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1.
Artigo em Chinês | MEDLINE | ID: mdl-36878503

RESUMO

Objective: To investigate the relationship between the long-non-coding RNA LINC00342 expression and the clinicopathological parameters of head and neck squamous cell carcinoma (HNSCC) and the biological function of LINC00342 in HNSCC cells. Methods: The expression level of LINC00342 in the HNSCC was analyzed using transcriptome sequencing data from TCGA (The Cancer Genome Atlas) database, and the expressions of LINC00342 in laryngeal squamous cell carcinoma tissues (LSCC) of 27 patients in the First Hospital of Shanxi Medical University were detected by transcriptome sequencing. The expression levels of LINC00342 in human embryonic lung diploid cells 2BS, HNSCC cell lines FD-LSC-1, CAL-27 and Detroit562 were determined by real-time quantitative polymerase chain reaction (qPCR). RNAi (RNA interference) was used for LINC00342 knockdown in HNSCC cell lines, and the changes of malignant phenotype in the tumor cells after LINC00342 knockdown were examined by cell counting kit-8 (CCK-8), colony formation, flow cytometry, transwell invasion and migration assays. Bioinformatics analysis was performed to construct a LINC00342-centered competing endogenous RNA (ceRNA) regulatory network, and GO (Gene Ontology) enrichment analysis was performed. Statistical analysis and graphing were performed using SPSS 25.0 software and GraphPad Prism 6 software. Results: Mean LINC00342 levels in HNSCC tissues and TCGA database were higher than that in normal control tissues, but with no significantly statistical difference (P=0.522). LINC00342 expression levels were positively correlated with cervical lymph node metastasis and pathological grade in patients with HNSCC, with higher expression in male patients than in female patients (P<0.05). Transcriptome sequencing analysis showed that mean expression level of LINC00342 in LSCC tissues of 27 patients was significantly higher than that in the paired adjacent normal mucosa tissues (t=1.56, P=0.036). LINC00342 expression was significantly upregulated in HNSCC cell lines FD-LSC-1, CAL-27 and Detroit562 (t-values of -12.17, -23.26 and -388.57, respectively; all P<0.001). Knockdown of LINC00342 by transfecting si-LINC00342-1 and si-LINC00342-2 inhibited HNSCC cell proliferation (t-values of 8.95 and 4.84, 2.70 and 5.55, 2.02 and 3.70, respectively), colony formation (t-values of 6.66 and 6.17, 7.38 and 11.65, 4.90 and 5.79, respectively), migration (t-values of 8.21 and 7.19, 5.76 and 6.46, 6.28 and 9.92, respectively) and invasion abilities (t-values of 9.29 and 10.25, 11.30 and 11.36, 8.02 and 8.66, respectively), but promoting apoptosis in cell lines FD-LSC-1 and CAL-27 (t-values of -2.21 and -5.83, -3.05 and -5.25 respectively) (all P-values<0.05). The LINC00342-centered ceRNA network consists of 10 downregulated microRNA and 647 upregulated mRNA nodes. GO analysis results indicated that LINC00342-regulated mRNAs were enriched in 22 biological processes, 32 molecular functions, and 12 cellular components. Conclusion: High level of LINC00342 is associated with the malignant progression of HNSCC. LINC00342 promotes the proliferation, migration, invasion, and antagonizes apoptosis of HNSCC cells, which serves as a potential molecular marker in HNSCC.


Assuntos
Neoplasias de Cabeça e Pescoço , RNA Longo não Codificante , Humanos , Feminino , Masculino , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , RNA Longo não Codificante/genética , Relevância Clínica , Células Epiteliais , Neoplasias de Cabeça e Pescoço/genética
2.
Eur Rev Med Pharmacol Sci ; 26(23): 8684-8692, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36524487

