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author:("An, jisheng")
1.  Oocyte spindle proteomics analysis leading to rescue of chromosome congression defects in cloned embryos 
Journal of proteome research  2010;9(11):6025-6032.
Embryos produced by somatic cell nuclear transfer (SCNT) display low term developmental potential. This is associated with deficiencies in spindle composition prior to activation and at early mitotic divisions, including failure to assemble certain proteins on the spindle. The protein-deficient spindles are accompanied by chromosome congression defects prior to activation and during the first mitotic divisions of the embryo. The molecular basis for these deficiencies and how they might be avoided are unknown. Proteomic analyses of spindles isolated from normal metaphase II (MII) stage oocytes and SCNT constructs, along with a systematic immunofluorescent survey of known spindle-associated proteins were undertaken. This was the first proteomics study of mammalian oocyte spindles. The study revealed four proteins as being deficient in spindles of SCNT embryos in addition to those previously identified; these were clathrin heavy chain (CLTC), aurora B kinase, dynactin 4, and casein kinase 1 alpha. Due to substantial reduction in CLTC abundance after spindle removal, we undertook functional studies to explore the importance of CLTC in oocyte spindle function and in chromosome congression defects of cloned embryos. Using siRNA knockdown we demonstrated an essential role for CLTC in chromosome congression during oocyte maturation. We also demonstrated rescue of chromosome congression defects in SCNT embryos at the first mitosis using CLTC mRNA injection. These studies are the first to employ proteomics analyses coupled to functional interventions to rescue a specific molecular defect in cloned embryos.
doi:10.1021/pr100827j
PMCID: PMC4316211  PMID: 20883044
somatic cell nuclear transfer; meiosis; oogenesis; gene expression; spindle assembly
2.  Loss of TLR3 aggravates CHIKV replication and pathology due to an altered virus-specific neutralizing antibody response 
EMBO Molecular Medicine  2014;7(1):24-41.
RNA-sensing toll-like receptors (TLRs) mediate innate immunity and regulate anti-viral response. We show here that TLR3 regulates host immunity and the loss of TLR3 aggravates pathology in Chikungunya virus (CHIKV) infection. Susceptibility to CHIKV infection is markedly increased in human and mouse fibroblasts with defective TLR3 signaling. Up to 100-fold increase in CHIKV load was observed in Tlr3−/− mice, alongside increased virus dissemination and pro-inflammatory myeloid cells infiltration. Infection in bone marrow chimeric mice showed that TLR3-expressing hematopoietic cells are required for effective CHIKV clearance. CHIKV-specific antibodies from Tlr3−/− mice exhibited significantly lower in vitro neutralization capacity, due to altered virus-neutralizing epitope specificity. Finally, SNP genotyping analysis of CHIKF patients on TLR3 identified SNP rs6552950 to be associated with disease severity and CHIKV-specific neutralizing antibody response. These results demonstrate a key role for TLR3-mediated antibody response to CHIKV infection, virus replication and pathology, providing a basis for future development of immunotherapeutics in vaccine development.
doi:10.15252/emmm.201404459
PMCID: PMC4309666  PMID: 25452586
Chikungunya virus; innate immunity; joint inflammation; neutralizing antibodies; TLR3
3.  High-density Lipoprotein Increases the Uptake of Oxidized Low Density Lipoprotein via PPARγ/CD36 Pathway in Inflammatory Adipocytes 
Aim: Previous studies have demonstrated that the dysregulated-secretion of adipokines by adipocytes may contribute to obesity-associated atherosclerosis (As) and high density lipoprotein (HDL) may protect against atherogenesis through multiple pathways. This study was to explore the effect of HDL on the oxLDL uptake in inflammatory adipocytes stimulated by endotoxin lipopolysaccharide (LPS) and the possible mechanism.
Methods and Results: 3T3-L1 adipocytes were cultured and induced to differentiation and maturation. Acute inflammation in adipocytes was induced by LPS (100 ng/ml) for 6 hours. The adipocytes were pretreated with HDL in various concentrations (10, 50, 100 μg/ml) for 16 hours or with specific PPARγ antagonist (GW9662, 10 μM) or agonist (Rosiglitazone, 10 μM) for 30 min before administration of LPS. The results showed that LPS significantly increased the release of inflammation-related adipokines, such as monocyte chemoattractant protein-1 (MCP-1), plasminogen activator inhibitor 1 (PAI-1), tumor necrosis factor-alpha (TNF-α), interleukin (IL)-8 and IL-6, while decreasing the release of leptin and adiponectin. Meanwhile, LPS reduced the uptake and degradation of 125I-oxLDL, and down-regulated the expression of PPARγ and CD36. Pretreatment with HDL dose-dependently affected the release of IL-8 and IL-6 and the reduced uptake and degradation of oxLDL of adipocytes stimulated by LPS, accompanied with marked upregulation of PPARγ and CD36 expression. Pretreatment with GW9662 markedly inhibited the upregulation of CD36 expression mediated by HDL (100 μg/ml), while the effects of Rosiglitazone were opposite to GW9662.
