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1.
Clin Transl Oncol ; 24(5): 909-918, 2022 May.
Article in English | MEDLINE | ID: mdl-35028929

ABSTRACT

PURPOSE: This study aimed to identify a diagnostic panel of serum microRNAs (miRNAs) for the early detection of bladder cancer (BC). METHODS: Serum samples were collected from 112 BC patients and 112 normal controls (NCs). A three-stage selection was conducted to identify differentially expressed miRNAs as candidates to construct the diagnostic panel. Further, to explore their potential roles in urothelial BC, bioinformatics analyses, including target genes prediction and functional annotation, were used. RESULTS: Six downregulated miRNAs (miR-1-3p, miR-30a-5p, miR-100-5p, miR-125b-5p, miR-143-3p, and miR-200c-3p) and one upregulated, miR-182-5p, in BC patients' serum were detected compared to NCs and were selected to establish the diagnostic panel. Based on a backward stepwise logistic regression analysis, miR-125b-5p, miR-182-5p, and miR-200c-3p comprehended the diagnostic panel [area under the curve (AUC) = 0.959, sensitivity = 91.67%, specificity = 92.5%]. CONCLUSION: The panel of three miRNAs had an excellent diagnostic capability, representing a potential non-invasive method for early BC detection.


Subject(s)
Carcinoma, Transitional Cell , MicroRNAs/genetics , Urinary Bladder Neoplasms , Biomarkers , Biomarkers, Tumor/genetics , Early Detection of Cancer , Female , Gene Expression Profiling/methods , Humans , Male , Urinary Bladder Neoplasms/diagnosis , Urinary Bladder Neoplasms/genetics
2.
Clin Transl Oncol ; 23(11): 2382-2393, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34075547

ABSTRACT

OBJECTIVES: High-risk human papillomavirus (HR-HPV) is an important risk factor for esophageal cancer. Macrophages constitute a crucial immune medium for regulating HPV-related tumors; however, the specific regulatory mechanisms remain unknown. Therefore, the purpose of our current study was to investigate the mechanism by which HPV16E6 regulates macrophages to promote the invasion and metastasis of esophageal cancer. METHODS: HPV16E6 infection was detected by polymerase chain reaction. Immunohistochemistry was used to verify the distribution of tumor-associated macrophages (TAMs) and MMP-9 expression in esophageal squamous cell carcinoma tissues (ESCCs), and cancer adjacent normal tissues (CANs) from Kazakh patients. ESCC cells were transfected with a plasmid over-expressing HPV16E6 and non-contact cocultured with macrophages. RESULTS: The infection rate of HPV16E6 in Kazakh ESCCs was clearly higher than that in CANs (P < 0.05). The density of CD163-positive TAMs was significantly positively correlated with HPV16E6 infection in ESCCs (P < 0.05). After coculturing macrophages and EC9706 cells transfected with the HPV16E6 plasmid, the phenotype of macrophages transformed into M2 macrophages. The migration and invasion ability of ESCC cells were higher in the HPV16E6-transfected and coculture group than in the HPV16E6 empty vector-transfected and non-cocultured HPV16E6-transfected groups (all P < 0.05). The density of M2-like TAMs in ESCCs was positively correlated with the level of MMP-9 expression. MMP-9 expression in the HPV16E6-ESCC coculture macrophages group was substantially higher than that in controls (all P < 0.05). CONCLUSIONS: HPV16 infection mediates tumor-associated macrophages to promote ESCC invasion and migration.


Subject(s)
Esophageal Neoplasms/pathology , Esophageal Squamous Cell Carcinoma/pathology , Human papillomavirus 16 , Oncogene Proteins, Viral/metabolism , Papillomavirus Infections/complications , Repressor Proteins/metabolism , Tumor-Associated Macrophages/pathology , Antigens, CD/metabolism , Antigens, Differentiation, Myelomonocytic/metabolism , Cell Differentiation , China/ethnology , Coculture Techniques , Esophageal Neoplasms/ethnology , Esophageal Neoplasms/virology , Esophageal Squamous Cell Carcinoma/ethnology , Esophageal Squamous Cell Carcinoma/virology , Humans , Matrix Metalloproteinase 9/metabolism , Neoplasm Invasiveness , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/ethnology , Phenotype , Receptors, Cell Surface/metabolism , Repressor Proteins/genetics , Tumor Microenvironment , Tumor-Associated Macrophages/metabolism , Tumor-Associated Macrophages/virology
3.
Genet Mol Res ; 15(4)2016 Nov 03.
Article in English | MEDLINE | ID: mdl-27819739

