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1.
Zhonghua Gan Zang Bing Za Zhi ; 31(7): 776-780, 2023 Jul 20.
Artigo em Chinês | MEDLINE | ID: mdl-37580265

RESUMO

Type II innate lymphoid cell (ILC2) is a newly identified innate immunological cell that belongs to the lymphocyte lineage in cell morphology, resides in the body's mucosal tissues, and has the dual functions of innate and adaptive immunity to promote tissue remodeling and repair after injury. Additionally, it is involved in the occurrence and development of a variety of liver diseases and plays an important role in maintaining the immunological homeostasis of the liver region. This article reviews the differentiation, development, and biological functions of ILC2, with particular attention to the research progress in liver diseases.


Assuntos
Imunidade Inata , Hepatopatias , Humanos , Linfócitos , Diferenciação Celular
2.
Braz J Med Biol Res ; 56: e12938, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37493775

RESUMO

Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.


Assuntos
Vacinas Bacterianas , Brucelose , Antígeno CTLA-4 , Brucelose/prevenção & controle , Brucella , Vacinas Bacterianas/imunologia , Antígeno CTLA-4/imunologia , Humanos , Animais , Epitopos/imunologia , Simulação de Acoplamento Molecular , Biologia Computacional , Proteínas de Bactérias/imunologia , Sequência de Aminoácidos , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/imunologia
3.
Braz. j. med. biol. res ; 56: e12938, 2023. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1447687

RESUMO

Brucellosis has become a global zoonotic disease, seriously endangering the health of people all over the world. Vaccination is an effective strategy for protection against Brucella infection in livestock in developed countries. However, current vaccines are pathogenic to humans and pregnant animals, which limits their use. Therefore, it is very important to improve the safety and immune protection of Brucella vaccine. In this study, different bioinformatics approaches were carried out to predict the physicochemical properties, T/B epitope, and tertiary structure of Omp2b and Omp31. Then, these two proteins were sequentially linked, and the Cytotoxic T lymphocyte associated antigen-4 (CTLA-4) variable region was fused to the N-terminal of the epitope sequence. In addition, molecular docking was performed to show that the structure of the fusion protein vaccine had strong affinity with B7 (B7-1, B7-2). This study showed that the designed vaccine containing CTLA-4 had high potency against Brucella, which could provide a reference for the future development of efficient brucellosis vaccines.

4.
Beijing Da Xue Xue Bao Yi Xue Ban ; 54(3): 505-510, 2022 Jun 18.
Artigo em Chinês | MEDLINE | ID: mdl-35701128

RESUMO

OBJECTIVE: To analyze the influenza vaccination trend of hospitalized elderly people (≥ 60 years old) in Beijing from 2013 to 2019. METHODS: The influenza vaccination status and hospitalization information of elderly people were extracted from the Beijing Elderly Influenza Vaccination database (2013-2019) and the Beijing Urban Employee Basic Medical Insurance database (2013-2019), se-parately. The influenza vaccine coverage rates and annual percentage change were calculated to compare the vaccination trends of elderly people hospitalized due to different diseases. The subjects in 2018-2019 influenza season were divided into different groups according to demographic status, health conditions and hospitalization outcomes to describe and compare the distribution of influenza vaccination rates. RESULTS: The influenza vaccine coverage rates among the elderly people hospitalized due to cardiovascular diseases, cerebrovascular diseases, respiratory diseases or diabetes mellitus were 14.6%, 13.4%, 13.4% and 11.8%, respectively. The influenza vaccination rate among those hospitalized for cardiovascular diseases remained the highest across six influenza seasons and those hospitalized for diabetes mellitus remained the lowest. The largest annual decline of influenza vaccine coverage rate was observed among the hospitalized elderly due to diabetes mellitus (-7.85%). The distribution of vaccinated population was significantly associated with age, gender, hospitalization outcome and comorbidities among the hospitalized elderly people with specific diseases in 2018-2019. Among the elderly people hospitalized due to four different diseases, the vaccination rate of the patients aged 70-79 years was higher than that of the other age groups and that of the patients aged 60-69 years was the lowest. Among the elderly people hospitalized due to respiratory diseases, the vaccination rate of men was higher than that of women, while the situation reversed among the elderly people hospitalized due to cardiovascular diseases and diabetes mellitus. Vaccination rates decreased among the older adults with poor hospitalization outcomes. Among the elderly people hospitalized due to diabetes mellitus, those with 0 comorbidity had the lowest vaccination rate (7.9%). CONCLUSION: The trend of influenza vaccine coverage rates among the elderly people in Beijing from 2013 to 2019 was downward. We should pay more attention to influenza vaccination in elderly people with diabetes mellitus and aged 60-69 years, and carry out more research on the protective effects of influenza vaccine to promote influenza vaccine coverage among people with chronic diseases.