Conclusions: HDL may increase oxLDL uptake of inflammatory adipocytes stimulated by LPS via upregulation of PPARγ/CD36 pathway, which may be a new mechanism of anti-atherosclerosis mediated by HDL.
doi:10.7150/ijbs.10258
PMCID: PMC4323365
Lipoprotein; Atherosclerosis; Inflammation; Lipopolysaccharide
4.  Biomass pyrolysis liquid to citric acid via 2-step bioconversion 
Microbial Cell Factories  2014;13(1):182.
Background
The use of fossil carbon sources for fuels and petrochemicals has serious impacts on our environment and is unable to meet the demand in the future. A promising and sustainable alternative is to substitute fossil carbon sources with microbial cell factories converting lignocellulosic biomass into desirable value added products. However, such bioprocesses require tolerance to inhibitory compounds generated during pretreatment of biomass. In this study, the process of sequential two-step bio-conversion of biomass pyrolysis liquid containing levoglucosan (LG) to citric acid without chemical detoxification has been explored, which can greatly improve the utilization efficiency of lignocellulosic biomass.
Results
The sequential two-step bio-conversion of corn stover pyrolysis liquid to citric acid has been established. The first step conversion by Phanerochaete chrysosporium (P. chrysosporium) is desirable to decrease the content of other compounds except levoglucosan as a pretreatment for the second conversion. The remaining levoglucosan in solution was further converted into citric acid by Aspergillus niger (A. niger) CBX-209. Thus the conversion of cellulose to citric acid is completed by both pyrolysis and bio-conversion technology. Under experimental conditions, levoglucosan yield is 12% based on the feedstock and the citric acid yield can reach 82.1% based on the levoglucosan content in the pyrolysis liquid (namely 82.1 g of citric acid per 100 g of levoglucosan).
Conclusion
The study shows that P. chrysosporium and A. niger have the potential to be used as production platforms for value-added products from pyrolyzed lignocellulosic biomass. Selected P. chrysosporium is able to decrease the content of other compounds except levoglucosan and levoglucosan can be further converted into citric acid in the residual liquids by A. niger. Thus the conversion of cellulose to citric acid is completed by both pyrolysis and bio-conversion technology.
doi:10.1186/s12934-014-0182-4
PMCID: PMC4299680  PMID: 25551193
Lignocellulosic biomass; Pyrolysis; Levoglucosan; Bio-conversion; Citric acid
5.  Analysis of codon usage bias of mitochondrial genome in Bombyx mori and its relation to evolution 
BMC Evolutionary Biology  2014;14(1):262.
Background
Synonymous codon usage bias (SCUB) is an inevitable phenomenon in organismic taxa, generally referring to differences in the occurrence frequency of codons across different species or within the genome of the same species. SCUB happens in various degrees under pressure from nature selection, mutation bias and other factors in different ways. It also attaches great significance to gene expression and species evolution, however, a systematic investigation towards the codon usage in Bombyx mori (B. mori) has not been reported yet. Moreover, it is still indistinct about the reasons contributing to the bias or the relationship between the bias and the evolution of B. mori.
Results
The comparison of the codon usage pattern between the genomic DNA (gDNA) and the mitochondrial DNA (mtDNA) from B. mori suggests that mtDNA has a higher level of codon bias. Furthermore, the correspondence analysis suggests that natural selection, such as gene length, gene function and translational selection, dominates the codon preference of mtDNA, while the composition constraints for mutation bias only plays a minor role. Additionally, the clustering results of the silkworm superfamily suggest a lack of explicitness in the relationship between the codon usage of mitogenome and species evolution.
Conclusions
Among the complicated influence factors leading to codon bias, natural selection is found to play a major role in shaping the high bias in the mtDNA of B. mori from our current data. Although the cluster analysis reveals that codon bias correlates little with the species evolution, furthermore, a detailed analysis of codon usage of mitogenome provides better insight into the evolutionary relationships in Lepidoptera. However, more new methods and data are needed to investigate the relationship between the mtDNA bias and evolution.
doi:10.1186/s12862-014-0262-4
PMCID: PMC4276022  PMID: 25515024
Bombyx mori; Synonymous codon usage bias; Genomic DNA; Mitochondrial DNA; Evolution
6.  A Phase I Study of Vorinostat in Combination with Bortezomib in Patients with Advanced Malignancies 
Investigational new drugs  2013;31(6):1539-1546.
Background
A phase I study to assess the maximum-tolerated dose (MTD), dose-limiting toxicity (DLT), pharmacokinetics (PK) and antitumor activity of vorinostat in combination with bortezomib in patients with advanced solid tumors.
Methods
Patients received vorinostat orally once daily on days 1–14 and bortezomib intravenously on days 1, 4, 8 and 11 of a 21-day cycle. Starting dose (level 1) was vorinostat (400 mg) and bortezomib (0.7 mg/m2). Bortezomib dosing was increased using a standard phase I dose-escalation schema. PKs were evaluated during cycle 1.