ABSTRACT

The differences in genetic backgrounds between deciduous and permanent teeth might contribute to the differences in developmental processes, histological characteristics, and tooth life cycles. Here, we attempted to identify significantly different modules between permanent and deciduous teeth via network and pathway analyses. We identified 291 differentially expressed genes (DEGs) between permanent and deciduous teeth using significance analysis of microarray methods. Co-expression networks of DEGs were constructed by weighted gene co-expression network analysis (WGCNA). Three pathways with a significant number of DEGs and P value <0.01 were identified. Integrated co-expression network and pathway (pathway and adjacent gene) analyses were used to extract three pathway-related modules: the calcium signaling pathway-related, ECM-receptor interaction pathway-related, and neuroactive ligand-receptor interaction pathway-related modules. We also attempted to analyze the topological centralities (degree, closeness, stress, and betweenness) of co-expression networks and modules. Four genes (TMEM229A, PPAPDC1A, LEPREL1, and GAD1) and three pathway-related modules that were significantly different in the deciduous and permanent teeth showed similar properties and good centralities. The relative expression levels of key genes were validated, and the differential expression of TMEM229A, LEPREL1, and GAD1 was confirmed by reverse transcription-polymerase chain reaction (P < 0.05). In conclusion, the results of this study may provide a greater understanding of the molecular pathogenesis and potential biomarkers of the progression from deciduous to permanent teeth.


Subject(s)
Gene Expression Profiling/methods , Gene Regulatory Networks , Tooth, Deciduous/metabolism , Gene Expression Regulation , Humans , Real-Time Polymerase Chain Reaction , Reproducibility of Results
4.
Genet Mol Res ; 15(3)2016 Jul 15.
Article in English | MEDLINE | ID: mdl-27525840

ABSTRACT

Hepatitis B virus (HBV) infection can cause HBV-related cirrhosis, liver failure, and hepatocellular carcinoma. At present, a hepatitis B surface antigen (HBsAg) blood test is the primary clinical and diagnostic marker for the identification of a chronic HBV infection. In the current study, we isolated a novel HBV mutant from a chronic HBV patient, capable of causing a false negative test result for most (7 of 8) commercial HBsAg ELISA kits. DNA sequencing of the HBsAg region of this HBV mutant revealed two novel mutation sites that resulted in a Thr-to-Met substitution at amino acid (aa) position 118 and a Lys to Asn substitution at aa position 122 of HBsAg. Moreover, a mutagenesis assay showed that the aa118 (Thr to Met) mutation was the leading cause of the false negative results from the HBsAg ELISA tests. The false negative result was restored, in that the mutation was correctly detected, when the Thr at aa position 118 of this mutated HBsAg was reconstituted. In conclusion, our study revealed a novel aa118 Met mutation of HBsAg HBV that will benefit the future development of HBV diagnosis.


Subject(s)
Hepatitis B Surface Antigens/genetics , Hepatitis B virus/genetics , Hepatitis B/blood , Mutation, Missense , Serologic Tests/standards , Enzyme-Linked Immunosorbent Assay/methods , Enzyme-Linked Immunosorbent Assay/standards , False Negative Reactions , HEK293 Cells , Hepatitis B/virology , Hepatitis B Surface Antigens/immunology , Hepatitis B virus/immunology , Humans , Serologic Tests/methods
5.
Braz J Med Biol Res ; 49(7)2016 Jul 04.
Article in English | MEDLINE | ID: mdl-27383119

ABSTRACT

The objective of this study was to perform a systematic review and meta-analysis to assess the effectiveness of proton pump inhibitors (PPI) for reflux disease in adult patients with laryngopharyngeal symptoms. A comprehensive search of Cochrane Library, EMBASE, Ovid EBM Reviews, and PubMed was performed for English-language literature about laryngopharyngeal reflux (LPR), in September 2014. The papers were filtered using pre-defined inclusion and exclusion criteria. Eight papers were identified and included in this meta-analysis. The sample comprised a pooled total of 370 patients, of which 210 and 160 patients took PPIs and placebo, respectively. The difference between PPIs and placebo groups in overall improvement of symptoms in adult patients with LPR was not statistically significant (RR=1.22; 95%CI=0.93-1.58; P=0.149). The difference in cough improvement was also not significant between PPIs and placebo groups (RR=0.65; 95%CI=0.30-1.41; P=0.279).