Assuntos
Doenças Cardiovasculares , Vacinas contra Influenza , Influenza Humana , Idoso , Pequim , Feminino , Hospitalização , Humanos , Influenza Humana/epidemiologia , Influenza Humana/prevenção & controle , Masculino , Pessoa de Meia-Idade , Vacinação
5.
Ann Oncol ; 28(9): 2264-2271, 2017 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-28633425

RESUMO

BACKGROUND: Mutations in the androgen receptor (AR) ligand-binding domain (LBD), such as F877L and T878A, have been associated with resistance to next-generation AR-directed therapies. ARN-509-001 was a phase I/II study that evaluated apalutamide activity in castration-resistant prostate cancer (CRPC). Here, we evaluated the type and frequency of 11 relevant AR-LBD mutations in apalutamide-treated CRPC patients. PATIENTS AND METHODS: Blood samples from men with nonmetastatic CRPC (nmCRPC) and metastatic CRPC (mCRPC) pre- or post-abiraterone acetate and prednisone (AAP) treatment (≥6 months' exposure) were evaluated at baseline and disease progression in trial ARN-509-001. Mutations were detected in circulating tumor DNA using a digital polymerase chain reaction-based method known as BEAMing (beads, emulsification, amplification and magnetics) (Sysmex Inostics' GmbH). RESULTS: Of the 97 total patients, 51 had nmCRPC, 25 had AAP-naïve mCRPC, and 21 had post-AAP mCRPC. Ninety-three were assessable for the mutation analysis at baseline and 82 of the 93 at progression. The overall frequency of detected AR mutations at baseline was 7/93 (7.5%) and at progression was 6/82 (7.3%). Three of the 82 (3.7%) mCRPC patients (2 AAP-naïve and 1 post-AAP) acquired AR F877L during apalutamide treatment. At baseline, 3 of the 93 (3.2%) post-AAP patients had detectable AR T878A, which was lost after apalutamide treatment in 1 patient who continued apalutamide treatment for 12 months. CONCLUSIONS: The overall frequency of detected mutations at baseline (7.5%) and progression (7.3%) using the sensitive BEAMing assay was low, suggesting that, based on this assay, AR-LBD mutations such as F877L and T878A are not common contributors to de novo or acquired resistance to apalutamide. CLINICALTRIALS.GOV IDENTIFIER: NCT01171898.


Assuntos
Antagonistas de Androgênios/uso terapêutico , Mutação Puntual , Neoplasias de Próstata Resistentes à Castração/tratamento farmacológico , Neoplasias de Próstata Resistentes à Castração/genética , Receptores Androgênicos/genética , Tioidantoínas/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , DNA Tumoral Circulante/genética , Humanos , Masculino , Pessoa de Meia-Idade
6.
Ann Oncol ; 27(1): 106-13, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26504153

RESUMO

BACKGROUND: Androgen receptor (AR) signaling and incomplete inhibition of estrogen signaling may contribute to metastatic breast cancer (MBC) resistance to a nonsteroidal aromatase inhibitor (NSAI; letrozole or anastrozole). We assessed whether combined inhibition of androgen biosynthesis with abiraterone acetate plus prednisone and estradiol synthesis with exemestane (E) may be of clinical benefit to postmenopausal patients with NSAI-pretreated estrogen receptor-positive (ER+) MBC. PATIENTS AND METHODS: Patients (N = 297) were stratified by the number of prior therapies for metastatic disease (0-1 versus 2) and by prior NSAI use (adjuvant versus metastatic), and randomized (1 : 1 : 1) to receive oral once daily 1000 mg abiraterone acetate plus 5 mg prednisone (AA) versus AA with 25 mg E (AAE) versus 25 mg E alone (E). Each treatment arm was well balanced with regard to the proportion of patients with AR-positive breast cancer. The primary end point was progression-free survival (PFS). Secondary end points included overall survival, clinical benefit rate, duration of response, and overall response rate. RESULTS: There was no significant difference in PFS with AA versus E (3.7 versus 3.7 months; hazard ratio [HR] = 1.1; 95% confidence interval [CI] 0.82-1.60; P = 0.437) or AAE versus E (4.5 versus 3.7 months; HR = 0.96; 95% CI 0.70-1.32; P = 0.794). Increased serum progesterone concentrations were observed in both arms receiving AA, but not with E. Grade 3 or 4 treatment-emergent adverse events associated with AA, including hypokalemia and hypertension, were less common in patients in the E (2.0% and 2.9%, respectively) and AA arms (3.4% and 1.1%, respectively) than in the AAE arm (5.8% for both). CONCLUSIONS: Adding AA to E in NSAI-pretreated ER+ MBC patients did not improve PFS compared with treatment with E. An AA-induced progesterone increase may have contributed to this lack of clinical activity. CLINICALTRIALSGOV: NCT01381874.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Ósseas/tratamento farmacológico , Neoplasias da Mama/tratamento farmacológico , Acetato de Abiraterona/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Androstadienos/administração & dosagem , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/mortalidade , Neoplasias Ósseas/secundário , Neoplasias da Mama/metabolismo , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Intervalo Livre de Doença , Feminino , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Pessoa de Meia-Idade , Pós-Menopausa , Modelos de Riscos Proporcionais , Receptores de Estrogênio/metabolismo , Resultado do Tratamento
7.
J Dent Res ; 95(4): 430-8, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26661713

