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1.  On-line monitoring of oxygen in Tubespin, a novel, small-scale disposable bioreactor 
Cytotechnology  2011;63(4):345-350.
A novel, optical sensor was fixed in a new type of disposable bioreactor, Tubespin, for the on-line (real-time) monitoring of dissolved oxygen concentrations during cell culture. The cell density, viability and volumetric mass transfer coefficient were also determined to further characterize the bioreactors. The kLa value of the Tubespin at standard conditions was 24.3 h−1, while that of a spinner flask was only 2.7 h−1. The maximum cell density in the Tubespin bioreactor reached 6 × 106 cells mL−1, which was two times higher than the cell density in a spinner flask. Furthermore, the dynamic dissolved oxygen level was maintained above 90% air-saturation in the Tubespin, while the value was only 1.9% in a spinner flask. These results demonstrate the competitive advantage of using the Tubespin system over spinner flasks for process optimization and scale-down studies of oxygen transfer and cell growth.
doi:10.1007/s10616-011-9361-x
PMCID: PMC3140833  PMID: 21574065
Tubespin; Spinner flask; Dissolved oxygen; kLa
2.  Enhancement of tumor cell death by combining cisplatin with an oncolytic adenovirus carrying MDA-7/IL-24 
Acta Pharmacologica Sinica  2009;30(4):467-477.
Aim:
The aim of this study was to creatively implement a novel chemo-gene-virotherapeutic strategy and further strengthen the antitumor effect in cancer cells by the combined use of ZD55-IL-24 and cisplatin.
Methods:
ZD55-IL-24 is an oncolytic adenovirus that harbors interleukin 24 (IL-24), which has a strong antitumor effect and was identified and evaluated by PCR, RT-PCR, and Western blot analysis. Enhancement of cancer cell death using a combination of ZD55-IL-24 and cisplatin was assessed in several cancer cell lines by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cytopathic effect (CPE) assay. Apoptosis induction by treatment with ZD55-IL-24 and/or cisplatin was detected in BEL7404 and SMMC7721 by morphological evaluation, apoptotic cell staining, and flow cytometry analysis. In addition, negative effects on normal cells were evaluated in the L-02 cell line using the MTT assay, the CPE assay, morphological evaluation, apoptotic cell staining, and flow cytometry analysis.
Results:
The combination of ZD55-IL-24 and cisplatin, which is superior to ZD55-IL-24, cisplatin, and ZD55-EGFP, as well as ZD55-EGFP plus cisplatin, resulted in a significantly increased effect. Most importantly, conjugation of ZD55-IL-24 with cisplatin had toxic effects equal to that of cisplatin and did not have overlapping toxicities in normal cells.
Conclusion:
This study showed that ZD55-IL-24 conjugated with cisplatin exhibited a remarkably increased cytotoxic and apoptosis-inducing effect in cancer cells and significantly reduced the toxicity in normal cells through the use of a reduced dose.
doi:10.1038/aps.2009.16
PMCID: PMC4002269  PMID: 19270721
cisplatin; MDA-7/IL-24; oncolytic adenovirus; apoptosis
3.  Genome-wide distribution comparative and composition analysis of the SSRs in Poaceae 
BMC Genetics  2015;16:18.
Background
The Poaceae family is of great importance to human beings since it comprises the cereal grasses which are the main sources for human food and animal feed. With the rapid growth of genomic data from Poaceae members, comparative genomics becomes a convinent method to study genetics of diffierent species. The SSRs (Simple Sequence Repeats) are widely used markers in the studies of Poaceae for their high abundance and stability.
Results
In this study, using the genomic sequences of 9 Poaceae species, we detected 11,993,943 SSR loci and developed 6,799,910 SSR primer pairs. The results show that SSRs are distributed on all the genomic elements in grass. Hexamer is the most frequent motif and AT/TA is the most frequent motif in dimer. The abundance of the SSRs has a positive linear relationship with the recombination rate. SSR sequences in the coding regions involve a higher GC content in the Poaceae than that in the other species. SSRs of 70-80 bp in length showed the highest AT/GC base ratio among all of these loci. The result shows the highest polymorphism rate belongs to the SSRs ranged from 30 bp to 40 bp. Using all the SSR primers of Japonica, nineteen universal primers were selected and located on the genome of the grass family. The information of SSR loci, the SSR primers and the tools of mining and analyzing SSR are provided in the PSSRD (Poaceae SSR Database, http://biodb.sdau.edu.cn/pssrd/).
Conclusions
Our study and the PSSRD database provide a foundation for the comparative study in the Poaceae and it will accelerate the study on markers application, gene mapping and molecular breeding.
doi:10.1186/s12863-015-0178-z
PMCID: PMC4333251
4.  Postoperative recurrence of desmoid tumors: clinical and pathological perspectives 
Background
The clinical features and the pathological changes of desmoid tumors were studied to point out the key factors affecting the recurrence.
