Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 49
Filtrar
1.
Zhonghua Yu Fang Yi Xue Za Zhi ; 55(2): 280-283, 2021 Feb 06.
Artigo em Chinês | MEDLINE | ID: mdl-34645194

RESUMO

This paper summarizes the development trend and characteristics of public opinion on health protection and disinfection strategies in the COVID-19 epidemic. The experience and deficiency of the strategies are discussed from the perspective of public opinion, and suggestions on how health protection and disinfection can help prevent and control infectious diseases are also put forward: to strengthen health protection and disinfection in key places and units; to evaluate health protection and disinfection effects as well as the transmission mechanism of virus in the environment; to establish a professional health protection and disinfection emergency science popularization mechanism and information release channel; to speed up the formulation and revision of health protection and disinfection standards related to the epidemic.

2.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 33(4): 359-364, 2021 Aug 30.
Artigo em Chinês | MEDLINE | ID: mdl-34505442

RESUMO

OBJECTIVE: To evaluate the impact of environmental and climatic factors on the distribution of suitable habitats of Haemaphysalis longicornis, and to predict the potential distribution of H. longicornis under different climate patterns in China. METHODS: Data pertaining to the distribution of H. longicornis were retrieved from public literatures. The effects of 19 climatic factors (annual mean temperature, annual mean temperature difference between day and night, isothermality, standard deviation of seasonal variation of temperature, maximum temperature of the warmest month, minimum temperature of the coldest month, temperature annual range, mean temperature of the wettest season, mean temperature of the driest season, mean temperature of the warmest season, mean temperature of the coldest season, annual mean precipitation, precipitation of the wettest month, precipitation of the driest month, coefficient of variance of precipitation, precipitation of the wettest season, precipitation of the driest season, precipitation of the warmest season and precipitation of the coldest season) and 4 environmental factors (elevation, slope, slope aspect and vegetation coverage) on the potential distribution of H. longicornis were assessed using the maximum entropy (MaxEnt) model based on the H. longicornis distribution data and climatic and environmental data, and the potential distribution of H. longicornis was predicted under the RCP 2.6 and 8.5 emissions scenarios. RESULTS: Among the environmental and climatic factors affecting the geographical distribution of H. longicornis in China, the factors contributing more than 10% to the distribution of H. longicornis mainly included the precipitation of the driest month (26.0%), annual mean temperature (11.2%), annual mean precipitation (10.0%) and elevation (24.2%). Under the current climate pattern, the high-, medium- and low-suitable habitats of H. longicornis are 1 231 900, 1 696 200 km2 and 1 854 400 km2 in China, respectively. The distribution of H. longicornis increased by 336 100 km2 and 367 300 km2 in 2050 and 2070 under the RCP 2.6 emissions scenario, and increased by 381 000 km2 and 358 000 km2 in 2050 and 2070 under the RCP 8.5 emissions scenario in China, respectively. CONCLUSIONS: Climatic and environmental factors, such as precipitation, temperature and elevation, greatly affect the distribution of H. longicornis in China, and the suitable habitats of H. longicornis may expand in China under different climate patterns in future.


Assuntos
Mudança Climática , Ecossistema , China , Clima , Previsões , Temperatura
3.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 33(3): 267-273, 2021 Jun 30.
Artigo em Chinês | MEDLINE | ID: mdl-34286528