Results
Twenty-three patients received 57 cycles of treatment on four dose levels ranging from bortezomib 0.7 mg/m2 to 1.5 mg/m2. The MTD was established at vorinostat 400 mg daily and bortezomib 1.3 mg/m2. DLTs consisted of grade 3 fatigue in three patients (1 mg/m2,1.3 mg/m2 and 1.5 mg/m2) and grade 3 hyponatremia in one patient (1.5 mg/m2). The most common grade 1/2 toxicities included nausea (60.9%), fatigue (34.8%), diaphoresis (34.8%), anorexia (30.4%) and constipation (26.1%). Objective partial responses were observed in one patient with NSCLC and in one patient with treatment-refractory soft tissue sarcoma. Bortezomib did not affect the PKs of vorinostat; however, the Cmax and AUC of the acid metabolite were significantly increased on day 2 compared with day 1.
Conclusions
This combination was generally well-tolerated at doses that achieved clinical benefit. The MTD was established at vorinostat 400 mg daily x 14 days and bortezomib 1.3 mg/m2 on days 1, 4, 8 and 11 of a 21-day cycle.
doi:10.1007/s10637-013-0029-6
PMCID: PMC3901262  PMID: 24114121
SAHA; vorinostat; PS-341; bortezomib; phase I
7.  Doxorubicin-eluting beads versus conventional transarterialchemoembolization for the treatment of hepatocellular carcinoma: a meta-analysis 
Objective: We conducted a meta-analysis to evaluate the efficacy and toxicity of DEB-TACE in the treatment of patients with intermediate-stage HCC. Methods: Studies published in PubMed, Embase and Web of Science, were systematically reviewed to identify those that assessed the efficacy and toxicity of DEB-TACE in the treatment of patients with HCC. Hazard ratio, risk ratioand 95% confidence intervalswere calculated, using a fixed-effects model or a random-effects model. Results: Nine studies with a total of 830 patients met the inclusion criteria were included in this study. DEB-TACE significantly improved overall survivaland progression free survival, and also increased objective response rateand disease control rate. However, in subgroup analyses, pooled results showed that, the survival benefits of DEB-TACE were not found in the randomized controlled trials, but were observed in Non-RCTs. The incidence of most common adverse events, including nausea, pain, fever, and fatigue, was not significant difference between the DEB-TACE group and conventional TACEgroup. Conclusions: Despite DEB-TACE significantly prolonged the survival and response rate in the patients with HCC, the conclusion about the survival benefits should be interpreted with caution, since these findings were only found in retrospective Non-RCTs, and not in prospective RCTs.
PMCID: PMC4276155  PMID: 25550897
DEB-TACE; hepatocellular carcinoma; efficacy; toxicity; meta-analysis
8.  Potential Role of Cyr61 Induced Degeneration of Human Müller Cells in Diabetic Retinopathy 
PLoS ONE  2014;9(10):e109418.
The degeneration of Müller cells has been recognized to involve in the pathogenesis of diabetic retinopathy. However, the mechanism is not yet clear. This study is to explore the potential role of Cyr61, a secreted signaling protein in extracellular matrix, in inducing human Müller cell degeneration in diabetic retinopathy (DR). Twenty patients with proliferative diabetic retinopathy (PDR) and twelve non-diabetic patients were recruited for this study. Vitreous fluid was collected during vitrectomy surgery for Cyr61 ELISA. Human Müller cell line MIO-M1 were cultured to be subconfluent, and then treated with glucose (0–20 mM) or Cyr61 (0–300 ng/ml). Cyr61 expression induced by increasing concentrations of glucose was evaluated by RT-qPCR and Western blot. Effects of Cyr61 on Müller cells viability, migration and apoptosis were observed by MTT assay, Transwell assay, and TUNEL assay. Vitreous Cyr61 levels were observed to be 8-fold higher in patients with PDR (3576.92±1574.58 pg/mL), compared with non-diabetic controls (436.14±130.69 pg/mL). Interestingly, the active PDR group was significantly higher than the quiescent PDR group (P<0.01). In retinal Müller cells culture, high glucose significantly and dose-dependently elevated Cyr61 expression at both mRNA and protein levels. Cyr61 at high concentrations dose-dependently inhibited the viability and migration of Müller cells. TUNEL assay further revealed that high concentration of Cyr61 significantly promoted the cell apoptosis. In conclusion, these findings demonstrated for the first time that the expression of Cyr61 was elevated by high glucose in Müller cells, and Cyr61 inhibited cell viability and migration while induced apoptosis, suggesting the potential role of Cyr61 in Müller cell degeneration. The elevated Cyr61 levels in vitreous fluid of PDR patients further support its role in diabetic retinopathy (DR).
doi:10.1371/journal.pone.0109418
PMCID: PMC4199605  PMID: 25329584
9.  [125I]SD-7015 reveals fine modalities of CB1 cannabinoid receptor density in the prefrontal cortex during progression of Alzheimer's disease 
Neurochemistry international  2011;60(3):286-291.