Methods
The clinical data and specimens of 56 patients who underwent desmoid tumor resection from 2003 to 2008 were reviewed. Possible clinical factors related to the postoperative recurrence were analyzed statistically. The specimens round the lesions were studied histopathologically.
Results
The overall recurrence rate was 39.3%. The postoperative recurrence rate of the patients with negative surgical margins and no tumor invasion of the major vessels and nerves was low (P < 0.05). However, the desmoid tumors could destroy the cortical bone and invade the medullary cavity.
Conclusions
Desmoid tumors were pathologically benign, which could extensively invade tissues around the lesions. The invasion of major vessels and nerves and quality of surgical margins are the key factors for the high postoperative recurrence rate.
doi:10.1186/s12957-015-0450-8
PMCID: PMC4329213
Histopathology; Immunohistochemistry; Tumor resection
5.  MicroRNA-185 Targets SOCS3 to Inhibit Beta-Cell Dysfunction in Diabetes 
PLoS ONE  2015;10(2):e0116067.
Diabetes is the most common and complex metabolic disorder, and one of the most important health threats now. MicroRNAs (miRNAs) are a group of small non-coding RNAs that have been suggested to play a vital role in a variety of physiological processes, including glucose homeostasis. In this study, we investigated the role of miR-185 in diabetes. MiR-185 was significantly downregulated in diabetic patients and mice, and the low level was correlated to blood glucose concentration. Overexpression of miR-185 enhanced insulin secretion of pancreatic β-cells, promoted cell proliferation and protected cells from apoptosis. Further experiments using in silico prediction, luciferase reporter assay and western blot assay demonstrated that miR-185 directly targeted SOCS3 by binding to its 3’-UTR. On the contrary to miR-185’s protective effects, SOCS3 significantly suppressed functions of β-cell and inactivated Stat3 pathway. When treating cells with miR-185 mimics in combination with SOCS3 overexpression plasmid, the inhibitory effects of SOCS3 were reversed. While combined treatment of miR-185 mimics and SOCS3 siRNA induced synergistically promotive effects compared to either miR-185 mimics or SOCS3 siRNA treatment alone. Moreover, we observed that miR-185 level was inversely correlated with SOCS3 expression in diabetes patients. In conclusion, this study revealed a functional and mechanistic link between miR-185 and SOCS3 in the pathogenesis of diabetes. MiR-185 plays an important role in the regulation of insulin secretion and β-cell growth in diabetes. Restoration of miR-185 expression may serve a potentially promising and efficient therapeutic approach for diabetes.
doi:10.1371/journal.pone.0116067
PMCID: PMC4319748  PMID: 25658748
7.  The Ctp Type IVb Pilus Locus of Agrobacterium tumefaciens Directs Formation of the Common Pili and Contributes to Reversible Surface Attachment 
Journal of Bacteriology  2014;196(16):2979-2988.
Agrobacterium tumefaciens can adhere to plant tissues and abiotic surfaces and forms biofilms. Cell surface appendages called pili play an important role in adhesion and biofilm formation in diverse bacterial systems. The A. tumefaciens C58 genome sequence revealed the presence of the ctpABCDEFGHI genes (cluster of type IV pili; Atu0216 to Atu0224), homologous to tad-type pilus systems from several bacteria, including Aggregatibacter actinomycetemcomitans and Caulobacter crescentus. These systems fall into the type IVb pilus group, which can function in bacterial adhesion. Transmission electron microscopy of A. tumefaciens revealed the presence of filaments, significantly thinner than flagella and often bundled, associated with cell surfaces and shed into the external milieu. In-frame deletion mutations of all of the ctp genes, with the exception of ctpF, resulted in nonpiliated derivatives. Mutations in ctpA (a pilin homologue), ctpB, and ctpG decreased early attachment and biofilm formation. The adherence of the ctpA mutant could be restored by ectopic expression of the paralogous pilA gene. The ΔctpA ΔpilA double pilin mutant displayed a diminished biovolume and lower biofilm height than the wild type under flowing conditions. Surprisingly, however, the ctpCD, ctpE, ctpF, ctpH, and ctpI mutants formed normal biofilms and showed enhanced reversible attachment. In-frame deletion of the ctpA pilin gene in the ctpCD, ctpE, ctpF, ctpH, and ctpI mutants caused the same attachment-deficient phenotype as the ctpA single mutant. Collectively, these findings indicate that the ctp locus is involved in pilus assembly and that nonpiliated mutants, which retain the CtpA pilin, are proficient in attachment and adherence.