RESUMO

OBJECTIVE: To identify the environmental factors affecting the geographical distribution of Rhipicephalus microplus in China, and to examine the impact of climate changes on the distribution of R. microplus in China. METHODS: The national and international publications pertaining to the geographical distribution of R. microplus in China were retrieved, and the geographical location was extracted. The suitable habitats of R. microplus and the dominant environmental factors affecting the distribution of suitable habitats of R. microplus were predicted in China based on the geographical data and environmental variables using the ArcGIS 10.7 software and the maximum entropy model. RESULTS: Among the main climatic factors affecting the geographical distribution of R. microplus in China, the factors contributing more than 10% to the suitable habitats of R. microplus mainly include the annual mean precipitation (38.2%), the average temperature of the coldest quarter (28.4%) and the precipitation of the driest month (14.2%). The current suitable habitats of R. microplus were mainly found in southern China, and the high-, medium- and low-suitable areas accounted for 8.6%, 13.1% and 10.5% of the total land area of China, respectively. The suitable habitats of R. microplus were predicted to increase by 399 800 km2 in China using the maximum entropy model under the RCP 4.5 emissions scenario in 2070, and the emerging suitable habitats were mainly distributed in Gansu Province, Ningxia Hui Autonomous Region, Qinghai Province, Hebei Province, Shaanxi Province, Liaoning Province, Inner Mongolia Autonomous Region, Shandong Province, Sichuan Province and Tibeten Autonomous Region. In addition, the suitable habitats of R. microplus were predicted to show an overall expansion towards northward from present to 2070. CONCLUSIONS: Climate changes affect the distribution of suitable habitats of R. microplus in China, and annual mean precipitation may be a key factor affecting the distribution of suitable habitats of R. microplus.


Assuntos
Mudança Climática , Rhipicephalus , Animais , China , Ecossistema , Temperatura
4.
Beijing Da Xue Xue Bao Yi Xue Ban ; 53(3): 485-490, 2021 Jun 18.
Artigo em Chinês | MEDLINE | ID: mdl-34145849

RESUMO

OBJECTIVE: To describe the distribution and trend of infantile epilepsy among infants under 36 months in Ningbo, Zhejiang Province. METHODS: Using the birth cohort design, we retrospectively collected the local born infants in Ningbo national health information platform from 2015 to 2019, and took the first visit of epilepsy in the electronic medical record of the platform as the new case. The incidence density and 95% confidence interval (CI) of epilepsy were estimated by Poisson distribution. RESULTS: From 2015 to 2019, a total of 294 900 children were born in Ningbo, with male accounting for 51.92%. The total person-years of observation were 595 300, while the median follow-up person-years was 2.31 [interquartile range (IQR): 1.90]. There were 575 new onset epilepsy patients during the whole observation period. The total number of visits was 2 599, with an average of 4.52. The total incidence density was 96.59/100 000 person-years (95%CI: 88.85-104.82). The median age of onset was 13 months (IQR: 15), 0-12 months old infants had the highest incidence density (102.18/100 000 person-years), 25-36 months old infants had the lowest incidence density (89.68/100 000 person-years), and the difference was not statistically significant (P>0.05). The incidence density of male was 97.58/100 000 person-years, female was 95.53/100 000 person-years, and the difference was not statistically significant (P>0.05). Fenghua was the highest (130.54/100 000 person-years, 95%CI: 94.47-175.83) and Ninghai was the lowest (66.44/100 000 person-years, 95%CI: 47.02-91. 19), with significant difference (P < 0.05). There was no significant difference in the incidence density in different birth years (P>0.05). There was significant difference in the incidence density between 0-12 months old infants in different calendar years (Ptrend < 0.05). In this age group, the incidence density was the lowest in 2015 (69.41/100 000 person-years, 95%CI: 41.79-108.39), and the highest in 2019 (225.61/100 000 person-years, 95%CI: 186.10-271.03). There was no significant difference in the incidence density between 13-24 and 25-36 months old infants in different calendar years (P>0.05). CONCLUSION: The incidence density of epilepsy in 0-36 months old infants in Ningbo City from 2015 to 2019 was low as a whole, and there was no difference in age group, gender, and year of birth. The incidence density of 0-12 months old infants increased with the year.