The cannabinoid type-1 receptor (CB1R) is one of the most abundant members of the G protein-coupled receptor family in the central nervous system. Once activated by their cognate ligands, endocannabinoids, CB1Rs generally limit the timing of neurotransmitter release at many cortical synapses. Prior studies have indicated the involvement of CB1R in neurodegeneration and in various neuronal insults, with an emphasis on their neuroprotective role. In the present study we used a novel selective CB1R radioligand to investigate regional variations in CB1R ligand binding as a factor of progressive Braak tau pathology in the frontal cortex of Alzheimer's disease (AD) patients. The frontal cortex was chosen for this study due to the high density of CB1Rs and their well-characterized involvement in the progression of AD. Post-mortem prefrontal cortex samples from AD patients from Braak stages I to VI and controls were subjected to CB1R autoradiography with [125I]SD-7015 as radioligand. Regional concentration of [125I]SD-7015, corresponding to, and thereby representing, regional CB1R densities, were expressed in fM/g_tissue. The results show that CB1R density inversely correlates with Braak tau pathology with the following tendency: controls
doi:10.1016/j.neuint.2011.11.004
PMCID: PMC4180663  PMID: 22222721
Alzheimer's disease; Braak classification; Endocannabinoid system; Molecular imaging; Biomarker; Human brain autoradiography; CB1R; [125I]SD7015
Arthritis and rheumatism  2013;65(10):2724-2736.
Objective
Arthrogenic alphaviruses such as Ross River virus (RRV) and chikungunya virus (CHIKV) circulate worldwide. This virus class causes debilitating illnesses characterised by arthritis, arthralgia, and myalgia. We previously identified macrophage migration inhibitory factor (MIF) as a critical inflammatory factor in the pathogenesis of alphaviral disease. Here, we characterise the role of CD74, a cell surface receptor of MIF, in both RRV- and CHIKV-induced alphavirus arthritides.
Methods
Mouse models of RRV and CHIKV infection were used to investigate the immunopathogenesis of arthritic alphavirus infection. The role of CD74 was assessed using histological analysis, real time PCR, flow cytometry and plaque assay.
Results
In comparison to wild-type mice, CD74−/− mice developed only mild clinical features and had low levels of tissue damage. Leukocyte infiltration, characterised predominantly by inflammatory monocytes and natural killer cells, was substantially reduced in infected tissues of CD74−/− mice, but production of pro-inflammatory cytokines and chemokines were not decreased. CD74 deficiency was associated with increased monocyte apoptosis, but had no effect on monocyte migratory capacity. Consistent with these findings, alphaviral infection resulted in a dose-dependent up-regulation of CD74 expression in human peripheral blood mononuclear cells and serum MIF levels were significantly elevated in humans with RRV or CHIKV infections.
Conclusion
We propose that CD74 regulates immune responses to alphaviral infection through effects on cellular recruitment and survival. These findings suggest that both MIF and CD74 play a critical role in mediating alphaviral disease and blocking these factors with novel therapeutic agents can substantially ameliorate pathology.
doi:10.1002/art.38090
PMCID: PMC3796577  PMID: 23896945
Brain research bulletin  2012;87(6):504-510.
Cannabinoid type-1 receptors (CB1Rs) modulate synaptic neurotransmission by participating in retrograde signaling in the adult brain. Increasing evidence suggests that cannabinoids through CB1Rs play an important role in the regulation of motor activities in the striatum. In the present study, we used human brain samples to examine the relationship between CB1R and dopamine receptor density in case of Parkinson’s disease (PD).
Post mortem putamen, nucleus caudatus and medial frontal gyrus samples obtained from PD patients were used for CB1R and dopamine D2/D3 receptor autoradiography. [125I]SD7015, a novel selective CB1R inverse agonist, developed by a number of the present co-authors, and [3H]raclopride, a dopamine D2/D3 antagonist, were used as radioligands. Our results demonstrate unchanged CB1R density in the putamen and nucleus caudatus of deceased PD patients, treated with levodopa (l-DOPA). At the same time dopamine D2/D3 receptors displayed significantly decreased density levels in case of PD putamen (control: 47.97 ± 10.00 fmol/g, PD: 3.73 ± 0.07 fmol/g (mean ± SEM), p < 0.05) and nucleus caudatus (control: 30.26 ± 2.48 fmol/g, PD: 12.84 ± 5.49 fmol/g, p < 0.0005) samples. In contrast to the putamen and the nucleus caudatus, in the medial frontal gyrus neither receptor densities were affected.
Our data suggest the presence of an unaltered CB1R population even in late stages of levodopa treated PD. This further supports the presence of an intact CB1R population which, in line with the conclusion of earlier publications, may be utilized as a pharmacological target in the treatment of PD. Furthermore we found discrepancy between a maintained CB1R population and a decreased dopamine D2/D3 receptor population in PD striatum. The precise explanation of this conundrum requires further studies with simultaneous examination of the central cannabinoid and dopaminergic systems in PD using higher sample size.
doi:10.1016/j.brainresbull.2012.02.012
PMCID: PMC4180092  PMID: 22421165
Parkinson’s disease; Endocannabinoid CB1 receptor; Dopamine D2/D3 receptor; Molecular imaging biomarker; Human brain autoradiography; Striatum
Scientific Reports  2014;4:6381.
