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OBJECTIVE: To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on ureteral injury repair via renal artery transplantation. METHODS: The left ureteral obstruction model was set up in 49 Balb/c mice by micro vascular clamp. The microscopic vascular clamp was taken out to lift the left ureteral obstruction after 10 days. The mice were randomly divided into an experimental group (n=25) and a control group (n=24). Balb/c mice BMSCs transfected by luciferase (Luc) were transplanted immediately through the renal artery after removing the microscopic vascular clamp from the experimental group; while mice in the control group was closed the incision after the microscopic vascular clamp was removed immediately and without BMSCs transplant. Magnetic resonance imaging (MRI) was used to scan the experimental mice. By measuring the left renal pelvis volume of the experimental mice at different time points and comparing the left ureter recanalization rate after removing left ureteral obstruction of the experimental group and the control group, we evaluated the repair effect of BMSCs on ureteral injury. RESULTS: The volume of the left renal pelvis in experimental mice became bigger obviously after the left ureter was obstructed (P<0.01). The left renal pelvis volume of the experimental group and the control group had no statistical significance 10 days after the left ureteral obstruction was set up (P=0.693). In the experimental group, the left ureter recanalization rate was higher than that in the control group, after removing the left ureteral obstruction (P=0.012). CONCLUSION: Transplantation through the renal artery can promote the restoration of ureteral injury in mice.
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Transplante de Células-Tronco Hematopoéticas/métodos , Obstrução Ureteral/terapia , Animais , Células-Tronco Hematopoéticas/citologia , Camundongos , Camundongos Endogâmicos BALB C , Artéria RenalRESUMO
Introduction: Root rot caused by the fungal pathogen Fusarium sp. poses significant challenges to tobacco cultivation in China, leading to major economic setbacks. The interplay between this pathogen and the wider soil microbial community remains poorly understood. Methods: High-throughput sequencing technology was utilized to evaluate soil prokaryotic, fungal, and protistan communities. We compared microbial communities in infected soils to those in healthy soils from the same field. Additionally, the influence of pH on the microbial communities was assessed. Results: Infected soils displayed elevated levels of soil nutrients but diminished observed richness across prokaryotic, fungal, and protistan groups. The pathogenic fungi Fusarium solani f sp. eumartii's abundance was notably increased in infected soils. Infection with F. solani significantly altered the soil's microbial community structure and interactions, manifested as a decrease in network scale and the number of keystone species. An evaluation of prokaryotes' role in F. solani's invasion revealed an increased number of connecting nodes in infected soils. Additionally, relationships between predatory protists and fungi were augmented, whereas predation on F. solani declined. Discussion: The study underscores the significance of comprehending the interactions among soil microorganisms and brings to light the susceptibility of soil microbial communities to pathogen invasion. It offers insights into the multifaceted relationships and potential vulnerabilities within the soil ecosystem in the context of Fusarium sp. invasion.
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As schizophrenia-like symptoms are produced by administration of phencyclidine (PCP), a noncompetitive antagonist of N-methyl-D-aspartate (NMDA) receptors, PCP-responsive genes could be involved in the pathophysiology of schizophrenia. We injected PCP to Wistar rats and isolated five different parts of the brain in 1 and 4 hr after the injection. We analyzed the gene expression induced by the PCP treatment of these tissues using the AGILENT rat cDNA microarray system. We observed changes in expression level in 90 genes and 21 ESTs after the treatment. Out of the 10 genes showing >2-fold expressional change evaluated by qRT-PCR, we selected 7 genes as subjects for the locus-wide association study to identify susceptibility genes for schizophrenia in the Japanese population. In haplotype analysis, significant associations were detected in combinations of two SNPs of BTG2 (P = 1.4 × 10(-6) ), PDE4A (P = 1.4 × 10(-6) ), and PLAT (P = 1 × 10(-3) ), after false discovery rate (FDR) correction. Additionally, we not only successfully replicated the haplotype associations in PDE4A (P = 6.8 × 10(-12) ) and PLAT (P = 0.015), but also detected single-point associations of one SNP in PDE4A (P = 0.0068) and two SNPs in PLAT (P = 0.0260 and 0.0104) in another larger sample set consisting of 2,224 cases and 2,250 controls. These results indicate that PDE4A and PLAT may be susceptibility genes for schizophrenia in the Japanese population.