Assuntos
Epilepsia , Pré-Escolar , Cidades , Epilepsia/epidemiologia , Feminino , Humanos , Incidência , Lactente , Recém-Nascido , Masculino , Estudos Retrospectivos
5.
Zhonghua Yu Fang Yi Xue Za Zhi ; 55(3): 424-427, 2021 Mar 06.
Artigo em Chinês | MEDLINE | ID: mdl-33730839

RESUMO

In this paper, the history and current situation of environmental health standardization in China are reviewed, and the experience and shortcomings in the process of environmental health standardization in China are analyzed, suggestions for the next step of environmental health standards are also put forward.


Assuntos
Saúde Ambiental , China , Humanos , Padrões de Referência
6.
Zhonghua Yu Fang Yi Xue Za Zhi ; 53(12): 1193-1197, 2019 Dec 06.
Artigo em Chinês | MEDLINE | ID: mdl-31795573

RESUMO

The Health Environment Promotion Campaigns (HEPCs) focus on the major environmental health issues and relevant factors of concern among the general public, and promote the achievement of the national health goal. Based on the summary and analysis of the background, key indicators, specific actions in different domains of the HEPCs, this paper proposes suggestions for scientifically implementing HEPCs from five aspects, namely, formulating implementation plans, establishing pilot areas, building comprehensive service platforms, improving the health literacy of residents and strengthening the development of protection technologies and standards.


Assuntos
Letramento em Saúde , Promoção da Saúde , Humanos
7.
Beijing Da Xue Xue Bao Yi Xue Ban ; 51(5): 893-899, 2019 Oct 18.
Artigo em Chinês | MEDLINE | ID: mdl-31624395

RESUMO

OBJECTIVE: To evaluate the change of cell surface CXC chemokine receptor 4 (CXCR4) expression of stem cells from apical papilla (SCAP) after the inhibition of endocytotic pathway, thus to provide experimental basis for the mechanism of SCAP migration. METHODS: The immunofluorescence analysis was conducted to examine the co-expression of CXCR4 and endocytotic compartments, including early endosomes, recycling endosomes and lysosomes in SCAP. Several Rab proteins were applied as markers of organelles in the endocytotic pathway, including Rab5 for early endosomes, Rab11A for recycling endosomes, and Lamp1 for lysosomes. The co-localization of CXCR4 with these endodontic compartments was further observed by proximity ligation assay (PLA). SCAP was treated with two kinds of endocytotic inhibitors, Blebbistatin and Dynasore, at a concentration of 80 µmol/L, respectively. The conditioning time was 1 hour. Flow cytometry was carried out to evaluate the proportion of SCAP that expressed CXCR4 on cell surface. The data were analysed by analysis of variance (ANOVA). RESULTS: The red staining of CXCR4 on immunofluorescence confocal microscopy predominantly overlapped with the green staining of Rab5 and Rab11A, and partly overlapped with Lamp1. It indicated that most CXCR4 molecules were located in early endosomes and recycling endosomes, and some were located in lysosomes. The PLA results revealed that the co-localizaiton of CXCR4 with endocytotic compartments could be observed in early endosomes, recycling endosomes and lysosomes. According to the results of flow cytometry, the proportion of SCAP that expressed CXCR4 on cell surface was as low as 0.13%±0.10%. After the inhibition of endocytosis by pretreating the cells with the following two inhibitors, Blebbistatin and Dynasore, the percentage of SCAP that positively expressed CXCR4 on cell surface was significantly increased to 13.34%±1.31% in Blebbistatin group and 4.03%±0.92% in Dynasore group (F=16.721, P<0.001). Moreover, the number of SCAP that expressed CXCR4 on cell surface in Blebbistatin group was significantly higher than that in Dynasore group (P<0.001). CONCLUSION: The inhibition of endocytotic pathway could increase the number of SCAP that expressed CXCR4 on cell surface, and provide potency for the migration of SCAP.


