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1.
Eur Rev Med Pharmacol Sci ; 24(15): 8057-8066, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32767332

RESUMO

OBJECTIVE: The aim of this study was to investigate the expression characteristics of MTMR2 in NK/T cell lymphoma (NKTCL), and to further study its relationship with clinical parameters and the prognosis of patients with NKTCL. In addition, the potential mechanisms of MTMR2 promoting the progression of NKTCL was further explored. MATERIALS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine MTMR2 level in peripheral blood of 45 patients with NK/T-cell lymphoma and 45 healthy volunteers. The interplay between MTMR2 expression and clinical indicators, as well as the prognosis of patients with NK/T-cell lymphoma was analyzed. Meanwhile, MTMR2 expression in NKTCL cell lines was verified by qRT-PCR. Subsequently, MTMR2 knockdown and the overexpression models were constructed using lentivirus in NKTCL cell lines, including SNK-6 and KHYG-1. Transwell invasion and cell wound healing assays were applied to analyze the effect of MTMR2 on the biological function of NKTCL cells. Finally, an in-depth study of the relationship between MTMR2 and JAK1 was conducted to explore the underlying mechanism. RESULTS: QRT-PCR results showed that the expression level of MTMR2 in the serum of patients with NKTCL was remarkably higher than that of healthy volunteers, and the difference was statistically significant (p<0.05). Compared with patients with low expression of MTMR2, patients with high expression of MTMR2 exhibited significantly higher incidence of distant metastasis and lower overall survival rate (p<0.05). The metastasis ability of NKTCL SNK-6 cells was remarkably attenuated in MTMR2 knockdown group when compared with the negative control sh-NC group (p<0.05). Meanwhile, the metastatic ability of NKTCL KHYG-1 cells in MTMR2 overexpressing group was remarkably enhanced when compared with the control NC group (p<0.05). The Luciferase reporter gene assay confirmed that MTMR2 could target JAK1, thereby jointly regulating the malignant progression of NKTCL. In addition, cell recovery experiment verified that JAK1 could partially reverse the enhanced metastatic ability of NKTCL cells induced by the overexpression of MTMR2. CONCLUSIONS: MTMR2 was highly expressed in NKTCL serum samples and cell lines, leading to high risk of distant metastasis and poor prognosis. In addition, MTMR2 might promote the malignant progression of NKTCL by regulating JAK1.


Assuntos
Janus Quinase 1/metabolismo , Linfoma Extranodal de Células T-NK/metabolismo , Proteínas Tirosina Fosfatases não Receptoras/metabolismo , Linhagem Celular , Feminino , Humanos , Linfoma Extranodal de Células T-NK/sangue , Linfoma Extranodal de Células T-NK/patologia , Masculino , Pessoa de Meia-Idade , Proteínas Tirosina Fosfatases não Receptoras/genética
2.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi ; 37(11): 873-876, 2019 Nov 20.
Artigo em Chinês | MEDLINE | ID: mdl-31826560

RESUMO

Nasal cancer has not been included in the current list of legal occupational diseases in China. There is also a lack of systematic and in-depth study on the relationship between nasal cancer and occupational exposure factors in China. In September 2018, the department for work and pensions of UK released the latest edition of the "List of diseases covered by industrial injuries disablement benefit", which lists nasal cancer and nasopharyngeal carcinoma associated with wood dust exposure on the UK's occupational disease list. In order to better protect the health of workers, the relationship between occupational wood dust exposure and nasal cancer is reviewed, which provides a reference for further revision and improvement of occupational disease catalogue.


Assuntos
Poeira , Neoplasias Nasais , Doenças Profissionais , Exposição Ocupacional , Madeira , China/epidemiologia , Humanos , Neoplasias Nasais/epidemiologia , Doenças Profissionais/epidemiologia , Exposição Ocupacional/estatística & dados numéricos
3.
Zhonghua Gan Zang Bing Za Zhi ; 27(6): 424-429, 2019 Jun 20.
Artigo em Chinês | MEDLINE | ID: mdl-31357757