RESUMO

OBJECTIVE: Understanding factors for prolonged operative time is essential for surgery. This study aims at identifying the factors related to prolonged ileostomy closure operation time. PATIENTS AND METHODS: The data of 88 patients who underwent ileostomy reversal in the Department of Gastrointestinal Surgery of Jiaxing First Hospital between January 2018 and October 2021 were retrospectively analyzed. Prolonged operation time was defined as operative time >65 minutes. The Chi-square test was used to compare data between patients with normal operative time (≤65 minutes) and patients with prolonged operative time. Univariate and multivariate logistic regression analyses were performed to identify the factors associated with prolonged ileostomy closure operation time. RESULTS: Among the 88 patients (mean age: 62.78 years), median ileostomy retention time was 127.50 (range: 61.00-1,192.00) days. The operation time ranged from 35.00 minutes to 125.00 minutes. Prolonged ileostomy closure time occurred in 41 (46.6%) patients. In univariate analysis, the factors associated with prolonged operation time were body mass index (BMI) ≥25 kg/m2, previous history of abdominal surgery, and manual suture. History of serious complications after the primary operation was associated with shorter operation time. In multivariate analysis, the independent risk factors for prolonged operation time were BMI ≥25 kg/m2 (OR = 4.552, 95% CI: 1.369-15.136, p = 0.013), previous history of abdominal surgery (OR = 4.377, 95% CI: 1.394-13.739, p = 0.011), and manual suture (OR = 3.941, 95% CI: 1.181-13.154, p = 0.026). CONCLUSIONS: Overweight, previous history of abdominal surgery, and manual suture appear to be risk factors for prolonged operative time for ileostomy closure.


Assuntos
Ileostomia , Complicações Pós-Operatórias , Humanos , Pessoa de Meia-Idade , Ileostomia/efeitos adversos , Estudos Retrospectivos , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Fatores de Risco , Anastomose Cirúrgica/efeitos adversos
3.
Animal ; 15(12): 100401, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34794097

RESUMO

In the egg production industry, trace elements are required as additional dietary supplements to play vital roles in performance and egg quality. Compared to inorganic microelements (ITs), appropriate dose of organic trace microelements (OTs) are environmentally friendly and sufficient to satisfy the needs of hens. In order to evaluate the extent to which low-dose OTs replace whole ITs, the effects of organic copper, zinc, manganese, and iron compound on the performance, eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens were investigated. A total of 1 080 57-week-old Jing Hong laying hens were assigned to five groups with six replicates of 36 layers each for an 8-week experimental period. The birds were fed either a basal diet (control treatment (CT)) or the basal diet supplemented with commercial levels of inorganic trace elements (IT 100%) or the equivalent organic trace elements at 20%, 30%, and 50% of the inorganic elements (OT 20%, OT 30%, and OT 50%, respectively). Results showed that compared with those in the CT treatment, feeding hens with inorganic or organic microelement diet had significant effects on the eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens (P < 0.05). The eggshell strength and ratio between OT 30%, OT 50%, and IT 100% were similar at weeks 4 and 8, and the eggshell thickness of these groups was also similar at weeks 6 and 8. At week 8, the eggshell colour in OT 50% was darker than that in IT 100%. The mineral content in the eggshells of OT 50% and IT 100% significantly increased (P < 0.001), with no significant difference in effective thickness, mammillary thickness, and mammillary knob width between groups. There were no differences in the malondialdehyde content, total antioxidant capacity, and total superoxide dismutase activity in serum between OT 30%, OT 50%, and IT100%. While the catalase activities, the interleukin-1ß, interleukin-10, immunoglobulin G, and immunoglobulin M concentrations in serum were not significantly different between OT 50% and IT 100%. The mineral contents in the faeces of the organic groups were considerably reduced compared with those in IT 100% (P < 0.001). In conclusion, dietary supplementation with 30-50% organic compound microelements has the potential to replace 100% inorganic microelements in the hen industry for improving eggshell quality, mineral deposition in the eggshell, antioxidant capacity, and immune function, and reducing emissions to the environment without negative effects on laying performance.


Assuntos
Casca de Ovo , Oligoelementos , Ração Animal/análise , Animais , Antioxidantes , Galinhas , Dieta/veterinária , Suplementos Nutricionais , Feminino , Imunidade , Minerais , Óvulo
4.
Zhonghua Fu Chan Ke Za Zhi ; 56(4): 264-270, 2021 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-33902238