Subject(s)
2-Pyridinylmethylsulfinylbenzimidazoles/therapeutic use , Laryngopharyngeal Reflux/drug therapy , Proton Pump Inhibitors/therapeutic use , Adult , Female , Humans , Male , Placebo Effect , Treatment Outcome
6.
Genet Mol Res ; 15(2)2016 May 13.
Article in English | MEDLINE | ID: mdl-27323021

ABSTRACT

The aim of this study was to evaluate the effects of small interfering RNA (siRNA)-inhibited expression of the Sal-like 4 (SALL4) gene on the proliferation, colony formation, and apoptosis of prostate cancer C4-2 cells. C4-2 cells were cultured and divided into a si-SALL4 group, a negative control siRNA group, and a blank control group. SALL4 mRNA levels and protein expression were detected by real-time polymerase chain reaction and western blot, respectively. Changes in the cell proliferation and colony formation capacities were observed by using the MTS colorimetric method and colony formation assay, respectively. The influence of SALL4 on apoptosis was assessed with flow cytometry, and the expression of apoptosis-related proteins B-cell lymphoma 2 (Bcl-2) and bcl-like-protein 4 (Bax) were detected by western blot. The si-SALL4 group had significantly lower mRNA and protein levels of SALL4 as well as decreased proliferation and colony formation capacities than the negative control group (P < 0.05). There were significantly more apoptotic cells in the si-SALL4 group compared to the negative control (P < 0.05), and the expression of Bcl-2 and Bax decreased and increased, respectively, after treatment with si-SALL4. Silencing SALL4 expression by using siRNA technology inhibited the proliferation and colony formation of C4-2 cells, and promoted apoptosis likely mediated by Bcl-2 and Bax expression. These results provide experimental basis for further elucidating the role of SALL4 in prostate cancer cells.


Subject(s)
Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , RNA, Small Interfering/administration & dosage , Transcription Factors/genetics , Apoptosis/genetics , Cell Line, Tumor , Cell Proliferation/genetics , Gene Silencing , Humans , Male , Prostatic Neoplasms/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/genetics , Transcription Factors/antagonists & inhibitors , Transcription Factors/biosynthesis , Transfection
7.
Genet Mol Res ; 15(2)2016 Apr 28.
Article in English | MEDLINE | ID: mdl-27173277

ABSTRACT

Gout is the most common form of inflammatory arthritis affecting men, and current evidence suggests that genetic factors contribute to its progression. As a previous study identified that WD40 repeat protein 1 (WDR1) is associated with gout in populations of European descent, we sought to investigate its relationship with this disease in the Han Chinese population. We genotyped six WDR1 single nucleotide polymorphisms in 143 gout cases and 310 controls using Sequenom MassARRAY technology. The SPSS 16.0 software was used to perform statistical analyses. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated by unconditional logistic regression, with adjustments for age and gender. In an analysis using an allelic model, we identified that the minor alleles of rs3756230 (OR = 0.64, 95%CI = 0.450-0.911, P = 0.013) and rs12498927 (OR = 1.377, 95%CI = 1.037-1.831, P = 0.027) were associated with gout risk. In addition, we found that the "A/A" genotype of rs12498927 was associated with increased risk of gout under codominant (OR = 2.22, 95%CI = 1.12- 4.40, P = 0.042) and recessive models (OR = 2.24, 95%CI = 1.20-4.17, P = 0.012). We also determined the "A/G" genotype of rs12498927 to be significantly associated with higher urea levels in gout patients (P = 0.017). Our data shed new light on the association between genetic variations in the WDR1 gene and gout susceptibility in the Han Chinese population.


