Publications by authors named "Yunyan He"

25 Publications

  • Page 1 of 1

Excellent Early Outcomes of Combined Chemotherapy With Arsenic Trioxide for Stage 4/M Neuroblastoma in Children: A Multicenter Nonrandomized Controlled Trial.

Oncol Res 2021 Sep 15;28(7):791-800. Epub 2021 Apr 15.

Pediatric Hematology/Oncology, Sun Yet-Sen Memorial Hospital, Sun Yet-Sen UniversityGuangzhouP.R. China.

This nonrandomized, multicenter cohort, open-label clinical trial evaluated the efficacy and safety of combined chemotherapy with arsenic trioxide (ATO) in children with stage 4/M neuroblastoma (NB). We enrolled patients who were newly diagnosed with NB and assessed as stage 4/M and received either traditional chemotherapy or ATO combined with chemotherapy according to their own wishes. Twenty-two patients were enrolled in the trial group (ATO combined with chemotherapy), and 13 patients were enrolled in the control group (traditional chemotherapy). Objective response rate (ORR) at 4 weeks after completing induction chemotherapy was defined as the main outcome, and adverse events were monitored and graded in the meantime. Data cutoff date was December 31, 2019. Finally, we found that patients who received ATO combined with chemotherapy had a significantly higher response rate than those who were treated with traditional chemotherapy (ORR: 86.36% vs. 46.16%, =0.020). Reversible cardiotoxicity was just observed in three patients who were treated with ATO, and no other differential adverse events were observed between the two groups. ATO combined with chemotherapy can significantly improve end-induction response in high-risk NB, and our novel regimen is well tolerated in pediatric patients. These results highlight the superiority of chemotherapy with ATO, which creates new opportunity for prolonging survival. In addition, this treatment protocol minimizes therapeutic costs compared with anti-GD2 therapy, MIBG, and proton therapy and can decrease the burden to families and society. However, we also need to evaluate more cases to consolidate our conclusion.
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http://dx.doi.org/10.3727/096504021X16184815905096DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8420893PMC
September 2021

Statistical mechanics of chromosomes: in vivo and in silico approaches reveal high-level organization and structure arise exclusively through mechanical feedback between loop extruders and chromatin substrate properties.

Nucleic Acids Res 2020 11;48(20):11284-11303

Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

The revolution in understanding higher order chromosome dynamics and organization derives from treating the chromosome as a chain polymer and adapting appropriate polymer-based physical principles. Using basic principles, such as entropic fluctuations and timescales of relaxation of Rouse polymer chains, one can recapitulate the dominant features of chromatin motion observed in vivo. An emerging challenge is to relate the mechanical properties of chromatin to more nuanced organizational principles such as ubiquitous DNA loops. Toward this goal, we introduce a real-time numerical simulation model of a long chain polymer in the presence of histones and condensin, encoding physical principles of chromosome dynamics with coupled histone and condensin sources of transient loop generation. An exact experimental correlate of the model was obtained through analysis of a model-matching fluorescently labeled circular chromosome in live yeast cells. We show that experimentally observed chromosome compaction and variance in compaction are reproduced only with tandem interactions between histone and condensin, not from either individually. The hierarchical loop structures that emerge upon incorporation of histone and condensin activities significantly impact the dynamic and structural properties of chromatin. Moreover, simulations reveal that tandem condensin-histone activity is responsible for higher order chromosomal structures, including recently observed Z-loops.
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http://dx.doi.org/10.1093/nar/gkaa871DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7672462PMC
November 2020

Polymer perspective of genome mobilization.

Mutat Res 2020 May - Dec;821:111706. Epub 2020 May 26.

Department of Biology, 623 Fordham Hall CB#3280, University of North Carolina, Chapel Hill, NC 27599-3280, United States. Electronic address:

Chromosome motion is an intrinsic feature of all DNA-based metabolic processes and is a particularly well-documented response to both DNA damage and repair. By using both biological and polymer physics approaches, many of the contributing factors of chromatin motility have been elucidated. These include the intrinsic properties of chromatin, such as stiffness, as well as the loop modulators condensin and cohesin. Various biological factors such as external tethering to nuclear domains, ATP-dependent processes, and nucleofilaments further impact chromatin motion. DNA damaging agents that induce double-stranded breaks also cause increased chromatin motion that is modulated by recruitment of repair and checkpoint proteins. Approaches that integrate biological experimentation in conjunction with models from polymer physics provide mechanistic insights into the role of chromatin dynamics in biological function. In this review we discuss the polymer models and the effects of both DNA damage and repair on chromatin motion as well as mechanisms that may underlie these effects.
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http://dx.doi.org/10.1016/j.mrfmmm.2020.111706DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7721199PMC
December 2020

Biomechanical behavior of endocrown restorations with different CAD-CAM materials: A 3D finite element and in vitro analysis.

J Prosthet Dent 2021 Jun 26;125(6):890-899. Epub 2020 May 26.