Assuntos
Endocitose , Receptores CXCR4 , Endossomos , Lisossomos , Células-Tronco
8.
Zhonghua Yu Fang Yi Xue Za Zhi ; 53(9): 865-870, 2019 Sep 06.
Artigo em Chinês | MEDLINE | ID: mdl-31474065

RESUMO

According to different epochs and development needs, a series of practices on environmental health and sanitary engineering were carried out, which played significant roles in promoting national economic and social developments and protecting the public health. This paper reviewed the main achievements in the past 70 years infields of patriotic health campaign, water sanitation and toilet improvement in rural areas, surveillance and investigation, health standard system, sanitary engineering equipment, stove improvement etc., and then proposed several prospects in the future.


Assuntos
Saúde Ambiental , População Rural , Engenharia Sanitária , Saneamento , China , Saúde Ambiental/normas , Saúde Ambiental/tendências , Humanos , Saúde Pública/normas , Engenharia Sanitária/tendências , Saneamento/normas , Saneamento/tendências , Toaletes/normas
9.
Zhonghua Yu Fang Yi Xue Za Zhi ; 53(7): 724-730, 2019 Jul 06.
Artigo em Chinês | MEDLINE | ID: mdl-31288345

RESUMO

Objective: To identify post-marketing active surveillance systems for vaccine safety around the world and understand their features and mechanisms, in order to provide guidance for vaccine administration activities in China. Methods: Following the steps of scoping review, literature about active surveillance system for vaccine safety and published by 30 June 2018 were identified by searching electronic databases, including PubMed, Scopus, and Cochrane Library. Grey literature were also sought by exploring relevant websites. Identified literature were screened according to eligibility criteria, and informative data from included literature were then charted. Framework Synthesis and Thematic Analysis were performed to integrate the charted data. Results: 97 pieces of literature were included for review, and 11 active surveillance systems for vaccine safety were identified, mostly located in developed countries. These systems were constructed by 3 types of organizations: administration departments, academic or research institutions, and health care providers. Their data sources included immunization registries, electronic medical records, claims data, case reports of adverse events following immunization electronic questionnaires, and epidemiologic study data. According to their operation procedures, these systems were grouped into 4 modes of active surveillance: Data Linkage, Investigator Network, Automatic Follow-up System, Studies Consortium. Conclusion: Practice of active surveillance for vaccine safety greatly varies across countries, with different conditions and advantages. It is suggested that developing countries should choose suitable mode of active surveillance considering their local situations.


Assuntos
Vigilância de Produtos Comercializados/métodos , Vacinas/efeitos adversos , China , Países em Desenvolvimento , Humanos
10.
Eur Rev Med Pharmacol Sci ; 22(20): 6912-6921, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30402857

RESUMO

OBJECTIVE: This study aims to explore whether homeobox A11 antisense RNA (HOXA11-AS) could regulate inflammation induced by diabetic arteriosclerosis (DAA) via PI3K/AKT pathway. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect expressions of HOXA11-AS and proinflammatory genes in carotid endarterectomy samples of symptomatic and asymptomatic atherosclerosis (AS) patients, diabetes mellitus (DM), and non-DM patients. The above-mentioned genes in DM animal model and non-DM animal model were also detected. We detected the expression of HOXA11-AS in vascular smooth muscle cells (VSMCs) treated with platelet-derived growth factor (PDGF) or PDGF inhibitor imatinib, respectively. Subsequently, we applied cell transfection technology to interfere with the expression of HOXA11-AS in VSMCs. In vascular endothelial cells (VECs) and VSMCs, we detected the effect of HOXA11-AS on the expressions of genes related to the proliferation, migration, and cell cycle. Then, VSMCs were treated with tumor necrosis factor-α (TNF-α), and the expression of HOXA11-AS was examined in VSMCs. The effect of HOXA11-AS on TNF-α-induced inflammation in VSMCs was detected as well. Finally, we analyzed the effect of HOXA11-AS on PDGF-induced activation of PI3K/AKT pathway in VSMCs and VECs. RESULTS: HOXA11-AS expression was markedly increased in carotid endarterectomy specimens of symptomatic AS patients compared to that of asymptomatic AS patients. Expression levels of HOXA11-AS and pro-inflammatory genes were significantly elevated in carotid endarterectomy specimens of DM patients. Similarly, HOXA11-AS expression was also significantly increased in carotid arteries of DM mice compared with that of non-DM mice. PDGF could upregulate HOXA11-AS expression in VSMCs, which was reversed by PDGF inhibitor imatinib. HOXA11-AS knockdown could reduce the expressions of the proliferation-associated gene (PCNA) and the cycle-related genes (p21, p53), and also inhibited the proliferation and migration of VSMCs induced by PDGF. HOXA11-AS was upregulated by TNF-α. HOXA11-AS knockdown remarkably downregulated expressions of inflammation-related genes in VSMCs induced by TNF-α. In VECs, low expression of HOXA11-AS can inhibit the expression of TNF-α-induced pro-inflammatory genes and PDGF-induced vascular inflammation-related genes. Low expression of HOXA11-AS inhibited PDGF-induced activation of PI3K/AKT pathway in VSMCs and VECs. CONCLUSIONS: HOXA11-AS may participate in DAA by activating the PI3K/AKT pathway to regulate inflammation in VSMCs and VECs.