Subject(s)
Gout/genetics , Microfilament Proteins/genetics , Polymorphism, Single Nucleotide , Adult , Case-Control Studies , China , Female , Gout/blood , Humans , Male , Middle Aged , Urea/blood
8.
Neotrop Entomol ; 45(4): 420-6, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27048172

ABSTRACT

In this study, the effect of host density, host, and parasitoid ages in choice and no-choice tests on the parasitism performance of Tetrastichus brontispae Ferriere, one of the major parasitoid of Brontispa longissima (Gestro), was investigated in the laboratory. The results revealed that an increased host density resulted in no increased parasitism of B. longissima by T. brontispae; the optimal host density was three host pupae per parasitoid when considering the costs for mass rearing. Moreover, parasitoid age was quite crucial for effective parasitism and affected the emergence rate. Although 2-h to 4-day-old parasitoids successfully parasitized the host pupae, younger parasitoids (within 2-day-old) presented higher parasitism capacity than older parasitoids. More importantly, both choice and no-choice tests confirmed that all host stages tested from 2-h to 4-day-old were suitable for T. brontispae parasitization, although 2-h to 2-day-old hosts were preferred. We also demonstrated that sex ratio, emergence rate, and egg to adult developmental time were not influenced by host density, parasitoid, and host age in both choice and no-choice tests. Our data will allow for more accurate prediction and interpretation on the parasitization by T. brontispae, supporting mass-production initiatives and mass release in programs of B. longissima.


Subject(s)
Coleoptera/parasitology , Hymenoptera/pathogenicity , Animals , Host-Parasite Interactions , Pupa , Wasps
9.
Clin Transl Oncol ; 18(5): 427-36, 2016 May.
Article in English | MEDLINE | ID: mdl-26370423

ABSTRACT

Oral squamous cell carcinoma (OSCC) is a common cause of cancer death. Despite decades of improvements in exploring new treatments and considerable advance in multimodality treatment, satisfactory curative rates have not yet been reached. The difficulty of early diagnosis and the high prevalence of metastasis associated with OSCC contribute to its dismal prognosis. In the last few decades the emerging data from both tumor biology and clinical trials led to growing interest in the research for predictive biomarkers. Non-coding RNAs (ncRNAs) are promising biomarkers. Among numerous kinds of ncRNAs, short ncRNAs, such as microRNAs (miRNAs), have been extensively investigated with regard to their biogenesis, function, and importance in carcinogenesis. In contrast to miRNAs, long non-coding RNAs (lncRNAs) are much less known concerning their functions in human cancers especially in OSCC. The present review highlighted the roles of miRNAs and newly discovered lncRNAs in oral tumorigenesis, metastasis, and their clinical implication.


Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/genetics , Mouth Neoplasms/genetics , RNA, Untranslated/genetics , Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/therapy , Humans , Mouth Neoplasms/diagnosis , Mouth Neoplasms/therapy , Prognosis
10.
Neotrop Entomol ; 45(1): 96-101, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26429578

ABSTRACT

The bean flower thrips, Megalurothrips usitatus (Bagrall) (Thysanoptera: Thripidae), is an important pest of legume crops in South China. Yellow, blue, or white sticky traps are currently recommended for monitoring and controlling thrips, but it is not known whether one is more efficient than the other or if selectivity could be optimized by trap color. We investigated the response of thrips and beneficial insects to different-colored sticky traps on cowpea, Vigna unguiculata. More thrips were caught on blue, light blue, white, and purple traps than on yellow, green, pink, gray, red, or black traps. There was a weak correlation on the number of thrips caught on yellow traps and survey from flowers (r = 0.139), whereas a strong correlation was found for blue traps and thrips' survey on flowers (r = 0.929). On commercially available sticky traps (Jiaduo®), two and five times more thrips were caught on blue traps than on white and yellow traps, respectively. Otherwise, capture of beneficial insects was 1.7 times higher on yellow than on blue traps. The major natural enemies were the predatory ladybird beetles (63%) and pirate bugs Orius spp. (29%), followed by a number of less representative predators and parasitoids (8%). We conclude the blue sticky trap was the best to monitor thrips on cowpea in South China.


