Publications by authors named "Yutian Liu"

31 Publications

Effects of perchlorate and exogenous T4 exposures on development, metamorphosis and endochondral ossification in Bufo gargarizans larvae.

Aquat Toxicol 2021 Nov 17;242:106036. Epub 2021 Nov 17.

College of Life Science, Shaanxi Normal University, Xi'an 710119, China. Electronic address:

Several endocrine-disrupting chemicals (EDCs) have been proven to interfere with the physiological function of thyroid hormone (TH), which affected growth and development. However, few studies have investigated the effects of EDCs on TH axis with consequence for skeletal development in amphibians. This study thus examined the potential role of perchlorate and T4 in growth, development and endochondral ossification during metamorphosis of Bufo gargarizans. Our studies showed that NaClO₄ treatment caused weight gain and delayed the developmental stage in B. gargarizans tadpoles, while T4 decreased body size and survival rate, accelerated metamorphic duration and increased the risk of early death. Histological sections suggested that NaClO₄ and T4 treatments caused damages to thyroid tissue, such as decreased thyroid gland size, follicle size, colloid area, the height of follicular epithelial cells and the number of follicles. In addition, the double skeletal staining and RT-qPCR showed that NaClO₄ and T4 treatments inhibited the endochondral ossification by regulating TH synthesis (TRs, Dios) and endochondral ossification-related genes (MMPs, Runxs, VEGFs and VEGFRs) expression levels, which might affect terrestrial locomotion and terrestrial life. Altogether, these thyroid injury and gene expression changes as caused by NaClO₄ and T4 may have an influence on development and endochondral ossification during the metamorphosis of amphibians.
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http://dx.doi.org/10.1016/j.aquatox.2021.106036DOI Listing
November 2021

A staging table of embryonic development for a viviparous (live-bearing) lizard Eremias multiocellata (Squamata: Lacertidae).

Reprod Fertil Dev 2021 Nov;33(14):782-797

Institute of Biochemistry and Molecular Biology, School of Life Science, Lanzhou University, 222 Tianshui South Road, Lanzhou 730000, China.

As the only viviparous reptile in China that has both temperature-dependent sex determination (TSD) and genetic-dependent sex determination (GSD) mechanisms, Eremias multiocellata is considered as an ideal species for studying the sex determination mechanism in viviparous lizards. However, studies on embryonic stage of viviparous lizards and morphological characteristics of each stage are limited. In the present study, the embryonic development process of E. multiocellata is divided into 15 stages (stages 28-42) according to the morphology of embryos. Embryos sizes are measured and continuous dynamic variation of some key features, including limbs, genitals, eyes, pigments, and brain scales are color imaged by a stereoscopic microscope. Furthermore, based on these morphological characteristics, we compare the similarities and differences in the embryonic development of E. multiocellata with other squamate species. Our results not only identified the staging table of E. multiocellata with continuous changes of external morphological characteristics but also developed a staging scheme for an important model species that provides a necessary foundation for study of sex determination in a viviparous lizard.
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http://dx.doi.org/10.1071/RD21082DOI Listing
November 2021

NRF2 signalling pathway: New insights and progress in the field of wound healing.

J Cell Mol Med 2021 Jun 18. Epub 2021 Jun 18.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

As one of the most common pathological processes in the clinic, wound healing has always been an important topic in medical research. Improving the wound healing environment, shortening the healing time and promoting fast and effective wound healing are hot and challenging issues in clinical practice. The nuclear factor-erythroid-related factor 2 (NFE2L2 or NRF2) signalling pathway reduces oxidative damage and participates in the regulation of anti-oxidative gene expression in the process of oxidative stress and thus improves the cell protection. Activation of the NRF2 signalling pathway increases the resistance of the cell to chemical carcinogens and inflammation. The signal transduction pathway regulates anti-inflammatory and antioxidant effects by regulating calcium ions, mitochondrial oxidative stress, autophagy, ferroptosis, pyroptosis and apoptosis. In this article, the role of the NRF2 signalling pathway in wound healing and its research progress in recent years are reviewed. In short, the NRF2 signalling pathway has crucial clinical significance in wound healing and is worthy of further study.
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http://dx.doi.org/10.1111/jcmm.16597DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8406474PMC
June 2021

The whole profiling and competing endogenous RNA network analyses of noncoding RNAs in adipose-derived stem cells from diabetic, old, and young patients.

Stem Cell Res Ther 2021 05 29;12(1):313. Epub 2021 May 29.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1277 Jiefang Avenue, Wuhan, 430022, China.

Background: Mesenchymal stem cells including adipose-derived stem cells (ASCs) have a considerable potential in the field of translational medicine. Unfortunately, multiple factors (e.g., older age, co-existing diabetes, and obesity) may impair cellular function, which hinders the overall effectiveness of autologous stem cell therapy. Noncoding RNAs-including microRNAs (miRNAs), long ncRNAs (lncRNAs), and circular RNAs (circRNAs)-have been shown to play important roles in stem cell biology. However, the overall diabetes-related and aging-related expression patterns and interactions of these RNAs in ASCs remain unknown.

Method: The phenotypes and functions of ASCs isolated from diabetic (D-ASCs), old (O-ASCs), and young (Y-ASCs) donors were evaluated by in vitro assays. We conducted high-throughput RNA sequencing (RNA-seq) in these ASCs to identify the differentially expressed (DE) RNAs. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and protein-protein interaction (PPI) analyses were performed to investigate mRNAs with significant differences among groups. The lncRNA- or circRNA-associated competing endogenous RNA (ceRNA) networks were constructed based on bioinformatics analyses and real-time polymerase chain reaction (RT-PCR) results. The miR-145-5p mimics were transfected into O-ASCs and verified by PCR.