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Nucleotídeo Cíclico Fosfodiesterase do Tipo 4/genética , Predisposição Genética para Doença , Fenciclidina/farmacologia , Esquizofrenia/enzimologia , Esquizofrenia/genética , Ativador de Plasminogênio Tecidual/genética , Adulto , Animais , Feminino , Estudo de Associação Genômica Ampla , Haplótipos/genética , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único/genética , Ratos , Ratos Wistar , Reprodutibilidade dos TestesRESUMO
BACKGROUND: Based on the glutamatergic dysfunction hypothesis for schizophrenia pathogenesis, we have been performing systematic association studies of schizophrenia with the genes involved in glutametergic transmission. We report here association studies of schizophrenia with SLC1A4, SLC1A5 encoding neutral amino acid transporters ASCT1, ASCT2, and SLC6A5, SLC6A9 encoding glycine transporters GLYT2, GLYT1, respectively. METHODS: We initially tested the association of 21 single nucleotide polymorphisms (SNPs) distributed in the four gene regions with schizophrenia using 100 Japanese cases-control pairs and examined allele, genotype and haplotype association with schizophrenia. The observed nominal significance were examined in the full-size samples (400 cases and 420 controls). RESULTS: We observed nominally significant single-marker associations with schizophrenia in SNP2 (P = 0.021) and SNP3 (P = 0.029) of SLC1A4, SNP1 (P = 0.009) and SNP2 (P = 0.022) of SLC6A5. We also observed nominally significant haplotype associations with schizophrenia in the combinations of SNP2-SNP7 (P = 0.037) of SLC1A4 and SNP1-SNP4 (P = 0.043) of SLC6A5. We examined all of the nominal significance in the Full-size Sample Set, except one haplotype with insufficient LD. The significant association of SNP1 of SLC6A5 with schizophrenia was confirmed in the Full-size Sample Set (P = 0.018). CONCLUSION: We concluded that at least one susceptibility locus for schizophrenia may be located within or nearby SLC6A5, whereas SLC1A4, SLC1A5 and SLC6A9 are unlikely to be major susceptibility genes for schizophrenia in the Japanese population.
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Sistema ASC de Transporte de Aminoácidos/genética , Proteínas da Membrana Plasmática de Transporte de Glicina/genética , Doença de Hartnup/genética , Polimorfismo Genético/genética , Esquizofrenia/genética , Alelos , Estudos de Casos e Controles , Éxons/genética , Feminino , Genótipo , Haplótipos , Doença de Hartnup/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Antígenos de Histocompatibilidade Menor , Esquizofrenia/diagnóstico , Esquizofrenia/epidemiologiaRESUMO
Ischemic heart disease within developed countries has been associated with high rates of morbidity and mortality. Cellbased cardiac repair is an emerging therapy for the treatment of cardiac diseases; however, a limited source of the optimal type of donor cell, such as an autologous cardiomyocyte, restricts clinical application. The novel therapeutic use of induced pluripotent stem cells (iPSCs) may serve as a unique and unlimited source of cardiomyocytes; however, iPSC contamination has been associated with teratoma formation following transplantation. The present study investigated whether cardiomyocytes from mouse fibroblasts may be reprogrammed in vitro with four cardiac transcription factors, including GATA binding protein 4, myocytespecific enhancer factor 2C, Tbox transcription factor 5, and heart and neural crest derivativesexpressed protein 2 (GMTH). Cardiacspecific markers, including αmyosin heavy chain (αMHC), ßMHC, atrial natriuretic factor, NK2 homeobox 5 and cardiac troponin T were observed within mouse fibroblasts reprogrammed with GMTH, which was reported to be more effective than GMT. In addition, Percoll density centrifugation enriched a population of ~72.4±5.5% αMHC+ induced cardiomyocytes, which retained the expression profile of cardiomyocyte markers and were similar to natural neonatal cardiomyocytes in welldefined sarcomeric structures. The findings of the present study provided a potential solution to myocardial repair via a cell therapy applying tissue engineering with minimized risks of immune rejection and tumor formation.