RESUMO

Objective: To investigate the clinical significance of the primary tumor size in patients with endometrial carcinoma (EC). Methods: A total of 385 patients with EC admitted to Peking University People's Hospital from January 2006 to December 2016 with complete follow up data were selected, whose tumor size data before biopsy were retrospectively studied. Results: (1) The mean diameter of the primary tumor was (3.6±1.8) cm (range: 1-15 cm). And 48 cases were 0-<2 cm, 78 cases were 2-<3 cm, 92 cases were 3-<4 cm, 73 cases were 4-<5 cm, 94 cases were ≥5 cm. The diameter of the tumor was associated with age <60 years old, premenopause, CA125≥35 kU/L, non-parturition, poor differentiation, stage Ⅲ-Ⅳ, depth of myometrial infiltration ≥1/2, cervical interstitial involvement, adnexal metastasis and lymph node metastasis (all P<0.05), but not associated with body mass index, hypertension, diabetes mellitus, pathology, lymph-vascular space invasion (all P>0.05). (2) Among the 334 patients underwent lymphadenectomy, 45 (13.5%, 45/334) cases with lymph node metastasis were observed. Stratified analysis showed that lymph node metastasis and recurrence rate of patients with EC gradually increased with the increase of tumor size (P<0.05). Adopting 2, 3, 4 and 5 cm as cut-off values of tumor size, there were significant differences in the rate of lymph node metastasis and recurrence among them observed (P<0.05), except for lymph node metastasis rate and recurrence rate when the cut-off value was 2 cm (P>0.05). (3) An receiver operating characteristic (ROC) curve analysis showed that a tumor diameter of 4.25 cm was the cut-off prognostic value to predict lymph node metastasis and recurrence of EC. Conclusions: Tumor diameter is significantly correlated with lymph node metastasis and recurrence in patients with EC. Tumor size should be considered in determining the scope of surgery and adjuvant therapy.


Assuntos
Neoplasias do Endométrio , Recidiva Local de Neoplasia , Neoplasias do Endométrio/patologia , Feminino , Humanos , Excisão de Linfonodo , Linfonodos/patologia , Linfonodos/cirurgia , Metástase Linfática , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Estudos Retrospectivos
5.
Eur Rev Med Pharmacol Sci ; 24(19): 9860-9868, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33090389

RESUMO

OBJECTIVE: The purpose of this study was to investigate the influences of micro ribonucleic acid (miR)-200b-5p on proliferation and apoptosis of ovarian cancer (OC) cells, and to explore its correlations with the target gene ATPase family, AAA domain containing 2 (ATAD2), and the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway. MATERIALS AND METHODS: Human ovarian fibroblasts (HOFs) or human OC cell lines (A2780) were cultured in vitro, and then, A2780 cells were separately transfected with miR-200b mimics or miR-NC or cultured with ATAD2-specific inhibitor BAY-850. Thereafter, the expression levels of miR-200b and ATAD2 messenger RNA (mRNA) were measured via qRT-PCR, and the proliferative capacity of cells was detected by CCK-8 assay. Next, the cell apoptosis was determined by means of flow cytometry and one-step TUNEL assay. Finally, the targeted regulatory relationship between miR-200b and ATAD2 was examined using a Luciferase reporter assay system, and the protein expressions were detected through Western blot (WB) assay. RESULTS: It was found that the expression level of miR-200b was remarkably lower (p<0.05), while the mRNA expression level of ATAD2 was notably higher (p<0.05) in A2780 cells than those in HOFs. The transfection with miR-200b mimics markedly reduced the mRNA expression level of ATAD2 (p<0.05) and the proliferative capacity (p<0.05) and increased the apoptosis rate (p<0.05) of A2780 cells. Besides, it was detected via the Luciferase reporter assay system that miR-200b inhibited ATAD2. BAY-850 significantly decreased the expression level of ATAD2 protein (p<0.05) and the proliferative capacity (p<0.05) but improved the apoptosis rate (p<0.05) of cells. Moreover, both miR-200b mimics and BAY-850 could distinctly repress the protein expression levels of PI3K and p-Akt of the PI3K/Akt signaling pathway (p<0.05) and enhance the expression of suppressor gene p53 (p<0.05). CONCLUSIONS: MiR-200b-5p can inhibit the proliferation and promote the apoptosis of OC cells through targeted inhibition of ATAD2 expression and regulation of the PI3K/Akt signaling pathway.


Assuntos
ATPases Associadas a Diversas Atividades Celulares/metabolismo , Proteínas de Ligação a DNA/metabolismo , MicroRNAs/metabolismo , Neoplasias Ovarianas/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , ATPases Associadas a Diversas Atividades Celulares/genética , Proliferação de Células , Células Cultivadas , Proteínas de Ligação a DNA/genética , Feminino , Humanos , MicroRNAs/genética , Neoplasias Ovarianas/patologia , Transdução de Sinais
6.
Zhonghua Yi Xue Za Zhi ; 100(17): 1315-1319, 2020 May 05.
Artigo em Chinês | MEDLINE | ID: mdl-32375439