Results: ASCs from diabetic and old donors showed inferior migration ability and increased cellular senescence. Furthermore, O-ASCs have decreased capacities for promoting endothelial cell angiogenesis and fibroblast migration, compared with Y-ASCs. The DE miRNAs, mRNAs, lncRNAs, and circRNAs were successfully identified by RNA-seq in O-ASCs vs. Y-ASCs and D-ASCs vs. O-ASCs. GO and KEGG analyses demonstrated that DE mRNAs were significantly enriched in aging and cell senescence terms separately. PPI networks revealed critical DE mRNAs in the above groups. RNAs with high fold changes and low p values were validated by PCR. ceRNA networks were constructed based on bioinformatics analyses and validated RNAs. Additionally, the lncRNA RAET1E-AS1-miR-145-5p-WNT11/BMPER axis was validated by PCR and correlation analyses. Finally, the overexpression of miR-145-5p was found to rejuvenate O-ASCs phenotype and augment the functionality of these cells.

Conclusion: Our research may provide insights regarding the underlying mechanisms of ASC dysfunction; it may also offer novel targets for restoring therapeutic properties in ASCs.
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http://dx.doi.org/10.1186/s13287-021-02388-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8164820PMC
May 2021

Roles of Multidrug Resistance Protein 4 in Microbial Infections and Inflammatory Diseases.

Authors:
Wei Liu Yutian Liu

Microb Drug Resist 2021 May 17. Epub 2021 May 17.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Numerous studies have reported the emergence of antimicrobial resistance during the treatment of common infections. Multidrug resistance (MDR) leads to failure of antimicrobial treatment, prolonged illness, and increased morbidity and mortality. Overexpression of multidrug resistance proteins (MRPs) as drug efflux pumps are one of the main contributions of MDR, especially multidrug resistance protein 4 (MRP4/ABCC4) in the development of antimicrobial resistance. The molecular mechanism of antimicrobial resistance is still under investigation. Various intervention strategies have been developed for overcoming MDR, but the effect is limited. Suppression of MRP4 may be an attractive therapeutic approach for addressing drug resistance. However, there are few reports on the involvement of MRP4 in antimicrobial resistance and inflammatory diseases. In this review, we introduced the function and regulation of MRP4, and then summarized the roles of MRP4 in microbial infections and inflammatory diseases as well as polymorphisms in the gene encoding this transporter. Further studies should be conducted on drug therapy targeting MRP4 to improve the efficacy of antimicrobial therapy. This review can provide useful information on MRP4 for overcoming antimicrobial resistance and anti-inflammatory therapy.
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http://dx.doi.org/10.1089/mdr.2020.0020DOI Listing
May 2021

Establishment of nitrous oxide (NO) dynamics model based on ASM3 model during biological nitrogen removal via nitrification.

Environ Technol 2020 Sep 24:1-11. Epub 2020 Sep 24.

Shandong Tong Yuan Design Group CO., LTD., Jinan, People's Republic of China.

Nitrous oxide (NO), as one of the six greenhouse gases, is mainly produced in the biological nitrogen removal process of wastewater treatment plants (WWTPs). Establishing the NO kinetic model can provide insight into the NO generation mechanism and regulate its production. This work uses Activated Sludge Model NO.3 (ASM3) as the basic framework, combines organic storage with endogenous respiration theory, and couples ammonia-oxidizing bacteria (AOB) denitrification pathway and the NHOH/NOH model to establish a kinetic model. Meanwhile, the Sequencing Batch Reactor (SBR) process with artificial simulated urban domestic sewage was used as the carrier; MATLAB and EXCEL software were used as tools to establish a model calculation programme. The simulated values obtained by substituting the operating conditions of the SBR process into the model and the measured values of the SBR process were analysed. The correlation coefficient () between the experimental values and simulated values obtained for the 5 components of COD, ammonia, nitrite, nitrate and total NO is 0.952, 0.996, 0.902, 0.991 and 0.956, respectively, which indicates that the NO kinetic model has great consistency, this further shows that the established model modelling mechanism is clear and accurate, and provides a new method for the NO dynamic model.
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http://dx.doi.org/10.1080/09593330.2020.1822447DOI Listing
September 2020

Clinical Validation of Two Recombinase-Based Isothermal Amplification Assays (RPA/RAA) for the Rapid Detection of African Swine Fever Virus.

Front Microbiol 2020 21;11:1696. Epub 2020 Jul 21.

National Reference Laboratory for African Swine Fever, National Surveillance and Research Center for Exotic Animal Diseases, National Surveillance and Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, China.

African swine fever (ASF), caused by African swine fever virus (ASFV), is a devastating infectious disease of domestic pigs and wild boars, and has tremendous negative socioeconomic impact on the swine industry and food security worldwide. It is characterized as a notifiable disease by World Organisation for Animal Health (OIE). No effective vaccine or treatment against ASF has so far been available. Early detection and rapid diagnosis are of potential significance to control the spread of ASF. Recombinase-based isothermal amplification assay, recombinase polymerase amplification (RPA) developed by TwistDx (Cambridge, United Kingdom) or recombinase-aided amplification (RAA) by Qitian (Wuxi, China), is becoming a molecular tool for the rapid, specific, and cost-effective identification of multiple pathogens. In this study, we aim to investigate if RPA/RAA can be a potential candidate for on-site, rapid and primary detection of ASFV. A panel of 152 clinical samples previously well-characterized by OIE-recommended qPCR was enrolled in this study, including 20 weak positive (Ct value ≥ 30) samples. This panel was consisted of different types, such as EDTA-blood, spleen, lung, lymph node, kidney, tonsil, liver, brain. We evaluated two recombinase-based isothermal amplification assays, RPA or RAA, by targeting the ASFV gene (p72), and validated the clinical performance in comparison with OIE real-time PCR. Our result showed that the analytical sensitivity of RPA and RAA was as 93.4 and 53.6 copies per reaction, respectively at 95% probability in 16 min, at 39°C. They were universally specific for all 24 genotypes of ASFV and no cross reaction to other pathogens including Classical swine fever virus (CSV), Foot-and-mouth disease virus (FMDV), Pseudorabies virus, Porcine circovirus 2 (PCV2), Porcine Reproductive and respiratory syndrome virus (PPRSV). The results on detection of various kinds of clinical samples indicated an excellent diagnostic agreement between RPA, RAA and OIE real-time PCR method, with the kappa value of 0.960 and 0.973, respectively. Compared to real-time PCR, the specificity of both RPA and RAA was 100% (94.40% ∼ 100%, 95% CI), while the sensitivity was 96.59% (90.36% ∼ 99.29%, 95% CI) and 97.73% (92.03% ∼ 99.72%, 95% CI), respectively. Our data demonstrate that the developed recombinase-based amplification assay (RPA/RAA), promisingly equipped with field-deployable instruments, offers a sensitive and specific platform for the rapid and reliable detection of ASFV, especially in the resource-limited settings for the purpose of screening and surveillance of ASF.
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http://dx.doi.org/10.3389/fmicb.2020.01696DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7385304PMC
July 2020

An extra insertion of tandem repeat sequence in African swine fever virus, China, 2019.