RESUMO

Objective: To explore the effect of substrate mechanical microenvironment and cell-cell interaction on differentiation of bone marrow mesenchymal stem cells (BMSCs), intrahepatic cellular function and phenotype. Methods: Bone marrow mesenchymal stem cells (BMSCs)-hepatocytes (HCs) and BMSCs-hepatic stellate cells (HSCs) were co-cultured on polyvinyl alcohol (PVA) hydrogel substrates at different stiffness (4.50 ± 0.47 kPa, 19.00 ± 3.51 kPa and 37.00 ± 2.09 kPa) by non-contact co-culture method. Furthermore, the effect of substrate mechanical microenvironment on BMSCs, HCs and HSCs and the activation and proliferation of HCs under different co-cultured condition was studied. A Student's t-test was used to compare the two groups. Results: The expression ofα-smooth muscle actin (α-SMA) and collagenα1- I (Col1A1) in BMSCs and HSCs cultured on its own increased with increase of substrate stiffness. After 72 h, the expression of albumin (ALB) of HCs on three stiff substrates was significantly higher than that of 24 and 48 h. Moreover, the expression of ALB of HCs increased with the increase of substrate stiffness. During the co-culture of BMSCs and HSCs, BMSCs of all three stiffness substrates promoted the expression ofα-SMA, Col1A1 in HSCs, but reduced the expression of PPARγin HSCs cells, thererby promoted the activation of HSCs, with apparent stiffness at 37 kPa. HSCs promoted the expression of ABL in BMSCs at three stiff substrates, but inhibited the expression of alpha-SMA and Col1A1 in BMSCs at 37 kPa, suggesting that co-culture had inhibited the differentiation of BMSCs myofibroblasts, and promoted the differentiation of hepatocyte-like cells, especially at high stiff substrates. In the co-culture of BMSCs and hepatic parenchymal cells, BMSCs had promoted the proliferation of hepatic parenchymal cells at 4.5 kPa. Further, hepatic parenchymal cells had inhibited the expression ofα-SMA in BMSCs, and promoted the expression of Alb, with inhibition of BMSCs differentiation towards myofibroblasts. Conclusion: The differentiation of BMSCs affects the substrate mechanical microenvironment, co-culture of HCs and HSCs. Simultaneously, affecting the function of hepatocytes in relation to the mechanical state of the substrates.


Assuntos
Células da Medula Óssea , Comunicação Celular , Técnicas de Cultura de Células , Diferenciação Celular , Células Estreladas do Fígado , Células-Tronco Mesenquimais , Animais , Células da Medula Óssea/citologia , Comunicação Celular/fisiologia , Técnicas de Cultura de Células/métodos , Células Cultivadas , Microambiente Celular/fisiologia , Células Estreladas do Fígado/citologia , Humanos , Células-Tronco Mesenquimais/citologia , Ratos , Ratos Sprague-Dawley
4.
Artigo em Chinês | MEDLINE | ID: mdl-30808141

RESUMO

Objective: To explore the clinical characteristics and treatment methods of esophageal foreign body. Method: The clinical data of 234 patients with esophageal foreign bodies admitted to our department from January 2015 to August 2018 were retrospectively analyzed, including course time, foreign body types, surgical methods, imaging manifestations and treatment related complications. Result: The diagnosis of esophageal foreign bodies was confirmed by esophageal CT or esophageal barium meal X-ray examination before operation in 234 patients. Course time varied from 3 hours to 7 days, and the jujube nucleus was the most common food-borne foreign body.223 patients underwent esophagoscopic exploration and foreign body removal under general intravenous anesthesia, 11 of them had no definite esophageal foreign body, 22 had esophageal perforation and periesophagitis. After removal of foreign body, the nasogastric feeding tube was inserted. About 10 days later, the nasogastric feeding tube was removed when they got healthy. Nine cases underwent cervical abscess incision and drainage under general anesthesia. The average postoperative hospital day was 11 days. Conclusion: The rigid esophagoscopy is a safe and effective method for the esophageal foreign bodies. And neck abscess incision must be necessary,when they suffered from esophageal perforation with neck abscess and other serious complications.


