techniques has resulted in disagreements regarding the most effective standards for clinical treatment. Current findings suggest that radius neck-to-humerus trochlea transposition offers a superior range of elbow movement and fewer complications than similar allograft and prosthetic techniques for patients with multiple metastatic cancers.
doi:10.1186/2047-783X-17-23
PMCID: PMC3464775  PMID: 22800611
Proximal ulna; Metastatic tumor; Reconstructive procedures; Elbow reconstruction; Ulnar reconstruction

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