doi:10.1128/JB.01670-14
PMCID: PMC4135632  PMID: 24914181
8.  Determination of royal jelly freshness by ELISA with a highly specific anti-apalbumin 1, major royal jelly protein 1 antibody*  
Major royal jelly protein 1 (MRJP1), designated apalbumin 1, has been regarded as a freshness marker of royal jelly (RJ). A MRJP1-specific peptide (IKEALPHVPIFD) identified by bioinformatics analysis of homologous members of the major royal protein family was synthesized and used to raise polyclonal anti-MRJP1 antibody (anti-SP-MRJP1 antibody). Western blot analysis showed that anti-SP-MRJP1 antibody only reacted with MRJP1 in RJ. In contrast, the previously reported antibody against recombinant MRJP1 (anti-R-MRJP1 antibody) reacted with other members of MRJP family in RJ. Enzyme-linked immunosorbent assay (ELISA) using anti-SP-MRJP1 antibody demonstrated that MRJP1 content in RJ stored at 40 °C significantly degraded by 37.3%, 55.9%, 58.0%, 60.6%, 65.7%, 72.7%, and 73.1% at 7, 14, 21, 28, 35, 42, and 49 d, respectively, when compared with MRJP1 content in fresh RJ (0 d). Optical density analysis of MRJP bands from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles demonstrated that the degradation of MRJP1, MRJP2, MRJP3, and MRJP5 in RJ was strongly and positively correlated with the period of storage (P<0.0001). Our results indicated anti-SP-MRJP1 antibody was highly specific for MRJP1, and ELISA using the antibody is a sensitive and easy-to-use method to determine the freshness and authenticity of RJ.
doi:10.1631/jzus.B1400223
PMCID: PMC4322426  PMID: 25644470
Freshness; Royal jelly; Major royal jelly protein 1 (MRJP1); Enzyme-linked immunosorbent assay (ELISA); High specific antibody
9.  Sustained release of vancomycin from novel biodegradable nanofiber-loaded vascular prosthetic grafts: in vitro and in vivo study 
This study describes novel biodegradable, drug-eluting nanofiber-loaded vascular prosthetic grafts that provide local and sustained delivery of vancomycin to surrounding tissues. Biodegradable nanofibers were prepared by first dissolving poly(D,L)-lactide-co-glycolide and vancomycin in 1,1,1,3,3,3-hexafluoro-2-propanol. The solution was then electrospun into nanofibers onto the surface of vascular prostheses. The in vitro release rates of the pharmaceutical from the nanofiber-loaded prostheses was characterized using an elution method and a high-performance liquid chromatography assay. Experimental results indicated that the drug-eluting prosthetic grafts released high concentrations of vancomycin in vitro (well above the minimum inhibitory concentration) for more than 30 days. In addition, the in vivo release behavior of the drug-eluting grafts implanted in the subcutaneous pocket of rabbits was also documented. The drug-eluting grafts developed in this work have potential applications in assisting the treatment of vascular prosthesis infection and resisting reinfection when an infected graft is to be exchanged.
doi:10.2147/IJN.S78675
PMCID: PMC4321605
drug-eluting prosthetic graft; vascular prosthesis infection; release characteristics
10.  An Assessment of Traditional Uighur Medicine in Current Xinjiang Region (China) 
Background
The main objectives of this study were to assess the current research and development of traditional Uighur medicine in Xinjiang (China), and to evaluate the promising pharmacological products of traditional Uighur medicine for further studies.
Materials and Methods
Traditional Uighur medicine data of medicine registry, patent, and academic publications was collected and analyzed.
Results
Data showed that, among the registered and studied traditional Uighur medicine, the main therapeutic areas of traditional Uighur medicine focused on skin disease, urogenital disease, rheumatism and digestive system disease. The representative traditional Uighur patent medicine included the following: BaixuanXiatare Tablets, Kaliziran Tincture and Vernoniaanthelmintica Injection (Psoriasis and vitiligo); Xi-payimazibiziLiquid (prostatitis); KursiKaknaq (urinary tract infection); Tongzhisurunjiang Capsules (anti-rheumatism medicine); HuganBuzure Granules (digestive system disease). Moreover, ten Uighur herbs were widely used, including: ResinaScammoniae, Folium FumicisDentati, HerbaDracocephali, Semen AmygdaliDulcis, HerbaChamomillae, FructusPimpinellaeanisi, Cortex Foeniculi, FructusVernoniae, FructusApii, and Radix AnacycliPyrethri.
Conclusion
This study concluded by indicating that traditional Uighur medicine with excellent curative effect should be screened in details for their phytochemical properties and pharmacological activity to discover new bioactive constituents.
PMCID: PMC4202637  PMID: 25435613
Traditional Uighur medicine; Traditional Uighur patent medicine; Uighur herb
11.  Direct evidence for kinetic effects associated with solar wind reconnection 
Scientific Reports  2015;5:8080.
Kinetic effects resulting from the two-fluid physics play a crucial role in the fast collisionless reconnection, which is a process to explosively release massive energy stored in magnetic fields in space and astrophysical plasmas. In-situ observations in the Earth's magnetosphere provide solid consistence with theoretical models on the point that kinetic effects are required in the collisionless reconnection. However, all the observations associated with solar wind reconnection have been analyzed in the context of magnetohydrodynamics (MHD) although a lot of solar wind reconnection exhausts have been reported. Because of the absence of kinetic effects and substantial heating, whether the reconnections are still ongoing when they are detected in the solar wind remains unknown. Here, by dual-spacecraft observations, we report a solar wind reconnection with clear Hall magnetic fields. Its corresponding Alfvenic electron outflow jet, derived from the decouple between ions and electrons, is identified, showing direct evidence for kinetic effects that dominate the collisionless reconnection. The turbulence associated with the exhaust is a kind of background solar wind turbulence, implying that the reconnection generated turbulence has not much developed.
doi:10.1038/srep08080
PMCID: PMC4308709  PMID: 25628139
12.  Family functioning, marital quality and social support in Chinese patients with epilepsy 
Background
The purpose of this study was to examine family functioning, marital quality, social support, and anxiety and depression in Chinese patients with Epilepsy (PWE) in comparison with healthy people.