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Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Reprogramação Celular , Fibroblastos/metabolismo , Fator de Transcrição GATA4/genética , Fatores de Transcrição MEF2/genética , Miócitos Cardíacos/metabolismo , Proteínas com Domínio T/genética , Animais , Fator Natriurético Atrial/genética , Fator Natriurético Atrial/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Biomarcadores/metabolismo , Fibroblastos/citologia , Fator de Transcrição GATA4/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Proteína Homeobox Nkx-2.5/genética , Proteína Homeobox Nkx-2.5/metabolismo , Lentivirus/genética , Lentivirus/metabolismo , Fatores de Transcrição MEF2/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Miócitos Cardíacos/citologia , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Cultura Primária de Células , Proteínas com Domínio T/metabolismo , Cauda/citologia , Cauda/metabolismo , Transdução Genética/métodos , Troponina T/genética , Troponina T/metabolismo , Miosinas Ventriculares/genética , Miosinas Ventriculares/metabolismoRESUMO
Traditional Chinese medicine has great potential to improve wound healing. ANBP, the mixture of 4 Chinese herbs- Agrimoniapilosa, Nelumbonucifera, Boswelliacarteri, and Pollen typhae-is effective in trauma treatment while its mechanism is still elusive. In this study, quantitative proteomics and bioinformatics analyses were performed to decipher the possible roles of ANBP in accelerated wound healing of mouse skin. Among all 3171 identified proteins, 90, 71, 80, and 140 proteins were found to be differently expressed in 6 hours, 3 days, 7 days, and 14 days ANBP-treated tissues compared with corresponding control tissues, respectively. The result showed that different biological processes and pathways were activated at different healing stages. At the early healing stage, ANBP treatment mainly affected several biological processes, including immune and defense response, vascular system restoration, hemostasis and coagulation regulation, lipid metabolism and signal transduction, while muscle tissue, hair, epidermis, extracellular matrix and tissue remodeling related activities were the major events in ANBP promoted later wound healing. This is the first quantitative proteome study of ANBP-treated wound tissues, which provide a new perspective for the mechanism of ANBP accelerated wound healing and is of guiding significance for clinical application of ANBP in trauma disorders cure.
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Medicamentos de Ervas Chinesas/uso terapêutico , Proteômica , Cicatrização/efeitos dos fármacos , Ferimentos e Lesões/tratamento farmacológico , Ferimentos e Lesões/patologia , Animais , Biópsia por Agulha , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Distribuição Aleatória , Valores de Referência , Sensibilidade e Especificidade , Pele/efeitos dos fármacos , Pele/patologia , Cicatrização/genética , Ferimentos e Lesões/genéticaRESUMO
Human trefoil factor 3 (hTFF3) is a small peptide of potential therapeutic value. The mechanisms underlying the transcriptional regulation of hTFF3 remain unclear. The purpose of this study was to identify the core functional elements for the self-induction action of hTFF3 and transcription factors. First, truncated promoters were constructed to identify the functional regions of the hTFF3 promoter. Next, point mutation, chromatin immunoprecipitation, RNA interference, and gene overexpression experiments were performed to analyze the transcriptional binding sites responsible for the self-induced transcription of hTFF3. Our results revealed the -1450 bp to -1400 bp fragment of the hTFF3 promoter was the functional region for the self-induction action of hTFF3. Bioinformatics analysis confirmed that a STAT3 binding site is present in the -1417 bp to -1409 bp region. Subsequently, site-directed mutagenesis analysis determined that this STAT3 binding site was critical for the self-induction effect of hTFF3. ChIP experiments confirmed that STAT3 binds to the hTFF3 promoter. STAT3 overexpression and knockdown experiments revealed that STAT3 enhanced the self-induction effect and the expression of hTFF3. This study confirmed that hTFF3 exhibits self-induction action, and that STAT3 is the key transcription factor to maintain the function of self-induction.