Precipitation variation on the Loess Plateau (LP) of China is not only important for rain-fed agriculture in this environmentally sensitive region, but also critical for the water and life securities over the whole Yellow River basin. Here we reconstruct high resolution precipitation variation on the western LP during the past 370 years by using two replicated, annually-laminated stalagmites. Spatial analysis suggests that the reconstruction can be also representative for the whole LP region. The precipitation variations show a significant quasi-50 year periodicity during the last 370 years, and have an important role in determining the runoff of the middle Yellow River. The main factor controlling the decadal scale variations and long-term trend in precipitation over this region is southerly water vapour transport associated with the Asian summer monsoon. The Pacific Decadal Oscillation is also an important influence on precipitation variation in this region, as it can affect the East Asian summer monsoon and the West Pacific Subtropical High.
doi:10.1038/srep06381
PMCID: PMC4165273  PMID: 25223372
Objective: This study aimed to investigate the effect of Katanin p60 on the neurite growth and collateral formation in the hippocampus. Methods: Gene cloning was performed to construct the Katanin p60 eukaryotic vector. The microtubule cutting effect and protein expression of Katanin p60 were investigated in 293T cells. Then, these vectors were transfected into hippocampal neurons of rats, and the effects of Katanin p60 on the neurite growth and collateral formation were observed. Results: In the present study, we successfully constructed Katanin p60-GFP recombinant plasmids. After transfecting into 293T cells, the Katanin p60 was over-expressed in these cells, the mesh-like structure of microtubules was disrupted, the residual microtubules circled the nucleus, the expression microtubule proteins reduced, and the tapered protrusions disappeared. In hippocampal neurons with Katanin p60 over-expression, the neural neurite growth was obvious, and a lot of dendrites arose from cell bodies. In cells without Katanin p60 expression, the neurites were small, and the number and length of dendrites reduced significantly when compared with Katanin p60 over-expressing cells (P < 0.05). In addition, in Katanin p60 over-expressing cells, the number of collaterals from the neurites and dendrites increased markedly when compared with cells without Katanin p60 expression (P < 0.05). Conclusion: Katanin p60 can promote the neurite growth and collateral formation of hippocampal neurons.
PMCID: PMC4211748  PMID: 25356098
Katanin; neurite; collateral; microtubule; neuron; hippocampus
DNA and Cell Biology  2013;32(5):243-251.
Improvement of blood flow and promotion of angiogenesis are important therapeutic measures for the treatment of ischemic peripheral vascular diseases. Since apolipoprotein (a) (apo (a)) is a glycoprotein with repetitive kringle domains exhibiting 75% to 98% structural homology with plasminogen (Plg), apo (a) may also have a negative effect on endothelial progenitor cell (EPC)-induced angiogenesis through Plg-like inhibitory effects on EPC proliferation, adhesion, migration, and angiogenesis. To evaluate the effect of apo (a) on EPCs-induced angiogenesis, EPCs were isolated from the bone marrow of apo (a) transgenic mice, wild-type litter mates, and normal mice. These cells were cultured without or with apo (a) before transplantation. Hindlimb ischemia models were surgically induced in mice, which then received an intravenous injection of 3×105 EPCs. At 3, 7, and 14 days post EPC transplantation, the adhesion, migration abilities, and capillary density in calf muscles were assessed. Results indicate that apo (a) significantly reduced the adhesion and migration abilities of EPCs. Furthermore, the tubule-like formation of EPCs on Matrigel gels was damaged. In vivo experiments showed the homing of EPCs to ischemic peripheral vascular, and the number of capillary vessels decreased significantly in apo(a) transgenic mice. This study demonstrated that apo (a) could attenuate the adhesion, migration, and homing abilities of EPCs and could impair the angiogenesis ability of EPCs.
Homing of endothelial progenitor cells to ischemic peripheral vascular sites and the number of capillaries were decreased in mice transgenic for Apo (a). This gene attenuated adhesion, migration, and homing abilities of EPCs and impaired angiogenesis.
doi:10.1089/dna.2013.1963
PMCID: PMC3651684  PMID: 23581552
Purpose
The stability of prepared infusions of the tumor necrosis factor (TNF)-α agent infliximab after storage for up to two weeks was investigated.
Methods
To determine the feasibility of liberalized expiration dating of infliximab (current recommendations call for the infusion of prepared doses within three hours), the stability of diluted infliximab stored in polyvinyl chloride (PVC) bags at 4 °C for up to 14 days was evaluated. A known quantity of TNF-α was combined with infliximab test samples in PVC bags for one hour; immediately after the reaction period and after 7 and 14 days of storage, the residual amount of TNF-α (an indirect measure of the drug's biological activity) was analyzed via a validated enzyme-linked immunosorbent assay (ELISA).