Methods
This case–control study included 42 PWE and 42 healthy controls. Participants completed the Zung’s self-rating depression scale, the Zung’s self-rating anxiety scale, the Chinese version of family cohesion and flexibility evaluation scales, the Chinese version of the marital inventory ENRICH, and the Chinese versions of the social support rating scale and perceived social support scale.
Results
PWE reported higher levels of anxiety and depression, and lower levels of family cohesion, marriage quality and social support compared with controls. Support within and outside the family was negatively associated with depression, however social support did not significantly predict depression in PWE. In patients, support within the family and emotional support predicted family cohesion and marriage quality. Instrumental support was negatively associated with anxiety in patients but positively associated with depression in healthy controls. Support within the family predicted family cohesion and marriage quality in both the control group and patient group, depression predicted family adaptation in both the control group and patient group, while support outside the family predicted marriage quality only in the patient group. Both emotional and instrumental support predicted family adaptation in the control group, and emotional support predicted family cohesion in patients.
Conclusions
PWE in China had higher levels of anxiety and depression, dissatisfaction with family functioning and marital life, and less social support compared with healthy controls. Emotional support within and outside families promoted family cohesion and marriage quality, depression decreased family adaptation, and instrumental support decreased anxiety of PWE. These findings suggest that enhancing family and emotional supports and decreasing depression could promote the family functioning and marital quality of PWE, and instrumental support may play a role in decreasing anxiety.
doi:10.1186/s12955-015-0208-6
PMCID: PMC4320606  PMID: 25627321
Family function; Marital quality; Epilepsy; Social support; Depression; Anxiety
13.  A comprehensive database of Duchenne and Becker muscular dystrophy patients (0–18 years old) in East China 
Background
Currently, there is no cure for Duchenne and Becker muscular dystrophies (DMD/BMD). However, clinical trials with new therapeutic strategies are being conducted or considered. A comprehensive database is critical for patient recruitment and efficacy evaluation. China has the largest population, yet, no comprehensive database for DMD/BMD is available. Our study registered the data of the DMD/BMD patients in East China.
Methods
A modified registry form of Remudy (http://www.remudy.jp/) was applied to Chinese DMD/BMD patients through the outpatient clinic at Children’s Hospital of Fudan University, Shanghai during the period of August 2011 to December 2013. The data included geographic distribution of patients, age at diagnosis, clinical manifestation, genetic analysis and treatment status.
Results
194 DMD and 35 BMD patients were registered. Most patients lived in East China, namely Jiangsu province, Anhui province, Zhejiang province, Jiangxi province, Shanghai, Fujian province and Shandong province. All individuals aged less than 18 years (age limit to a children’s hospital). Diagnosis was made for a majority of patients during the age of 3–4 (16.6%) and 7–8 (14.8%) years old. Exon deletion was the most frequent genetic mutations (65.5% and 74.3%) followed by point mutations (14.4% and 11.4%), duplications (9.8% and 8.6%) and small insertion/deletion (9.3% and 2.9%) for DMD and BMD, respectively. 82.5% of DMD registrants were ambulatory, and all the BMD registrants were able to walk. 26.3% of DMD registrants have been treated with steroids. Cardiac functions were examined for 46.4% DMD boys and 45.7% BMD boys and respiratory functions were examined for 18.6% DMD boys and 14.3% BMD boys. Four boys with abnormal cardiac function were prescribed for treatment with cardiac medicine. 33.2% of DMD patients are eligible for exon skipping therapy, and among them 9.2% and 4.3% patients are eligible for skipping exon 51 and 53, respectively.
Conclusions
The database is the first linking accurate genetic diagnosis with clinical manifestation and treatment status of dystrophinopathy patients in East China. It provides comprehensive information essential for further patient management, especially for promotion of international cooperation in developing experimental therapies such as exon skipping and read-through of nonsense mutations targeting a subgroup of DMD patient population.
doi:10.1186/s13023-014-0220-7
PMCID: PMC4323212  PMID: 25612904
Duchenne and Becker muscular dystrophy; The CHFU database; Patient management
14.  Emergence of differentially regulated pathways associated with the development of regional specificity in chicken skin 
BMC Genomics  2015;16(1):22.
Background
Regional specificity allows different skin regions to exhibit different characteristics, enabling complementary functions to make effective use of the integumentary surface. Chickens exhibit a high degree of regional specificity in the skin and can serve as a good model for when and how these regional differences begin to emerge.