RESUMO

Diabetes mellitus is closely related to oral-complicated diseases by oxidative stress. This study investigates whether cellular myeloblastosis (c-myb) could protect human dental pulp cells against glucose oxidative stress and regulate autophagy activity for pulp vitality. Diabetes mellitus was induced by streptozotocin in Sprague-Dawley rats, and their pulp tissue in teeth was analyzed in terms of pulp cavity and molecules by hematoxylin and eosin and immunohistochemistry staining. Human dental pulp cells were serially subcultured and treated with glucose oxidase in the presence of elevated glucose to generate glucose oxidative stress. The replication-deficient adenovirus c-myb and small interfering RNA c-myb were introduced for c-myb expression. The pulp tissue from the diabetic rats was structurally different from normal tissue in terms of narrow pulp capacity, reduced c-myb, and dentinogenesis molecules. Glucose oxidase treatment decreased c-myb and dentinogenesis molecules (bone morphogenetic protein 2 and 7, dentin matrix protein 1, and dentin sialophosphoprotein) in human dental pulp cells. However, overexpression of c-myb by adenovirus c-myb increased dentinogenesis, autophagy molecules (autophagy protein 5, microtubule-associated protein 1A/1B-light chain 3, and Beclin-1), and cell survival via p-AMPK/AKT signaling even with glucose oxidative stress. In contrast, the lack of c-myb decreased the above molecules and cell survival by downregulating p-AMPK/AKT signaling. The results indicate that diabetes leads to irreversible damage to dental pulp, which is related to downexpression of autophagy via the p-AMPK/AKT pathway by decline of c-myb. The findings of this study provide a new insight that c-myb could ameliorate autophagy activity and that it is applicable for monitoring complicated diseases of dental pulp. The involvement of c-myb in pulp pathology could serve a therapeutic target in oral-complicated diseases.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia/fisiologia , Polpa Dentária/citologia , Diabetes Mellitus Experimental/metabolismo , Glucose/metabolismo , Estresse Oxidativo/fisiologia , Proteínas Proto-Oncogênicas c-myb/fisiologia , Animais , Western Blotting , Células Cultivadas , Dentinogênese/efeitos dos fármacos , Glucose Oxidase/farmacologia , Humanos , Imuno-Histoquímica , Masculino , RNA Interferente Pequeno/metabolismo , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Transfecção
8.
J Dent Res ; 94(5): 729-37, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25744070

RESUMO

This study investigated the relevance between pulp vitality and autophagy in aged human dental pulp cells (HDPCs) and whether peroxisome proliferator-activated receptor gamma (PPARγ) affects autophagy regulation for homeostasis in the aging progress. In vivo experiments were used in human and Sprague-Dawley rat teeth obtained from young and adult individuals. Aging- and autophagy-related molecules were determined by immunohistochemistry and hematoxylin and eosin staining. HDPCs were serially subcultured until spontaneously arrested for in vitro aging, and the replication deficiency adenovirus was introduced for PPARγ overexpression. Subsequently, the effect of PPARγ on regulation of autophagy molecules, mitochondria activity, and cell viability was assessed using Western blotting, confocal microscopy, and the MTT assay, respectively. In adult pulp tissue, autophagy molecules (autophagy protein 5, microtubule-associated protein 1A/1B light chain, and Beclin-1) were increased, but aging-related (PPARγ and heme oxygenase 1 [HO-1]) and dentinogenesis (dentin sialophosphoprotein and dentin matrix acidic phosphoprotein) molecules were decreased. In aged HDPCs, autophagy and intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 were increased, while PPARγ and HO-1 were decreased. Under stimulation with lipopolysaccharide, autophagy- and aging-related molecules were differentially expressed between young and aged cells. PPARγ induced HO-1 and autophagy molecules but reduced inflammatory molecules in aged cells. In addition, PPARγ activated strong mitochondrial activity and cell viability in aging cells. Inhibition of HO-1 by tin protoporphyrin IX exacerbated autophagy and mitochondrial activity as well as cell viability in young cells. This study indicates that PPARγ maintains pulp homeostasis through the regulation of autophagy molecules during the life span of HDPCs.


