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1.  Complete Genome Sequence of a Novel Genotype of Squash Mosaic Virus Infecting Squash in Spain 
Genome Announcements  2015;3(1):e01583-14.
The complete genome sequence of a new isolate of squash mosaic virus (SqMV) infecting squash plants in Spain was obtained using deep sequencing of small RNAs. The low nucleotide sequence identities, with only 87 to 88% for RNA1 and 84 to 86% for RNA2 to known SqMV isolates, suggested that this isolate belongs to a novel genotype.
PMCID: PMC4333673  PMID: 25676773
3.  Chitosan/siRNA Nanoparticles Targeting Cyclooxygenase Type 2 Attenuate Unilateral Ureteral Obstruction-induced Kidney Injury in Mice 
Theranostics  2015;5(2):110-123.
Cyclooxygenase type 2 (COX-2) plays a predominant role in the progression of kidney injury in obstructive nephropathy. The aim of this study was to test the efficacy of chitosan/small interfering RNA (siRNA) nanoparticles to knockdown COX-2 specifically in macrophages to prevent kidney injury induced by unilateral ureteral obstruction (UUO). Using optical imaging techniques and confocal microscopy, we demonstrated that chitosan/siRNA nanoparticles accumulated in macrophages in the obstructed kidney. Consistent with the imaging data, the obstructed kidney contained a higher amount of siRNA and macrophages. Chitosan-formulated siRNA against COX-2 was evaluated on RAW macrophages demonstrating reduced COX-2 expression and activity after LPS stimulation. Injection of COX-2 chitosan/siRNA nanoparticles in mice subjected to three-day UUO diminished the UUO-induced COX-2 expression. Likewise, macrophages in the obstructed kidney had reduced COX-2 immunoreactivity, and histological examination showed lesser tubular damage in COX-2 siRNA-treated UUO mice. Parenchymal inflammation, assessed by tumor necrosis factor-alpha (TNF-α) and interleukin 6 mRNA expression, was attenuated by COX-2 siRNA. Furthermore, treatment with COX-2 siRNA reduced heme oxygenase-1 and cleaved caspase-3 in UUO mice, indicating lesser oxidative stress and apoptosis. Our results demonstrate a novel strategy to prevent UUO-induced kidney damage by using chitosan/siRNA nanoparticles to knockdown COX-2 specifically in macrophages.
PMCID: PMC4278998  PMID: 25553102
Cyclooxygenase type 2; siRNA; chitosan; unilateral ureteral obstruction; mice.
4.  Osteoblast inhibition by chemokine cytokine ligand3 in myeloma-induced bone disease 
Cancer Cell International  2014;14(1):132.
Multiple myeloma is a hematologic malignancy characterized by the accumulation of monoclonal plasma cells in the bone marrow. A common manifestation of the disease is myeloma bone disease (MBD), which is caused by increased osteoclastic bone resorption and decreased bone formation. The chemokine cytokine ligand 3 (CCL3) is a pro-inflammatory protein and chemokine that stimulates osteoclasts in MBD. However, little is known about the effect of CCL3 on osteoblasts (OB).
The OBs are induced from patients with MBD and healthy donors, cultured in vitro, and identified by histochemistry. The effects of CCL3 and CCL3 antibody on the OBs in vitro are observed. The CCL3 receptor (CCR1), osteocalcin (OCN), runt-related transcription factor 2 (Runx2), and osterix (Osx) are detected using flow cytometry, enzyme-linked immunosorbent assay, and real-time PCR.
Proliferation and osteogenic potential of the OB in patients with MBD are suppressed. Moreover, the CCR1 expression is significantly higher in patients with MBD than in normal controls. The OCN level, quantity of calcium nodules, and Runx2 and Osx levels decrease after CCL3 stimulation, which indicates that CCL3 inhibits OB function. Furthermore, CCL3 antibody partially restores OB activity through the upregulation of the OCN, Runx2, and Osx.