Assuntos
Perfuração Esofágica , Corpos Estranhos , Abscesso , Perfuração Esofágica/diagnóstico , Perfuração Esofágica/cirurgia , Esofagoscopia , Corpos Estranhos/diagnóstico , Corpos Estranhos/cirurgia , Humanos , Intubação Gastrointestinal , Estudos Retrospectivos
5.
Eur Rev Med Pharmacol Sci ; 21(24): 5668-5676, 2017 12.
Artigo em Inglês | MEDLINE | ID: mdl-29272001

RESUMO

OBJECTIVE: To determine whether the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR). PATIENTS AND METHODS: In this experiment, we selected epithelial cells from normal esophageal mucosa as the negative control group, and the ESCC EC109 and TE-1 cell strain as the observation group. Real-time PCR and Western-blotting were used to detect the expression of EP2, EGFR and phosphorylated EGFR (p-EGFR). The pre-treatment of ESCC cell strains was carried out using Butaprost (special agonist of PGE2 and EP2) and RNAi of EP2, and we observed the expression of EP2, EGFR, and p-EGFR. WST-8 (CCK-8) was applied for the detection of the cell proliferation rate. The transwell invasion experiment was conducted for the detection of the invasion capability of cells. The expression of MMP-9 (matrix metalloproteinase-9), VEGF (vascular endothelial growth factor), pro-inflammatory factors (IL-6 and TNF-α) in the cell supernatant were detected using ELISA. RESULTS: The high mRNA and protein expression of EP2, EGFR, and p-EGFR were found in the EC109 and TE-1 cell strains in the observation group, which were higher than those in the control group (p < 0.05). After the intervention of PGE2, EP2 expression was decreased and the p-EGFR expression was increased (p < 0.05). There was no variation found in the expression of EGFR (p > 0.05). After cells were intervened using Butaprost, the expressions of EP2 and p-EGFR were increased (p < 0.05), and there were no changes identified in the expression of EGFR (p > 0.05). After the intervention of RNAi, the expression of EP2 and p-EGFR was decreased (p < .05), and no changes were identified in the expression of EGFR (p > 0.05). After the intervention of PGE2 and Butaprost, great increases were seen in the cell proliferation rate, invasion capability, and the expression of MMP-9, VEGF, IL-6, and TNF-α in EC109 and TE-1 cell strains (p < 0.05), however, the intervention of RNAi could reduce above indexes (p < 0.05). CONCLUSIONS: Through cell experiments, we verified that the combination of prostaglandin E2 (PGE2) and EP2, the subtype receptor of PGE2, could trans-activate the epidermal growth factor receptor (EGFR) to regulate the proliferation and invasion capability of esophageal squamous cell carcinoma (ESCC) cells, and secrete and express multiple cytokines, thus discovering the pathological mechanism of inflammation to carcinoma transition in the occurrence of ESCC, and providing the experimental evidence for the search of new target in the treatment of ESCC. ESCC cells can highly express the receptor subtype EP2 of PGE2 that can transactivate the EGFR, through which PGE2 is involved in the transition mechanism from inflammation to cancer.


Assuntos
Alprostadil/análogos & derivados , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/patologia , Receptores de Prostaglandina E Subtipo EP2/fisiologia , Ativação Transcricional , Alprostadil/farmacologia , Linhagem Celular Tumoral , Receptores ErbB/genética , Humanos
6.
Zhonghua Yi Xue Za Zhi ; 97(42): 3320-3323, 2017 Nov 14.
Artigo em Chinês | MEDLINE | ID: mdl-29141378

RESUMO

Objective: To investigate the protein and mRNA expression of KIF2A in ovarian cancer, and to investigate the migration and invasion ability changes in ovarian cancer cell line HO-8910 transfected by KIF2A-siRNA. Methods: Immunohistochemical method was used to detect the expression of KIF2A in 30 cases of ovarian cancer and 20 cases of ovarian normal tissues. The expression of KIF2A mRNA was detected by RT-PCR. The mRNA and protein expression of KIF2A in cell line HO-8910 was detected by RT-PCR and Western blot after transfected by KIF2A-siRNA in vitro. After the transfection, the cell migration and invasion ability were observed by scratch test and transwell experiments. Result: The expression of KIF2A mRNA and protein in HO-8910 was significantly lower than that in normal ovarian tissue (P<0.05). The capacities of migration and invasion of HO-8910 was suppressed notably after the knockdown of KIF2A (P<0.05). Conclusion: KIF2A gene expression was increased in ovarian cancer, and knockdown of KIF2A gene can inhibit the migration and invasion of ovarian cancer cells. It suggested that KIF2A gene may be a new target for the development of ovarian cancer.