Results
We used developing feather and scale regions in embryonic chickens as a model to gauge the differences in their molecular pathways. We employed cosine similarity analysis to identify the differentially regulated and co-regulated genes. We applied low cell techniques for expression validation and chromatin immunoprecipitation (ChIP)-based enhancer identification to overcome limited cell availabilities from embryonic chicken skin.
We identified a specific set of genes demonstrating a high correlation as being differentially expressed during feather and scale development and maturation. Some members of the WNT, TGF-beta/BMP, and Notch family known to be involved in feathering skin differentiation were found to be differentially regulated. Interestingly, we also found genes along calcium channel pathways that are differentially regulated. From the analysis of differentially regulated pathways, we used calcium signaling pathways as an example for further verification. Some voltage-gated calcium channel subunits, particularly CACNA1D, are expressed spatio-temporally in the skin epithelium. These calcium signaling pathway members may be involved in developmental decisions, morphogenesis, or epithelial maturation. We further characterized enhancers associated with histone modifications, including H3K4me1, H3K27ac, and H3K27me3, near calcium channel-related genes and identified signature intensive hotspots that may be correlated with certain voltage-gated calcium channel genes.
Conclusion
We demonstrated the applicability of cosine similarity analysis for identifying novel regulatory pathways that are differentially regulated during development. Our study concerning the effects of signaling pathways and histone signatures on enhancers suggests that voltage-gated calcium signaling may be involved in early skin development. This work lays the foundation for studying the roles of these gene pathways and their genomic regulation during the establishment of skin regional specificity.
Electronic supplementary material
The online version of this article (doi:10.1186/s12864-014-1202-9) contains supplementary material, which is available to authorized users.
doi:10.1186/s12864-014-1202-9
PMCID: PMC4326372  PMID: 25612663
Development; Chicken; Skin; Cosine similarity; Calcium; Histone modifications
15.  Biodegradable Vancomycin-eluting Poly[(d,l)-lactide-co-glycolide] Nanofibres for the Treatment of Postoperative Central Nervous System Infection 
Scientific Reports  2015;5:7849.
The incidence of postoperative central nervous system infection (PCNSI) is higher than 5%–7%. Successful management of PCNSI requires a combined therapy of surgical debridement and long-term antibiotic treatment. In this study, Duraform soaked in a prepared bacterial solution was placed on the brain surface of rats to induce PCNSI. Virgin poly[(d,l)-lactide-co-glycolide] (PLGA) nanofibrous membranes (vehicle-control group) and vancomycin-eluting PLGA membranes (vancomycin-nanofibres group) were implanted. The wound conditions were observed and serial brain MRI and pathology examinations were performed regularly. PCNSI was consistently induced in a single, simple step. In the vehicle-control group, most rats died within 1 week, and the survival rate was low (odds ratio = 0.0357, 95% confidence interval = 0.0057–0.2254). The wounds and affected cerebral tissues necrosed with purulence and increased in mass from the resulting PCNSI volumes. Initially, the mean PCNSI volumes showed no significant difference between the two groups. The PCNSI volume in the rats in the vancomycin-nanofibres group significantly decreased (P < 0.01), and the wound appearance was excellent. Pathologic examinations revealed that the necrosis and leukocyte infiltration area decreased considerably. The experimental results suggest that vancomycin-eluting PLGA nanofibres are favourable candidates for treating PCNSI after surgical debridement.
doi:10.1038/srep07849
PMCID: PMC4297986  PMID: 25597553
16.  A Reduced Order Model for Whole-Chip Thermal Analysis of Microfluidic Lab-on-a-Chip Systems 
Microfluidics and nanofluidics  2013;16(1-2):369-380.
This paper presents a Krylov subspace projection-based Reduced Order Model (ROM) for whole microfluidic chip thermal analysis, including conjugate heat transfer. Two key steps in the reduced order modeling procedure are described in detail, including (1) the acquisition of a 3D full-scale computational model in the state-space form to capture the dynamic thermal behavior of the entire microfluidic chip; and (2) the model order reduction using the Block Arnoldi algorithm to markedly lower the dimension of the full-scale model. Case studies using practically relevant thermal microfluidic chip are undertaken to establish the capability and to evaluate the computational performance of the reduced order modeling technique. The ROM is compared against the full-scale model and exhibits good agreement in spatiotemporal thermal profiles (<0.5% relative error in pertinent time scales) and over three orders-of-magnitude acceleration in computational speed. The salient model reusability and real-time simulation capability renders it amenable for operational optimization and in-line thermal control and management of microfluidic systems and devices.
doi:10.1007/s10404-013-1210-0
PMCID: PMC3891786  PMID: 24443647
17.  Gene order rearrangement of the M gene in the rabies virus leads to slower replication 
Virusdisease  2014;25(3):365-371.