CCL3 contributes to the OB/OC imbalance by inhibiting OB differentiation and function in MBD.
PMCID: PMC4268854  PMID: 25520585
Chemokine cytokine ligand 3; Myeloma bone disease; Osteoblast; Runx2; Osterix
5.  Understanding the Role of Tbx1 as a Candidate Gene for 22q11.2 Deletion Syndrome 
Current allergy and asthma reports  2013;13(6):10.1007/s11882-013-0384-6.
22q11.2 deletion syndrome (22q11.2DS) is caused by a commonly occurring microdeletion on chromosome 22. Clinical findings include cardiac malformations, thymic and parathyroid hypoplasia, craniofacial dysmorphisms, and dental defects. These phenotypes are due mainly to abnormal development of the pharyngeal apparatus. Targeted deletion studies in mice and analysis of naturally occurring mutations in humans have implicated Tbx1 as a candidate gene for 22q11.2DS. Tbx1 belongs to an evolutionarily conserved T-box family of transcription factors, whose expression is precisely regulated during embryogenesis, and it appears to regulate the proliferation and differentiation of various progenitor cells during organogenesis. In this review, we discuss the mechanisms of Tbx1 during development of the heart, thymus and parathyroid glands, as well as during formation of the palate, teeth, and other craniofacial features.
PMCID: PMC3840116  PMID: 23996541
22q11.2 Deletion syndrome; DiGeorge syndrome; Velocardiofacial syndrome; Tbx1; Pharyngeal apparatus; Cardiovascular; Pharyngeal arch artery; Odontogenesis; Palatogenesis; Proliferation; Craniofacial; Thymus; Parathyroid; Differentiation; Candidate gene
6.  Prevalence, Risk, and Correlates of Posttraumatic Stress Disorder across Ethnic and Racial Minority Groups in the U.S 
Medical care  2013;51(12):1114-1123.
We assess whether posttraumatic stress disorder (PTSD) varies in prevalence, diagnostic criteria endorsement, and type and frequency of traumatic events (PTEs) among a nationally representative U.S. sample of 5071 non-Latino whites, 3264 Latinos, 2178 Asians, 4249 African Americans, and 1476 Afro-Caribbeans.
PTSD and other psychiatric disorders were evaluated using the World Mental Health-Composite International Diagnostic Interview (WMH-CIDI) in a national household sample that oversampled ethnic/racial minorities (n=16,238) but was weighted to produce results representative of the general population.
Asians have lower prevalence rates of probable lifetime PTSD while African Americans have higher rates as compared to non-Latino whites, even after adjusting for type and number of exposures to traumatic events, and for sociodemographic, clinical and social support factors. Afro-Caribbeans and Latinos seem to demonstrate similar risk to non-Latino whites, adjusting for these same covariates. Higher rates of probable PTSD exhibited by African Americans and lower rates for Asians, as compared to non-Latino whites, do not appear related to differential symptom endorsement, differences in risk or protective factors or differences in types and frequencies of PTEs across groups.
There appears to be marked differences in conditional risk of probable PTSD across ethnic/racial groups. Questions remain about what explains risk of probable PTSD. Several factors that might account for these differences are discussed as well as the clinical implications of our findings. Uncertainty of the PTSD diagnostic assessment for Latinos and Asians requires further evaluation.
PMCID: PMC3922129  PMID: 24226308
Posttraumatic Stress Disorders across racial and ethnic minority groups; diagnosis
7.  Increased Na+/Ca2+ Exchanger Activity Promotes Resistance to Excitotoxicity in Cortical Neurons of the Ground Squirrel (a Hibernator) 
PLoS ONE  2014;9(11):e113594.