Professor, Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, PR China. Electronic address:

Statement Of Problem: The performance of endocrowns fabricated with different types of computer-aided design and computer-aided manufacturing (CAD-CAM) materials is unclear.

Purpose: The purpose of this finite element analysis (FEA) and in vitro study was to compare and evaluate the stress distribution, failure probability, and fracture resistance of endodontically treated teeth restored with endocrowns from CAD-CAM milling blocks including ceramic, polymer-infiltrated ceramic (PICN), and composite resin.

Material And Methods: An endodontically treated first mandibular molar restored with an endocrown was modeled by using a CAD software program and imported into an FEA software program. The model was duplicated and received restorations made from CAD-CAM blocks: Vita Suprinity (VS), IPS e.max CAD (EMX), Vita Enamic (VE), Lava Ultimate (LU), and Grandio blocs (GR). Stress distributions under axial and oblique loading were analyzed. The Weibull function was combined with the FEA results to predict long-term failure probability. The mechanical failure behavior of endocrowns manufactured with these materials was tested by using a universal testing machine. Load-to-failure was recorded, and fractured specimens were subjected to fractography. The data were analyzed by 1-way ANOVA and the post hoc Tukey test (α=.05).

Results: The models of GR and LU exhibited a more even stress distribution. The Weibull analysis revealed that 5 models performed in a similar manner under normal occlusal forces, while LU and VE models achieved the highest probabilities during clenching. The fracture loads of GR (3808 ±607 N) were significantly higher than those of other materials (P<.05). More favorable failure modes were observed in the GR and VE groups. Fractography showed a greater probability of compression curls and arrest lines in the endocrowns of VE, LU, and GR groups.

Conclusions: When restoring endodontically treated teeth, endocrown fabricated with composite resin exhibited a more uniform stress distribution and higher fracture resistance. More evidence from long-term clinical studies is needed to verify this effect.
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http://dx.doi.org/10.1016/j.prosdent.2020.03.009DOI Listing
June 2021

Anastrozole plus fulvestrant vs. anastrozole alone for hormone receptor-positive advanced breast cancer: a meta-analysis of randomized controlled trials.

Breast Cancer Res Treat 2020 Apr 1;180(2):269-278. Epub 2020 Feb 1.

Department of Thoracic Surgery, The Second Affiliated Hospital of Nanchang University, 1 Minde Road, Nanchang, 330006, China.

Background: For patients with hormone receptor (HR)-positive advanced breast cancer, whether the combination of anastrozole and fulvestrant is more effective than anastrozole alone is controversial. Our meta-analysis aimed to compare the efficacy and safety of the two therapies.

Methods: We retrieved relevant studies in Embase, the Cochrane Library, Ovid MEDLINE, PubMed, ScienceDirect, Web of Science, Scopus, and Google Scholar. The primary outcomes were overall survival (OS) and progression-free survival (PFS). The secondary outcomes were the disease control rate (DCR), the objective response rate (ORR), and adverse events (AEs).

Results: Five articles based on 4 randomized controlled trials containing 2146 patients were identified in our meta-analysis. The combination group had better efficacy in the endpoints of OS (hazard ratio [HR] 0.86; 95% confidence interval [CI] 0.74-0.99, p = 0.03) and PFS (HR 0.87; 95% CI 0.77-0.97, p = 0.02). Regarding the ORR, DCR, total AEs and grade 3-5 AEs, we found no difference between the two treatments. The combination group showed a clearly higher rate of treatment discontinuations (95% CI 1.05-3.60, p = 0.03) and AEs leading to death (95% CI 1.12-9.11, p = 0.03). The subgroup analysis of AEs showed an increased incidence of extremity or muscle pain, hematologic effects, gastrointestinal disorders, and hot flashes in the combination group.

Conclusions: For HR-positive advanced breast cancer patients, the combination of anastrozole and fulvestrant appears to be superior to anastrozole alone in extending PFS and OS, despite relatively serious AEs.
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http://dx.doi.org/10.1007/s10549-020-05551-3DOI Listing
April 2020

Construction and Analysis of a Long Non-Coding RNA (lncRNA)-Associated ceRNA Network in β-Thalassemia and Hereditary Persistence of Fetal Hemoglobin.

Med Sci Monit 2019 Sep 21;25:7079-7086. Epub 2019 Sep 21.

Department of Pediatrics, First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, Nanning, Guangxi, China (mainland).