The matrix protein (M) is one of only five genes in the RV genome and is an important multifunctional protein. Besides to allow for the release of newly replicated virions pairing with G, the M protein also functions in virus replication, pathogenicity, and host cell apoptosis. The goal of present study is to generate recombinant viruses with M gene rearranged, thus laying the foundation for further exploring what will happen when the gene for M is relocated on the RV single-strand RNA. We used rHEP-Flury, an attenuated virus that remains virulent for less than 3 days in sucking mice, to reshuffle the M gene, using an approach that leaves the other viral nucleotide sequence intact. Two viruses with translocated M genes (N1M2 and N1M4) were recovered from each of the rearranged cDNAs, whose gene order is 3′-N-M-P-G-L-5′ and 3′-N-P-G-M-L-5′ respectively. The growth dynamics of these viruses showed slower replication than the wild-type virus in multiple-step growth curves, but they can grow to a comparable titer in tests of single-step growth curves. Further experimentation with these rearranged viruses will provide insights into the relationships between genome structure and virus phenotypes.
Electronic supplementary material
The online version of this article (doi:10.1007/s13337-014-0220-1) contains supplementary material, which is available to authorized users.
doi:10.1007/s13337-014-0220-1
PMCID: PMC4188201
Gene rearrangement; Matrix protein; Rabies virus; Reverse genetics
18.  Off to the Organelles - Killing Cancer Cells with Targeted Gold Nanoparticles 
Theranostics  2015;5(4):357-370.
Gold nanoparticles (AuNPs) are excellent tools for cancer cell imaging and basic research. However, they have yet to reach their full potential in the clinic. At present, we are only beginning to understand the molecular mechanisms that underlie the biological effects of AuNPs, including the structural and functional changes of cancer cells. This knowledge is critical for two aspects of nanomedicine. First, it will define the AuNP-induced events at the subcellular and molecular level, thereby possibly identifying new targets for cancer treatment. Second, it could provide new strategies to improve AuNP-dependent cancer diagnosis and treatment.
Our review summarizes the impact of AuNPs on selected subcellular organelles that are relevant to cancer therapy. We focus on the nucleus, its subcompartments, and mitochondria, because they are intimately linked to cancer cell survival, growth, proliferation and death. While non-targeted AuNPs can damage tumor cells, concentrating AuNPs in particular subcellular locations will likely improve tumor cell killing. Thus, it will increase cancer cell damage by photothermal ablation, mechanical injury or localized drug delivery. This concept is promising, but AuNPs have to overcome multiple hurdles to perform these tasks. AuNP size, morphology and surface modification are critical parameters for their delivery to organelles. Recent strategies explored all of these variables, and surface functionalization has become crucial to concentrate AuNPs in subcellular compartments.
Here, we highlight the use of AuNPs to damage cancer cells and their organelles. We discuss current limitations of AuNP-based cancer research and conclude with future directions for AuNP-dependent cancer treatment.
doi:10.7150/thno.10657
PMCID: PMC4329500
Gold nanoparticles; AuNPs; cancer cell imaging
19.  Rapid and Sensitive Detection of Listeria ivanovii by Loop-Mediated Isothermal Amplification of the smcL Gene 
PLoS ONE  2014;9(12):e115868.
A loop-mediated isothermal amplification (LAMP) assay for rapid and sensitive detection of the L. ivanovii strains had been developed and evaluated in this study. Oligonucleotide primers specific for L. ivanovii species were designed corresponding to smcL gene sequences. The primers set comprise six primers targeting eight regions on the species-specific gene smcL. The LAMP assay could be completed within 1 h at 64°C in a water bath. Amplification products were directly observed by the Loopamp Fluorescent Detection Reagent (FD) or detected by agarose gel electrophoresis. Moreover, the LAMP reactions were also detected by real-time measurement of turbidity. The exclusivity of 77 non-L. ivanovii and the inclusivity of 17 L. ivanovii were both 100% in the assay. Sensitivity of the LAMP assay was 250 fg DNA and 16 CFU per reaction for detection of L. ivanovii in pure cultures and simulated human stool. The LAMP assay was 10 and 100-fold more sensitive than quantitative PCR (qPCR) and conventional PCR assays,respectively. When applied to human stool samples spiked with low level (8 CFU/0.5 g) of L. ivanovii strains, the new LAMP assay described here achieved positive detection after 6 hours enrichment. In conclusion, the new LAMP assay in this study can be used as a valuable, rapid and sensitive detection tool for the detection of L. ivanovii in field, medical and veterinary laboratories.