Ground squirrel, a hibernating mammalian species, is more resistant to ischemic brain stress than rat. Gaining insight into the adaptive mechanisms of ground squirrels may help us design treatment strategies to reduce brain damage in patients suffering ischemic stroke. To understand the anti-stress mechanisms in ground squirrel neurons, we studied glutamate toxicity in primary cultured neurons of the Daurian ground squirrel (Spermophilus dauricus). At the neuronal level, for the first time, we found that ground squirrel was more resistant to glutamate excitotoxicity than rat. Mechanistically, ground squirrel neurons displayed a similar calcium influx to the rat neurons in response to glutamate or N-methyl-D-aspartate (NMDA) perfusion. However, the rate of calcium removal in ground squirrel neurons was markedly faster than in rat neurons. This allows ground squirrel neurons to maintain lower level of intracellular calcium concentration ([Ca2+]i) upon glutamate insult. Moreover, we found that Na+/Ca2+ exchanger (NCX) activity was higher in ground squirrel neurons than in rat neurons. We also proved that overexpression of ground squirrel NCX2, rather than NCX1 or NCX3, in rat neurons promoted neuron survival against glutamate toxicity. Taken together, our results indicate that ground squirrel neurons are better at maintaining calcium homeostasis than rat neurons and this is likely achieved through the activity of ground squirrel NCX2. Our findings not only reveal an adaptive mechanism of mammalian hibernators at the cellular level, but also suggest that NCX2 of ground squirrel may have therapeutic value for suppressing brain ischemic damage.
PMCID: PMC4240599  PMID: 25415196
8.  Activation of autophagy and paraptosis in retinal ganglion cells after retinal ischemia and reperfusion injury in rats 
Glaucoma is a neurodegenerative disease characterized by elevated intraocular pressure (IOP), which causes retinal ischemia and progressive neuronal death. Retinal ischemia/reperfusion (RIR) injury is a common clinical condition representing the main cause of irreversible visual field defects in humans. The aim of this study was to investigate whether non-apoptotic forms of programmed cell death (PCD) have an effect on RIR injury in an experimental model that replicates features of acute hypertensive glaucoma and to explore the possible underlying mechanisms. The activation of autophagy was investigated in retinal ganglion cells (RGCs) following RIR in comparison with a control group, using immunofluorescence against microtubule-associated protein 1 light chain 3 (LC3). RIR injury increased LC3 expression in the cytoplasm of RGCs in the ganglion cell layer (GCL) 6 h after the insult, and the increased expression was sustained throughout the experimental period. Following RIR insult, the number of neurons in the GCL significantly decreased. Ultra-structural analyses showed that double- or multiple-membrane autophagosomes were markedly accumulated in the cytoplasm of RGCs following IOP elevation. Since there are no known markers for paraptosis, its identification was based on morphological criteria. Electron microscopy (EM) analysis revealed severe structural alterations associated with cytoplasmatic vacuolization within the 6 h after RIR injury and RGC death. EM also revealed that vacuoles were derived predominantly from the progressive swelling of the endoplasmic reticulum (ER) and/or mitochondria in RGCs after RIR injury. The results provide novel evidence implicating an important role of autophagy and paraptosis in the pathogenesis of RIR injury. Autophagy and paraptosis take place during developmental cell death in the nervous system as well as in certain cases of neurodegeneration. Therefore, targeting autophagy and paraptosis could have therapeutic potential for the prevention of glaucoma involving RIR injury.
PMCID: PMC4280957  PMID: 25574219
autophagy; paraptosis; retinal ischemia/reperfusion injury; retinal ganglion cells
9.  Long-Term Effects of Musical Training and Functional Plasticity in Salience System 
Neural Plasticity  2014;2014:180138.