BACKGROUND Higher fetal hemoglobin (HbF) levels can ameliorate the clinical severity of ß-thalassemia. The use of integrative strategies to combine results from gene microarray expression profiling, experimental evidence, and bioinformatics helps reveal functional long noncoding RNAs (lncRNAs) in ß-thalassemia and HbF induction. MATERIAL AND METHODS In a previous study, a microarray profiling was performed of 7 individuals with high HbF levels and 7 normal individuals. Thirteen paired samples were used for validation. lncRNA NR_001589 and uc002fcj.1 were chosen for further research. The quantitative reverse transcription-PCR was used to detect the expression levels of 2 lncRNAs. The Spearman correlation test was employed. The nuclear and cytoplasmic distribution experiment in K562 cells was used to verify the subcellular localization of 2 lncRNAs. Potential relationships among lncRNAs, predicted microRNAs (miRNAs), and target gene HBG1/2 were based on competitive endogenous RNA theory and bioinformatics analysis. RESULTS Average expression levels of NR_001589 and uc002fcj.1 were significantly higher in the high-HbF group than in the control group. A positive correlation existed between NR_001589, uc002fcj.1, and HbF. The expression of NR_001589 was in both the cytoplasm and the nucleus, mostly (77%) in the cytoplasm. The expression of uc002fcj.1 was in both the cytoplasm and the nucleus; the cytoplasmic proportion was 43% of the total amount. A triple lncRNA-miRNA-mRNA network was established. CONCLUSIONS Novel candidate genetic factors associated with the HBG1/2 expression were identified. Further functional investigation of NR_001589 and uc002fcj.1 can help deepen the understanding of molecular mechanisms in ß-thalassemia.
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http://dx.doi.org/10.12659/MSM.915946DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6767942PMC
September 2019

Using microarray analysis to identify genes and pathways that regulate fetal hemoglobin levels.

Ann Hum Genet 2020 01 8;84(1):29-36. Epub 2019 Aug 8.

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, Nanning, Guangxi Province, China.

Increased levels of fetal hemoglobin (HbF: α2γ2) can ameliorate the clinical severity of the β-hemoglobinopathies. Microarray analysis represents a powerful approach to identify novel genetic factors regulating the γ-globin gene. Gene expression profiling was previously performed on 14 individuals with high or normal HbF levels to identify the genetic factors that control γ-globin gene expression. To obtain more accurate and reliable results, our results were combined with public microarray dataset GSE22109 deposited in the Gene Expression Omnibus database. Annotation of case versus control samples was taken directly from the microarray documentation. The differentially expressed genes (DEGs) were obtained and were deeply analyzed by bioinformatics methods. Combined with our own chip expression data, potential genes HBE1, TFRC, and CSF2 were selected out for subsequent qRT-PCR validation. A total of 184 DEGs were identified from GSE22109 and the protein-protein interaction network was constructed. Gene set enrichment analysis showed that the hematopoietic cell lineage pathway overlaps in the two datasets. HBE1, CSF2, and TFRC were confirmed by qRT-PCR. Our results suggest novel candidate genes and pathways associated with the γ-globin gene expression.
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http://dx.doi.org/10.1111/ahg.12346DOI Listing
January 2020

Three-Dimensional Thermodynamic Simulation of Condensin as a DNA-Based Translocase.

Methods Mol Biol 2019 ;2004:291-318

Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.

Chromatin dynamics and organization can be altered by condensin complexes. In turn, the molecular behavior of a condensin complex changes based on the tension of the substrate to which condensin is bound. This interplay between chromatin organization and condensin behavior demonstrates the need for tools that allows condensin complexes to be observed on a variety of chromatin organizations. We provide a method for simulating condensin complexes on a dynamic polymer substrate using the polymer dynamics simulator ChromoShake and the condensin simulator RotoStep. These simulations can be converted into simulated fluorescent images that are able to be directly compared to experimental images of condensin and fluorescently labeled chromatin. Our pipeline enables users to explore how changes in condensin behavior alters chromatin dynamics and vice versa while providing simulated image datasets that can be directly compared to experimental observations.
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http://dx.doi.org/10.1007/978-1-4939-9520-2_21DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6904244PMC
March 2020

Identification of novel pathogenic F13A1 mutation and novel NBEAL2 gene missense mutation in a pedigree with hereditary congenital factor XIII deficiency.

Gene 2019 Jun 29;702:143-147. Epub 2019 Mar 29.

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, No.6, Shuangyong Road, Qingxiu District, Nanning, Guangxi Province, 530021, PR China. Electronic address:

The genetic defects of a 12-year-old patient with factor XIII deficiency (FXIIID) and eight pedigree members suspected with FXIIID were studied. Clinical diagnosis, pedigree investigation, phenotypic study and genetic analysis were performed. DNA sequence analysis revealed that the proband had a novel deletion mutation of F13A1 gene (NM_000129: exon 12: c.1652delC: p.Thr551LysfsTer26) which he inherited from both the parents who were heterozygous for the same 1652delC deletion. This frameshift (p.Thr551LysfsTer26) led in homozygous form to severe FXIIID. Additionally, a homozygous missense mutation of NBEAL2 gene (NM_015175: exon 13: c.1367C > T: p.Ala456Val) was identified in the proband. Again, the mutation was inherited from both the parents who were heterozygous for the same c.1367C > T novel mutation. Other members of the pedigree were also revealed to be heterozygous for the same proband's F13A1 and NBEAL2 genes mutations. We first report a pedigree with pathogenic F13A1 gene mutation and a novel mutant NBEAL2 gene.
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http://dx.doi.org/10.1016/j.gene.2019.03.067DOI Listing
June 2019

A novel PKLR gene mutation identified using advanced molecular techniques.