Virus Genes 2019 Dec 27;55(6):843-847. Epub 2019 Sep 27.

China Animal Health and Epidemiology Center, Qingdao, China.

On 7 March 2019, African swine fever in a domestic pig farm was detected in Guangxi Province of China. The phylogenetic analysis showed that its causative strain contained two tandem repeat sequence insertions in the intergenic region between the I73R and the I329L genes, and was different from previously reported strains in China and other countries.
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http://dx.doi.org/10.1007/s11262-019-01704-9DOI Listing
December 2019

Mechanism of Caulophyllum robustum Maxim against rheumatoid arthritis using LncRNA-mRNA chip analysis.

Gene 2020 Jan 12;722:144105. Epub 2019 Sep 12.

Pharmaceutical College, Guangdong Pharmaceutical University, Guangzhou 510224, China. Electronic address:

Background: Caulophyllum robustum Maxim (CRM) is a medicinal compound of the Northeast and is commonly used in China for the treatment of rheumatic pain and rheumatoid arthritis (RA). A preliminary study found that CRM has good anti-inflammatory, analgesic and immunosuppressive effects. However, the specific links and targets for its function remain unclear. Our study aimed to provide a mechanism for the action of Caulophyllum robustum Maxim extraction (CRME) against RA and to establish a method for studying disease treatment using Chinese medicine.

Methods: The 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) method was used to detect the toxicity of CRME in L929 cells, and the concentration ranges of the blank, model, and CRME drug groups were determined. Differentially expressed long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) were identified between the three groups. Gene Ontology (GO) and pathway enrichment analyses were performed to analyze the biological functions and pathways of the differentially expressed genes. Expression of Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2 and Egr1 in the blank, model and drug groups was detected by quantitative real-time PCR (qRT-PCR), and the role of CRME on the above factors was determined to ensure consistency with the chip data.

Results: A total of 329 significantly upregulated genes and 141 downregulated genes were identified between the blank and model groups. A total of 218 significantly upregulated genes and 191 downregulated genes were identified between the CRME drug group and model group. CRME has a significant role in multiple pathways involved in the occurrence and development of RA. Additionally, Hist1h2bj, Hist1h2ba, Zfp36, Ccl3, Cxcl2, and Egr1 were observed in modules of the lncRNA-mRNA weighted co-expression network, consistent with the chip data.

Conclusions: CRME has regulatory effects on inflammatory factors, the histone family, chemokines and their ligands that are related to RA-related cytokines, the RA pathway, the TNF signaling pathway, the Toll receptor-like signaling pathway, the chemokine signaling pathways and other pathways are related to the course of RA.
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http://dx.doi.org/10.1016/j.gene.2019.144105DOI Listing
January 2020

Differentiated human adipose-derived stromal cells exhibit the phenotypic and functional characteristics of mature Schwann cells through a modified approach.

Cytotherapy 2019 09 25;21(9):987-1003. Epub 2019 Jul 25.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address:

Background Aims: Tissue engineering technology is a promising therapeutic strategy in peripheral nerve injury. Schwann cells (SCs) are deemed to be a vital component of cell-based nerve regeneration therapies. Many methods for producing SC-like cells derived from adipose-derived stromal cells (ADSCs) have been explored, but their phenotypic and functional characteristics remain unsatisfactory.

Methods: We investigated whether human ADSCs can be induced to differentiate into mature and stable SC-like cells with the addition of insulin, progestero``ne and glucocorticoids. The phenotypic and functional characteristics of new differentiated ADSCs (modified SC-like cells) were evaluated by real-time quantitative polymerase chain reaction, enzyme-linked immunosorbent assay and immunocytochemistry in vitro. Cells loaded into collagen sponge biomaterials were implanted around transected sciatic nerves with a 10-mm gap in vivo. The axon regrowth and functional recovery of the regenerated nerves were assessed by immunohistochemistry and Walking footprint analysis.

Results: After differentiation induction, the modified SC-like cells showed significantly up-regulated levels of S100B and P0 and enhanced proliferative and migratory capacities. In addition, the modified SC-like cells showed increased secretion of neurotrophic factors, and their functional characteristics were maintained for more than 3 weeks after removing the induction reagents. The modified SC-like cells exhibited significantly enhanced axon regrowth, myelination and functional recovery after sciatic nerve injury.

Conclusions: Overall, the results suggest that this modified induction method can induce human ADSCs to differentiate into cells with the molecular and functional properties of mature SCs and increase the promotion of peripheral nerve regeneration.
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http://dx.doi.org/10.1016/j.jcyt.2019.04.061DOI Listing
September 2019

Exosomes from human adipose-derived stem cells promote sciatic nerve regeneration via optimizing Schwann cell function.