Assuntos
Envelhecimento/fisiologia , Autofagia/fisiologia , Polpa Dentária/citologia , Homeostase/fisiologia , PPAR gama/fisiologia , Adulto , Idoso , Animais , Proteínas Reguladoras de Apoptose/análise , Autofagia/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Células Cultivadas , Inibidores Enzimáticos/farmacologia , Proteínas da Matriz Extracelular/análise , Heme Oxigenase-1/análise , Heme Oxigenase-1/antagonistas & inibidores , Humanos , Molécula 1 de Adesão Intercelular/análise , Lipopolissacarídeos/farmacologia , Metaloporfirinas/farmacologia , Proteínas Associadas aos Microtúbulos/análise , Pessoa de Meia-Idade , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/fisiologia , PPAR gama/análise , Fosfoproteínas/análise , Protoporfirinas/farmacologia , Ratos , Ratos Sprague-Dawley , Sialoglicoproteínas/análise , Molécula 1 de Adesão de Célula Vascular/análise , Adulto Jovem
9.
Oncogene ; 32(18): 2282-91, 2291.e1-7, 2013 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-22797075

RESUMO

Cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is expressed in the epithelial cells of a wide range of organs/tissues from which most cancers are derived. Although accumulating reports have indicated the association of cancer incidence with genetic variations in CFTR gene, the exact role of CFTR in cancer development and the possible underlying mechanism have not been elucidated. Here, we report that CFTR expression is significantly decreased in both prostate cancer cell lines and human prostate cancer tissue samples. Overexpression of CFTR in prostate cancer cell lines suppresses tumor progression (cell growth, adhesion and migration), whereas knockdown of CFTR leads to enhanced malignancies both in vitro and in vivo. In addition, we demonstrate that CFTR knockdown-enhanced cell proliferation, cell invasion and migration are significantly reversed by antibodies against either urokinase plasminogen activator (uPA) or uPA receptor (uPAR), which are known to be involved in various malignant traits of cancer development. More interestingly, overexpression of CFTR suppresses uPA by upregulating the recently described tumor suppressor microRNA-193b (miR-193b), and overexpression of pre-miR-193b significantly reverses CFTR knockdown-enhanced malignant phenotype and abrogates elevated uPA activity in prostate cancer cell line. Finally, we show that CFTR gene transfer results in significant tumor repression in prostate cancer xenografts in vivo. Taken together, the present study has demonstrated a previously undefined tumor-suppressing role of CFTR and its involvement in regulation of miR-193b in prostate cancer development.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , MicroRNAs/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Animais , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/genética , Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Regulação para Baixo , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Masculino , Camundongos , Camundongos Nus , Dados de Sequência Molecular , Neoplasias da Próstata/metabolismo , Receptores de Ativador de Plasminogênio Tipo Uroquinase/genética , Receptores de Ativador de Plasminogênio Tipo Uroquinase/imunologia , Receptores de Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Ativador de Plasminogênio Tipo Uroquinase/genética , Ativador de Plasminogênio Tipo Uroquinase/imunologia , Ensaios Antitumorais Modelo de Xenoenxerto
10.
J Biomed Nanotechnol ; 8(3): 361-71, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22764405

RESUMO

Cancer gene therapy involves the replacement of missing or altered genes with healthy ones. In this paper, we have proposed tumor suppressor gene-carrying superparamagnetic iron oxide nanoparticles (SPIONs) for anti-cancer gene therapy. Thermally crosslinked SPIONs (TCL-SPIONs) were conjugated with branched polyethylenimine (PEI 1800 Da) by EDC-NHS chemistry for p53 plasmid DNA delivery. The morphology of the bPEI conjugated TCL-SPIONs (bPEI-TCL-SPION) and pDNA-loaded bPEI-TCL-SPION nanoparticles was measured using transmission electron microscopy (TEM). The particle sizes of the pDNA-loaded bPEI-TCL-SPION nanoparticles were also confirmed by dynamic light scattering, and ranged from 100 to 130 nm, depending on the molar charge ratio. The fluorescently labeled pDNA was complexed with bPEI-TCL-SPION and its intracellular internalization was investigated using confocal microscopy. The p53 plasmid-loaded bPEI-TCL-SPION nanoparticles achieved significantly higher p53 tumor suppressor gene expression and cellular viability compared to positive controls. The expressed wild-type p53 protein suppressed tumor cell proliferation as compared to the mutant control. When transgene expression of the p53 tumor suppressor gene was evaluated at the mRNA level and quantified using real-time PCR, the results were highly dependent on the molar charge ratio (N/P) as well as the cancer cell type. SPIONs internalized within cancer cells were tracked by magnetic resonance (MR) imaging. It was concluded that bPEI-TCL-SPION could be used as efficient gene delivery carriers that can be tracked by MR imaging.