Assuntos
Arteriosclerose/genética , Angiopatias Diabéticas/etiologia , Proteínas de Homeodomínio/genética , Animais , Ciclo Celular/genética , Proliferação de Células/genética , Células Cultivadas , Regulação para Baixo , Células Endoteliais/metabolismo , Humanos , Inflamação/metabolismo , Camundongos , Miócitos de Músculo Liso/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fator de Crescimento Derivado de Plaquetas/administração & dosagem , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/genética , Regulação para Cima
11.
Eur Rev Med Pharmacol Sci ; 19(19): 3723-30, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26502863

RESUMO

OBJECTIVE: Rat models of hypobaric hypoxia-induced pulmonary hypertension are commonly used in studies of chronic mountain sickness, while there are few researches specially focusing on these rats model. This study aims to exploring possible pathogenesis of hypobaric hypoxia-induced pulmonary hypertension by experimenting on hypobaric hypoxia-induced PH rat models at different simulate- altitudes. MATERIALS AND METHODS: 32 healthy male SD rats were randomly divided into six groups of different degree and time period of hypobaric hypoxia. The mean pulmonary arterial pressure (m PAP), right ventricular pressure (RVSP), the right ventricle (RV), left ventricular (LV), ventricular septal (S), the right ventricular hypertrophy index (RVHI) [calculated under the formula of RV / (LV + S)], hematoxylin-eosin staining, elastic fibers staining, the ratio of the thickness of vascular wall to its outer diameter (MT%), the ratio of the cross-sectional area of the middle vascular wall to the total vascular cross-sectional area (MA%); the α-SMA, and the Ki6 expressions were detected to evaluated the pulmonary hypertension. RESULTS: There were significant differences of the mPAP, RVSP and RVHI value between the hypobaric hypoxia groups and the control group (p < 0.05). The mPAP, RVSP, RVHI, MT%, MA%, α-SMA, and Ki6 of rats in model groups at an altitude of 3KM were higher than those of the control group, which raised gradually with the number of weeks increasing. The mPAP, RVSP, RV / (LV + S) value, MT%, MA%, α-SMA, and Ki67 of the 5KM-4W group were significantly higher than those of the control group (p < 0.05). CONCLUSIONS: Rat models with pulmonary hypertension at different altitudes have been successfully established by automatic adjusting hypobaric hypoxia chamber. Exposure to a low oxygen environment at a simulate-altitude of 3 km for 8 weeks have caused the pathological remodeling of pulmonary vascular walls and pulmonary hypertension, and further led to a series of pathological changes, including right ventricular hypertrophy. This model is easy to be replicated with good reproducibility and provides evidence for clinical trial of drugs.