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Regiões Promotoras Genéticas , Fator de Transcrição STAT3/metabolismo , Transcrição Gênica , Fator Trefoil-3/genética , Pareamento de Bases/genética , Linhagem Celular Tumoral , Análise Mutacional de DNA , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Mutação/genética , Reprodutibilidade dos Testes , Ativação Transcricional/genéticaRESUMO
BACKGROUND: The glutamatergic dysfunction hypothesis of schizophrenia suggests that genes involved in glutametergic transmission are candidates for schizophrenic susceptibility genes. We have been performing systematic association studies of schizophrenia with the glutamate receptor and transporter genes. In this study we report an association study of the excitatory amino acid transporter 2 gene, SLC1A2 with schizophrenia. METHODS: We genotyped 100 Japanese schizophrenics and 100 controls recruited from the Kyushu area for 11 single nucleotide polymorphism (SNP) markers distributed in the SLC1A2 region using the direct sequencing and pyrosequencing methods, and examined allele, genotype and haplotype association with schizophrenia. The positive finding observed in the Kyushu samples was re-examined using 100 Japanese schizophrenics and 100 controls recruited from the Aichi area. RESULTS: We found significant differences in genotype and allele frequencies of SNP2 between cases and controls (P = 0.013 and 0.008, respectively). After Bonferroni corrections, the two significant differences disappeared. We tested haplotype associations for all possible combinations of SNP pairs. SNP2 showed significant haplotype associations with the disease (P = 9.4 x 10-5, P = 0.0052 with Bonferroni correction, at the lowest) in 8 combinations. Moreover, the significant haplotype association of SNP2-SNP7 was replicated in the cumulative analysis of our two sample sets. CONCLUSION: We concluded that at least one susceptibility locus for schizophrenia is probably located within or nearby SLC1A2 in the Japanese population.
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Sistema X-AG de Transporte de Aminoácidos/genética , Polimorfismo Genético/genética , Esquizofrenia/genética , Simportadores/genética , Adulto , Transportador 2 de Aminoácido Excitatório , Feminino , Frequência do Gene/genética , Predisposição Genética para Doença , Genótipo , Proteínas de Transporte de Glutamato da Membrana Plasmática , Haplótipos/genética , Humanos , Japão/etnologia , Desequilíbrio de Ligação/genética , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Esquizofrenia/etnologiaRESUMO
Human trefoil factor 3 (hTFF3) is a small-molecule peptide with potential medicinal value. Its main pharmacological function is to alleviate gastrointestinal mucosal injuries caused by various factors and promote the repair of damaged mucosa. However, how its transcription is regulated is not yet known. The aim of this study was to clone the hTFF3 gene promoter region, identify the core promoter and any transcription factors that bind to the promoter, and begin to clarify the regulation of its expression. The 5' flanking sequence of the hTFF3 gene was cloned from human whole blood genomic DNA by PCR. Truncated promoter fragments with different were cloned and inserted into the pGL3-Basic vector to determine the position of the core hTFF3 promoter. Transcription element maintaining basic transcriptional activity was assessed by mutation techniques. Protein-DNA interactions were analyzed by chromatin immunoprecipitation (ChIP). RNA interference and gene over-expression were performed to assay the effect of transcription factor on the hTFF3 expression. The results showed that approximately 1,826 bp of the fragment upstream of hTFF3 was successfully amplified, and its core promoter region was determined to be from -300 bp to -280 bp through analysis of truncated mutants. Mutation analysis confirmed that the sequence required to maintain basic transcriptional activity was accurately positioned from -300 bp to -296 bp. Bioinformatic analysis indicated that this area contained a Sp1 binding site. Sp1 binding to the hTFF3 promoter was confirmed by ChIP experiments. Sp1 over-expression and interference experiments showed that Sp1 enhanced the transcriptional activity of the hTFF3 promoter and increased hTFF3 expression. This study demonstrated that Sp1 plays an important role in maintaining the transcription of hTFF3.