RESUMO

Objective: To investigate the expression of T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) in patients with chronic heart failure and its modulatory activity on T lymphocytes. Methods: Eighty-six patients with chronic heart failure (CHF group) who were hospitalized in Department of Cardiology, the First Affiliated Hospital of Zhengzhou University between January and October 2018 were enrolled in the study. Meanwhile, thirty-two healthy controls (HC group) who received healthy examination were also selected. Peripheral blood mononuclear cells were isolated, and TIM-3 expression of CD4(+)and CD8(+)T cells was investigated by flow cytometry. CD4(+)and CD8(+)T cells were purified, and were stimulated by anti-TIM-3 antibody. Interferon-γ (IFN-γ), interleukin (IL)-4, IL-10, IL-35, IL-17, and IL-22 expressions in the supernatants of cultured CD4(+)T cells and tumor necrosis factor-α (TNF-α) and IFN-γ expressions in the supernatants of cultured CD8(+) T cells were measured by enzyme-linked immunosorbent assay. mRNA expressions of T-bet, GATA-3, FoxP3, and RORγt in CD4(+)T cells and perforin and granzyme B in CD8(+)T cells were semi-quantified by real-time PCR. Student t test or paired t test was used for comparisons between the two groups. Results: TIM-3(+) CD4(+) T cell percentage significantly increased in CHF group than that of HC group (3.47%±1.06% vs 0.92%±0.27%, P<0.001). TIM-3(+)CD8(+)T cell percentage also notably elevated in CHF group compared to HC group (6.12%±1.91% vs 1.77%±0.63%, P<0.001). CD4(+)T and CD8(+)T cells were dysfunctional in chronic heart failure. The levels of IFN-γ, IL-17, and IL-22 secreted by purified CD4(+) T cells significantly reduced in CHF group, while IL-10 and IL-35 expressions elevated in CHF group (all P<0.05). The relative mRNA expression levels of T-bet and RORγt in CD4(+) T cells remarkably decreased in CHF group than those of HC group (all P<0.01), while relative expression level of FoxP3 mRNA increased in CHF group (1.93±0.88 vs 0.97±0.28, P=0.031). The levels of TNF-α and IFN-γ produced by purified CD8(+)T cells notably reduced in CHF group than those of HC group (all P<0.05), and relative mRNA expression levels of perforin and granzyme B also decreased in CHF group (all P<0.05). The levels of anti-TIM-3 antibody stimulation-produced IFN-γ, IL-17, and IL-22 increased (all P<0.05) but IL-35 secretion reduced [(61±13) ng/ml vs (72±17) ng/ml, P=0.029] by CD4(+)T cells in CHF group. The relative mRNA expression levels of T-bet and RORγt also elevated in response to anti-TIM-3 antibody stimulation (all P<0.05). Anti-TIM-3 antibody stimulation promoted TNF-α and IFN-γ production by CD8(+)T cells in CHF group (all P<0.01). The relative mRNA expression levels of perforin and granzyme B also increased (all P<0.05). Conclusion: TIM-3 was increasingly expressed in T cells from patients with chronic heart failure, and might take part in the regulation of T cell dysfunction in chronic heart failure.


Assuntos
Insuficiência Cardíaca , Linfócitos T CD4-Positivos , Linfócitos T CD8-Positivos , Doença Crônica , Humanos , Imunoglobulinas , Leucócitos Mononucleares , Mucina-3
7.
Artigo em Chinês | MEDLINE | ID: mdl-29775017

RESUMO

Objective:The aim of this study is to analysis the correlation between nasal and eye symptom and psychological status in patients with allergic rhinitis, in order to provide a basis for improving the mental health of patients. Method:The nasal symptom score scale, eye symptom score scale and Kessler 10 were used in the study, and the survey data was analyzed. Result:At least one of 147 patients had a nasal symptom at a rate of 100% (147/147); at least one ocular symptom was 86.4% (127/147). Of the 147 patients, 29 had K10 scores more than 21.The nasal symptom including nasal congestion, itching, eye pain, eye swelling, tearing positively correlated with K10 total score. Conclusion:The clinical staffs must pay attention to the mental health status of AR patients, strengthen the assessment of nasal symptoms, eye symptoms and mental status, and give targeted intervention to patients with poor mental health.


Assuntos
Oftalmopatias/complicações , Transtornos Mentais/complicações , Rinite Alérgica/fisiopatologia , Humanos , Saúde Mental , Obstrução Nasal , Nariz , Rinite Alérgica/complicações
8.
Clin Transl Oncol ; 19(9): 1154-1160, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28357632