Subject(s)
Insect Control/instrumentation , Thysanoptera , Vigna , Animals , China
11.
Rev. bras. pesqui. méd. biol ; Braz. j. med. biol. res;49(7): e5149, 2016. tab, graf
Article in English | LILACS | ID: biblio-951688

ABSTRACT

The objective of this study was to perform a systematic review and meta-analysis to assess the effectiveness of proton pump inhibitors (PPI) for reflux disease in adult patients with laryngopharyngeal symptoms. A comprehensive search of Cochrane Library, EMBASE, Ovid EBM Reviews, and PubMed was performed for English-language literature about laryngopharyngeal reflux (LPR), in September 2014. The papers were filtered using pre-defined inclusion and exclusion criteria. Eight papers were identified and included in this meta-analysis. The sample comprised a pooled total of 370 patients, of which 210 and 160 patients took PPIs and placebo, respectively. The difference between PPIs and placebo groups in overall improvement of symptoms in adult patients with LPR was not statistically significant (RR=1.22; 95%CI=0.93-1.58; P=0.149). The difference in cough improvement was also not significant between PPIs and placebo groups (RR=0.65; 95%CI=0.30-1.41; P=0.279).


Subject(s)
Humans , Male , Female , Adult , 2-Pyridinylmethylsulfinylbenzimidazoles/therapeutic use , Proton Pump Inhibitors/therapeutic use , Laryngopharyngeal Reflux/drug therapy , Placebo Effect , Treatment Outcome
12.
Genet Mol Res ; 14(4): 16444-9, 2015 Dec 09.
Article in English | MEDLINE | ID: mdl-26662442

ABSTRACT

Pteroceltis tatarinowii (Ulmaceae) is a scientifically and economically important temperate deciduous tree that is endemic to China. In the present study, 12 P. tatarinowii polymorphic microsatellite loci were developed using the tailed primer-M13-simple sequence repeats (TP-M13-SSR) biotin-capture method. The number of alleles per locus ranged from 2 to 10, with an average of 6.58. The observed and expected heterozygosity ranged from 0.208 to 0.958 and from 0.198 to 0.858, with average values of 0.703 and 0.710, respectively. The markers isolated in this study represent a favorable tool for further analyses of the population genetic structure and evolutionary history of this relic tree.


Subject(s)
Microsatellite Repeats , Polymorphism, Genetic , Ulmaceae/genetics , Genetic Loci , Genetics, Population , Nucleotide Motifs
13.
Genet Mol Res ; 14(4): 16771-81, 2015 Dec 14.
Article in English | MEDLINE | ID: mdl-26681023

ABSTRACT

Dysregulation of microRNA (miR) is often associated with cancer development and progression. Aberrant expression of miR-134 has been found in some types of cancer. However, its expression and function in osteosarcoma remain unclear. The aim of this study was to explore the effects of miR-134 in osteosarcoma tumorigenesis and development. The expression level of miR-134 was quantified by real-time reverse transcription-polymerase chain reaction in human osteosarcoma cell lines and tissues. The effects of miR-134 on MG-63 cell phenotypes and tumorigenicity in vivo were observed using flow cytometry, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, transwell invasion, migration, and scratch migration assays. MiR-134 was significantly downregulated in osteosarcoma cell lines and clinical specimens. Decreased miR-134 expression was significantly associated with large tumor size, positive distant metastasis, and advanced clinical stage. Low miR-134 expression in osteosarcoma was an independent predictor of poor survival. Overexpression of miR-134 inhibited MG-63 cell proliferation, invasion, and migration, promoted cell apoptosis in vitro, and suppressed tumorigenicity in vivo. These findings indicate that miR-134 may act as a tumor suppressor in osteosarcoma and could serve as a novel therapeutic agent for miRNA-based therapy.


Subject(s)
Bone Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Genes, Tumor Suppressor , MicroRNAs/genetics , Osteosarcoma/genetics , Adolescent , Adult , Animals , Biomarkers , Bone Neoplasms/mortality , Bone Neoplasms/pathology , Cell Line, Tumor , Disease Models, Animal , Female , Humans , Kaplan-Meier Estimate , Male , Mice , Neoplasm Metastasis , Neoplasm Staging , Osteosarcoma/mortality , Osteosarcoma/pathology , Prognosis , Xenograft Model Antitumor Assays , Young Adult
14.
West Indian Med J ; 65(1): 229-231, 2015 Oct 29.
Article in English | MEDLINE | ID: mdl-26716808

ABSTRACT

Cytomegalovirus (CMV) infection has been associated with ulcerative colitis (UC). The prevalence of CMV infection in UC patients ranges from 10% to 16%. It is particularly high in the patients with steroidrefractory UC and those treated with immunosuppressants. However, synchronous onset of CMV colitis and UC in an immunocompetent patient is rare. It was originally described in 1990 and since then sixteen cases have been reported, as far as we are aware. Here, we present a case of CMV colitis and UC synchronously developing in an elderly immunocompetent woman. She was diagnosed through tissue immunohistochemistry and successfully treated with intravenous ganciclovir. This case demonstrates that in patients with severe active UC, even with new onset of the disease, CMV infection needs to be ruled out before initiating an aggressive immunosuppressive therapy.