Assuntos
Movimento Celular , Cinesinas/metabolismo , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Ovarianas/metabolismo , Transfecção , Carcinoma Epitelial do Ovário , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Microtúbulos , Invasividade Neoplásica , RNA Interferente Pequeno
8.
Genet Mol Res ; 15(3)2016 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-27525937

RESUMO

Currently, the relationship between the trinucleotide repeat containing 9 (TNRC9) rs3803662 C>T polymorphism and risk of breast cancer (BC) is uncertain. Here, we attempted to obtain a more accurate assessment of this association by conducting a meta-analysis of all eligible case-control investigations, comprising 44,820 cases and 58,316 controls. A comprehensive search was performed to identify all suitable studies involving the TNRC9 rs3803662 polymorphism and BC risk. Pooled odds ratios (ORs) and 95% confidence intervals (95%CIs) were estimated using fixed- or random-effect models. Heterogeneity, publication bias, and sensitivity analyses were also carried out. We found that the variant T allele of rs3803662 C>T greatly increases BC risk (CT vs CC: OR = 1.14, 95%CI = 1.07-1.22, P < 0.001; TT vs CC: OR = 1.38, 95%CI = 1.25-1.53, P < 0.001; CT/TT vs CC: OR = 1.19, 95%CI = 1.11-1.28, P < 0.001; TT vs CT/CC: OR = 1.28, 95%CI = 1.19-1.38, P < 0.001). Stratified analysis based on ethnicity also revealed a markedly increased risk in Asian and Caucasian populations. Moreover, studies with hospital-based control groups showed elevated risk under the four genetic models employed, as did those using population-based controls, except under heterozygote comparison. The TNRC9 rs3803662 C>T polymorphism is greatly related to increased risk of BC, in both Asian and Caucasian populations.


Assuntos
Neoplasias da Mama/genética , Receptores de Progesterona/genética , Proteínas Reguladoras de Apoptose , Estudos de Casos e Controles , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Heterozigoto , Proteínas de Grupo de Alta Mobilidade , Humanos , Polimorfismo de Nucleotídeo Único , Risco , Transativadores
9.
Bioengineered ; 7(4): 235-40, 2016 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-27459596

RESUMO

Thermal denaturation of lysozymes was studied as a function of protein concentration, phosphate buffer concentration, and scan rate using differential scanning calorimetry (DSC), which was then analyzed by the isoconversional method. The results showed that lysozyme thermal denaturation was only slightly affected by the protein concentration and scan rate. When the protein concentration and scan rate increased, the denaturation temperature (Tm) also increased accordingly. On the contrary, the Tm decreased with the increase of phosphate buffer concentration. The denaturation process of lysozymes was accelatated and the thermal stability was reduced with the increase of phosphate concentration. One part of degeneration process was not reversible where the aggregation occurred. The other part was reversible. The apparent activation energy (Ea) was computed by the isoconversional method. It decreased with the increase of the conversion ratio (α). The observed denaturation process could not be described by a simple reaction mechanism. It was not a process involving 2 standard reversible states, but a multi-step process. The new opportunities for investigating the kinetics process of protein denaturation can be supplied by this novel isoconversional method.


Assuntos
Muramidase/química , Fosfatos/química , Desnaturação Proteica , Proteínas/química , Soluções Tampão , Varredura Diferencial de Calorimetria , Temperatura Alta , Concentração de Íons de Hidrogênio , Modelos Teóricos
10.
Phys Chem Chem Phys ; 15(24): 9498-502, 2013 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-23685875

RESUMO

The reactivity of sp(2) carbon materials is studied using the adsorption and dissociation of O2 on graphene and graphene oxide as model systems. The reactions on the basal plane, zigzag and armchair edges of graphene and graphene oxide with different oxygen-containing groups are calculated using first principles calculations. Two Brønsted-Evans-Polanyi relationships are identified and an electron delocalization model is suggested to understand the general trend of reactivity for sp(2) carbon materials.