RESUMO

BACKGROUND: Several studies have found benefits of radiotherapy for adrenal metastasis from hepatocellular carcinoma (HCC). However, the efficacy, safety and outcome issues have not yet been fully addressed. Therefore, we performed this study to further elucidate the feasibility and outcome of radiotherapy in treating adrenal metastasis from HCC. METHODS: We retrospectively analyzed 81 patients with adrenal metastasis from HCC between 2001 and 2015. Eighteen patients received helical tomotherapy and 63 patients received conventional radiotherapy, including two-dimensional (2-D) or three-dimensional conformal radiotherapy (3-D CRT). The median radiation dose was 50 Gy (range 26-64 Gy) with median fraction size of 2.0 Gy (range 2.0-5.0 Gy). Tumor responses, adverse effects, patient outcomes and prognostic factors were analyzed. RESULTS: An objective response (complete and partial response) was achieved in 55.6% patients. The helical tomotherapy group showed higher objective response rate than the conventional radiotherapy group (P = 0.031). The major adverse effects were anorexia (51.8%), nausea (41.9%), and fatigue (35.8%). Similar toxicity profile occurred in the 2-D, 3-D CRT and helical tomotherapy groups. The overall survival (OS) rate at 1, 2 and 5 years was 59.9, 35.0, and 12.9%, respectively, with a median survival of 15 months. Patients who received helical tomotherapy achieved a better OS compared to the conventional radiotherapy group (P = 0.047). However, multivariate analysis indicated that radiotherapy technique was not an independent prognostic factor for patient outcome. CONCLUSION: These results suggest that radiotherapy offers a noninvasive approach in controlling adrenal metastasis from HCC with promising local control and acceptable tolerability.


Assuntos
Neoplasias das Glândulas Suprarrenais/radioterapia , Neoplasias das Glândulas Suprarrenais/secundário , Carcinoma Hepatocelular/radioterapia , Carcinoma Hepatocelular/secundário , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/radioterapia , Neoplasias das Glândulas Suprarrenais/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/mortalidade , Feminino , Humanos , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/mortalidade , Masculino , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Resultado do Tratamento
9.
Artigo em Chinês | MEDLINE | ID: mdl-29871273

RESUMO

As a common chronic disease, allergic rhinitis (AR) not only damage the physical health of the patients, but also bring psychological distress to the patients. Thus the comprehensive treatment should include both medical and mental health interventions as needed. In order to draw more attention to the mental status of the patients with AR and provide the basis for the researchers to choose reasonable AR psychological assessment tools, in this article, we evaluated the commonly used scales for the psychological status of AR patients, and the reason and significance of the psychological assessment in patients with AR were also reviewed.


Assuntos
Ansiedade/psicologia , Depressão/psicologia , Rinite Alérgica/psicologia , Estresse Psicológico , Ansiedade/complicações , Doença Crônica , Depressão/complicações , Humanos , Escalas de Graduação Psiquiátrica , Rinite , Rinite Alérgica/complicações , Inquéritos e Questionários
10.
Artigo em Chinês | MEDLINE | ID: mdl-29774679

RESUMO

SummaryAllergic rhinitis is a non-infectious disease with high risk, which seriously influenced on human health of body and mind. Therefore the life quality of patients with allergic rhinitis is worthy of attention. In this paper, the commonly used allergic rhinitis life quality questionnaires and selection principle have been introduced, and hope that the appropriate questionnaires for patients would be selected for the study of the life quality.


Assuntos
Qualidade de Vida , Rinite Alérgica Perene/psicologia , Rinite Alérgica Sazonal/psicologia , Humanos , Perfil de Impacto da Doença , Inquéritos e Questionários/normas
11.
Zhonghua Wai Ke Za Zhi ; 54(7): 498-503, 2016 Jul 01.
Artigo em Chinês | MEDLINE | ID: mdl-27373474

RESUMO

OBJECTIVE: To investigate the safety and effectiveness of laparoscopic reoperation for patients with gastroesophageal reflux disease (GERD) recurred form previous anti-reflux surgery. METHODS: Totally 19 patients received laparoscopic reoperation for symptomatic and anatomic recurred GERD in Department of Gastroesophageal Reflux Disease, Rocket Force General Hospital from January 2008 to September 2015 were retrospectively analyzed. There were 12 male and 7 female patients. The average reoperation age was (48±14) years, the average duration of reoperation from original ones was (43±38) months. The patients underwent preoperative barium, endoscopy, manometry and 24-hour pH studies. Laparoscopic hiatal hernia repair plus fundoplication was carried out for reoperation. Gastroesophageal reflux related symptoms (reflux, heartburn, chest pain, chough, wheezing, chest tightness and globus sensation) before and after surgery were compared by a questionnaire. The patients' medication consumption, complications and satisfaction of the reoperation were investigated as well. The repeated measures analysis of variance was used for statistical comparison of data preoperatively and postoperatively. RESULTS: No major complication and death occurred. Six cases (32%) had complications such as diarrhea, increased passing wind, flatulence, dysphagia and abdominal pain. The GERD related symptom score of reflux, heartburn, chest pain, chough, wheezing, chest tightness and globus sensation all significantly decreased (F: 25.0 to 56.7; P: 0.000 to 0.001) after the reoperation, with 68% good outcome of all the patients. After a follow-up of (33±22) months after reoperation, 1 case had partial recurrence at the 3(rd) month after reoperation. For all the patients, 12 cases felt very satisfied or satisfied with the reoperation. CONCLUSION: Laparoscopic reoperation is generally effective with acceptable morbidity rates for patients with esophageal and extraesophageal symptoms recurred form previous hiatal repair and (or) fundoplication.