Musicians undergoing long-term musical training show improved emotional and cognitive function, which suggests the presence of neuroplasticity. The structural and functional impacts of the human brain have been observed in musicians. In this study, we used data-driven functional connectivity analysis to map local and distant functional connectivity in resting-state functional magnetic resonance imaging data from 28 professional musicians and 28 nonmusicians. Compared with nonmusicians, musicians exhibited significantly greater local functional connectivity density in 10 regions, including the bilateral dorsal anterior cingulate cortex, anterior insula, and anterior temporoparietal junction. A distant functional connectivity analysis demonstrated that most of these regions were included in salience system, which is associated with high-level cognitive control and fundamental attentional process. Additionally, musicians had significantly greater functional integration in this system, especially for connections to the left insula. Increased functional connectivity between the left insula and right temporoparietal junction may be a response to long-term musical training. Our findings indicate that the improvement of salience network is involved in musical training. The salience system may represent a new avenue for exploration regarding the underlying foundations of enhanced higher-level cognitive processes in musicians.
PMCID: PMC4247966  PMID: 25478236
10.  Complete Genome Sequence of an Emerging Genotype of Tobacco Streak Virus in the United States 
Genome Announcements  2014;2(6):e01138-14.
We report here the complete genome sequence of an emerging genotype of tobacco streak virus (TSV) infecting zucchini squash in Florida (TSV_FL13-07), obtained using deep sequencing of short RNAs (sRNAs) and validation by Sanger sequencing. TSV_FL13-07 shares only <90% sequence identity in all three genomic RNAs to several known U.S. isolates.
PMCID: PMC4223465  PMID: 25377714
11.  A low-toxic site-directed mutant of Clostridium perfringens ε-toxin as a potential candidate vaccine against enterotoxemia 
Human Vaccines & Immunotherapeutics  2013;9(11):2386-2392.
Clostridium perfringens epsilon toxin (ETX), one of the most potent toxins known, is a potential biological weapon; therefore, the development of an effective vaccine is important for preventing intoxication or disease by ETX. In this study, genetically detoxified epsilon toxin mutants were developed as candidate vaccines. We used site-directed mutagenesis to mutate the essential amino acid residues (His106, Ser111 and Phe199). Six site-directed mutants of ETX (mETXH106P, mETXS111H, mETXS111Y, mETXF199H, mETXF199E, mETXS111YF199E) were generated and then expressed in Escherichia coli. Both mETXF199E and mETXH106P with low or non-cytotoxicity that retained their immunogenicity were selected to immunize mice 3 times, and the mouse survival data were recorded after challenging with recombinant wild-type ETX. mETXF199E induces the same protection as mETXH106P, which was reported previously as a promising toxin mutant for vaccine, and both of them could protect immunized mice against a 100× LD50 dose of active wild-type recombinant ETX. This work showed that mETXF199E is another promising candidate vaccine against enterotoxemia and other diseases caused by ETX.
PMCID: PMC3981848  PMID: 23835363
Clostridium perfringens; cytotoxicity; enterotoxemia; site-directed mutagenesis; toxin mutant; vaccine; ε-toxin
12.  Expression Analysis of Immune Related Genes Identified from the Coelomocytes of Sea Cucumber (Apostichopus japonicus) in Response to LPS Challenge 
The sea cucumber (Apostichopus japonicus) occupies a basal position during the evolution of deuterostomes and is also an important aquaculture species. In order to identify more immune effectors, transcriptome sequencing of A. japonicus coelomocytes in response to lipopolysaccharide (LPS) challenge was performed using the Illumina HiSeq™ 2000 platform. One hundred and seven differentially expressed genes were selected and divided into four functional categories including pathogen recognition (25 genes), reorganization of cytoskeleton (27 genes), inflammation (41 genes) and apoptosis (14 genes). They were analyzed to elucidate the mechanisms of host-pathogen interactions and downstream signaling transduction. Quantitative real-time polymerase chain reactions (qRT-PCRs) of 10 representative genes validated the accuracy and reliability of RNA sequencing results with the correlation coefficients from 0.88 to 0.98 and p-value <0.05. Expression analysis of immune-related genes after LPS challenge will be useful in understanding the immune response mechanisms of A. japonicus against pathogen invasion and developing strategies for resistant markers selection.