11.
Plant Dis ; 97(11): 1514, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30708454

RESUMO

Liquorice (root of Glycyrrhiza uralensis Fisch.) is an important Chinese traditional medicine for many ailments (4). From 2002, severe outbreaks of root rot occurring on cultivated G. uralensis plants in Ningxia, China, have affected the yield and quality of liquorice and been considered as a major threat to commercial production of liquorice. Approximately 30% of the plants die from this disease in Ningxia every year. The disease, mainly affecting 2- to 4-year-old G. uralensis seedlings, begins with brown rot of root tips or lateral roots followed by internal decay of taproots during June to August every year. The infected plants are wilted with chlorotic foliage and easily pulled up from the soil. Root rot is clearly visible as a severe brown discoloration of vascular tissue along taproots. In severe cases, white mycelia are clearly visible on the surface of diseased roots and roots are decomposed. Isolations from diseased roots were made on potato dextrose agar (PDA) amended with streptomycin sulfate. Isolates (n = 78) were recovered from symptomatic roots (n = 105) and pure cultures were established by the single spore method. The two most frequently isolated fungi were transferred to potato sucrose agar and identified as Fusarium solani (61.5%) and F. oxysporum (30.8%) (1). The monophialides bearing microconidia of F. solani are long when compared to those of F. oxysporum. Genomic DNA of strains F. solani G013 and F. oxysporum FLR were extracted from mycelia with the cetyltrimethylammonium bromide (CTAB) method. Primers EF1-728F and EF1-986R were used to amplify the translation elongation factor-1α (TEF-1α) gene (2). The TEF-1α sequences of F. solani G013 (GenBank Accession No. AB777258) and F. oxysporum FLR (AB777257) shared 99 and 100% similarity with F. solani isolate NRRL52790 and F. oxysporum isolate NRRL 38328, respectively. Pathogenicity tests with one representative isolate of each species were conducted in the greenhouse on 1-month-old potted G. uralensis seedlings (12 plants per treatment). Isolates of the tested fungi were transferred to PDA and incubated in darkness at 24 ± 1°C for 7 days. Plant taproots about 5 cm below the soil surface were wounded with a sterile needle and five 5-mm-diameter fungal disks on the margin of colony were taken and firmly placed on the wounded location of each taproot with tinfoil; wounded taproots of seedlings inoculated with sterile PDA disks were used as controls (3). Root rot was assessed 2 months after inoculation. F. solani G013 and F. oxysporum FLR produced root rot symptoms on inoculated plants that were the same as those observed in field plants, and the fungi were reisolated from roots with typical symptoms. Control plants inoculated with sterile PDA disks remained asymptomatic, and no pathogen was isolated from them. To our knowledge, this is the first report of root rot caused by F. solani and F. oxysporum on G. uralensis in China. Effective control strategies are needed to minimize losses. References: (1) C. Booth. The Genus Fusarium. Commonwealth Mycological Institute, Farnham Royal, 1971. (2) I. Carbone and L. M. Kohn. Mycologia 91:553, 1999. (3) M. Guo et al. Plant Dis. 96:909, 2012. (4) T. Wu et al. Am. Assoc. Pharm. Sci. J. 13:1, 2011.