15.
Genet Mol Res ; 14(4): 14181-8, 2015 Nov 13.
Article in English | MEDLINE | ID: mdl-26600475

ABSTRACT

RNA-Seq technology has been widely applied to transcriptomics, genomics, molecular marker development, and functional gene studies. In the genome, microsatellites are simple sequence repeats (SSR) with a high degree of polymorphism that are used as DNA markers in many molecular genetic studies. Using traditional methods such as magnetic bead enrichment, only a few microsatellite markers have been isolated. Coilia nasus is an anadromous, small-to-moderately sized fish species that is famous as an important fishery resource. Here, we have identified a large number of microsatellites from the fish brains by using Illumina sequencing. About 20 million Illumina reads were assembled into 148,845 unigenes. A total of 13,038 SSR motifs were identified via analysis of 3,958,293,117 (3.96 Gb) nucleotides to produce a comprehensive transcript dataset for the C. nasus brain, including mono-, di-, tri-, tetra-, and penta-repeat motifs. The most abundant type of repeat motif was di-nucleotide (42.97%), followed by mono-nucleotide (38.86%), tri-nucleotide (16.21%), tetra-nucleotide (1.83%), and penta-nucleotide (0.05%) repeat units, which is similar to the results obtained in studies in other species. These data provide a base of sequence information to improve molecular-assisted markers to study C. nasus genetic diversity.


Subject(s)
Fishes/genetics , Sequence Analysis, RNA/methods , Animals , Expressed Sequence Tags , Gene Expression Profiling/methods , Genetic Markers/genetics , Genetic Variation , Genome , Microsatellite Repeats , Molecular Sequence Annotation , Polymorphism, Genetic , Transcriptome
16.
Genet Mol Res ; 14(4): 12394-405, 2015 Oct 16.
Article in English | MEDLINE | ID: mdl-26505389

ABSTRACT

We determined the potential for induced pluripotent stem (iPS) cells to differentiate into nucleus pulposus (NP)-like cells in mice. iPS cells were generated from tail-tip fibroblasts. We used a pellet culture model with the aim of determining the applicability of iPS cell-based therapy to intervertebral disc degeneration (IVD). The cell pellet was cultured in an NP cell basal medium comprising Dulbecco's modified Eagle's medium supplemented with transforming growth factor beta 1, dexamethasone, ascorbate-2-phosphate, and 1% ITS-Premix. The pellet was evaluated by quantitative reverse transcription polymerase chain reaction, immunohistochemical staining, and biochemical composition. The differentiation of iPS cells into NP cells was demonstrated by the protein and mRNA expression levels of proteoglycan, collagen II, aggrecan, and CD24. Furthermore, increased hydroxyproline content and dimethylmethylene blue staining demonstrated that the collagen II and glycosaminoglycan content in the NP cells increased with time. We have shown that cultured mouse iPS cells can be induced to differentiate into NP cells. Such proof-of-concept opens up the possibility of producing patient-specific NP cells in a relatively simple and straightforward manner with high efficiency. We are confident that such cells could be immediately useful for the study of IVD disease.


Subject(s)
Induced Pluripotent Stem Cells/cytology , Aggrecans/metabolism , Animals , CD24 Antigen/metabolism , Cell Differentiation/physiology , Cells, Cultured , Collagen Type II/metabolism , Glycosaminoglycans/metabolism , Immunohistochemistry , Intervertebral Disc Degeneration/metabolism , Intervertebral Disc Displacement/metabolism , Karyotype , Mice , Proteoglycans/metabolism , Regenerative Medicine
17.
Genet Mol Res ; 14(3): 10619-29, 2015 Sep 09.
Article in English | MEDLINE | ID: mdl-26400293