PMCID: PMC4264123  PMID: 25421239
sea cucumber (Apostichopus japonicus); differentially expressed genes; quantitative real-time polymerase chain reaction
13.  Quantitative proteomic analysis of thylakoid from two microalgae (Haematococcus pluvialis and Dunaliella salina) reveals two different high light-responsive strategies 
Scientific Reports  2014;4:6661.
Under high light (HL) stress, astaxanthin-accumulating Haematococcus pluvialis and β-carotene-accumulating Dunaliella salina showed different responsive patterns. To elucidate cellular-regulating strategies photosynthetically and metabolically, thylakoid membrane proteins in H. pluvialis and D. salina were extracted and relatively quantified after 0 h, 24 h and 48 h of HL stress. Proteomic analysis showed that three subunits of the cytochrome b6/f complex were greatly reduced under HL stress in H. pluvialis, while they were increased in D. salina. Additionally, the major subunits of both photosystem (PS) II and PSI reaction center proteins were first reduced and subsequently recovered in H. pluvialis, while they were gradually reduced in D. salina. D. salina also showed a greater ability to function using the xanthophyll-cycle and the cyclic photosynthetic electron transfer pathway compared to H. pluvialis. We propose a reoriented and effective HL-responsive strategy in H. pluvialis, enabling it to acclimate under HL. The promising metabolic pathway described here contains a reorganized pentose phosphate pathway, Calvin cycle and glycolysis pathway participating in carbon sink formation under HL in H. pluvialis. Additionally, the efficient carbon reorientation strategy in H. pluvialis was verified by elevated extracellular carbon assimilation and rapid conversion into astaxanthin.
PMCID: PMC4205843  PMID: 25335577
14.  Age-Triggered and Dark-Induced Leaf Senescence Require the bHLH Transcription Factors PIF3, 4, and 5 
Molecular Plant  2014;7(12):1776-1787.
Numerous lines of evidence have suggested an involvement of phytochromes in the regulation of leaf senescence, but the related signaling pathway and physiological mechanisms are poorly understood. In this study, we demonstrated that PIF3, 4, and 5, the master regulators of light signaling, modulate age-triggered and dark-induced senescence and particularly PIF4 positively regulates leaf senescence by directly targeting genes related to chlorophyll degradation and chloroplast activity maintaining as well as ethylene biosynthesis.
Leaf senescence can be triggered and promoted by a large number of developmental and environmental factors. Numerous lines of evidence have suggested an involvement of phytochromes in the regulation of leaf senescence, but the related signaling pathways and physiological mechanisms are poorly understood. In this study, we initially identified phytochrome-interacting factors (PIFs) 3, 4, and 5 as putative mediators of leaf senescence. Mutations of the PIF genes resulted in a significantly enhanced leaf longevity in age-triggered and dark-induced senescence, whereas overexpressions of these genes accelerated age-triggered and dark-induced senescence in Arabidopsis. Consistently, loss-of-function of PIF4 attenuated dark-induced transcriptional changes associated with chloroplast deterioration and reactive oxygen species (ROS) generation. ChIP–PCR and Dual-Luciferase assays demonstrated that PIF4 can activate chlorophyll degradation regulatory gene NYE1 and repress chloroplast activity maintainer gene GLK2 by binding to their promoter regions. Finally, dark-induced ethylene biosynthesis and ethylene-induced senescence were both dampened in pif4, suggesting the involvement of PIF4 in both ethylene biosynthesis and signaling pathway. Our study provides evidence that PIF3, 4, and 5 are novel positive senescence mediators and gains an insight into the mechanism of light signaling involved in the regulation of leaf senescence.
PMCID: PMC4261840  PMID: 25296857
phytochrome-interacting factor (PIF); leaf senescence; chloroplast deterioration; NYE1/SGR1; GLK2; ethylene.
15.  Application safety evaluation of the radio frequency identification tag under magnetic resonance imaging 
Radio Frequency Identification(RFID) has been widely used in healthcare facilities, but it has been paid little attention whether RFID applications are safe enough under healthcare environment. The purpose of this study is to assess the effects of RFID tags on Magnetic Resonance (MR) imaging in a typical electromagnetic environment in hospitals, and to evaluate the safety of their applications.