Pediatr Transplant 2018 03 18;22(2). Epub 2018 Jan 18.

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, Nanning, Guangxi Province, China.

This study's purposes were to diagnose intractable hemolytic anemia and to provide guiding treatment for the affected family members. We performed NGS in a panel of 600 genes for blood diseases on a patient with obscure hemolytic anemia and her parents. We confirmed the diagnosis of pyruvate kinase deficiency, identified a novel homozygous mutation of the PKLR gene (NM_000298: exon 6: c.T941C: p.I314T), and ruled out other blood diseases in the Chinese family. Furthermore, amniotic fluid was taken from the mother during the second trimester, and DNA was extracted to analyze the type of PKLR gene mutation. The proband received cord blood and bone marrow from the second child of the mother for hematopoietic stem cell transplantation and achieved normal hematopoiesis. The genetic characterization analysis and genotype-phenotype correlation study of PKLR gene suggested that NGS was an effective method to confirm the molecular diagnosis of intractable hemolytic anemia. The identification of the mutation aided in prenatal diagnosis in the second pregnancy and the effective clinical management of the affected family.
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http://dx.doi.org/10.1111/petr.13143DOI Listing
March 2018

ARHGAP18 is a novel gene under positive natural selection that influences HbF levels in β-thalassaemia.

Mol Genet Genomics 2018 Feb 5;293(1):207-216. Epub 2017 Oct 5.

Department of Pediatrics Surgery, The First Affiliated Hospital of Guangxi Medical University, No. 6, Shuangyong Road, Qingxiu District, Nanning, 530021, Guangxi, People's Republic of China.

Foetal haemoglobin (HbF) plays a dominant role in ameliorating the morbidity and mortality of β-thalassaemia. A better understanding of the loci and genes involved in HbF expression would be beneficial for the treatment of β-thalassaemia major. However, the genes associated with HbF expression remain largely unknown. In this study, we first explored large-scale data sets and examined the human genome for evidence of positive natural selection to screen out single nucleotide polymorphisms (SNPs). A genetic analysis of HbF levels was conducted in a Chinese cohort of patients with β-thalassaemia to confirm the bioinformatics results. A total of 1141 subjects with β-thalassaemia were recruited. The results showed that the SNP rs11759328 in the ARHGAP18 gene was significantly associated with HbF levels (Ρ = 5.1 × 10). ARHGAP18 belongs to the RhoGAP family and controls angiogenesis, cellular morphology and motility. Second, after determining that ARHGAP18 was highly expressed in the human K562 cell line, we used lentiviral-mediated small interfering RNA to knock down ARHGAP18 expression and subsequently assessed cell proliferation and apoptosis using cell proliferation assays and flow cytometry, respectively. ARHGAP18 downregulation in K562 cells significantly increased HBG1/2 expression and apoptosis, but proliferation was not significantly affected in vitro. Our data suggest that ARHGAP18, which was located by the SNP rs11759328 via positive selection, plays a potential role in regulating HbF expression in β-thalassaemia and may be a promising therapeutic target. Knockout studies of ARHGAP18 warrant further investigation into its aetiology in HbF.
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http://dx.doi.org/10.1007/s00438-017-1377-2DOI Listing
February 2018

Tissue-specific extracellular matrix promotes myogenic differentiation of human muscle progenitor cells on gelatin and heparin conjugated alginate hydrogels.

Acta Biomater 2017 10 17;62:222-233. Epub 2017 Aug 17.

Wake Forest Institute for Regenerative Medicine, 391 Technology Way, Winston-Salem, NC, USA; Hypertension and Vascular Research Center, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, USA; Center on Diabetes, Obesity, and Metabolism, Wake Forest School of Medicine, Medical Center Boulevard, Winston-Salem, NC, USA. Electronic address:

Myogenic differentiation, cell fusion, and myotube formation of skeletal muscle progenitor cells (SMPCs) have key roles during skeletal muscle development and repair. However, after isolation from living tissue and transition to culture dishes, SMPCs gradually lose their function and stop propagating due to the absence of extracellular matrix (ECM). Despite encouraging results of experiments using ECM components in cell culture for maintenance and propagation of some tissue types, the benefits of this approach on SMPC culture are limited, because the bioactive molecules and proteins instantly release and are degraded during culture. In this study, we developed a novel approach to enhance the proliferation and differentiation of human skeletal muscle progenitor cells (hSMPCs) in vitro with skeletal muscle ECM in combination with a modified alginate hydrogel conjugated with gelatin and heparin (Alg-G-H) as a substrate. This Alg-G-H substrate, together with skeletal muscle ECM, significantly enhanced cell expansion, differentiation, and maturation of hSMPCs compared with individual substrata (i.e. gelatin, Matrigel®, or ECM alone). In Western-blot and immunocytochemical analyses, the Alg-G-H-ECM predominantly enhanced expression of skeletal myogenesis markers (MyoD, Myf5, Myogenin, Desmin and Myosin) and myotube formation in hSMPCs. This study demonstrated that combining Alg-G-H substrates with skeletal muscle ECM modulated homeostasis of cell proliferation, differentiation, and maturation of hSMPCs by releasing signaling molecules and growth factors. This technique could be a cost-effective tool for in vitro skeletal muscle cell differentiation and maturation, with potential applications in tissue regeneration and drug development.