J Cell Physiol 2019 12 23;234(12):23097-23110. Epub 2019 May 23.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Human adipose-derived stem cells (ASCs) have a potential for the treatment of peripheral nerve injury. Recent studies demonstrated that stem cells can mediate therapeutic effect by secreting exosomes. We aimed to investigate the effect of human ASCs derived exosomes (ASC-Exos) on peripheral nerve regeneration in vitro and in vivo. Our results showed after being internalized by Schwann cells (SCs), ASC-Exos significantly promoted SC proliferation, migration, myelination, and secretion of neurotrophic factors by upregulating corresponding genes in vitro. We next evaluated the efficacy of ASC-Exo therapy in a rat sciatic nerve transection model with a 10-mm gap. Axon regeneration, myelination, and restoration of denervation muscle atrophy in ASC-Exos treated group was significantly improved compared to vehicle control. This study demonstrates that ASC-Exos effectively promote peripheral nerve regeneration via optimizing SC function and thereby represent a novel therapeutic strategy for regenerative medicine and nerve tissue engineering.
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http://dx.doi.org/10.1002/jcp.28873DOI Listing
December 2019

A natural product enhances apoptosis via mitochondria/caspase-mediated pathway in HeLa cells.

J Cell Biochem 2019 10 17;120(10):16811-16823. Epub 2019 May 17.

Shanghai Key Laboratory of Chemical Biology, School of Pharmacy, East China University of Science and Technology, Shanghai, China.

Cervical cancer is the fourth most lethal human malignancy and the leading cause of death among females around the world. Many antitumor agents have microbial origins. 5'-epi-SPA-6952A is a new 24-membered macrolide isolated from the cultured broth of Streptomyces diastatochromogenes. Therefore, we studied the activity and molecular mechanism of 5'-epi-SPA-6952A in human cervical carcinoma HeLa cell. The results showed that 5'-epi-SPA-6952A significantly inhibited cell proliferation and migration. In addition, 5'-epi-SPA-6952A obviously increased the production of intracellular reactive oxygen species and DNA damage in HeLa cells. Moreover, nuclear shrinkage of cells, decrease in mitochondrial membrane potential, and upregulation of Bax/Bcl-2 ratio resulted in the release of cytochrome c, and activation of caspase-9/3 was observed in HeLa cells treated with 5'-epi-SPA-6952A, which means it enhanced the intrinsic mitochondrial apoptosis. Besides, DNA-damage associated proteins poly (ADP-ribose) polymerase (PARP) and p53 were also studied, and the expressions of cleaved-PARP and p53 were drastically increased in HeLa cells treated with 5'-epi-SPA-6952A. Furthermore, we confirmed that 5'-epi-SPA-6952A affected the survival of HeLa cells by blocking cell cycle progression in the G1 phase. Taken together, the results shows that 5'-epi-SPA-6952A significantly inhibited HeLa cells proliferation via intrinsic mitochondrial apoptosis, cell cycle arrest, and blocking cell migration.
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http://dx.doi.org/10.1002/jcb.28939DOI Listing
October 2019

Microarray analyses of lncRNAs and mRNAs expression profiling associated with diabetic peripheral neuropathy in rats.

J Cell Biochem 2019 09 26;120(9):15347-15359. Epub 2019 Apr 26.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Diabetic peripheral neuropathy (DPN) is considered to be the most frequent neuropathic complication of diabetes, and severely affects the quality of life of patients. Long noncoding RNAs (lncRNAs) participate in various pathophysiological processes and associate with many diseases. However, the exact impact of lncRNAs on DPN remains obscure. To discover a potential connection, a microarray study was conducted to analyze the expression profiling of lncRNAs and messenger RNAs (mRNAs) in dorsal root ganglia (DRG) from streptozotocin-induced diabetic rats with DPN. As a result, 983 lncRNAs and 1357 mRNAs were aberrantly expressed compared with control samples. Using bioinformatics analyses, we identified 558 Gene Ontology terms and 94 Kyoto Encyclopedia of Genes and Genomes pathways to be significantly enriched. Additionally, the signal-net analysis indicated that integrin receptors, including Itgb3, Itgb1, Itgb8, and Itga6, might be important players in network regulation. Furthermore, the lncRNA-mRNA network analysis showed dynamic interactions between the dysregulated lncRNAs and mRNAs. This is the first study to present an overview of lncRNA and mRNA expressions in DRG tissues from DPN rats. Our results indicate that these differentially expressed lncRNAs may have crucial roles in pathological processes of DPN by regulating their coexpressed mRNAs. The data may provide novel targets for future studies, which should focus on validating their roles in the progression of DPN.
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http://dx.doi.org/10.1002/jcb.28802DOI Listing
September 2019

Identification of competitive endogenous RNAs network in breast cancer.

Cancer Med 2019 05 1;8(5):2392-2403. Epub 2019 Apr 1.

Department of Biochemistry and Molecular Biology, Mudanjiang Medical University, Mudanjiang, China.

Background: MiRNAs can regulate gene expression directly or indirectly, and long noncoding RNAs as competing endogenous RNA (ceRNAs) can bind to miRNAs competitively and affect mRNA expression. The ceRNA network is still unclear in breast cancer. In this study, a ceRNA network was constructed, and new treatment and prognosis targets and biomarkers for breast cancer were explored.

Methods: A total of 1 096 cancer tissues and 112 adjacent normal tissues to cancer from the TCGA database were used to screen out significant differentially expressed mRNAs (DEMs), lncRNAs (DELs), and miRNAs (DEMis) to construct a ceRNA network. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were used to predict potential functions. Survival analysis was performed to predict which functions were significant for prognosis.

Results: From the analysis, 2 139 DEMs, 1 059 DELs, and 84 DEMis were obtained. Targeting predictions for DEMis-DELs and DEMis-DEMs can yield 26 DEMs, 90 DELs, and 18 DEMis. We performed GO enrichment analysis, and the results showed that the upregulated DEMs were involved in nucleosomes, extracellular regions, and nucleosome assembly, while the downregulated DEMs were mainly involved in Z disk, muscle contraction, and structural constituents of muscle. KEGG pathway analysis was performed on all DEMs, and the pathways were enriched in retinol metabolism, steroid hormone biosynthesis, and tyrosine metabolism. Through survival analysis of the ceRNA network, we identified four DEMs, two DELs, and two DEMis that were significant for poor prognosis.