Assuntos
Refluxo Gastroesofágico , Transtornos de Deglutição , Feminino , Fundoplicatura , Humanos , Laparoscopia , Masculino , Manometria , Pessoa de Meia-Idade , Recidiva , Reoperação , Estudos Retrospectivos , Inquéritos e Questionários
12.
Genet Mol Res ; 15(2)2016 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-27420937

RESUMO

Recombinant human anti-tumor necrosis factor (TNF)-α scFv-Fc was expressed in TKO mutant Arabidopsis thaliana seeds using plant-specific codons. Immunoblotting using a human IgG1 antibody detected the expression of anti-TNF-α proteins in plants. Results from qRT-PCR analysis demonstrated that the time of harvest significantly affected the protein yield and quality. Our results indicate that the Phaseolus vulgaris ß-phaseolin promoter directed anti-TNF-α scFv-Fc expression in A. thaliana seeds, with a maximum yield obtained at 20-days of development. Although the yield of anti-TNF-α scFv-Fc protein was not very high, accumulation of recombinant proteins in seeds is an attractive and simple method that can be used to purify biologically active anti-TNF-α scFv-Fc.


Assuntos
Arabidopsis/genética , Fragmentos Fc das Imunoglobulinas/genética , Sementes/metabolismo , Anticorpos de Cadeia Única/genética , Transgenes , Fator de Necrose Tumoral alfa/imunologia , Arabidopsis/metabolismo , Fragmentos Fc das Imunoglobulinas/imunologia , Fragmentos Fc das Imunoglobulinas/metabolismo , Proteínas Recombinantes , Sementes/genética , Anticorpos de Cadeia Única/imunologia , Anticorpos de Cadeia Única/metabolismo
13.
Genet Mol Res ; 15(2)2016 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-27420939

RESUMO

Glycine betaine is an important quaternary ammonium compound that is produced in response to several abiotic stresses in many organisms. The synthesis of glycine betaine requires the catalysis of betaine aldehyde dehydrogenase (BADH), which can convert betaine aldehyde into glycine betaine in plants, especially in halotolerant plants. In this study, we isolated the full-length cDNA of BADH from Suaeda corniculata (ScBADH) using reverse transcriptase-polymerase chain reaction and rapid amplification of cDNA ends. Next, we analyzed the expression profile of ScBADH using real-time PCR. The results showed that ScBADH expression was induced in the roots, stems, and leaves of S. corniculata seedlings under salt and drought stress. Next, ScBADH was overexpressed in Arabidopsis, resulting in the transgenic plants exhibiting enhanced tolerance over wild-type plants under salt and drought stress. We then analyzed the levels of glycine betaine and proline, as well as superoxide dismutase (SOD) activity, during salt stress in WT and transgenic Arabidopsis. The results indicated that overexpression of ScBADH produced more glycine betaine and proline, and increased SOD activity under NaCl treatment. Our results suggest that ScBADH might be a positive regulator in plants during the response to NaCl.


Assuntos
Betaína-Aldeído Desidrogenase/genética , Chenopodiaceae/genética , Proteínas de Plantas/genética , Betaína/metabolismo , Betaína-Aldeído Desidrogenase/metabolismo , Chenopodiaceae/enzimologia , Clonagem Molecular , Secas , Regulação da Expressão Gênica de Plantas , Glicina/metabolismo , Proteínas de Plantas/metabolismo , Prolina/metabolismo , Salinidade , Estresse Fisiológico , Superóxido Dismutase/metabolismo
14.
Br J Dermatol ; 174(3): 542-52, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26498849