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Peptídeos/genética , Regiões Promotoras Genéticas/genética , Linhagem Celular , Imunoprecipitação da Cromatina , Humanos , Plasmídeos/genética , RNA Interferente Pequeno/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator Trefoil-3RESUMO
OBJECTIVE: To study the influence of heat stimulation on expression of coxsackie-adenovirus receptor (CAR) in keratinocytes (KCs) of mouse skin and the effect of CAR on production of cell growth factors by dendritic epidermal T lymphocytes (DETCs). METHODS: (1) Twenty BALB/c mice were divided into heat stimulation group (HS) and control group (C) according to the random number table, with 10 mice in each group. Mice in group HS were inflicted with scald milder than superficial-thickness by dressing wet hot gauze, which had been soaked in 100°C hot water for 3 min, in the hair removed area on the back for 1 to 3 s, while mice in group C were sham injured by dressing a wet gauze which had been soaked in water of room temperature for 3 min in the hair removed area on the back for 1 to 3 s. Square full-thickness skin specimens measuring 2.0 cm × 2.0 cm in size were obtained from the center of the bare skin. The expression of CAR in skin tissue sections were detected by immunohistochemistry staining. The mRNA and protein expression levels of CAR in skin tissue sections were respectively determined by real-time fluorescent quantitation RT-PCR and Western blotting. (2) KCs were isolated and cultured from full-thickness skin obtained from the trunk of 2 fetal BALB/c mice, and they were divided into 2 groups according to the random number table, with 5 wells in each group. The cells in group HS and group C were respectively cultured in 42°C and 37°C, 5% CO2 incubator for 1 h, and then all the cells were cultured in 37 °, 5% CO2 incubator for 6 h. The apoptosis of the cells and their expression of CAR were detected by flow cytometer. (3) Five BALB/c mice were sacrificed, and full-thickness skin was obtained from the trunk. The DETCs were divided into 7 groups according to the random number table after being isolated and purified from the skin specimens. Cells in group C were cultured without any stimulation, and cells in the 0.5, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/L CAR groups were respectively cultured with corresponding concentration of recombinant mice CAR nutrient solutions, with 5 wells in each group. The contents of insulin-like growth factor I (IGF-I) and keratinocyte growth factor (KGF) were determined with ELISA. Data were processed with independent samples t test and one-way analysis of variance. RESULTS: (1) The immunohistochemistry staining showed that there was mild positive staining in the skin tissue sections of mice in group C, while the positive staining was more obvious in group HS. The positive staining was mainly located in KCs, hair follicles, and sweat gland epithelial cells, while no positive staining was observed in fibroblasts. The mRNA expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.157 ± 0.027 and 0.773 ± 0.029. There was statistically significant difference between them (t = 3.052, P < 0.01). The protein expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.23 ± 0.09 and 0.89 ± 0.14. There was statistically significant difference between them (t = 2.556, P < 0.05). (2) The apoptosis rates of KCs in group C and group HS were respectively (5.7 ± 1.3)% and (7.4 ± 1.7)% (t = 0.464, P > 0.05). The expression rates of CAR in KCs in group C and group HS were respectively (48 ± 6)% and (80 ± 8)%. There was statistically significant difference between them (t = 2.585, P < 0.05). (3) The contents of IGF-Iin culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (23.1 ± 1.8), (22.5 ± 2.1), (31.2 ± 2.5), (39.7 ± 2.3), (61.8 ± 3.5), (45.1 ± 2.8), and (29.0 ± 2.0) µg/L. There was statistically significant difference among 7 groups (F = 3.414, P < 0.05). The contents of KGF in culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (131 ± 9), (217 ± 12), (355 ± 21), (563 ± 21), (535 ± 34), (292 ± 20), and (245 ± 10) ng/L. There was statistically significant difference among 7 groups (F = 5.063, P < 0.01). CONCLUSIONS: The expression of CAR in KCs would rise after HS. The optimum CAR concentration to increase IGF-I and KGF production in DETCs is low.
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Queimaduras/metabolismo , Proteína de Membrana Semelhante a Receptor de Coxsackie e Adenovirus/metabolismo , Queratinócitos/metabolismo , Linfócitos T/metabolismo , Animais , Células Cultivadas , Feminino , Fator 7 de Crescimento de Fibroblastos/metabolismo , Temperatura Alta , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pele/citologiaRESUMO
OBJECTIVES: The regulatory mechanism of microRNA (miRNA) within macrophage innative response to Mycobacterium tuberculosis infection is not clear yet. METHODS: The expression profile of cellular miRNAs during Mycobacterium bovis BCG infection was analyzed by using microarray. The expression of miR-146a was evaluated in alveolar macrophages (AMs) of bronchoalveolar lavage solution from pulmonary tuberculosis (PTB) patients and healthy volunteers respectively. Inhibitor experiment and promoter analysis were used to investigate the pathway involved in the induction of miR-146a. Examination of miR-146a function in macrophages was performed by overexpression and inhibition of miR-146a. RESULTS: Among the altered miRNAs, 10 were downregulated whereas 8 were upregulated in M. bovis BCG-infected macrophage. MiR-146a was high expressed in cultured macrophage respond to M. bovis BCG but decreased in AMs of PTB patients, and stated a negative correlation with degree of smear-positive. Nuclear factor-κB pathway was required for the induction of miR-146a. Overexpression of miR-146a results in significant reduction of PTGS2 and enhanced the killing ability of THP-1 cells to intracellular M. bovis BCG, and miR-146a negatively regulated TNF-α release in feedback manner. CONCLUSIONS: Our findings suggest an important role of miR-146a in M. bovis BCG infection that helps to fine-tune the inflammation response of MTB infection.