Conclusions: This study suggested that constructing a ceRNA network and performing survival analysis on the network could screen out new significant treatment and prognosis targets and biomarkers.
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http://dx.doi.org/10.1002/cam4.2099DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6536941PMC
May 2019

Microvesicles from human adipose stem cells promote wound healing by optimizing cellular functions via AKT and ERK signaling pathways.

Stem Cell Res Ther 2019 01 31;10(1):47. Epub 2019 Jan 31.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, No. 1277 Jiefang Avenue, Wuhan, 430022, China.

Background: Human adipose stem cells (ASCs) have emerged as a promising treatment paradigm for skin wounds. Recent works demonstrate that the therapeutic effect of stem cells is partially mediated by extracellular vesicles, which comprise exosomes and microvesicles. In this study, we investigate the regenerative effects of isolated microvesicles from ASCs and evaluate the mechanisms how ASC microvesicles promote wound healing.

Methods: Adipose stem cell-derived microvesicles (ASC-MVs) were isolated by differential ultracentrifugation, stained by PKH26, and characterized by electron microscopy and dynamic light scattering (DLS). We examined ASC-MV effects on proliferation, migration, and angiogenesis of keratinocytes, fibroblasts, and endothelial cells both in vitro and in vivo. Next, we explored the underlying mechanisms by gene expression analysis and the activation levels of AKT and ERK signaling pathways in all three kinds of cells after ASC-MV stimulation. We then assessed the effect of ASC-MVs on collagen deposition, neovascularization, and re-epithelialization in an in vivo skin injury model.

Results: ASC-MVs could be readily internalized by human umbilical vein endothelial cells (HUVECs), HaCAT, and fibroblasts and significantly promoted the proliferation, migration, and angiogenesis of these cells both in vitro and in vivo. The gene expression of proliferative markers (cyclin D1, cyclin D2, cyclin A1, cyclin A2) and growth factors (VEGFA, PDGFA, EGF, FGF2) was significantly upregulated after ASC-MV treatment. Importantly, ASC-MVs stimulated the activation of AKT and ERK signaling pathways in those cells. The local injection of ASC-MVs at wound sites significantly increased the re-epithelialization, collagen deposition, and neovascularization and led to accelerated wound closure.

Conclusions: Our data suggest that ASC-MVs can stimulate HUVEC, HaCAT, and fibroblast functions. ASC-MV therapy significantly accelerates wound healing, and the benefits of ASC-MVs may due to the involvement of AKT and ERK signaling pathways. This illustrates the therapeutic potential of ASC-MVs which may become a novel treatment paradigm for cutaneous wound healing.
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http://dx.doi.org/10.1186/s13287-019-1152-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6357421PMC
January 2019

Genome comparison of African swine fever virus China/2018/AnhuiXCGQ strain and related European p72 Genotype II strains.

Transbound Emerg Dis 2019 May 25;66(3):1167-1176. Epub 2019 Feb 25.

China Animal Health and Epidemiology Center, Qingdao, Shandong, China.

African swine fever was introduced into China in August 2018 and led to high mortality in domestic pigs. We reported the genome characterization of the China/2018/AnhuiXCGQ strain mainly based on next-generation sequencing and comparison with related European p72 Genotype II strains. The genome was 189,393 bp long, encoding 181 open reading frames. Pair-wise genome sequence comparison revealed 54-107 variation sites between China/2018/AnhuiXCGQ and the other genotype II virulent strains contributing to the change of expression or alteration of amino acid residues in 10-38 genes. China/2018/AnhuiXCGQ strain shared the highest similarity with POL/2015/Podlaskie strain. Phylogenetic analysis based on a 125 kb long conserved central region revealed that the China/2018/AnhuiXCGQ strain and four European genotype II strains were grouped into three clusters. This study expanded our knowledge on the genetic diversity and evolution of ASFV and provided valuable information for diagnosis improvement and vaccine development.
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http://dx.doi.org/10.1111/tbed.13124DOI Listing
May 2019

Infection of African swine fever in wild boar, China, 2018.

Transbound Emerg Dis 2019 May 15;66(3):1395-1398. Epub 2019 Jan 15.

China Animal Health and Epidemiology Center, Qingdao, China.

On 16 November 2018, a wild boar infected with African swine fever was reported in China. The phylogenetic analysis showed that its causative strain belonged to the p72 genotype II, CD2v serogroup 8 and contained no additional tandem repeat sequences between the I73R and the I329L protein genes, which was different from previously reported strains in China.
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http://dx.doi.org/10.1111/tbed.13114DOI Listing
May 2019

Development of reverse genetics system for small ruminant morbillivirus: Rescuing recombinant virus to express Echinococcus granulosus EG95 antigen.

Virus Res 2019 02 14;261:50-55. Epub 2018 Dec 14.

OIE Reference Laboratory for Peste des Petits Ruminants, National Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, No.369 Nanjing Road, Qingdao, Shandong 266032, China. Electronic address:

Peste des petits ruminants and cystic hydatidosis may be simultaneously endemic in a given area. Their pathogens are small ruminant morbillivirus (SRMV) and Echinococcus granulosus (E. granulosus), respectively. The SRMV, formerly called peste-des-petits-ruminants virus (PPRV), is classified into the genus Morbillivirus in the family Paramyxoviridae. This virus is an ideal vaccine vector to deliver immunogenic proteins. In this study, a reverse genetics system was developed to rescue a recombinant SRMV (Nigeria 75/1 strain) expressing E. granulosus EG95 antigen in vitro. The recombinant SRMV, albeit replicating more slowly than its parental virus, could effectively express the EG95 antigen in cells by analyses of Western blot, indirect immunofluorescence and mass spectrometry. An EG95 subunit vaccine has been widely used for prevention of cystic hydatidosis in some areas of China. The EG95-expressing SRMV, if proven to induce effective immune responses against both diseases in a future animal experiment, would become a potential candidate of bivalent vaccine.
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http://dx.doi.org/10.1016/j.virusres.2018.12.008DOI Listing
February 2019

Molecular Characterization of African Swine Fever Virus, China, 2018.