Results
The mean ± S.D. amount of TNF-α consumed by infliximab was calculated to be 24.5 ± 5.6 pg/mL at baseline, 29.0 ± 4.4 pg/mL at 7 days, and 24.8 ± 17.3 pg/mL at 14 days. At all evaluated time points, ELISA results indicated that 19–24% of the original TNF-α had been consumed by infliximab (mean ± S.D. consumption: 19.6% ± 4.5% at baseline, 23.2% ± 3.5% at 7 days, and 19.8% ± 13.8% at 14 days).
Conclusion
Infliximab, when prepared at a concentration of 400 μg/mL in 0.9% sodium chloride injection, incurred no loss of biological activity when stored for up to 14 days at 4 °C in PVC bags. Changing infliximab preparation practices may improve clinic efficiency by reducing patient dissatisfaction with long wait times for infusions and avoiding costly waste.
doi:10.2146/ajhp100116
PMCID: PMC3987119  PMID: 22899746
Immunology and cell biology  2013;91(5):368-376.
Previous studies using MCPIP1/Zc3h12a-deficient mice suggest that MCPIP1 is an important regulator of inflammation and immune homeostasis. However, the characterization of the immunological phenotype of MCPIP1-deficient mice has not been detailed. In this study, we performed evaluation through histological, flow cytometric, ELISA and real-time PCR analysis and found that targeted disruption of MCPIP1 gene leads to fatal, highly aggressive, and widespread immune-related lesions. In addition to previously observed growth retardation, splenomegaly, lymphoadenopathy, severe anemia and premature death, MCPIP1-deficient mice showed disorganization of lymphoid organs, including spleen, lymph nodes and thymus, and massive infiltration of lymphocytes, macrophages and neutrophils into many other non-lymphoid organs, primarily in lungs and liver. Flow cytometric analysis found significant increase in activated and differentiated T cells in peripheral blood and spleen of MCPIP1-deficient mice. Moreover, heightened production of inflammatory cytokines from activated macrophages and T cells were observed in MCPIP1-deficient mice. Interestingly, treatment of MCPIP1-deficient mice with antibiotics resulted in significant improvement of life-span and a decrease in inflammatory syndrome. Taken together, these results suggest a prominent role for MCPIP1 in the control of inflammation and immune homeostasis.
doi:10.1038/icb.2013.11
PMCID: PMC3932977  PMID: 23567898
MCPIP1; autoimmune disease; inflammation; cytokine; lymphocyte
ABSTRACT
The prevalence of Hypoderma spp. in yaks grazed in the east of Qinghai province was investigated in 2008. In this area, the prevalence in young yaks (1- to 3-year-old) was very high at 82.2–98.7%, whilst in adult yaks (4-year-old and older), the prevalence was 42.4–50.6%. The seasonal development and migration pattern of Hypoderma larvae in yak bodies was found to be similar for different locations in this area. The numbers of first, second and third instar larvae detected in yak bodies peaked in October, December and March, respectively. Different doses of ivermectin (125 to 500 µg/kg body weight) almost completely dewormed the larvae from yaks, suggesting that using a quarter of the prescribed dose (500 µg/kg body weight) was effective. In October of each year between 2009 and 2012, ivermectin (125 µg/kg body weight) was administered to a total of 562,995 yaks grazed in four counties in Qinghai province, and the pevalence of Hypoderma larval infection in yaks was reduced to 0.5–1.0%.
doi:10.1292/jvms.13-0299
PMCID: PMC3982821  PMID: 24107486
China; Hypoderma; ivermectin; Qinghai; yak
Collapsin response mediator proteins (CRMPs) have been reported to control axonal guidance during neuronal development and degeneration. Among these proteins, CRMP-5 has been indicated to play an important role in growth cone development. However, the mechanisms underlying the linkage between growth cone development and the cytoskeleton remain to be elucidated. Here, we report that CRMP-5 interacts with tubulin to mediate growth cone development in cultured hippocampal neurons. We found that CRMP-5 physically interacted with tubulin in the growth cones of developing neurons. CRMP-5 colocalized with tubulin in lamellipodia in HEK293 cells and in the growth cones of cultured hippocampal neurons. Genetic silencing of CRMP-5 using RNA interference led to abnormal growth cone morphology in neurons. Overexpression of CRMP-5 led to significantly increased filopodial formation and enlarged growth cones. These results suggest that CRMP-5 interacts with tubulin to regulate growth cone dynamics, thus complying with the restrictive intracellular guidance cues.
PMCID: PMC3902242  PMID: 24482690
CRMP-5; tubulin; growth cone; hippocampal neuron
Scientific Reports  2014;4:3657.
The electrical transport properties of graphene are greatly influenced by its environment. Owing to its high dielectric constant, strontium titanate (STO) is expected to suppress the long-range charged impurity scattering and consequently to enhance the mobility. However, the absence of such enhancement has caused some controversies regarding the scattering mechanism. In graphene devices transferred from SiO2 to STO using a newly developed technique, we observe a moderate mobility enhancement near the Dirac point, which is the point of charge neutrality achieved by adjusting the Fermi level. While bulk STO is not known as a ferroelectric material, its surface was previously reported to exhibit an outward displacement of oxygen atoms and ferroelectric-like dipole moment. When placed on STO, graphene shows strong and asymmetric hysteresis in resistivity, which is consistent with the dipole picture associated with the oxygen displacement. The hysteretic response of the surface dipole moment diminishes the polarizability, therefore weakens the ability of STO to screen the Coulomb potential of the impurities.
doi:10.1038/srep03657
PMCID: PMC3888981  PMID: 24413768
PLoS ONE  2013;8(12):e83462.