ABSTRACT

Biofilm-forming bacteria are highly resistant to antibiotics, host immune defenses, and other external conditions. The formation of biofilms plays a key role in colonization and infection. To explore the mechanism of biofilm formation, mutant strains of Proteus vulgaris XC 2 were generated by Tn5 random transposon insertion. Only one biofilm defective bacterial species was identified from among 500 mutants. Inactivation of the glpC gene coding an anaerobic glycerol-3-phosphate dehydrogenase subunit C was identified by sequence analysis of the biofilm defective strain. Differences were detected in the growth phenotypes of the wild-type and mutant strains under pH, antibiotic, and organic solvent stress conditions. Furthermore, we observed an increase in the phagocytosis of the biofilm defective strain by the mouse macrophage RAW264.7 cell line compared to the wild-type strain. This study shows that the glpC gene plays an important role in biofilm formation, in addition to imparting pH, organic solvent, and antibiotic tolerance, and defense against phagocytosis to Proteus sp. The results further clarified the mechanism of biofilm formation at the genomic level, and indicated the importance of the glpC gene in this process. This data may provide innovative therapeutic measures against P. vulgaris infections; furthermore, as an important crocodile pathogen, this study also has important significance in the protection of Chinese alligators.


Subject(s)
Bacterial Proteins/genetics , Biofilms/growth & development , Glycerolphosphate Dehydrogenase/genetics , Proteus Infections/veterinary , Proteus vulgaris/genetics , Proteus vulgaris/immunology , Adaptation, Physiological/immunology , Alligators and Crocodiles/microbiology , Animals , Bacterial Proteins/immunology , Biofilms/drug effects , Cell Line , Cyclohexanes/pharmacology , DNA Transposable Elements , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Glycerolphosphate Dehydrogenase/immunology , Hexanes/pharmacology , Hydrogen-Ion Concentration , Immune Evasion , Macrophages/microbiology , Mice , Mutation , Proteus Infections/microbiology , Proteus Infections/pathology , Proteus vulgaris/drug effects , Proteus vulgaris/isolation & purification , Recombinant Proteins/genetics , Recombinant Proteins/immunology
18.
Genet Mol Res ; 14(3): 9034-44, 2015 Aug 07.
Article in English | MEDLINE | ID: mdl-26345835

ABSTRACT

In this study, we identified myogenic regulatory factors (MRFs) and analyzed the correlation between MRFs and meat quality in rainbow trout. The MyoD1a and MyoD1b genes were cloned from rainbow trout using a homology cloning method. Introns 1 and 2 in the MyoD1a and MyoD1b genes were cloned and submitted to GenBank (accession Nos. FJ623462 and FJ793566). Polymorphisms of MyoD1a and MyoD1b genes were analyzed using single-strand conformation polymorphism and sequencing, respectively. Two single nucleotide polymorphisms were detected in the MyoD1 gene, located at 129G→A in exon 1 and 37 G→A in exon 2. The 37 G→A mutation in exon 2 induced the R185K amino acid change in the polypeptide chain. Seven single nucleotide polymorphisms in the MyoD2 gene were detected, including 218T→C, 224T→C, 242A→C, 246T→A, 248T→C, 305T→C, and 329C→T. The 246T→A mutation in exon 1 induced the R83K change in the polypeptide chain. In the S3 fragment, meat quality traits of genotypes AA and AB significantly differed from those of genotype BB (P < 0.05). In the S5 fragment, meat quality traits of the genotypes AA and AC were significantly different from the genotypes BB and BC (P < 0.05). These results indicate that the MyoD1a and MyoD1b genes have an important influence on meat quality or were linked to the major genes in these strains. These genes can be used to control muscle fiber traits in rainbow trout, and the mutations in the S3 and S5 fragments can be used as molecular markers for selecting rainbow trout with better meat quality traits.