Statement Of Significance: Alginate based biomaterials are commonly used in tissue engineering and regenerative medicine field, however, the inefficient sequestration of growth factors restricted its utilization. In this study, a novel alginate based substrates was produced covalently modified with gelatin and heparin, in order to capture more effective cytokines and proteins in the culture milieu, keep homeostasis for cell survival and tissue regeneration with growth factor sequestration and long-term release capacities. Combining with skeletal muscle derived ECM, the modified Alginate-Gelatin-Heparin gel could most effectively mimic the tissue specific microenvironment to support skeletal muscle progenitor cells proliferation, differentiation and myotube formation. Additionally, the economical and practical features will make it more promising in high-throughput application for regenerative medicine research.
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http://dx.doi.org/10.1016/j.actbio.2017.08.022DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8151673PMC
October 2017

A Novel Variant with Positive Natural Selection Influenced Hb A Levels in Chinese Individuals with β-Thalassemia.

Hemoglobin 2017 May 5;41(3):193-197. Epub 2017 Sep 5.

a Department of Pediatrics , The First Affiliated Hospital of Guangxi Medical University , Nanning , Guangxi Province , People's Republic of China.

β-Thalassemia (β-thal) is the most common inherited hemolytic anemia worldwide. Elevated Hb A is a mark of β-thal carriers. The aim of this study was to identify the pathogenic variants associated with the Hb A levels. One thousand and thirty β-thal carriers were recruited for this study. Using positive natural expression quantitative trait loci (eQTL) analysis, a significant variant was selected. Genotyping for the rs231841 polymorphism was performed by the Sequenom MassARRAY IPLEX platform. All genetic association analyses were performed with the PLINK program. The linear regression analysis showed that rs231841 in the intron region of the potassium voltage-gated channel subfamily Q member 1 (KCNQ1) gene on chromosome 11p15 was significantly associated with Hb A levels. The presence of the C allele was associated with elevated Hb A levels. Our results suggest that rs231841 on the KCNQ1 gene with positive natural selection is related to Hb A levels in Chinese β-thal carriers, and KCNQ1 is probably associated with the expression of the β-like globin gene cluster.
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http://dx.doi.org/10.1080/03630269.2017.1358177DOI Listing
May 2017

Novel mutations in patients with hereditary red blood cell membrane disorders using next-generation sequencing.

Gene 2017 Sep 8;627:556-562. Epub 2017 Jul 8.

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, N0.6, Shuangyong Road, Qingxiu District, Nanning, Guangxi Province 530021, PR China. Electronic address:

To diagnose and investigate the genotype-phenotype relationship in intractable hereditary red blood cell (RBC) membrane cases, we have utilized next-generation sequencing (NGS) to develop a high-throughput, highly sensitive assay. Three unrelated families including 15 individuals were analysed with a panel interrogating 600 genes related to haematopathy disorders. Where possible, inheritance patterns of pathogenic mutations were determined by sequencing the relatives. We identified 2 novel mutations in ANK1 (Y216X and E142X) responsible for hereditary spherocytosis (HS) that were stop-gain single nucleotide variants (SNVs). Furthermore, a novel SPTA1 mutation (H54P) was identified; it is a nonsynonymous SNV and is associated with hereditary elliptocytosis (HE). In addition, patients who also carried erythropoiesis gene mutations showed more severe disease phenotype. The NGS panel provides a fast and accurate method for molecular diagnosis in patients with intractable hereditary RBC membrane disorders. An approach integrating medical history, clinical and molecular testing, and pedigree analysis is beneficial for these patients and families.
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http://dx.doi.org/10.1016/j.gene.2017.07.009DOI Listing
September 2017

Genome-wide analysis of aberrantly expressed lncRNAs and miRNAs with associated co-expression and ceRNA networks in β-thalassemia and hereditary persistence of fetal hemoglobin.

Oncotarget 2017 Jul;8(30):49931-49943

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Guangxi Zhuang Autonomous Region, Nanning 530021, China.