Assuntos
Doença da Altitude/complicações , Hipóxia Celular/fisiologia , Hipertensão Pulmonar/fisiopatologia , Artéria Pulmonar/fisiopatologia , Doença da Altitude/fisiopatologia , Animais , Modelos Animais de Doenças , Hipóxia/fisiopatologia , Pulmão/patologia , Masculino , Ratos , Ratos Sprague-Dawley
12.
Genet Mol Res ; 13(2): 3940-6, 2014 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-24938604

RESUMO

We investigated the clinical significance of RUNX3 gene expression in human pancreatic carcinoma. Five samples of pancreatic tissues and 30 samples of pancreatic cancer tissues and paracancerous tissues were collected. RUNX3 expression was detected by real-time PCR and immunohistochemistry. The relationships between clinicopathological findings and the expression of RUNX3 were analyzed. The relative quantification level of RUNX3 mRNA expression in human pancreatic carcinoma tissues and paracancerous tissues was 2.60 (0.42-12.82) and 1.02 (0.19-3.58), respectively (P < 0.05). The percentage of positive cells expressing RUNX3 protein in human pancreatic tissues and paracancerous tissues was 45.5 ± 26.2 and 6.9 ± 6.0%, respectively (P < 0.01). The high RUNX3 group (N = 9) with 45.5% or more of the cancer cells staining for RUNX3 and the low RUNX3 group (N = 21) with less than 45.5% cancer cells staining for RUNX3. Low expression of RUNX3 correlated significantly with an advanced TNM stage (χ(2) = 6.897, P = 0.045), lymph node metastasis (χ(2) = 4.739, P = 0.029) and neural invasion (χ(2) = 5.44, P = 0.020). On the other hand, no association could be found between RUNX3 expression and clinicopathological variables including age, gender, tumor location, tumor size, tumor differentiation or the serum concentration of CEA and CA199. The expression of RUNX3 in pancreatic cancer tissues was obviously higher than that in the paracancerous tissues. Low expression of RUNX3 may have an important role in aggressiveness, lymph node metastasis and neural invasion in pancreatic cancer. In pancreatic carcinoma tissues, low expression of RUNX3 may indicate a poor prognosis.


Assuntos
Adenocarcinoma/genética , Subunidade alfa 3 de Fator de Ligação ao Core/biossíntese , Neoplasias Pancreáticas/genética , Prognóstico , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Subunidade alfa 3 de Fator de Ligação ao Core/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias Pancreáticas/patologia
13.
Gene Ther ; 20(2): 201-14, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22436965

RESUMO

Achieving transgene integration into preselected genomic sites is currently one of the central tasks in stem cell gene therapy. A strategy to mediate such targeted integration involves site-specific endonucleases. Two genomic sites within the MBS85 and chemokine (C-C motif) receptor 5 (CCR5) genes (AAVS1 and CCR5 zinc-finger nuclease (CCR5-ZFN) sites, respectively) have recently been suggested as potential target regions for integration as their disruption has no functional consequence. We hypothesized that efficient transgene integration maybe affected by DNA accessibility of endonucleases and therefore studied the transcriptional and chromatin status of the AAVS1 and CCR5 sites in eight human induced pluripotent stem (iPS) cell lines and pooled CD34+ hematopoietic stem cells (HSCs). Matrix chromatin immunoprecipitation (ChIP) assays demonstrated that the CCR5 site and surrounding regions possessed a predominantly closed chromatin configuration consistent with its transcriptional inactivity in these cell types. In contrast, the AAVS1 site was located within a transcriptionally active region and exhibited an open chromatin configuration in both iPS cells and HSCs. To show that the AAVS1 site is readily amendable to genome modification, we expressed Rep78, an AAV2-derived protein with AAVS1-specific endonuclease activity, in iPS cells after adenoviral gene transfer. We showed that Rep78 efficiently associated with the AAVS1 site and triggered genome modifications within this site. On the other hand, binding to and modification of the CCR5-ZFN site by a ZFN was relatively inefficient. Our data suggest a critical influence of chromatin structure on efficacy of site-specific endonucleases used for genome editing.