Subject(s)
Genetic Association Studies , Meat , MyoD Protein/genetics , Oncorhynchus mykiss/genetics , Animals , Base Sequence , Exons , Genotype , Oncorhynchus mykiss/growth & development , Phenotype , Polymorphism, Single Nucleotide
19.
Genet Mol Res ; 14(2): 6852-8, 2015 Jun 18.
Article in English | MEDLINE | ID: mdl-26125893

ABSTRACT

The aim of this study was to explore the mRNA levels of tumor necrosis factor-α (TNF-α), vessel endothelial growth factor (VEGF), and matrix metalloproteinase-3 (MMP-3) in synovial tissues in ankylosing spondylitis (AS), and to analyze the functions of these proteins in the differentiation of AS synovial tissue fibroblasts into osteoblasts (OB) and osteoclasts. Synovial tissue samples from 22 AS patients and 22 normal individuals were collected. In situ hybridization was utilized to detect TNF-α, VEGF, and MMP-3 transcripts. After counting numbers of positive cells, Spearman analysis was used to determine the correlation between transcriptional levels of the three mRNAs and the AS disease activity index (BASDAI) and the C-response protein (CRP) levels. With the addition of TNF-α, VEGF, or both factors into cultured normal synovial fibroblasts, osteocalcin (bone gla protein, BGP) secretion levels were compared. We found that expression of TNF-α, VEGF, and MMP-3 was identified exclusively in the disease group. mRNA levels were significantly positively correlated with BASDAI (r = 0.42, 0.38, and 0.47, respectively; P < 0.05) and CRP (r = 0.44, 0.34, and 0.47 respectively; P < 0.05) scores. The secretion level of BGP in normal synovial fibroblasts increased progressively with increasing concentrations of VEGF or TNF-α (P < 0.01 compared to levels before treatment). Furthermore, co-incubation using both VEGF and TNF-α significantly elevated BGP levels compared to the single addition of VEGF or TNF-α (P < 0.01). These results suggest TNF-α, VEGF, and MMP-3 might directly participate in the differentiation of fibroblasts into OBs.


Subject(s)
Fibroblasts/metabolism , Matrix Metalloproteinase 3/metabolism , Osteoblasts/metabolism , Osteoclasts/metabolism , Spondylitis, Ankylosing/genetics , Tumor Necrosis Factor-alpha/metabolism , Vascular Endothelial Growth Factor A/metabolism , Adult , C-Reactive Protein/genetics , C-Reactive Protein/metabolism , Case-Control Studies , Cell Differentiation , Female , Fibroblasts/pathology , Gene Expression , Humans , Male , Matrix Metalloproteinase 3/genetics , Osteoblasts/pathology , Osteocalcin/genetics , Osteocalcin/metabolism , Osteoclasts/pathology , Osteogenesis/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Spondylitis, Ankylosing/metabolism , Spondylitis, Ankylosing/pathology , Synovial Membrane/metabolism , Synovial Membrane/pathology , Tumor Necrosis Factor-alpha/genetics , Vascular Endothelial Growth Factor A/genetics
20.
Genet Mol Res ; 14(3): 7964-75, 2015 Jul 17.
Article in English | MEDLINE | ID: mdl-26214478

ABSTRACT

Grapevine downy mildew, caused by Plasmopara viticola, is a devastating disease that results in considerable economic losses as well as environmental damage through the repeated application of fungicides. The nucleotide-binding site leucine-rich repeat gene family functions in plant immunoactivity against various pathogens and pests. In this study, the 5' and 3' ends of the resistance gene homology fragment RGA5 were obtained by rapid amplification of cDNA ends. The 4282-base pair full-length cDNA was obtained using gene-specific primers, and the corresponding 1335-amino acid protein sequence contained characteristic nucleotide-binding site leucine-rich repeat domains of plant resistance proteins, including the toll-interleukin receptor type region. Expression of RGA5 during P. viticola infection and abiotic stress was investigated using quantitative real-time polymerase chain reaction. The results showed that treatment with P. viticola and 4 abiotic stimuli (salicyclic acid, methyl-jasmonate, abscisic acid, H2O2) significantly induced RGA5 within 12 days of inoculation. Therefore, RGA5 may play a critical role in protecting grapevines against P. viticola via signaling pathways involving these molecules.


Subject(s)
Disease Resistance/genetics , Genes, Plant , Plant Diseases/genetics , Plant Diseases/microbiology , Vitis/genetics , Vitis/microbiology , Amino Acid Sequence , Gene Expression Regulation, Plant , Molecular Sequence Data , Peronospora/physiology , Plant Proteins/chemistry , Plant Proteins/genetics , Protein Structure, Tertiary , Real-Time Polymerase Chain Reaction , Up-Regulation/genetics
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