12.
Dis Esophagus ; 25(1): 10-6, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21595780

RESUMO

Muscularis externa of mouse esophagus is composed of two skeletal muscle layers in the adult. But less attention is paid to the histogenesis of the muscularis externa of the esophagus, and controversies still exist about the developmental process and the spatio-temporal expression characteristics of muscle-specific proteins during the development of esophageal muscularis externa. To further probe into the developmental pattern of muscularis externa of the mouse esophagus and the expression characteristics of different muscle-specific proteins, immunohistochemical and terminal deoxyribonucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP)-digoxigenin nick-end labeling apoptotic staining methods are used to investigate the expression patterns of different muscle-specific proteins and to elucidate the relationship of these protein expressions with the development of muscularis externa of the mouse esophagus. Thus, an understanding of the developing esophageal muscularis externa may be important for developing therapeutic strategies for the treatment of human esophagus diseases. Serial sections of mouse embryos from embryonic day (ED) 12 to ED18, and full-length esophagi from postnatal first to 5th day were stained with monoclonal antibodies against α-smooth muscle actin (α-SMA), α-sarcomerical actin (α-SCA), desmin, and monoclonal anti-skeletal myosin (MHC), while apoptosis was determined using the terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling assay. The expression of α-SMA was started at ED12. During the development of ED14-ED15, α-SMA positive cells were seen extending from the walls of left three, four, and six arch arteries toward the dorsal wall of esophagus. Stronger expression of α-SCA and desmin could be detected at ED14 and ED15, expression intensity in caudal segment and inner layer was stained stronger than that of cranial segment and outer layer, but after ED16, strong expression of α-SCA and desmin was found in the outer layer of muscularis externa. Expression of MHC was first detected in the outer layer of cranial segment of muscularis externa at ED17. At ED18, MHC had extended to the level of thyroid gland, staining intensity in the outer layer and cranial segment was stronger than that of inner layer and caudal segment. One to five days after birth, the thickness of the esophageal muscle layer was obviously increased. Most of the muscle cells in the cranial segment of esophagus showed strong expression of α-SCA and clear cross striations at higher magnification. With progression toward the caudal segment, expression intensity of α-SCA became weaker, but the expression intensity of desmin was the same at different levels of esophagus. The muscle fibers were arranged densely with high expression of MHC in the cranial segment. During the development of esophageal muscularis externa, few apoptotic cells were observed. α-SMA, α-SCA, desmin, and MHC show different expression patterns. The differentiation of outer layer of esophageal muscularis externa is quicker than that of inner layer, and the caudal segment is quicker than that of the cranial segment. Besides, apoptosis may not participate in the development of esophageal muscularis externa. The smooth muscle cells from arch arteries may participate in the development of esophageal muscularis externa.


Assuntos
Actinas/metabolismo , Esôfago/anatomia & histologia , Esôfago/embriologia , Desenvolvimento Muscular , Músculo Esquelético/embriologia , Animais , Anticorpos Monoclonais/metabolismo , Apoptose , Desmina/metabolismo , Esôfago/metabolismo , Feminino , Masculino , Camundongos , Músculo Esquelético/citologia , Músculo Esquelético/metabolismo , Miosinas de Músculo Esquelético/imunologia , Miosinas de Músculo Esquelético/metabolismo , Fatores de Tempo
13.
J Cell Mol Med ; 14(6A): 1328-37, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19432814

RESUMO

Hepatoma-derived growth factor (HDGF) is a novel mitogenic growth factor that has been implicated in many different carcinomas. Its role in keloid biology has not yet been investigated. The present study is aimed at examining the role of HDGF in keloid pathogenesis. Immunohistochemical staining and Western blot analyses were used to examine in vivo localization and expression of HDGF in keloid and normal skin tissue. This was followed by the detection of HDGF expression in fibroblasts cultured in vitro and fibroblasts exposed to serum. To investigate the effect of epithelial-mesenchymal interactions, a two-chamber system was employed in which keratinocytes on membrane inserts were co-cultured with the fibroblasts. HDGF expression levels in all cell extracts and conditioned media were assayed through Western blot analysis. In another set of experiments, the effect of exogenous recombinant HDGF on keloid fibroblasts (KF) and normal fibroblasts (NF) was examined. Cell proliferation was assessed by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and by quantifying proliferating cell nuclear antigen (PCNA) expression. Downstream targets of HDGF were identified by detecting their expression through Western blot analysis. Our results indicate that there was an increase in HDGF expression in the dermis of keloid compared with normal skin tissue. The application of serum and epithelial-mesenchymal interactions did not seem to have any effect on intracellular HDGF expression levels. However, co-culturing keloid keratinocytes with KFs resulted in increased HDGF secretion when compared with monoculture or normal controls. Furthermore, treatment with exogenous recombinant HDGF was found to increase the proliferation of KFs, activate the extracellular signal-regulated kinase (ERK) pathway and up-regulate the secretion of vascular endothelial growth factor (VEGF).