A Magphan phantom was used to simulate the imaging objects, while active RFID tags were placed at different distances (0, 4, 8, 10 cm) from the phantom border. The phantom was scanned by using three typical sequences including spin-echo (SE) sequence, gradient-echo (GRE) sequence and inversion-recovery (IR) sequence. The quality of the image was quantitatively evaluated by using signal-to-noise ratio (SNR), uniformity, high-contrast resolution, and geometric distortion. RFID tags were read by an RFID reader to calculate their usable rate.
RFID tags can be read properly after being placed in high magnetic field for up to 30 minutes. SNR: There were no differences between the group with RFID tags and the group without RFID tags using SE and IR sequence, but it was lower when using GRE sequence.Uniformity: There was a significant difference between the group with RFID tags and the group without RFID tags using SE and GRE sequence. Geometric distortion and high-contrast resolution: There were no obvious differences found.
Active RFID tags can affect MR imaging quality, especially using the GRE sequence. Increasing the distance from the RFID tags to the imaging objects can reduce that influence. When the distance was longer than 8 cm, MR imaging quality were almost unaffected. However, the Gradient Echo related sequence is not recommended when patients wear a RFID wristband.
PMCID: PMC4175226  PMID: 25187420
RFID tags; MRI sequence; Electromagnetic interference; Imaging quality
17.  Moyamoya syndrome associated with Graves’ disease: a case series study 
The aim of this study was to describe the clinical and radiological findings of patients with moyamoya syndrome and Graves’ disease. Possible mechanisms predisposing these individuals to ischemic stroke are discussed.
We retrospectively analyzed 12 consecutive patients with both moyamoya syndrome and Graves’ disease. Moyamoya vasculopathy was diagnosed by digital subtract angiography or magnetic resonance angiography (MRA). The clinical characteristics, laboratory data, vascular radiological characteristics and outcome were reported.
All patients were female and mean age was 33.33±12.65 years. Stenosis or occlusion of bilateral terminal internal carotid artery and/or proximal anterior/middle cerebral arteries was found in nine patients. Among them, three patients displayed asymmetrical stenosis. In addition, there were three patients with probable unilateral moamoya syndrome. Eleven patients presented with ischemic stroke and/or transient ischemic attack (TIA) and one with dizziness. Thyroid function tests demonstrated elevated thyroid hormone levels and suppressed thyroid stimulating hormone levels in all the patients with ischemic events. All patients received anti-thyroid therapy and two had recurrent ischemic attack after drug withdrawal.
Moyamoya syndrome associated Graves’ disease often presented with asymmetric stenosis or occlusion. We hypothesize cerebrovascular hemodynamic changes due to thyrotoxicosis contribute to the ischemic events.
PMCID: PMC4200642  PMID: 25333052
Moyamoya syndrome; Graves’ disease; thyrotoxicosis; treatment
18.  Luminal thrombosis in middle cerebral artery occlusions: a high-resolution MRI study 
Background and purpose
High signals within occluded vessels on T1-weighted fat-suppressed images (HST1) are highly suggestive of luminal thrombosis. We sought to investigate the feasibility of in vivo identification of luminal thrombosis in middle cerebral artery (MCA) occlusions using high-resolution magnetic resonance imaging (HR-MRI).
We retrospectively reviewed the HR-MRI data of 25 patients with unilateral symptomatic MCA occlusion. HST1 were defined as an area of high signal within the cross-section of occluded MCA, the intensity of which was >150% of the signal of adjacent muscles. The prevalence of HST1 and their relationship to infarct sizes and infarct patterns were analyzed.