Assuntos
Dextranos , Iminas/química , Imageamento por Ressonância Magnética/métodos , Nanopartículas de Magnetita , Nanocápsulas/química , Neoplasias Experimentais/genética , Plasmídeos/genética , Polietilenos/química , Proteína Supressora de Tumor p53/genética , Animais , Linhagem Celular Tumoral , Meios de Contraste , Perfilação da Expressão Gênica/métodos , Genes Supressores , Humanos , Camundongos , Neoplasias Experimentais/patologia , Plasmídeos/administração & dosagem
11.
Dig Liver Dis ; 38(8): 605-8, 2006 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16824812

RESUMO

BACKGROUND: Perforation and bleeding are major complications associated with gastric endoscopic mucosal resection. Evident perforation during endoscopic mucosal resection can be managed by endoscopic clipping. However, management of microperforation is not well established. PATIENT AND METHOD: From January 2002 to June 2004, 109 early gastric cancers and 300 adenomas were treated with endoscopic mucosal resection. Iatrogenic perforations occurred in 4.16% (n=17) patients. Following exclusion of four evident perforations, microperforation was observed in 3.18% (n=13) patients. The clinical features of microperforation in patients were retrospectively reviewed. RESULTS: In a total of 13 microperforation cases, 2 patients were managed surgically. The remaining patients successfully recovered without surgical management. In the case of 11 patients without surgery, 7 experienced abdominal pain, which required analgesics, 2 patients experienced mild discomfort and 2 patients experienced no symptoms. A body temperature above 37.5 degrees C was observed in 9.1% (n=1) patients and leucocytosis above 9000 microL-1 was in 72.7% (n=8) patients. The mean duration of nasogastric tube drainage was 2.36+/-1.03 days, of fasting 4.18+/-1.17 days, of intravenous antibiotics 5.55+/-1.44 days and of hospitalisation 7.45+/-1.04 days. CONCLUSION: Microperforation induced by gastric endoscopic mucosal resection can be managed successfully using a non-surgical approach including fasting, nasogastric tube drainage and intravenous antibiotics.


Assuntos
Mucosa Gástrica/cirurgia , Gastroscopia/efeitos adversos , Perfuração Intestinal/etiologia , Perfuração Intestinal/terapia , Dor Abdominal/etiologia , Dor Abdominal/terapia , Adenoma/cirurgia , Idoso , Idoso de 80 Anos ou mais , Analgésicos/uso terapêutico , Antibacterianos/uso terapêutico , Drenagem , Jejum , Feminino , Seguimentos , Mucosa Gástrica/patologia , Hemorragia Gastrointestinal/etiologia , Hemorragia Gastrointestinal/terapia , Hospitalização , Humanos , Intubação Gastrointestinal , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Neoplasias Gástricas/cirurgia , Resultado do Tratamento
13.
Cell Mol Life Sci ; 61(16): 2083-91, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15316657

RESUMO

HAb18G/CD147 is a heavily glycosylated protein containing two immunoglobulin superfamily domains. Our previous studies have indicated that overexpression of HAb18G/CD147 enhances metastatic potentials in human hepatoma cells by disrupting the regulation of store-operated Ca2+ entry by nitric oxide (NO)/cGMP. In the present study, we investigated the structure-function of HAb18G/CD147 by transfecting truncated HAb18G/CD147 fragments into human 7721 hepatoma cells. The inhibitory effect of HAb18G/CD147 on 8-bromo-cGMP-regulated thapsigargin-induced Ca2+ entry was reversed by the expression of either C or N terminus truncated HAb18G/CD147 in T7721deltaC and T7721deltaN cells, respectively. The potential effect of HAb18G/CD147 on metastatic potentials, both adhesion and invasion capacities, of hepatoma cells was abolished in T7721deltaC cells, but not affected in T7721deltaN cells. Release and activation of matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were found to be enhanced by the expression of HAb18G/CD147, and this effect was abolished by both truncations. Thapsigargin significantly enhanced release and activation of MMPs (MMP-2 and MMP-9) in non-transfected 7721 cells, and this effect was negatively regulated by SNAP. However, no effects of thapsigargin or SNAP were observed in T7721 cells, and expression of HAb18G/CD147 enhanced secretion and activation of MMPs at a stable and high level. Taken together, these results suggest that both ectodomain and intracellular domains of HAb18G/CD147 are required to mediate the effect of HAb18G/CD147 on the secretion and activation of MMPs and metastasis-related processes in human hepatoma cells by disrupting the regulation of NO/cGMP-sensitive intracellular Ca2+ mobilization although each domain may play different roles.