doi:10.1371/journal.pone.0115868
PMCID: PMC4280119  PMID: 25549337
20.  Detection of the electromechanical delay and its components during voluntary isometric contraction of the quadriceps femoris muscle 
Electromechanical delay (EMD) was described as a time elapsed between first trigger and force output. Various results have been reported based on the measurement method with observed inconsistent results when the trigger is elicited by voluntary contraction. However, mechanomyographic (MMG) sensor placed far away on the skin from the contracting muscle was used to detect muscle fiber motion and excitation-contraction (EC) coupling which may give unreliable results. On this basis, the purpose of this study was to detect EMD during active muscle contraction whilst introducing an ultrafast ultrasound (US) method to detect muscle fiber motion from a certain depth of the muscle. Time delays between onsets of EMG-MMG, EMG-US, MMG-FORCE, US-FORCE, and EMG-FORCE were calculated as 20.5 ± 4.73, 28.63 ± 6.31, 19.21 ± 6.79, 30.52 ± 8.85, and 49.73 ± 6.99 ms, respectively. Intrarater correlation coefficient (ICC) was higher than MMG when ultrafast US was used for detecton of the Δt EMG-US and Δt US-FORCE, ICC values of 0.75 and 0.70, respectively. Synchronization of the ultrafast ultrasound with EMG and FORCE sensors can reveal reliable and clinically useful results related to the EMD and its components when muscle is voluntarily contracted. With ultrafast US, we detect onset from the certain depth of the muscle excluding the tissues above the muscle acting as a low-pass filter which can lead to inaccurate time detection about the onset of the contracting muscle fibers. With this non-invasive technique, understanding of the muscle dynamics can be facilitated.
doi:10.3389/fphys.2014.00494
PMCID: PMC4274888  PMID: 25566091
electromechanical delay; excitation-contraction coupling; series elastic components; contractile components; rectus femoris muscle; transient ultrasound
21.  Proinflammatory effects and molecular mechanisms of interleukin-17 in intestinal epithelial cell line HT-29 
World Journal of Gastroenterology : WJG  2014;20(47):17924-17931.
AIM: To evaluate the proinflammatory effects and molecular mechanisms of interleukin (IL)-17 in intestinal epithelial cell line HT-29.
METHODS: HT-29 cells were cultured with IL-17, tumor necrosis factor (TNF)-α, or the combination of both IL-17 and TNF-α. Real-time PCR and Western blot were used to measure the gene expression levels of neutrophil chemokines CXCL1, CXCL2, CXCL5, CXCL6, IL-8 and TH-17 cell chemokine CCL20, the phosphorylation levels of p38 and TNF-α, and the expression level of IL-8, after using the p38 inhibitor in HT-29 cells. The stable Act1 knockdown HT-29 cell line was established to further test the phosphorylation changes of p38, after using IL-17 and TNF-α.
RESULTS: After HT-29 cells were cultured with IL-17 and TNF-α, the expression levels of neutrophil chemokines (CXCL1, CXCL2, CXCL5, CXCL6, IL-8) and Th17 chemokine (CCL20) significantly improved (24.96 ± 2.53, 28.47 ± 2.87, 38.08 ± 2.72, 33.47 ± 2.41, 31.7 ± 2.38, 44.37 ± 2.73, respectively), and the differences were all statistically significant (P < 0.01). Western blot results showed that IL-17 obviously enhanced the phosphorylation level of p38, which was induced by TNF-α. Compared with the control group, the expression level of IL-8 significantly declined (9.47 ± 1.36 vs 3.06 ± 0.67, P < 0.01) when TH-29 cells were cultured with IL-17 and TNF-α. p38 inhibition assay showed that the p38 pathway played an essential role in the inflammatory response induced by IL-17. p38 phosphorylation levels could not be changed after using IL-17 and TNF-α in the stable Act1 knockdown HT-29 cell line.
CONCLUSION: IL-17 significantly promoted the gene expression levels of TNF-α-induced neutrophil chemokines and Th17 cell chemokine. It is obvious that IL-17 and TNF-α have synergistic effects on p38.
doi:10.3748/wjg.v20.i47.17924
PMCID: PMC4273142  PMID: 25548490
IL-17; HT-29; TNF-α; Inflammatory bowel disease
22.  Cerebellar cryptococcosis characterized by a space-occupying lesion in an immunocompetent non-HIV patient 
Central nervous system (CNS) cryptococcosis is an opportunistic fungal infection that typically occurs in patients with reduced immunological function, such as patients with AIDS, patients receiving organ transplants, or patients receiving corticosteroid and immunosuppressive therapy. CNS cryptococcosis rarely occurs in immunocompetent patients. CNS cryptococcosis is characterized by meningitis and encephalitis and occasionally forms isolated granulomas. Isolated cerebellar cryptococcoma is a rare condition, especially in immunocompetent patients, and the misdiagnosis rate is high. A definite diagnosis must be based on pathology. To raise awareness of this disease, the clinical data of a patient with cryptococcomas in the right side of the cerebellum are reported.