The implications of lncRNAs regarding fetal hemoglobin (HbF) induction in hemoglobin disorders remain poorly understood. In this study, microarray analysis was performed to profile lncRNAs, miRNAs and mRNAs in individuals with hereditary persistence of fetal hemoglobin (HPFH), β-thalassemia carriers with high HbF levels and healthy controls. The results show aberrant expression of 862 lncRNAs, 568 mRNAs and 63 miRNAs in the high-HbF group compared with the control group. Altered NR_001589, NR_120526, T315543, miR-486-3p, miR-19b-1-5p and miR-20a-3p expression was confirmed by quantitative reverse transcription-polymerase chain reaction, and Spearman correlation coefficients revealed significant positive correlations with HbF. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses showed the hematopoietic cell lineage and apoptosis to be most significantly dysregulated in HbF induction. We analyzed coding genes near the lncRNAs and constructed a coding-noncoding co-expression network. Based on the results, lncRNAs likely contribute to increased HbF levels by activating expression of HBE1 and hematopoietic cell lineage-inducible molecules and by inhibiting that of apoptosis-inducible molecules. Finally, through construction of a competing endogenous RNA network, we found that 6 lncRNAs could bind competitively with miR-486-3p, resulting in increased HbF levels. Taken together, our findings provide new insights into the mechanisms of HbF induction and potentially provide new targets for the treatment of β-thalassemia major.
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http://dx.doi.org/10.18632/oncotarget.18263DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5564818PMC
July 2017

The prevalence of thalassemia in mainland China: evidence from epidemiological surveys.

Sci Rep 2017 04 19;7(1):920. Epub 2017 Apr 19.

Department of Pediatrics, the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China.

Comprehensive data regarding the epidemiology and prevalence of thalassemia in mainland China are lacking. To assess the prevalence of thalassemia, we performed a meta-analysis including 16 articles published from 1981 to 2015. The overall prevalence of α-thalassemia, β-thalassemia and α + β-thalassemia was 7.88%, 2.21% and 0.48%, respectively. Trends in thalassemia prevalence in mainland China were not steady; a prevalence map based on a geographic information system (GIS) showed that the geographic distribution of thalassemia was highest in the south of China and decreased from south to north. Additionally, the most common α- and β-globin gene mutation was -- and CD41/42, respectively. The current study provides valuable information regarding epidemiology and intervention and supports the planning, implementation and management of prevention programmes for public health.
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http://dx.doi.org/10.1038/s41598-017-00967-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5430438PMC
April 2017

Development and evaluation of enzyme-linked immunosorbent assay of nucleic acid sequence-based amplification for diagnosis of invasive aspergillosis.

AMB Express 2016 Dec 6;6(1):91. Epub 2016 Oct 6.

Department of Laboratory Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, People's Republic of China.

Invasive aspergillosis (IA) is a life-threatening infection in immunocompromised patients, rapid and sensitive detection of Aspergillus from clinical samples has been a major challenge in the early diagnosis of IA. An enzyme-linked immunosorbent assay of nucleic acid sequence-based amplification (NASBA-ELISA) was developed to fulfil the need for the efficient diagnosis of these infections. The primers targeting 18S rRNA were selected for the amplification of Aspergillus RNA by the isothermal digoxigenin (DIG)-labeling NASBA process. The DIG-labeled RNA amplicons were hybridized with a specific biotinylated DNA probe immobilized on streptavidin-coated microtiter plate. The hybrids were colorimetrically detected by the addition of an anti-DIG antibodies linked to ALP and substrate (disodium 4-nitrophenyl phosphate). The detection limit of the Aspergillus NABSA-ELISA system was 1 CFU and the RNA in non-target bacteria or fungus was not amplified. The performance of this NASBA-ELISA compared to RT-PCR and galactomannan (GM) was evaluated by testing blood samples from 86 patients at high risk for IA. The sensitivity of NASBA-ELISA, RT-PCR and GM-ELISA was 80.56 % (95 % CI 63.98-91.81), 72.22 % (95 % CI 54.81-85.80), 58.33 % (95 % CI 40.76-74.49), respectively, and the specificity was 80.00 % (95 % CI 66.28-89.97), 84.00 % (95 % CI 70.89-92.83), 82.00 % (95 % CI 68.56-91.42). The efficiency of the three methods in various combinations was also evaluated. Combination of NASBA-ELISA and GM-ELISA testing achieved perfect specificity (100 %; 95 % CI 92.89-100) and perfect positive predictive value (100 %; 95 % CI 83.16-100). The best sensitivity (97.22 %; 95 % CI 85.47-99.93) and the highest Youden index (0.652) were obtained by testing with both NASBA and RT-PCR in parallel. In conclusion, the NASBA-ELISA assay consists of an alternative process for large-scale samples detection with semi-quantitative results and provides good clinical performance without resorting to expensive equipment. This assay makes it possible for the NASBA based RNA diagnosis to become a routine work in laboratories in less developed countries with fewer resources.
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http://dx.doi.org/10.1186/s13568-016-0266-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5053951PMC
December 2016

Inhibiting the growth of methicillin-resistant Staphylococcus aureus in vitro with antisense peptide nucleic acid conjugates targeting the ftsZ gene.

Int J Infect Dis 2015 Jan 5;30:1-6. Epub 2014 Nov 5.

Department of Clinical Laboratory, the First Affiliated Hospital of Chongqing Medical University, 1 You Yi Road, Yuzhong District, Chongqing 40016, China.