Quantifying elemental carbon (EC) content in geological samples is challenging due to interferences of crustal, salt, and organic material. Thermal/optical analysis, combined with acid pretreatment, represents a feasible approach. However, the consistency of various thermal/optical analysis protocols for this type of samples has never been examined. In this study, urban street dust and soil samples from Baoji, China were pretreated with acids and analyzed with four thermal/optical protocols to investigate how analytical conditions and optical correction affect EC measurement. The EC values measured with reflectance correction (ECR) were found always higher and less sensitive to temperature program than the EC values measured with transmittance correction (ECT). A high-temperature method with extended heating times (STN120) showed the highest ECT/ECR ratio (0.86) while a low-temperature protocol (IMPROVE-550), with heating time adjusted for sample loading, showed the lowest (0.53). STN ECT was higher than IMPROVE ECT, in contrast to results from aerosol samples. A higher peak inert-mode temperature and extended heating times can elevate ECT/ECR ratios for pretreated geological samples by promoting pyrolyzed organic carbon (PyOC) removal over EC under trace levels of oxygen. Considering that PyOC within filter increases ECR while decreases ECT from the actual EC levels, simultaneous ECR and ECT measurements would constrain the range of EC loading and provide information on method performance. Further testing with standard reference materials of common environmental matrices supports the findings. Char and soot fractions of EC can be further separated using the IMPROVE protocol. The char/soot ratio was lower in street dusts (2.2 on average) than in soils (5.2 on average), most likely reflecting motor vehicle emissions. The soot concentrations agreed with EC from CTO-375, a pure thermal method.
doi:10.1371/journal.pone.0083462
PMCID: PMC3866270  PMID: 24358286
Volatile components from Exocarpium Citri Grandis (ECG) were, respectively, extracted by three methods, that is, steam distillation (SD), headspace solid-phase microextraction (HS-SPME), and solvent extraction (SE). A total of 81 compounds were identified by gas chromatography-mass spectrometry including 77 (SD), 56 (HS-SPME), and 48 (SE) compounds, respectively. Despite of the extraction method, terpenes (39.98~57.81%) were the main volatile components of ECG, mainly germacrene-D, limonene, 2,6,8,10,14-hexadecapentaene, 2,6,11,15-tetramethyl-, (E,E,E)-, and trans-caryophyllene. Comparison was made among the three methods in terms of extraction profile and property. SD relatively gave an entire profile of volatile in ECG by long-time extraction; SE enabled the analysis of low volatility and high molecular weight compounds but lost some volatiles components; HS-SPME generated satisfactory extraction efficiency and gave similar results to those of SD at analytical level when consuming less sample amount, shorter extraction time, and simpler procedure. Although SD and SE were treated as traditionally preparative extractive techniques for volatiles in both small batches and large scale, HS-SPME coupled with GC/MS could be useful and appropriative for the rapid extraction and qualitative analysis of volatile components from medicinal plants at analytical level.
doi:10.1155/2013/918406
PMCID: PMC3855974  PMID: 24349825
Open Biology  2013;3(11):130100.
The HIV-1 viral infectivity factor (Vif) neutralizes cell-encoded antiviral APOBEC3 proteins by recruiting a cellular ElonginB (EloB)/ElonginC (EloC)/Cullin5-containing ubiquitin ligase complex, resulting in APOBEC3 ubiquitination and proteolysis. The suppressors-of-cytokine-signalling-like domain (SOCS-box) of HIV-1 Vif is essential for E3 ligase engagement, and contains a BC box as well as an unusual proline-rich motif. Here, we report the NMR solution structure of the Vif SOCS–ElonginBC (EloBC) complex. In contrast to SOCS-boxes described in other proteins, the HIV-1 Vif SOCS-box contains only one α-helical domain followed by a β-sheet fold. The SOCS-box of Vif binds primarily to EloC by hydrophobic interactions. The functionally essential proline-rich motif mediates a direct but weak interaction with residues 101–104 of EloB, inducing a conformational change from an unstructured state to a structured state. The structure of the complex and biophysical studies provide detailed insight into the function of Vif's proline-rich motif and reveal novel dynamic information on the Vif–EloBC interaction.
doi:10.1098/rsob.130100
PMCID: PMC3843819  PMID: 24225024
HIV-1 viral infectivity factor; SOCS-box domain; ElonginBC; NMR; solution structure
PLoS ONE  2013;8(9):e69512.