The average time from stroke onset to HR-MRI examination was 9±6 days. There were 18 (72%) occluded vessels with HST1 on HR-MRI. HST1 were observed in 5/7 patients with a large territory infarct (≥1/3 MCA distribution) and 13/18 patients without (P=0.37). In the patients with non-large territory infarcts, the presence of HST1 was similar in those with and without border zone infarcts (9/13 vs. 4/5, P=0.42).
It’s feasible to in vivo identify luminal thrombosis in occluded MCA. HR-MRI is a potentially powerful tool for investigating the mechanisms of stroke due to MCA occlusions.
PMCID: PMC4200643  PMID: 25333050
Middle cerebral artery occlusion (MCA occlusion); magnetic resonance imaging; thrombosis; cerebral infarct
19.  Low awareness of stroke guidelines and preference for Chinese herbs in community physicians: a national survey in China 
Physicians’ adherence to stroke guidelines is becoming a critical part of public stroke care system. The objective of this national survey was to examine Chinese physicians’ awareness of the guidelines in secondary stroke prevention.
This is a non-commercial and no-incentive internet survey. Respondents were asked to perform a self-examination of 13 questions regarding their stroke practice. Their awareness of stroke guidelines, preference for Chinese traditional herbs (CTH), and patients’ expense for stroke treatment were surveyed and compared between physicians from community and from tertiary hospitals using univariate analysis and logistic regression.
A total of 8,581 physicians (70.1% from community hospitals) responded to the survey. Only 32.1% physicians considered risk factors control necessary for stroke. For the treatments of symptomatic carotid stenosis, only 10.4% physicians selected carotid endarterectomy and anti-platelet plus controlling stroke risk factors. Only 21.45% physicians selected warfarin anticoagulation for stroke patients with atrial fibrillation. In contrast, a high percentage (64.56%) of physicians had positive attitude towards CTH. Compared with those from tertiary hospitals, community physicians were more likely unaware of the guidelines and preferred CTH. Those who prescribed CTH reported more patients’ cost (P<0.001, OR 1.78, 95% CI, 1.55-2.04) than who didn’t.
There is a very low awareness of stroke guidelines in Chinese community physicians. A well-organized continuing stroke-guidelines education should be an essential part of public stroke-care system in China. Also, more well-designed clinical trials are required to establish the safety and effectiveness of CTH.
PMCID: PMC4200646  PMID: 25333051
Guideline adherence; stroke; China; community health workers; Chinese traditional medicine
20.  Gene profiling analysis for patients with oral verrucous carcinoma and oral squamous cell carcinoma 
Oral verrucous carcinoma (OVC) is one malignant tumor which was carved out from the oral squamous cell carcinoma (OSCC). However, the clinical and pathological features as well as the treatment strategies of OVC are different from OSCC. Here, global transcript abundance of tumor tissues from five patients with primary OVC and six patients with primary OSCC including their matched adjacently normal oral mucosa were profiled using the Affymetrix HGU133 Plus 2.0. Ingenuity Systems IPA software was used to analyse the gene function and biological pathways. There were 109 differentially expressed genes (more than 2-fold) between OVC and the adjacently normal tissue, among them 66 were up-regulated and 43 were down-regulated; 1172 differentially expressed genes (more than 2-fold) between OSCC and the adjacently normal tissue, among them 608 were up-regulated and 564 were down-regulated. There were 39 common differentially expressed genes in OVC and OSCC compared with their matched normal oral mucosa, among them 22 up-regulated and 17 down-regulated, and 8 of them different between OVC and OSCC. In addition, the gene expression profile was further validated by quantitative real-time PCR (Q-RT-PCR) analysis for four of those 39 selected genes.
PMCID: PMC4132153  PMID: 25126189
Oral verrucous carcinoma; oral squamous cell carcinoma; biological pathways; expression profile; quantitative real-time PCR
21.  Applications of RNA interference high-throughput screening technology in cancer biology and virology 
Protein & Cell  2014;5(11):805-815.
RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function. Large-scale gene silencing using RNAi high-throughput screening (HTS) has opened an exciting frontier to systematically study gene function in mammalian cells. This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight. Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.