Emerg Infect Dis 2018 11 17;24(11):2131-2133. Epub 2018 Nov 17.

On August 3, 2018, an outbreak of African swine fever in pigs was reported in China. We subjected a virus from an African swine fever-positive pig sample to phylogenetic analysis. This analysis showed that the causative strain belonged to the p72 genotype II and CD2v serogroup 8.
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http://dx.doi.org/10.3201/eid2411.181274DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6199985PMC
November 2018

Comprehensive analysis of differentially expressed microRNAs and mRNAs in dorsal root ganglia from streptozotocin-induced diabetic rats.

PLoS One 2018 17;13(8):e0202696. Epub 2018 Aug 17.

Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Diabetic peripheral neuropathy is a common complication associated with diabetes mellitus with a pathogenesis that is incompletely understood. By regulating RNA silencing and post-transcriptional gene expression, microRNAs participate in various biological processes and human diseases. However, the relationship between microRNAs and the progress of diabetic peripheral neuropathy still lacks a thorough exploration. Here we used microarray microRNA and mRNA expression profiling to analyze the microRNAs and mRNAs which are aberrantly expressed in dorsal root ganglia from streptozotocin-induced diabetic rats. We found that 37 microRNAs and 1357 mRNAs were differentially expressed in comparison to non-diabetic samples. Bioinformatics analysis indicated that 399 gene ontology terms and 29 Kyoto Encyclopedia of Genes and Genomes pathways were significantly enriched in diabetic rats. Additionally, a microRNA-gene network evaluation identified rno-miR-330-5p, rno-miR-17-1-3p and rno-miR-346 as important players for network regulation. Finally, quantitative real-time polymerase chain reaction analysis was used to confirm the microarray results. In conclusion, this study provides a systematic perspective of microRNA and mRNA expression in dorsal root ganglia from diabetic rats, and suggests that dysregulated microRNAs and mRNAs may be important promotors of peripheral neuropathy. Our results may be the underlying framework of future studies regarding the effect of the aberrantly expressed genes on the pathophysiology of diabetic peripheral neuropathy.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202696PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6097669PMC
February 2019

Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of nucleos(t)ide analogues.

Liver Int 2016 09 24;36(9):1284-94. Epub 2016 Mar 24.

Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Background & Aims: Multidrug resistance protein 4 (MRP4) has been associated with nucleos(t)ide analogue (NA) antiretroviral therapy failure, though is unclear if MRP4 is also correlated with the failure of anti-hepatitis B virus (HBV) therapy.

Methods: Multidrug resistance protein 4 expression in human peripheral blood mononuclear cells (PBMCs), liver tissues and human hepatoma cell lines was detected by real-time polymerase chain reaction (PCR), western blotting and immunohistochemistry assays. Supernatant and intracellular HBV DNA levels of MRP4-overexpressing or silenced HepG2.4D14 (wild-type) and HepG2.A64 (entecavir-resistant mutant) cells were measured by quantitative PCR. NA concentrations and HBV mutational analysis were assessed by liquid chromatography/mass spectrometry assays and DNA sequencing. Multivariate analysis was used to assess predictive factors for treatment failure.

Results: High expression of MRP4 was found in hepatoma cell lines and PBMCs, and up- or down-regulation of MRP4 expression altered the susceptibility of cells to NAs. MRP inhibitors increased NA intracellular accumulation and decreased extracellular levels. Moreover, MRP4 expression in PBMCs was correlated with that in paired liver tissues. Furthermore, multivariate analysis showed MRP4 mRNA expression to be an independent predictor of NA treatment failure.

Conclusions: Multidrug resistance protein 4 is a critical protein associated with the antiviral efficacy of NAs, and combination therapy of NA and MRP inhibitors could reduce the dosage for long-term NA use. This is the first report to demonstrate that MRP4 expression is an important factor predicting treatment failure in chronic hepatitis B patients and will provide a potential therapeutic target against HBV.
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http://dx.doi.org/10.1111/liv.13104DOI Listing
September 2016

A Modified Approach to Inducing Bone Marrow Stromal Cells to Differentiate into Cells with Mature Schwann Cell Phenotypes.

Stem Cells Dev 2016 Feb 29;25(4):347-59. Epub 2016 Jan 29.

1 Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology , Wuhan, China .

Marrow stromal cells (MSCs) can be induced to differentiate into Schwann-like cells under classical induction conditions. However, cells derived from this method are unstable, exhibiting a low neurotrophin expression level after the induction conditions are removed. In Schwann cell (SC) culture, progesterone (PROG) enhances neurotrophic synthesis and myelination, specifically regulating the expression of the myelin protein zero (P0)- and peripheral myelin protein 22 (PMP22)-encoding genes by acting in concert or in synergy with insulin and glucocorticoids (GLUCs). In the present study, we investigated whether combined PROG, GLUC, and insulin therapy induced MSCs to differentiate into modified SC-like cells with phenotypes similar to those of mature SCs. After being cultured for 2 weeks in modified differentiation medium, the modified SC-like cells showed increased expression of P0 and PMP22. In addition, morphological and phenotypic characterizations were conducted over a period of over 2 weeks, and functional characteristics persisted for more than 3 weeks after the induction reagents were withdrawn. The transplantation of green fluorescent protein-labeled, modified SC-like cells into transected sciatic nerves with a 10-mm gap significantly increased the proliferation of the original SCs and improved axon regeneration and myelination compared with original BM-SCs. Immunostaining for P0 revealed that more of the transplanted modified SC-like cells retained the phenotypic characteristics of SCs. Taken together, these results reveal that the combined application of PROG, GLUC, and insulin induces MSCs to differentiate into cells with phenotypic, molecular, and functional properties of mature SCs.
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http://dx.doi.org/10.1089/scd.2015.0295DOI Listing
February 2016

Allelic variants of PRNP in 16 Chinese local sheep breeds.

Arch Virol 2014 Aug 19;159(8):2141-4. Epub 2014 Mar 19.