Assuntos
Antígenos CD/metabolismo , Antígenos de Neoplasias/metabolismo , Cálcio/metabolismo , Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , Basigina , Carcinoma Hepatocelular/patologia , Humanos , Neoplasias Hepáticas/patologia , Inibidores de Metaloproteinases de Matriz , Metaloproteinases da Matriz/metabolismo , Metástase Neoplásica , Estrutura Terciária de Proteína , Tapsigargina/farmacologia
14.
Biol Reprod ; 68(1): 302-8, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12493726

RESUMO

The luminal fluid microenvironment of the uterus is important for sperm capacitation and embryo development. In an attempt to understand the possible role of Na(+)/H(+) exchangers (NHEs) in uterine function, the mRNAs of different NHE isoforms as well as their subcellular localization (apical versus basolateral) and functional activity were investigated in mouse endometrial epithelial cells using reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemistry, and intracellular pH (pH(i)) measurement techniques. The presence of NHE1, NHE2, and NHE4, but not NHE3 mRNAs were revealed by RT-PCR. Immunostaining showed that NHE1, NHE2, and NHE4 were present in both apical and basolateral membranes. The pH(i) recovery from intracellular acidification was Na(+)-dependent; however, the rate of pH(i) recovery depending on basolateral Na(+) was 12.4 times faster than that depending on apical Na(+). The Na(+)-dependent rate of pH(i) recovery was also inhibited by amiloride, indicating H(+) extrusion through NHEs; however, the amiloride sensitivity of the apical membrane was less than that of the basolateral membrane, suggesting the involvement of different types of NHEs in the two membranes. The results indicate that the basolaterally located NHE1, NHE2, and NHE4, in addition to participating in the homeostatic control of intracellular pH, may play a role in H(+) extrusion in order to achieve transepithelial HCO(3)(-) secretion. The apically located NHEs may be involved in mediating Na(+) absorption as alternatives of or complementary to epithelial Na(+) channels.


Assuntos
Endométrio/metabolismo , Trocadores de Sódio-Hidrogênio/genética , Trocadores de Sódio-Hidrogênio/metabolismo , Animais , Sequência de Bases , DNA/genética , Epitélio/metabolismo , Feminino , Expressão Gênica , Concentração de Íons de Hidrogênio , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos ICR , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
15.
Med Phys ; 29(11): 2606-10, 2002 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-12462727

RESUMO

The Enhanced Dynamic Wedge (EDW) presents many advantages over the physical wedge. However, in order to calculate monitor units (MUs) necessary to deliver a certain dose at a certain point, EDW factors (EDWFs) need to be determined. In this work, based on analysis of the golden segmented treatment table (GSTT) and the MU fraction model, an empirical analytic formula has been developed to calculate EDW factors for symmetric and asymmetric fields. This formalism is an extension of the MU fraction model. However in comparison with previous studies [J. P. Gibbons, Med. Phys. 25, 1411-1418 (1998) and M. Miften et al., Med. Dosim. 25, 81-86 (2000)], this formula is simpler, and easier to use. It is applicable to EDW fields of different sizes, wedge angles and different photon energies. For 6 and 18 MV beams from a Varian 21EX accelerator with 7 EDW angles (Varian Oncology Systems, Palo Alto, CA), more than 250 measured EDWFs for symmetric and asymmetric fields with different off-axis distances and field sizes were compared with model calculations. Results show that 80% and 98% of calculated EDWFs match corresponding measured values to within 0.5% and 1.0%, respectively, the maximum deviation being 1.3%.