Assuntos
Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Queloide/etiologia , Queloide/metabolismo , Proliferação de Células/efeitos dos fármacos , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Derme/efeitos dos fármacos , Derme/enzimologia , Derme/patologia , Ativação Enzimática/efeitos dos fármacos , Epitélio/efeitos dos fármacos , Epitélio/patologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Fibroblastos/patologia , Humanos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Queloide/enzimologia , Queloide/patologia , Mesoderma/efeitos dos fármacos , Mesoderma/patologia , Modelos Biológicos , Transporte Proteico/efeitos dos fármacos , Proteínas Recombinantes/farmacologia , Soro , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo
15.
Cancer Immunol Immunother ; 55(8): 910-7, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16193335

RESUMO

Identifying appropriate tumor antigens is critical to the development of successful specific cancer immunotherapy. Serological analysis of tumor antigens by a recombinant cDNA expression library (SEREX) allows the systematic cloning of tumor antigens recognized by the spontaneous autoantibody repertoire of cancer patients. We applied SEREX to the cDNA expression library of cell line HMy2, which led to the isolation of six known characterized genes and 12 novel genes. Known genes, including ring finger protein 167, KLF10, TPT1, p02 protein, cDNA FLJ46859 fis, and DNMT1, were related to the development of different tumors. Bioinformatics was performed to predict 12 novel MMSA (multiple myeloma special antigen) genes. The prediction of tumor antigens provides potential targets for the immunotherapy of patients with multiple myeloma (MM) and help in the understanding of carcinogenesis. Crude lysate ELISA methodology indicated that the optical density value of MMSA-3 and MMSA-7 were significantly higher in MM patients than in healthy donors. Furthermore, SYBR Green real-time PCR showed that MMSA-1 presented with a high number of copy messages in MM. In summary, the antigens identified in this study may be potential candidates for diagnosis and targets for immunotherapy in MM.


Assuntos
Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Biologia Computacional , Mieloma Múltiplo/genética , Mieloma Múltiplo/imunologia , Sequência de Bases , Linhagem Celular Tumoral , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Biblioteca Gênica , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína Tumoral 1 Controlada por Tradução
16.
Zhongguo Yao Li Xue Bao ; 19(4): 347-51, 1998 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10375782

RESUMO

AIM: To study the genetic basis of N-acetylatransferase polymorphism in Chinese. METHODS: Genotypes in 120 healthy Han volunteers from 19 provinces of China were assayed. The 3 common mutant alleles (M1, M2, M3) and one normal wild-type (WT) allele of the N-acetyltransferase (NAT2) gene were detected by allele-specific polymerase chain reaction technique. RESULTS: The NAT2 allele frequencies in 120 Chinese (WT = 0.625, M1 = 0.0458, M2 = 0.188, M3 = 0.142) were different (P < 0.01). The NAT2 genotype distribution for all detected combinations of NAT2 alleles in 120 Chinese subjects was consisitent with Hardy-Weinberg equilibrium (chi 2 = 7.27, nu = 8, 0.7 > P > 0.5). Fifty subjects (41.7%) were homozygous wildtypes, 50 subjects (41.7%) were heterozygous mutants, and 20 subjects (16.7%) were homozygous mutants. CONCLUSION: The lower frequency of mutant M1 allele compared with that of Caucasians explains the low frequency of slow acylators in Chinese.


Assuntos
Acetiltransferases/genética , Adolescente , Adulto , Idoso , Alelos , Povo Asiático , China , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo Genético
18.
Yao Xue Xue Bao ; 32(8): 607-11, 1997 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-11596311

RESUMO

An HPLC method for the determination of caffeine metabolites in urine was established. Shim Pack CLC-ODS column (5 microns) was eluted with the mobile phase of methanol--acetonitrile--0.05% acetic acid = 12:1:87 (v/v) at a flow rate of 1.2 ml.min-1, and the ultraviolet absorbance was monitored at 280 nm. The 13 caffeine metabolites and caffeine were well separated and the concentrations of the five metabolites, AFMU, 1U, 1X, 17U, and 17X, were determined. The recoveries of the five metabolites were above 87%, the inter- and intra-day variations were less than 3%. The concentrations of the five metabolites in 120 volunteers were determined. The ratios of the metabolites were employed for the assessment of CYP1A2, NAT, and XO enzymes successfully.