China Animal Health and Epidemiology Center, No. 369, Nanjing Road, Qingdao, 266032, Shandong, People's Republic of China.

Here, polymorphisms of the ovine prion protein gene were analyzed in 486 Chinese sheep from 16 main local breeds. Polymorphisms R or H at codons 154 and four polymorphisms at codon 171 encoding Q, R, H, or K were identified. The A/V polymorphism at codon 136 was not observed, and all sheep were homozygous for A at this position. In addition, ten polymorphisms at codons 21, 101, 112, 127, 138, 141, 143, 146, 153 and 189 were detected. The predominant Q allele occurred at codon 171, with a high frequency of 88.68 %, implying a risk of scrapie in China.
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http://dx.doi.org/10.1007/s00705-014-2048-9DOI Listing
August 2014

Common genetic variants in the microRNA biogenesis pathway are associated with malignant peripheral nerve sheath tumor risk in a Chinese population.

Cancer Epidemiol 2013 Dec 12;37(6):913-6. Epub 2013 Jun 12.

Hand Surgery Department, Wuhan Union Hospital, Huazhong University of Science and Technology, Wuhan, China. Electronic address:

Purpose: The role of microRNAs (miRNAs) in tumorigenesis has been well established. Genetic variants in the miRNA biogenesis pathway genes may modify cancer development and survival by affecting the miRNA biogenesis. Our aim is to investigate the association of polymorphisms in the miRNA biogenesis pathway genes and malignant peripheral nerve sheath tumor (MPNST) risk among neurofibromatosis type 1 (NF1) patients.

Methods: A case-control study was performed to analyze 53 SNPs in 11 miRNA biogenesis pathway genes in 356 patients (200 patients with NF1 and 156 patients with both NF1 and MPNST) in China. Association analysis was performed in an additive genetic model by logistics regression.

Results: Four SNPs (DDX5 rs1991401, OR=1.79, 95% CI, 1.34-2.38, P=7.90 × 10(-5); DROSHA rs10719, OR=1.64, 95% CI, 1.23-2.20, P=8.76 × 10(-4); AGO2 rs7005286, OR=0.48, 95% CI, 0.32-0.72, P=3.46 × 10(-4); GEMIN4 rs7813, OR=0.50, 95% CI, 0.34-0.72, P=2.65 × 10(-4)) were significantly associated with MPNST risk. A strong gene-dose effect with increased MPNST risk (P for trend<0.001) was observed.

Conclusions: Genetic variants in the miRNA biogenesis pathway genes may modify MPNST risk both individually and jointly.
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http://dx.doi.org/10.1016/j.canep.2013.05.003DOI Listing
December 2013

Identification of serum microRNAs in genome-wide serum microRNA expression profiles as novel noninvasive biomarkers for malignant peripheral nerve sheath tumor diagnosis.

Med Oncol 2013 Jun 13;30(2):531. Epub 2013 Mar 13.

Hand Surgery Department, WuHan Union Hospital, Huazhong University of Science and Technology, Wuhan 430030, Hubei, China.

Recent studies found that serum microRNAs (miRNAs) could serve as stable and noninvasive biomarkers for disease diagnosis. We used genome-wide serum miRNA expression analysis to investigate the role of serum miRNAs in distinguishing malignant peripheral nerve sheath tumor (MPNST) patients with and without neurofibromatosis type 1 (NF1) from NF1 patients. A total of 100 patients with NF1, 93 sporadic MPNST patients, and 71 NF1 MPNST patients were enrolled in this two-stage, case-control study. Solexa sequencing was used to screen for miRNAs that expressed differentially in three pooled serum samples from 10 NF1 patients, 10 sporadic MPNST patients, and 10 NF1 MPNST patients. The detected serum miRNAs then were validated in 90 patients with NF1, 83 patients with sporadic MPNST, and 61 NF1 MPNST patients by individual quantitative reverse transcriptase polymerase chain reaction assays. Eight serum miRNAs altered more than fivefold by Solexa sequencing between MPNST patients (with and without NF1) and NF1 patients. MiR-801 and miR-214 increased both in sporadic MPNST patients and NF1 MPNST patients when compared with NF1 patients. The sensitivity and the specificity of sporadic MPNST detection by the two-miRNA signature were 0.747 and 0.856, respectively. MiR-24 was only significantly up-regulated in NF1 MPNST patients. The combination of the three miRNAs (MiR-801, miR-214, and miR-24) could distinguish NF1 MPNST patients from NF1 patients with a sensitivity of 0.820 and a specificity of 0.844. The serum-based miRNA expression profiles could serve as novel noninvasive biomarkers in sporadic MPNST and NF1 MPNST detection.
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http://dx.doi.org/10.1007/s12032-013-0531-xDOI Listing
June 2013

Diabetes and hypertension have become leading causes of CKD in Chinese elderly patients: a comparison between 1990-1991 and 2009-2010.

Int Urol Nephrol 2012 Aug 31;44(4):1269-76. Epub 2012 May 31.

Department of Nephrology, China-Japan Friendship Hospital, No. 2 Yinghua Street, Chaoyang District, Beijing 100029, People's Republic of China.

Background: It is unclear whether any significant changes in the epidemiology of chronic kidney disease (CKD), possibly resulting from the exceedingly rapid economic development and the aggressive prevention and treatment of kidney diseases in the Chinese population, have occurred over the past two decades.

Methods: The medical records of 173 CKD patients from 1990-1991 and 956 patients from 2009-2010 were retrospectively analyzed.

Results: In patients from 1990-1991, CKD occurred at an earlier age, with severely decreased kidney function. On average, these patients progressed to end-stage renal disease 4.5 years faster. Diabetes and hypertension have become the most common causes of CKD in elderly patients in 2009-2010 period, representing 39.5 % and 24.2 % of all CKD causes, respectively. In the 1990-1991 period, CKD was characterized by a high frequency of severe anemia in stage 4 and 5 patients. The mean arterial pressure in patients with CKD stage 4 and stage 5 decreased significantly in. Although the number of dyslipidemia patients in stage 4 and 5 was significantly higher in the 2009-2010 period, there were no substantial differences in the average levels of serum triglycerides and total cholesterol in the two cohorts. Hypoproteinemia also became less common than before. The occurrence of cardiovascular events was significantly reduced in stage 4 and stage 5 patients in 2009-2010.