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Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno , Macrófagos/imunologia , MicroRNAs/biossíntese , Mycobacterium bovis/imunologia , Tuberculose Pulmonar/imunologia , Células Cultivadas , Ciclo-Oxigenase 2/biossíntese , Humanos , Macrófagos/microbiologia , Mycobacterium bovis/isolamento & purificação , Tuberculose Pulmonar/microbiologia , Fator de Necrose Tumoral alfa/biossínteseRESUMO
The aim of this study was to investigate the effects of heat stress on the expression of the coxsackievirus and adenovirus receptor (CAR) in mouse skin keratinocytes. Twenty BALB/c mice were randomly divided into two groups: the sham heat (control) and scald groups. Skin specimens were obtained 6 h after the treatments. Changes in the expression of CAR in skin keratinocyte samples were detected by immunohistochemistry, quantitative polymerase chain reaction and western blotting. In an in vitro assay, mouse skin keratinocytes were cultured and randomly divided into two groups: the normal control and heat stress groups. Six hours subsequently, the changes in CAR expression in the two groups were estimated by flow cytometry to determine the differences between the two groups. Heat stress significantly increased the expression of CAR in the mouse skin keratinocytes (P<0.05). The upregulation of CAR in mouse keratinocytes in burn wounds may be beneficial for restoring healing in organisms.
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BACKGROUND: Chronic kidney disease (CDK) is a worldwide health problem, but there is currently no effective treatment that can completely cure this disease. Recently, studies with mesenchymal stem cells (MSCs) on treating various renal diseases have shown breakthroughs. This study is to observe the homing features of MSCs transplanted via kidney artery and effects on renal fibrosis in a reversible unilateral ureteral obstruction (R-UUO) model. METHODS: Thirty-six Balb/c mice were divided into UUO group, UUO-MSC group, and sham group randomly, with 12 mice in each group. The MSCs had been infected by a lentiviral vector to express stably the luciferase reporter gene and green fluorescence protein genes (Luc-GFP-MSC). Homing of MSCs was tracked using in vivo imaging system (IVIS) 1, 3, 14, and 28 days after transplantation. Imaging results were verified by detecting GFP expression in frozen section under a fluorescence microscope. E-cadherin, α-SMA, TGF-ß1, and TNF-α mRNA expression in all groups at 1 and 4 weeks after transplantation were analyzed by quantitative PCR. RESULTS: Transplanted Luc-GFP-MSCs showed increased Luciferase expression 3 days after transplantation. The expression decreased from 7 days, weakened thereafter and could not be detected 14 days after transplantation. Quantitative PCR results showed that all gene expressions in UUO group and UUO-MSC group at 1 week had no statistical difference, while at 4 weeks, except TGF-ß expression (P > 0.05), the expression of E-cadherin, α-SMA, and TNF-α in the above two groups have statistical difference (P < 0.01). CONCLUSION: IVIS enables fast, noninvasive, and intuitive tracking of MSC homing in vivo. MSCs can be taken home to kidney tissues of the diseased side in R-UUO model, and renal interstitial fibrosis can be improved as well.