Na+:K+:2Cl- cotransporter 1 (NKCC1) has been implicated in mediating ischemia-, trauma- or ammonia-induced astrocyte swelling/brain edema in mammals. This study aimed to determine the effects of ammonia or terrestrial exposure on ammonia concentrations in the plasma and brain, and the mRNA expression and protein abundance of nkcc/Nkcc in the brain, of the swamp eel Monopterusalbus. Ammonia exposure led to a greater increase in the ammonia concentration in the brain of M. albus than terrestrial exposure. The brain ammonia concentration of M. albus reached 4.5 µmol g-1 and 2.7 µmol g-1 after 6 days of exposure to 50 mmol l-1 NH4Cl and terrestrial conditions, respectively. The full cDNA coding sequence of nkcc1b from M. albus brain comprised 3276 bp and coded for 1092 amino acids with an estimated molecular mass of 119.6 kDa. A molecular characterization indicated that it could be activated through phosphorylation and/or glycosylation by osmotic and/or oxidative stresses. Ammonia exposure for 1 day or 6 days led to significant decreases in the nkcc1b mRNA expression and Nkcc1b protein abundance in the brain of M. albus. In comparison, a significant decrease in nkcc1b mRNA expression was observed in the brain of M. albus only after 6 days of terrestrial exposure, but both 1 day and 6 days of terrestrial exposure resulted in significant decreases in the protein abundance of Nkcc1b. These results are novel because it has been established in mammals that ammonia up-regulates NKCC1 expression in astrocytes and NKCC1 plays an important role in ammonia-induced astrocyte swelling and brain edema. By contrast, our results indicate for the first time that M. albus is able to down-regulate the mRNA and protein expression of nkcc1b/Nkcc1b in the brain when confronted with ammonia toxicity, which could be one of the contributing factors to its extraordinarily high brain ammonia tolerance.
doi:10.1371/journal.pone.0069512
PMCID: PMC3777983  PMID: 24069137
Biomedical Reports  2013;1(5):766-770.
This study was conducted to investigate the effects of quercetin on the expression of cyclin-dependent kinase (CDK4) mRNA and protein in A549 lung epithelial tumor cells infected by H1N1. First, the Thiazolyl Blue Tetrazolium Bromide (MTT) method was used to determine H1N1 virulence, quercetin cytotoxicity and inhibition of the cytopathic effect of H1N1 on A549 cells by quercetin. Subsequently, 100 TCID50 H1N1 was used to infect A549 cells for 2 h prior to culture in maintenance media containing 10 mg/l quercetin. After 4, 12, 24 and 48 h of culture, the cells were collected and total RNA and protein were extracted. Fluorescent quantitative polymerase chain reaction and western blot analysis were then performed to assess the expression of CDK4 mRNA and protein. The experiment demonstrated that the TCID50 of H1N1 in A549 cells was 10−4.75, the maximum non-toxic concentration of quercetin in A549 cells was 30–60 mg/l and the minimum effective concentration of quercetin for the inhibition of the H1N1 cytopathic effect on A549 cells was 10 mg/l. The results indicated that quercetin may significantly inhibit CDK4 mRNA and protein overexpression caused by H1N1 within 4–48 h. In conclusion, quercetin may protect against H1N1 infection by effectively reducing the mRNA and protein expression of CDK4 caused by H1N1 infection.
doi:10.3892/br.2013.141
PMCID: PMC3917044  PMID: 24649026
quercetin; cyclin-dependent kinase 4; A549 cells; H1N1
Fertility and Sterility  2012;98(1):200-206.
Objective
To examine whether spontaneous oocyte activation is determined by genetic differences and interacted with culture environment.
Design
Experimental Study.
Setting
Temple University School of Medicine.
Animals
C57BL/6, DBA/2, C3H/HeJ, and A/J strains, along with reciprocal F1 hybrid female mice (5–6 weeks).
Intervention(s)
Immature oocytes from different mouse strains were collected and cultured in different maturation conditions including different serum, serum replacement, bovine serum albumin (BSA) and follicle stimulation hormone (FSH).
Main Outcome Measure(s)
The emission of first polar body, pronucleus formation, meiotic arrest, spontaneous activation, and expression of maturation regulators.
Result(s)
Oocytes from C57BL/6 mice display a high rate of delayed first meiotic division and spontaneous activation after the first meiotic division with in vitro maturation (IVM), and the second meiosis with in vivo maturation (VVM) following superovulation. Spontaneous activation with IVM is sensitive to culture environment. Oocytes spontaneously activated during the first meiotic division with IVM have unusual paired tetrad chromosomes with slight connections at centromeres, whereas oocytes activated in vivo display haploidization from the second meiosis. Spontaneous activation is also seen in F1 hybrid oocytes, indicating a dominant trait from C57BL/6. Delayed meiosis was associated with reduced cylcin B and securin expression.
Conclusion(s)
Both mouse strain and culture environment have a significant effect on the incidence of meiotic defects and spontaneous activation. Reduced expression of meiotic regulators may underlie this effect.
doi:10.1016/j.fertnstert.2012.03.060
PMCID: PMC3389194  PMID: 22584025
oocyte; meiosis; in vitro maturation; spontaneous activation; assisted reproduction

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