Background: The increasing emergence of clinical infections caused by methicillin-resistant Staphylococcus aureus (MRSA) challenges existing therapeutic options and highlights the need to develop novel treatment strategies. The ftsZ gene is essential to bacterial cell division.

Methods: In this study, two antisense peptide nucleic acids (PNAs) conjugated to a cell-penetrating peptide were used to inhibit the growth of MRSA. PPNA1, identified with computational prediction and dot-blot hybridization, is complementary to nucleotides 309-323 of the ftsZ mRNA. PPNA2 was designed to target the region that includes the translation initiation site and the ribosomal-binding site (Shine-Dalgarno sequence) of the ftsZ gene. Scrambled PPNA was constructed with mismatches to three bases within the antisense PPNA1 sequence.

Results: PPNA1 and PPNA2 caused concentration-dependent growth inhibition and had bactericidal effects. The minimal bactericidal concentrations of antisense PPNA1 and PPNA2 were 30μmol/l and 40μmol/l, respectively. The scrambled PPNA had no effect on bacterial growth, even at higher concentrations, confirming the sequence specificity of the probes. RT-PCR showed that the antisense PPNAs suppressed ftsZ mRNA expression in a dose-dependent manner.

Conclusion: Our results demonstrate that the potent effects of PNAs on bacterial growth and cell viability were mediated by the down-regulation or even knock-out of ftsZ gene expression. This highlights the utility of ftsZ as a promising target for the development of new antisense antibacterial agents to treat MRSA infections.
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http://dx.doi.org/10.1016/j.ijid.2014.09.015DOI Listing
January 2015

Retrospective comparison of nucleic acid sequence-based amplification, real-time PCR, and galactomannan test for diagnosis of invasive aspergillosis.

J Mol Diagn 2014 Sep 13;16(5):584-590. Epub 2014 Aug 13.

Department of Laboratory Medicine, First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

Invasive aspergillosis is a life-threatening infection in immunocompromised patients, and treating these infections at an early stage is often crucial for a favorable outcome. Early diagnosis, however, remains challenging. We performed a retrospective comparison of three methods: real-time quantitative PCR (qPCR), nucleic acid sequence-based amplification (NASBA), and galactomannan enzyme-linked immunosorbent assay (GM-ELISA); these detect circulating Aspergillus DNA, RNA, and galactomannan, respectively. Blood samples from 80 patients at high risk for invasive aspergillosis were tested by each assay. The sensitivity of NASBA, qPCR, and GM-ELISA was 76.47% (95% CI, 58.4-88.6%), 67.65% (95% CI, 49.4-82.0%), and 52.94% (95% CI, 35.4-69.8%), respectively, and the specificity was 80.43% (95% CI, 65.6-90.1%), 89.13% (95% CI, 75.6-95.9%), and 80.43% (95% CI, 65.6-90.1%), respectively. We also evaluated the efficiency of the three tests in various combinations. Perfect specificity (100%; 95% CI, 90.4-100%) and perfect positive predictive value (100%; 95% CI, 77.1-100%) were achieved by combining NASBA and qPCR testing in series. Testing with both NASBA and qPCR in parallel was the most sensitive and had the highest Youden index. Our data support the great potential of NASBA and qPCR, singly or in combination, for diagnosis of invasive aspergillosis in high-risk populations.
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http://dx.doi.org/10.1016/j.jmoldx.2014.05.001DOI Listing
September 2014

Inhibition of gene expression and growth of multidrug-resistant Acinetobacter baumannii by antisense peptide nucleic acids.

Mol Biol Rep 2014 Nov 5;41(11):7535-41. Epub 2014 Aug 5.

Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, No. 1 Youyi Road, Yuzhong District, Chongqing, 400016, People's Republic of China.

Acinetobacter baumannii causes common and severe community- and hospital-acquired infections. The increasing emergence of multidrug-resistant (MDR) and pan-drug resistant A. baumannii has limited the therapeutic options, highlighting the need for new therapeutic strategies. The goal of this study was to investigate whether antisense peptide nucleic acids (PNAs) could mediate gene-specific inhibition effects in MDR A. baumannii. We described a screening strategy based on computational prediction and dot hybridization for identifying potential inhibitory PNAs, and evaluated the in vitro growth inhibition potency of two PNAs conjugated to the (KFF)3K peptide (pPNA1 and pPNA2), both of which targeted the growth essential gene gyrA of A. baumannii. Both pPNAs showed strong inhibition effects on bacterial growth and gyrA mRNA expression in a dose-dependent manner. The lowest inhibitory and bactericidal concentration were 5 and 10 μM, respectively. Combination of the two pPNAs showed superimposed effect other than synergistic effect. Control PNAs without (KFF)3K peptide conjugation or with mismatched antisense sequence had no inhibition effects on bacterial growth or mRNA expression. Our study suggests that anti-gyrA pPNAs can efficiently inhibit gene expression and bacterial growth, and has the potential as a new therapeutic option for MDR A. baumannii.
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http://dx.doi.org/10.1007/s11033-014-3643-2DOI Listing
November 2014

Investigation of mechanism and molecular epidemiology of linezolid-resistant Enterococcus faecalis in China.