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Fibrose/patologia , Nefropatias/patologia , Transplante de Células-Tronco Mesenquimais/métodos , Obstrução Ureteral/terapia , Animais , Células Cultivadas , Fibrose/terapia , Nefropatias/terapia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJECTIVE: To observe the effects of glutamine combined with ulinastatin on inflammatory response of patients with severe burn injury. METHODS: Sixty patients with severe burn injury admitted to our burn wards from January 2010 to December 2011 conforming to the study criteria were divided into control group (C, n = 20), glutamine group (G, n = 20), and glutamine combined with ulinastatin group (G + U, n = 20) according to the random number table. Another 10 healthy volunteers were chosen as normal control group (NC). Isonitrogenous and isocaloric nutrition supports were given to patients in groups C, G, and G + U from post burn day (PBD) 2. 0.3 g/kg protein in the form of glutamine dipeptide was given to patients in group G for 10 days. 0.3 g/kg protein was given to patients in group G + U for 10 days with the same amount of glutamine dipeptide as that in group G, followed by intravenous injection of 100 kU ulinastatin (once per 8 hours) for 7 days during 10 days. The nitrogen concentration of 24 h urine was determined with Kieldahl nitrogen determination method, and nitrogen balance was calculated one day before treatment and ten days after treatment. Meanwhile, the levels of D-lactate in serum was determined by colorimetric method, the levels of diamine oxidase (DAO), TNF-α, and IL-6 by enzyme-linked immunosorbent assay, and LPS level by kinetic turbidimetric assay with TAL. Above-mentioned indexes were also examined in group NC. The wound healing rate on PBD 30, total hospital stay days, and the incidence of burn sepsis of all burn patients were recorded. Data were processed with one-way analysis of variance, LSD test, t test, and chi-square test. RESULTS: Compared with that in group C [(-5.40 ± 1.67) g/d], nitrogen balance in group G was significantly increased ten days after treatment [(-1.35 ± 0.59) g/d, P < 0.01]. The serum levels of D-lactate, DAO, LPS, TNF-α, and IL-6 in group G ten days after treatment were significantly lower than those in group C (P < 0.05 or P < 0.01). No statistically significant difference was observed in nitrogen balance and the serum levels of D-lactate, DAO between group G + U and group G (P values all above 0.05). The serum levels of LPS, TNF-α, and IL-6 in group G + U ten days after treatment were respectively (0.167 ± 0.064) EU/mL, (43 ± 14) pg/mL, (139 ± 23) pg/mL, which were significantly lower than those in group G [(0.240 ± 0.079) EU/mL, (59 ± 8) pg/mL, (195 ± 31) pg/mL, respectively, P < 0.05 or P < 0.01]. The would healing rate on PBD 30 and total hospital stay days in group G were respectively higher and shorter than those in group C (P values all below 0.01), but no statistically significant difference in the incidence of burn sepsis was found between them (P > 0.05). The would healing rate on PBD 30 in group G+U [(96 ± 4)%] was enhanced, and total hospital stay days [(41 ± 4) d] were lowered than those in group G [(88 ± 7)%, (49 ± 5)d, P values all below 0.01]. The incidence of burn sepsis of patients in group G + U (5%) was significantly lower than that in group C (35%, χ(2) = 6.234, P < 0.05). CONCLUSIONS: Glutamine combined with ulinastatin treatment can alleviate damage to intestine after severe burn injury, lower the serum level of inflammatory cytokines, promote wound healing, and reduce the incidence of burn sepsis.
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Queimaduras/sangue , Queimaduras/tratamento farmacológico , Glutamina/uso terapêutico , Glicoproteínas/uso terapêutico , Adolescente , Adulto , Feminino , Humanos , Interleucina-6/sangue , Ácido Láctico/sangue , Lipopolissacarídeos/sangue , Masculino , Fator de Necrose Tumoral alfa/sangue , Cicatrização , Adulto JovemRESUMO
Based on the glutamatergic dysfunction hypothesis for schizophrenia pathogenesis, we have been performing systematic association studies of schizophrenia with the glutamate receptor and transporter genes. We report here association studies of schizophrenia with three glutamate transporter genes SLC1A1, SLC1A3, and SLC1A6 encoding the glutamate transporters EAAT3, EAAT1, and EAAT4, respectively. We initially performed the screening of the total 25 single nucleotide polymorphisms (SNPs) distributed in the three gene regions using 100 out of 400 Japanese cases and 100 out of 420 Japanese controls. After controlling the false discovery rate (FDR) at level 0.05, we observed significant associations of schizophrenia with a genotype of SNP4 (rs2097837, P = 0.007) and with haplotypes of SNP2-SNP5 (P = 7.5 x 10(-5)) and SNP3-SNP5 (P = 9.0 x 10(-4)) in the SLC1A6 region. The haplotype of SNP2-SNP5 of SLC1A6 even showed marginally significant association with the disease in the full-size sample (400 cases and 420 controls, P = 0.031). We concluded that at least one susceptibility locus for schizophrenia may be located within or nearby SLC1A6, whereas SLC1A1 and SLC1A3 are unlikely to be major susceptibility genes for schizophrenia in the Japanese population.