PMCID: PMC4225462  PMID: 24952721
RNA interference (RNAi); short interfering RNA (siRNA); short hairpin RNA (shRNA); high-throughput screening; cancer; virology
22.  Applications of RNA interference high-throughput screening technology in cancer biology and virology 
Protein & Cell  2014;5(11):805-815.
RNA interference (RNAi) is an ancient intra-cellular mechanism that regulates gene expression and cell function. Large-scale gene silencing using RNAi high-throughput screening (HTS) has opened an exciting frontier to systematically study gene function in mammalian cells. This approach enables researchers to identify gene function in a given biological context and will provide considerable novel insight. Here, we review RNAi HTS strategies and applications using case studies in cancer biology and virology.
PMCID: PMC4225462  PMID: 24952721
RNA interference (RNAi); short interfering RNA (siRNA); short hairpin RNA (shRNA); high-throughput screening; cancer; virology
23.  Chitosan Hydrogel as siRNA vector for prolonged gene silencing 
The periodontitis is one of the most prevalent diseases with alveolar resorption in adult people and is the main cause of the tooth loss. To investigate the possibility for protecting the loss of alveolar bone in periodontal diseases, a RNAi-based therapeutic strategy is applied for silencing RANK signaling using thermosensitive chitosan hydrogel as siRNA reservoir and vector.
The thermosensitive chitosan hydrogel was formed from solution (PH = 7.2, at 4°C) at 37°C within 8 minutes. The degradation rates of hydrogel were ~50% and 5% (W remaining/W beginning) in the presence and absence of lysozyme, respectively, over a period of 20 days. The concurrent cumulative in vitro release of Cy3-labeled siRNA from the hydrogel was 50% and 17% over 14 days, with or without lysozyme digestion, respectively. High cell viability (>88%) was maintained for cells treated with hydrogel loaded with RANK specific siRNA and RANK knockdown was prolonged for up to 9 days when cells were incubated with siRNA/hydrogel complex. In vivo release of siRNA was investigated in a subcutaneous delivery setup in mice. The fluorescent signal from siRNA within hydrogel was remained for up to 14 days compared to less than one day for siRNA alone.
Chitosan hydrogel can potentially serve as a suitable reservoir and vector for local sustained delivery of siRNA in potential therapy.
PMCID: PMC4104730  PMID: 24946934
24.  Possible prognostic significance of p53, cyclin D1 and Ki-67 in the second primary malignancy of patients with double primary malignancies 
Patients with two types of primary cancers are rare. In this study, we investigated the expression of p53, cyclin D1, and Ki-67 in the second primary malignancy. Tissue samples were obtained from the second primary cancer site of 43 patients who met the diagnostic criteria for double primary cancer. p53, cyclin D1 and Ki-67 were determined using immunohistochemistry. Categorical variables were compared using the Chi-squared test; correlation between data scores and histology was calculated using the Spearman’s rank-order correlation. The expression rates of p53, cyclin D1 and Ki-67 in the second primary malignancy site were 60.5%, 30.2% and 65.1% respectively. p53 expression showed statistically significant association with tumor occurrence interval, pathological grading and nodal metastasis (p < 0.05). Positive correlation was detected between the expression of cyclin D1 and Ki-67 and the expression of p53 (r = 0.313, p = 0.041; r = 0.319, p = 0.037, respectively). High-expressing p53 or cyclin D second primary malignancies were associated with decreased overall survival (p = 0.040 and p = 0.043, respectively). Ki-67 expression levels did not exhibit statistically significant differences in survival. In conclusion, elevated protein expression of p53, cyclin D1 and Ki-67 in the second primary malignancy is an indicator of more aggressive malignant behavior of the secondary tumor. These markers may have prognostic value in the clinical setting.
PMCID: PMC4129009  PMID: 25120774
Double primary malignancies; second primary malignancy; immunohistochemistry; p53; cyclin D1; Ki-67

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