Infect Genet Evol 2014 Aug 9;26:14-9. Epub 2014 May 9.

Department of Clinical Laboratory, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.

Enterococcus is a major cause of important nosocomial infections. Linezolid, the first member of an entirely new class of antibiotics (oxazolidinones), is effective against serious infections caused by Enterococcus. However, resistance to linezolid has been discovered throughout the world rapidly. From 2011 to 2013, nine linezolid-resistant E. faecalis isolates were collected and the possible mechanisms of linezolid resistance, including mutations in domain V of 23S rRNA genes and in ribosomal proteins L3 and L4, and the multiresistance gene cfr, were investigated. Furthermore, an epidemiological survey of the nine linezolid-resistant E. faecalis isolates was performed by pulsed field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and DiversiLab. The three methods were compared to evaluate their merits and demerits, respectively. We failed to find the resistance mechanisms that have been revealed in recent years by PCR and sequencing analysis in the linezolid-resistant E. faecalis. Epidemiological investigation suggested that a small-scale outbreak of linezolid-resistant E. faecalis emerged in neurosurgery ICU from March to May of 2013. DiversiLab was a reliable typing tool and a suitable alternative to PFGE because it was as discriminatory as PFGE and better than MLST.
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http://dx.doi.org/10.1016/j.meegid.2014.05.001DOI Listing
August 2014

MEK1 promotes YAP and their interaction is critical for tumorigenesis in liver cancer.

FEBS Lett 2013 Dec 7;587(24):3921-7. Epub 2013 Nov 7.

Department of Clinical Laboratory Medicine, Shanghai Tenth People's Hospital of Tongji University, Shanghai 200072, China; Department of Laboratory Medicine, Chongqing Zhongshan Hospital, Chongqing 400013, China.

Mitogen-activated protein kinase kinase 1 (MAP2K1/MEK1) as well as Yes-associated protein (YAP), the downstream effector of Hippo signaling pathway, is linked to hepatocarcinogenesis. However, little is known about whether and how MEK1 interacts with YAP. In this study, we find that MEK1-YAP interaction is critical for liver cancer cell proliferation and maintenance of transformed phenotypes both in vitro and in vivo. Moreover, MEK1 and YAP proteins are closely correlated in human liver cancer samples. Mechanistically, inhibition of MEK1 by both PD98059 and U0126 as well as RNAi reduces beta-transducin repeat containing E3 ubiquitin protein ligase (BTRC), which acts as a potential endogenous YAP protector.
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http://dx.doi.org/10.1016/j.febslet.2013.10.042DOI Listing
December 2013

Analysis of the rs35959442 polymorphism in Hb E/β-thalassemia in Guangxi Province of the Republic of China.

Hemoglobin 2012 22;36(2):166-9. Epub 2011 Dec 22.

Department of Pediatrics, the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, Republic of China.

Hb E [β26(B8)Glu→Lys]/β-thalassemia (β-thal) is a worldwide inherited disorder. We determined the phenotype of 65 unrelated Hb E/β-thal subjects and 70 healthy individuals in the Guangxi Province of the Republic of China (ROC). Single nucleotide polymorphism (SNP) rs35959442 in HBS1L analysis was performed using the polymerase chain reaction (PCR)/restriction enzyme method. The data suggested that the frequency of the rs35959442 polymorphism was relatively high in patients with Hb E/β-thal in Guangxi Province, ROC, when associated with Hb F augmentation.
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http://dx.doi.org/10.3109/03630269.2011.644650DOI Listing
December 2012

Influences of genetic variation on fetal hemoglobin.

Pediatr Hematol Oncol 2011 Nov;28(8):708-17

Department of Pediatrics, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China.

Fetal hemoglobin (HbF) plays a dominant role in ameliorating morbidity and mortality of hemoglobinopathies. The authors performed a replicated study following the genome-wide association study (GWAS) guidelines to identify the genetic mechanics that influence HbF. The authors recruited and phenotyped 312 unrelated β-thalassemia subjects. Single-nucleotide polymorphism (SNP) analysis was performed by using polymerase chain reaction (PCR)/restriction enzymes. Four independent regions of interest were identified: HBS1L-MYB intergenic region, BCL11A locus, β-globin gene cluster, and the CSNK2A1 gene. There were 10 SNPs associated with HbF levels. In addition, haplotypes of HBS1L-MYB and BCL11A were identified and showed association with HbF production. Three independent regions, including HBS1L-MYB intergenic region, BCL11A locus, and β-globin gene cluster, were associated with HbF levels. This study can significantly improve the GWAS findings in Chinese cohorts and is useful for further research in the field of common predictors of the erythropoiesis.
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http://dx.doi.org/10.3109/08880018.2011.616573DOI Listing
November 2011
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