RESUMO

BACKGROUND: Cold atmospheric plasma (CAP) has shown promise for wound healing, although little is understood of the underpinning mechanisms. Little has been reported so far of its potential use in the treatment of immune-mediated diseases such as psoriasis. OBJECTIVES: To study CAP-induced cell death and cytokine release in human keratinocytes as a first assessment of possible CAP use for psoriasis. METHODS: Using a CAP generator free of energetic ions, we observed its effects on keratinocytes in terms of morphology, cell viability and apoptosis, intracellular and mitochondrial reactive oxygen species (ROS), lysosomal integrity and mitochondrial membrane potential; and on secretion and expression of eight cytokines at protein and gene levels. RESULTS: CAP-induced reduced cell viability, apoptotic death and production of intracellular and mitochondrial ROS in dose-dependent manner. Mitochondrial dysfunction and lysosomal leakage were found in CAP-treated cells. It also induced release of interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-α and vascular endothelial growth factor (VEGF), and enhanced the mRNA expression of IL-1ß, IL-6, IL-8, IL-10, TNF-α, interferon-γ and VEGF. By contrast, IL-12 declined monotonically. CONCLUSIONS: The results suggest that with appropriate control of its dose, physical plasma could induce cell death via apoptotic pathways and enable simultaneous reduction in IL-12. These effects may be used to suppress keratinocyte hyperproliferation and to target T-cell activation to control amplification of inflammation. This provides an initial basis for further studies of CAP as a potential therapeutic option for inflammatory and immune-related diseases in dermatology, including psoriasis.


Assuntos
Morte Celular/fisiologia , Citocinas/biossíntese , Queratinócitos/metabolismo , Plasma , Psoríase/terapia , Caspase 3/metabolismo , Caspase 9/metabolismo , Sobrevivência Celular/fisiologia , Células Cultivadas , Humanos , Peróxido de Hidrogênio/metabolismo , Queratinócitos/patologia , Lisossomos/fisiologia , Potencial da Membrana Mitocondrial/fisiologia , Mitocôndrias/química , Nitratos/metabolismo , Nitritos/metabolismo , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo
15.
Genet Mol Res ; 14(3): 7395-405, 2015 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-26214418

RESUMO

The regulatory region of the ribulose-1,5-bisphosphate carboxylase small subunit gene SRS4 from soybean (Glycine max) was cloned using TAIL-PCR and general PCR, and named the rbcS promoter. The promoter was fused with the GUS gene and introduced into Nicotiana tabacum via Agrobacterium-mediated leaf disk transformation. In 4-week-old transgenic tobacco plants, the highest GUS expression levels were observed in the leaves, GUS activity was 7.13- and 7.40-fold higher in leaves than in stems and roots, respectively. Moreover, GUS activity was stimulated by light. In conclusion, spatial and light regulation of the soybean rbcS promoter was observed in N. tabacum, thus illustrating a leaf-specific and light-induced promoter.


Assuntos
Glycine max/genética , Nicotiana/genética , Ribulose-Bifosfato Carboxilase/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Regulação da Expressão Gênica de Plantas , Luz , Dados de Sequência Molecular , Folhas de Planta/genética , Plantas Geneticamente Modificadas/genética , Reação em Cadeia da Polimerase/métodos , Regiões Promotoras Genéticas , Glycine max/enzimologia , Nicotiana/enzimologia
16.
Eur Rev Med Pharmacol Sci ; 18(7): 1085-91, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24763891

RESUMO

OBJECTIVES: Psoriasis is a chronic inflammatory skin disease characterized by excessive proliferation of keratinocytes. Fibroblast growth factor 10 (FGF10) acts as a growth factor for keratinocyte proliferation. The aim of this study is to investigate whether FGF10 blockage, a new monoclonal antibody against FGF10 we generated, could mitigate topical propranolol-induced psoriasis-like lesions in guinea pigs. MATERIALS AND METHODS: The monoclonal anti-FGF10 was generated by a routine method and purified by affinity chromatography. The effect of FGF10 and anti-FGF10 on human keratinocyte HaCaT cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The back of the ears of individual guinea pigs was topically exposed to 5% propranolol emulsion to induce psoriasis-like lesions and randomly treated topically with phosphate buffered saline (PBS), hydrocortisone butyrate, or different doses of anti-FGF10. The pathologic changes and the degrees of inflammation in the auricular areas of individual animals were examined histologically. RESULTS: Characterization revealed that anti-FGF10 had a purity of 90% and a titer of 1:12800. We found that FGF10 stimulated HaCaT cell proliferation while treatment with different doses of anti-FGF10 inhibited FGF10-induced cell proliferation in a dose-dependent manner (100, 200 ng/ml, p < 0.05 vs. control; 400, 800, 1600 ng/ml, p < 0.01 vs. control). Compared to PBS-treated psoriatic animals, treatment with anti-FGF10, like hydrocortisone butyrate, greatly inhibited the severity of psoriasis-like lesions by reducing the Baker's scores, the thickness of epidermis, and the numbers of monocyte infiltrates in the dermis of animals. CONCLUSIONS: The newly generated anti-FGF10 monoclonal antibody inhibited the proliferation of human keratinocytes in vitro and mitigated inflammation and pathogenic changes in propranolol-induced psoriasis-like lesions in animals. Therefore, these findings may provide a proof of principle that blockage of FGF-10 may inhibit psoriasis-related inflammation.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Fator 10 de Crescimento de Fibroblastos/imunologia , Queratinócitos/efeitos dos fármacos , Psoríase/tratamento farmacológico , Administração Tópica , Animais , Anticorpos Monoclonais/farmacologia , Linhagem Celular , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Feminino , Cobaias , Humanos , Masculino , Camundongos Endogâmicos BALB C , Propranolol , Psoríase/induzido quimicamente , Psoríase/patologia , Pele/patologia
17.
Colorectal Dis ; 15(9): e488-95, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23810105