Assuntos
Algoritmos , Radiometria/métodos , Planejamento da Radioterapia Assistida por Computador/métodos , Radioterapia de Alta Energia/métodos , Fótons , Controle de Qualidade , Radiometria/normas , Dosagem Radioterapêutica , Planejamento da Radioterapia Assistida por Computador/normas , Sensibilidade e Especificidade
16.
Biol Reprod ; 66(6): 1846-52, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12021071

RESUMO

The present study investigated the involvement of Na+-HCO3- cotransporter in mediating cAMP-stimulated HCO3- secretion across the cultured mouse endometrial epithelium using the short-circuit current (I(SC)) technique and intracellular pH measurement. Forskolin stimulated a rise in the I(SC), 55.6% and 52.1% of which could be reduced by the removal of extracellular Cl- or by eliminating the contribution of Cl- secretion by bumetanide, an inhibitor of Na+-K+-2Cl- cotransporter, respectively. More than 80% reduction in the forskolin-induced I(SC) was obtained when both Cl- and HCO3- in the bath were removed or in HCO3--free solution with bumetanide, indicating that the I(SC) depended on both Cl- and HCO3-. The presence of the Na+ channel-blocker amiloride in the apical solution did not reduce the forskolin-induced I(SC); however, the I(SC) could be abolished by removing Na+ from the bathing solution, suggesting that the Cl-- and HCO3--dependent I(SC) was also dependent on basolateral Na+. The forskolin-stimulated I(SC) could be reduced 43.6% by removal of HCO3- and 47.9% by a Na+-HCO3--cotransporter inhibitor, dihydrogen-4,4'-didsothiocyanostilbene-2,2'-disulfonic acid (H2DIDS). The inhibitory effect of H2DIDS was observed in Cl--free solution, but not when HCO3- was removed, thus confirming its effect on HCO3--dependent transport. Intracellular pH measurements demonstrated that the recovery from cellular acidification depended on the presence of both basolateral Na+ and HCO3-, further indicating the involvement of Na+-HCO3- cotransporter. Reverse transcription-polymerase chain reaction experiments confirmed the expression of Na+-HCO3- cotransporter in the mouse endometrium. The results suggest that basolaterally located Na+-HCO3- cotransporter is involved in mediating cAMP-stimulated HCO3- secretion across the mouse endometrial epithelium.


Assuntos
Ácido 4,4'-Di-Isotiocianoestilbeno-2,2'-Dissulfônico/análogos & derivados , Bicarbonatos/metabolismo , AMP Cíclico/farmacologia , Endométrio/fisiologia , Simportadores de Sódio-Bicarbonato/fisiologia , Ácido 4,4'-Di-Isotiocianoestilbeno-2,2'-Dissulfônico/farmacologia , Animais , Bicarbonatos/farmacologia , Células Cultivadas , Cloretos/farmacologia , Colforsina/farmacologia , Condutividade Elétrica , Endométrio/efeitos dos fármacos , Endométrio/metabolismo , Epitélio/efeitos dos fármacos , Epitélio/fisiologia , Feminino , Expressão Gênica , Concentração de Íons de Hidrogênio , Camundongos , Camundongos Endogâmicos ICR , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sódio/farmacologia , Simportadores de Sódio-Bicarbonato/antagonistas & inibidores , Simportadores de Sódio-Bicarbonato/genética
17.
Bioorg Med Chem Lett ; 11(24): 3107-10, 2001 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-11720853

RESUMO

P(1)-Citronellyl-P(2)-alpha-D-pyranosyl pyrophosphates containing alpha-D-N-acetylglucoseaminyl, alpha-D-glucosyl, and alpha-D-N-acetylmuramyl carbohydrates were synthesized and used in substrate specificity studies of the Escherichia coli MurG enzyme. Oxalyl chloride activation of citronellyl phosphate for coupling to alpha-D-pyranose-1-phosphates resulted in markedly improved yields over traditional Khorana-Moffatt and diphenyl chlorophosphate activation strategies.


Assuntos
Proteínas da Membrana Bacteriana Externa , Difosfatos/síntese química , Escherichia coli/enzimologia , N-Acetilglucosaminiltransferases/metabolismo , Difosfatos/metabolismo , Espectroscopia de Ressonância Magnética , Espectrometria de Massas por Ionização por Electrospray , Especificidade por Substrato
18.
J Biol Chem ; 276(50): 46870-7, 2001 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-11591720