Assuntos
Cafeína/metabolismo , Uracila/urina , Ácido Úrico/análogos & derivados , Cafeína/urina , Cromatografia Líquida de Alta Pressão , Humanos , Teofilina/urina , Uracila/análogos & derivados , Ácido Úrico/urina , Xantinas/urina
19.
Yao Xue Xue Bao ; 32(11): 813-8, 1997 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-11596199

RESUMO

Caffeine was used as a metabolic probe to measure, in 120 healthy volunteers, the activities of three enzymes, deduced to be N-acetyltransferase(NAT2), CYP1A2 and xanthine oxidase (XO). The caffeine metabolites of 5-acetylamino-6-formylamino-3-methyluracil (AFMU), 1-methylxanthine(1X), 1-methyluric acid(1U), 1, 7-dimethylxanthine(17X), and 1, 7-dimethyluric acid(17U) in urine were determined with HPLC after 4-5 hours of caffeine drink. The ratios of AFMU/1X or AFMU/(AFMU + 1X + 1U), (AFMU + 1X + 1U)/17X or (AFMU + 1X + 1U)/17U, and 1U/1X or 1U/(1X + 1U) were used as the index of NAT2, CYP1A2, and XO activities respectively. Frequency distribution analysis of the metabolic ratios of NAT2 indicated two distinct group with 20 slow acetylators and 100 rapid acetylators. Similar CYP1A2 activity was found in Chinese compared with European volunteers. Frequency analysis of CYP1A2 indicated the log normal distribution in 120 Chinese. The CYP1A2 index was much higher in smokers than that in nonsmokers. But no obvious difference was observed between young and old volunteers. The XO index also showed log normal distribution and has the similar value compared with European volunteers. The concentration variations of 1X and 1U in young volunteers were much lower than that in old volunteers.


Assuntos
Arilamina N-Acetiltransferase/metabolismo , Cafeína/metabolismo , Citocromo P-450 CYP1A2/metabolismo , Uracila/análogos & derivados , Ácido Úrico/análogos & derivados , Xantina Oxidase/metabolismo , Adolescente , Adulto , Idoso , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Teofilina/urina , Uracila/urina , Ácido Úrico/urina , Xantinas/urina
20.
Shi Yan Sheng Wu Xue Bao ; 29(2): 101-7, 1996 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-9387788

RESUMO

In order to know whether terminal monosaccharide of human sperm antigen is involved in stimulating antisperm immune response (especialy IgA production) in mucosal immune system, 33 IgA, 12 IgG and 35 IgM monoclonal antibodies (MAbs) all obtained by intragastrointestinal immunization were used in the present study. The molecular weight (MW) of sperm antigens reactive with MAbs were dectected by Western blotting. The MW range of sperm antigens to the above monclonal IgA, IgM and IgG class, except 12 MAbs not reactive with blotted sperm antigen on nitrocellulose strip, is 10-89 KDa, 11-75 KDa and 12-94 KDa respectively. The sperm antigens were separately blocked and digested with 5 lectins and 4 glycosidases. After that, the capacity of human sperm antigens bind with the above antibodies were determined by ELISA. One or more terminal monosaccharides of sperm antigens are involved with the reaction of most MAbs tested. The loss of terminal alpha-fucose, alpha-N-acetylgalactosamine, alpha-N-acetylglucosamine, mannose and beta-galactose but neuraminic acid in the antigenic determinant of human sperm were found to seriously affect the IgA binding with the corresponding sperm antigen. The binding of IgG with sperm antigen was obviously damaged by the removing of terminal alpha-fucose, alpha-N-acetylgalactosamine and alpha-mannose. The results suggest that the terminal monosaccharides of the sperm antigens play an important role in stimulation of antisperm IgA and other antibodies produced by using intragastrointestinal immunization.


Assuntos
Anticorpos Monoclonais/imunologia , Epitopos Imunodominantes/química , Imunoglobulina A/imunologia , Espermatozoides/imunologia , Reações Antígeno-Anticorpo , Antígenos/imunologia , Humanos , Masculino , Monossacarídeos/imunologia , Monossacarídeos/isolamento & purificação
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