RESUMO

AIM: Confocal laser endomicroscopy (CLE) is a recently developed technique used to image colorectal neoplasia. Trials have shown varied results when it is compared with conventional colonoscopy. A meta-analysis was performed to determine the diagnostic accuracy of CLE in the detection of colorectal neoplasia. METHOD: A search was performed for studies assessing the accuracy of CLE in colorectal neoplasia. Studies comparing CLE diagnostic accuracy with conventional endoscopy in the detection of colorectal neoplasia were included. Exclusion criteria included case reports or case series, reviews, duplicate reports or insufficient data in the paper. Seventy-eight titles came up in the initial search and six studies were selected. These were subjected to a meta-analysis. In all, 284 patients with 1030 lesions were included. Each patient underwent conventional colonoscopy and CLE. Per-lesion sensitivity and specificity with 95% CI were calculated. RESULTS: In the individual studies, the sensitivity ranged from 33.3% to 100% and specificity from 71.6% to 99.4%. The weighted and total pooled result (random effects model) for sensitivity was 81% (95% CI 77-85) and for specificity was 88% (95% CI 85-90). The area under the weighted symmetric summary receiver operating curve was 0.9186. In the endoscope-based CLE subgroup, the sensitivity was 82% (95% CI 69-91) and specificity was 94% (95% CI 91-96). In the probe-based CLE subgroup, the sensitivity was 81% (95% CI 76-85) and the specificity was 75% (95% CI 69-81). CONCLUSION: CLE, using either the endoscope-based CLE or probe-based CLE technique, has high sensitivity and specificity. It could therefore be considered as an alternative endoscopic method to distinguish neoplastic from non-neoplastic lesions.


Assuntos
Adenocarcinoma/diagnóstico , Adenoma/diagnóstico , Colonoscopia/métodos , Neoplasias Colorretais/diagnóstico , Microscopia Confocal/métodos , Adenocarcinoma/patologia , Adenoma/patologia , Neoplasias Colorretais/patologia , Detecção Precoce de Câncer/métodos , Humanos , Sensibilidade e Especificidade
18.
Okajimas Folia Anat Jpn ; 75(4): 167-72, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9871400

RESUMO

Wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) injection into the midbrain periaqueductal gray (PAG) resulted in heavy accumulation of retrogradely labeled neurons in the lateral habenular nucleus (LHb) bilaterally. When the left femoral artery was persistently ligated for 2 h, the expression of Fos-like immunoreactivity (FOS-LI) was also found bilaterally in the LHb. In the present study, by combining the retrograde labeling method of injecting WGA-HRP into the PAG and the immunohistochemical staining of the FOS-LI neurons in the LHb induced by occlusion of the femoral artery, it was demonstrated that there are neurons containing both HRP labeling and FOS-LI. These neurons appeared to be located mainly in the medial part and posterior half of the LHb, and constitute about 24% of the total number of all labeled cells.


Assuntos
Arteriopatias Oclusivas/metabolismo , Artéria Femoral/metabolismo , Habenula/metabolismo , Neurônios Eferentes/metabolismo , Substância Cinzenta Periaquedutal/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Animais , Vias Eferentes/citologia , Vias Eferentes/metabolismo , Habenula/citologia , Membro Posterior/irrigação sanguínea , Imuno-Histoquímica , Neurônios Eferentes/citologia , Substância Cinzenta Periaquedutal/citologia , Ratos , Ratos Wistar , Conjugado Aglutinina do Germe de Trigo-Peroxidase do Rábano Silvestre/administração & dosagem
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