RESUMO

The present study examined the effect of hepatoma-associated antigen HAb18G (homologous to CD147) expression on the NO/cGMP-regulated Ca(2+) mobilization and metastatic process of human hepatoma cells. HAb18G/CD147 cDNA was transfected into human 7721 hepatoma cells to obtain a cell line stably expressing HAb18G/CD147, T7721, as demonstrated by Northern blot and immunocytochemical studies. 8-Bromo-cGMP (cGMP) inhibited the thapsigargin-induced Ca(2+) entry in a concentration-dependent manner in 7721 cells. The cGMP-induced inhibition was abolished by an inhibitor of protein kinase G, KT5823 (1 microm). However, expression of HAb18G/CD147 in T7721 cells decreased the inhibitory response to cGMP. A similar concentration-dependent inhibitory effect on the Ca(2+) entry was observed in 7721 cells in response to a NO donor, (+/-)-S-nitroso-N-acetylpenicillamine (SNAP). The inhibitory effect of SNAP on the thapsigargin-induced Ca(2+) entry was significantly reduced in HAb18G/CD147-expressing T7721 cells, indicating a role for HAb18G/CD147 in NO/cGMP-regulated Ca(2+) entry. Experiments investigating metastatic potentials demonstrated that HAb18G/CD147-expressing T7721 cells attached to the Matrigel-coated culture plates and invaded through Matrigel-coated permeable filters at the rate significantly greater than that observed in 7721 cells. Both the attachment and invasion rates could be suppressed by SNAP, and the inhibitory effect of SNAP could be reversed by NO inhibitor, N(G)-nitro-l-arginine methyl ester. The sensitivity of the attachment and invasion rates to cGMP was significantly reduced in T7721 cells as compared with 7721 cells when cells were pretreated with thapsigargin. The difference in the sensitivity between the two cells could be abolished by a Ca(2+) channel blocker, Ni(2+) (3 mm). These results suggest that HAb18G/CD147 enhances metastatic potentials in human hepatoma cells by disrupting the regulation of store-operated Ca(2+) entry by NO/cGMP.


Assuntos
Antígenos CD , Antígenos de Neoplasias , Antígenos de Superfície , Proteínas Aviárias , Proteínas Sanguíneas , Carbazóis , Carcinoma Hepatocelular/metabolismo , GMP Cíclico/análogos & derivados , Indóis , Neoplasias Hepáticas/metabolismo , Glicoproteínas de Membrana/metabolismo , Glicoproteínas de Membrana/fisiologia , Penicilamina/análogos & derivados , Alcaloides/farmacologia , Basigina , Northern Blotting , Cálcio/metabolismo , Adesão Celular , Linhagem Celular , Movimento Celular , Colágeno/química , GMP Cíclico/metabolismo , Proteínas Quinases Dependentes de GMP Cíclico/antagonistas & inibidores , DNA Complementar/metabolismo , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Inibidores Enzimáticos/farmacologia , Humanos , Imuno-Histoquímica , Laminina/química , NG-Nitroarginina Metil Éster/farmacologia , Metástase Neoplásica , Níquel/metabolismo , Óxido Nítrico/metabolismo , Penicilamina/farmacologia , Proteoglicanas/química , Transdução de Sinais , Tapsigargina/farmacologia , Fatores de Tempo , Transfecção , Células Tumorais Cultivadas
19.
Cell Biol Int ; 25(10): 993-5, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11589615

RESUMO

This study aimed to investigate cGMP-regulated store-operated Ca(2+)entry in human 7721 hepatoma cells. [Ca(2+)](i)was measured using Fura2/AM. After incubation of the cells with 4 microm thapsigargin, Ca(2+)entry was evoked by application of 1 mMm Ca(2+)to extracellular solution and was blocked by 3 m m Ni(2+), indicating the presence of store-operated Ca(2+)entry in human 7721 hepatoma cell line. Application of 8-Br-cGMP reduced the [Ca(2+)](i)in hepatoma 7721 cells by 80%. These data demonstrated for the first time that store-operated Ca(2+)entry pathway is present in human hepatoma cells, which is regulated by cGMP.


Assuntos
Canais de Cálcio/metabolismo , Cálcio/metabolismo , Carcinoma Hepatocelular/metabolismo , GMP Cíclico/análogos & derivados , GMP Cíclico/fisiologia , Neoplasias Hepáticas/metabolismo , ATPases Transportadoras de Cálcio/antagonistas & inibidores , GMP Cíclico/farmacologia , Inibidores Enzimáticos/farmacologia , Humanos , Níquel/farmacologia , Tapsigargina/farmacologia , Células Tumorais Cultivadas
20.
Urol Clin North Am ; 28(1): 53-60, viii, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11277069

RESUMO

The pediatric laparoscopic nephrectomy/nephroureterectomy literature is reviewed. The authors' method for pediatric nephrectomy/nephroureterectomy is presented, as well as the clinical experience from Washington University.


Assuntos
Nefropatias/cirurgia , Laparoscópios , Nefrectomia/instrumentação , Ureter/cirurgia , Refluxo Vesicoureteral/cirurgia , Adolescente , Criança , Pré-Escolar , Desenho de Equipamento , Feminino , Humanos , Lactente , Masculino , Complicações Pós-Operatórias/etiologia , Grampeadores Cirúrgicos , Técnicas de Sutura/instrumentação
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