Conclusions: Diabetes and hypertension have increased and become the leading causes of CKD in elderly Chinese patients. Improvement in the control of CKD complications has been significant, mainly regarding anemia, hypertension, dyslipidemia, and hypoproteinemia.
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http://dx.doi.org/10.1007/s11255-012-0194-0DOI Listing
August 2012

[Analysis of monoclonal antibody binding sites in ovine prion protein].

Sheng Wu Gong Cheng Xue Bao 2009 Mar;25(3):348-53

National Diagnostic Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao 266032, China.

Binding sites of five monoclonal antibodies were obtained by reinforceable method of overlapping recombinant prion protein and synthetic peptide. Overlapping peptides of PrP core were expressed in Escherichia coli by insertion of serial PCR amplicons of ovine PrP gene fragments into pET32a. The expressed fusion peptides were then tested for the binding activity to PrP monoclonal antibodies in Western blotting. The binding sites of 5 monoclonal antibodies of ovine PrP were located respectively as follows: 2H3 in 199 aa-213 aa, 4C6, 5F11 and 7F11 in 139 aa-168 aa and 7F1 in 214 aa-227 aa. There oligo peptides were synthesized and used in ELISA test for more accurate localization of the binding sites. The binding sites of 4C6, 5F11 and 7F11 were further confirmed to be in 149 aa-158 aa. This conclusion may contribute to the research for pathogenesis and diagnostic method of scrapie and bovine transmissible spongiform encephalopathy.
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March 2009

Peste des petits ruminants virus in Tibet, China.

Emerg Infect Dis 2009 Feb;15(2):299-301

China Animal Health and Epidemiology Center, Qingdao, People's Republic of China.

Serologic and molecular evidence indicates that peste des petits ruminants virus (PPRV) infection has emerged in goats and sheep in the Ngari region of southwestern Tibet, People's Republic of China. Phylogenetic analysis confirms that the PPRV strain from Tibet is classified as lineage 4 and is closely related to viruses currently circulating in neighboring countries of southern Asia.
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http://dx.doi.org/10.3201/eid1502.080817DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2657621PMC
February 2009

Treatment of thoracolumbar vertebrate fracture by transpedicular morselized bone grafting in vertebrae for spinal fusion and pedicle screw fixation.

J Huazhong Univ Sci Technolog Med Sci 2008 Jun 19;28(3):322-6. Epub 2008 Jun 19.

Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

To enhance the fusion of graft bone in thoracolumbar vertebrae and minimize the postoperative loss of correction, short-segment pedicle screw fixation was reinforced with posterior morselized bone grafting in vertebrae for spinal fusion in patients with thoracolumbar vertebrate fractures. Seventy patients with thoracolumbar vertebrate fractures were treated by short-segment pedicle screw fixation and were randomly divided into two groups. Fractures in group A (n=20) were reinforced with posterior morselized bone grafting in vertebrae for spinal fusion, while patients group B (n=50) did not receive the morselized bone grafting for bone fusion. The two groups were compared in terms of kyphotic deformity, anterior vertebral height, instrument failure and neurological functions after the treatment. Frankel grading system was used for the evaluation of neurological evaluation and Denis scoring scale was employed for pain assessment. The results showed that the kyphosis correction was achieved in both group A and group B (group A: 6.4 degree; group B: 5.4 degree)/ At the end of follow-up, kyphosis correction was maintained in group A but lost in group B (P=0.0001). Postoperatively, greater anterior height was achieved in group A than in group B (P<0.01). During follow-up study, anterior vertebral height was maintained only in Group A (P<0.001). Both group A and group B showed good Denis pain scores (P1 and P2) but group A outdid group B in terms of control of severe and constant pain (P4 and P5). By Frankel criteria, the changes in neurological functions in group A was better than those of group B (P<0.001). It is concluded that reinforcement of short-segment pedicle fixation with morselized bone grafting for the treatment of patients with thoracolumbar vertebrae fracture could achieve and maintain kyphosis correction, and it may also increase and maintain anterior vertebral height. Morselized bone grafting in vertebrae offers immediate spinal stability in patients with thoracolumbar vertebrate fractures, decreases the instrument failure and provides better postoperative pain control than without the morselized bone grafting.
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http://dx.doi.org/10.1007/s11596-008-0321-4DOI Listing
June 2008

Development of one-step real-time RT-PCR assay for detection and quantitation of peste des petits ruminants virus.

J Virol Methods 2008 Mar;148(1-2):232-6

China Animal Health and Epidemiology Center, Qingdao, Shandong 266032, China.

In this study, a rapid and specific TaqMan-based, one-step real-time quantitative reverse transcription PCR (qRT-PCR) has been described for the detection of peste des petits ruminants virus (PPRV). Primers and probe were designed based on the nucleocapsid protein gene sequence. The real-time qRT-PCR assay was able to detect PPRV isolates from very distinct geographical areas (Africa, Middle East and Asia). The specificity of the assay was assessed by including rinderpest virus and other morbillivirus RNAs but none of these tested positive in the assay. The analytical sensitivity of the real-time qRT-PCR assay was achieved through the construction of an in-house PPRV cRNA for the generation of a standard curve. The detection limit of the assay was found to be 8.1 RNA copies per reaction mixture. The assay had excellent intra- and inter-assay reproducibility. In total 30 field samples were screened for the presence of PPRV by conventional RT-PCR in parallel with qRT-PCR. The detection rate increased from 46.7% to 73.3% by use of the real-time qRT-PCR. The real-time qRT-PCR assay described in this report allows the rapid, specific and sensitive laboratory detection of PPRV in tissue samples from field cases.
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http://dx.doi.org/10.1016/j.jviromet.2007.12.003DOI Listing
March 2008
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