doi:10.2147/NDT.S75432
PMCID: PMC4277240  PMID: 25552907
cryptococcosis; cerebellar; central nervous system; magnetic resonance imaging; immunosuppressive
23.  BCL6 is a negative prognostic factor and exhibits pro-oncogenic activity in ovarian cancer 
Background: Dysregulation of BCL6 plays critical oncogenic roles and facilitates tumorigenesis in various malignancies. However, whether the aberrant expression of BCL6 in ovarian carcinoma is associated with malignancy, metastasis or prognosis remains unknown. Our study aimed to investigate the expression of BCL6 in ovarian carcinoma and its possible correlation with clinicopathological features as well as patient survival to reveal its biological effects in ovarian tumor progression. Methods: Immunochemistry analysis was performed in 105 cases of ovarian carcinoma covering the histological types of serous, endometrioid and clear cell. Spearman analysis was used to calculate the correlation between pathological parameters and the expression of BCL6. Kaplan–Meier method and Cox proportional hazards analysis were used to analyze the disease-specific survival (DSS) and disease-free survival (DFS). We also assessed whether overexpression and knockdown of BCL6 influence in vitro cell proliferation, cell cycle progression, as well as tumor cell invasion and migration. Results: The expression of BCL6 was higher in all three major kinds of ovarian cancer in comparison with paratumorous epithelium. BCL6 expression was tightly correlated with FIGO staging, lymph node metastasis and recurrence. Higher expression of BCL6 led to a significantly poorer DSS and DFS and multivariate analysis revealed that BCL6 was an independent risk factor of DSS and DFS. Enforced overexpression of BCL6 in ovarian tumor cells stimulated proliferation by inducing G1–S transition, and promoted tumor cell invasion and migration. Conversely, RNA interference–mediated silencing BCL6 expression inhibited proliferation by altered cell cycle progression and reduced the ability of the cells to migrate, and invade the extracellular matrix in culture. Conclusions: Our study suggests that the inappropriate activation of BCL6 predicts poor prognosis and promotes tumor progression in ovarian carcinoma. Targeting BCL6 could be a novel therapeutic choice for treating ovarian carcinoma patients.
PMCID: PMC4300693  PMID: 25628935
BCL6; ovarian carcinoma; prognosis; proliferation; invasion
24.  Prognostic significance of osteopontin in patients with lung cancer: a meta-analysis 
Both plasma/serum/pleural effusion osteopontin concentration (PSPO) and tumor tissue osteopontin expression (TTO) have recently been reported to be involved in the prognosis of lung cancer. In this study, we performed a meta-analysis to demonstrate the association between PSPO/TTO and survival in patients with lung cancer. We searched in PubMed, EMBASE, Cochrane library, Web of Science and Chinese Biomedical database (CBM) for relevant literatures. Stata 12.0 was applied to pool the eligible studies and synthesize hazard ratios (HRs) and its corresponding 95% confidence interval (CI). For PSPO, a total of 8 studies with 1000 patients were included in final analysis. Combined HR suggested high PSPO predicted an unfavorable overall survival (OS) (HR=1.52, 95% CI: 1.13-2.05) and progress-free survival (PFS) (HR=1.73, 95% CI: 1.35-2.21). For TTO, 5 studies with a total of 747 patients were employed in final analysis. Pooled HR indicated that elevated TTO was associated with poor OS (HR=2.16, 95% CI: 1.65-2.83) and disease/relapse-free survival (D/RFS) (HR=2.36, 95% CI: 1.79-3.12). Subgroup analysis was performed to explore the causes of heterogeneity. Publication bias by begg’s test was not statistically significant. Sensitivity analysis showed that the pooled results were robust. This study revealed that both high TTO and PSPO are associated with poor prognosis in patients with lung cancer.
PMCID: PMC4307405  PMID: 25663958
Osteopontin; lung cancer; prognosis; meta-analysis
25.  Protective effect of epimedium combined with oligomeric proanthocyanidins on exercise-induced renal ischemia-reperfusion injury of rats 
Objective: This paper studied the protective effect and mechanism of epimedium combined with oligomeric proanthocyanidins on exercise-induced renal ischemia-reperfusion injury of rats. Methods: In the experiment, the rats were given exhaustive swimming training and then their blood urea nitrogen (BUN) and other biochemical indexes were measured after they were given gastric perfusion with 6.01 g/kg doze of epimedium and 50 mg/kg doze of oligomeric proanthocyanidins for 56 days. Results: The result indicated that 8 weeks of over training led to ischemia-reperfusion injury of rats. Moreover, their kidney tissues were significantly changed pathologically and renal functions drastically damaged. BUN and serum creatinine increased and EOM group (P < 0.05), OPCOM group (P < 0.05) and EOPCOM group (P < 0.01) were lower than OM group. EOPCOM group was lower than OPCOM group. SOD activity decreased, EOM group (P < 0.05), OPCOM group (P < 0.05), EOPCOM group (P < 0.01) higher than OM group, and EOPCOM group (P < 0.05) higher than OPCOM group. The content of MDA increased, EOM group (P < 0.05), OPCOM group (P < 0.05), EOPCOM group (P < 0.01) lower than OM group, and EOPCOM group (P < 0.05) lower than OPCOM group. Conclusion: Both epimedium and oligomeric proanthocyanidins can boost SOD activity, clean oxygen radicals, clean and alleviate peroxidation of lipids, which exert protection on exercise-induced renal ischemia-reperfusion. The two combined yield a much better result.
PMCID: PMC4307546  PMID: 25664099
Epimedium; oligomeric proanthocyanidins; exercise-induced ischemia-reperfusion

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