Assuntos
Cromatina/química , Marcação de Genes , Genoma Humano , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Transgenes , Antígenos CD34/genética , Antígenos CD34/metabolismo , Cromatina/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Dependovirus/genética , Endodesoxirribonucleases/genética , Endodesoxirribonucleases/metabolismo , Loci Gênicos , Vetores Genéticos , Células-Tronco Hematopoéticas/química , Humanos , Células-Tronco Pluripotentes Induzidas/química , Proteína Fosfatase 1/genética , Proteína Fosfatase 1/metabolismo , Receptores CCR5/genética , Receptores CCR5/metabolismo , Transcrição Genética , Proteínas Virais/genética , Proteínas Virais/metabolismo , Dedos de Zinco/genética
14.
Scand J Immunol ; 68(1): 12-21, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18482205

RESUMO

Monoclonal antibody (mAb) has been widely applied in the treatment of human diseases, especially in malignant tumours. However, most antibodies produced in mouse by hybridoma technology might induce severe human anti-mouse reactions. We had reported a murine mAb CAb-1 of therapeutic interest for its specifically binding to a cell surface glycoprotein of human colon cancer. Here, we attempted to generate a reconstituted human-mouse chimeric Fab (cFab) of CAb-1 in vitro to reduce its antigenicity and increase its capacity of penetration. First, the genes of heavy and light chain variable region (VH, VL) of CAb-1 were cloned. Then, the chimeric light chain (cL) and Fd (cFd) were constructed and expressed in Escherichia coli. Finally, the reconstituted cFab was obtained by gradient dialysis of the mixture of cFd and cL. SDS-PAGE and western blot analysis showed the reconstituted cFab with a recovery rate of 70.2% when the initial total concentration of cL and cFd proteins to be 100 microg/ml. The reconstituted cFab maintained the affinity and specificity to colon cancer cells compared with its parental antibody as determined by immunostaining analysis, FACS and ELISA. Our results established a foundation for further application of the cFab in diagnosis and treatment of colon cancer.


Assuntos
Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Antígenos de Neoplasias/imunologia , Neoplasias do Colo/imunologia , Fragmentos Fab das Imunoglobulinas/genética , Fragmentos Fab das Imunoglobulinas/imunologia , Animais , Especificidade de Anticorpos , Western Blotting , Quimera , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Genes Sintéticos , Vetores Genéticos , Humanos , Hibridomas , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
15.
Scand J Immunol ; 65(5): 435-43, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17444954

RESUMO

The hepatocellular carcinoma-associated antigen HAb18G/CD147, a member of CD147 family, could promote tumour invasion and metastasis via inducing the secretion of matrix metalloproteinases (MMP). Anti-CD147 monoclonal antibodies (MoAb) have exhibited obvious inhibitory effect on MMP induction. However, none of the epitopes of these MoAb has been reported. We previously prepared five MoAb against HAb18G/CD147, named HAb18, 3B3, 1B3, 5A5 and 4D2. To map the epitopes of these MoAb, a series of truncated fragments of extracellular region of HAb18G/CD147 was expressed in Escherichia coli and the MoAb-binding affinity to these fragments was examined with an enzyme-linked immunosorbent assay and Western blot. The residues (39)LTCSLNDSATEV(50), (36)KILLTCS(42) and (22)AAGTVFTTVEDL(33) were determined to be the epitopes of HAb18, 3B3 and 1B3, respectively, which were further proved by a dot-blot analysis with synthesized peptides and bioinformatics epitope prediction. The binding regions of MoAb 5A5 and 4D2 were located at residues E(120)-R(203). Then we constructed and expressed full-length HAb18G/CD147 and truncated HAb18G/CD147 without residues A(22)-V(50) in COS-7 cells. Gelatin zymography and Boyden chamber assay showed that the COS-7 cells expressing truncated HAb18G/CD147 failed to induce MMP production and enhance the cells' invasive potential, compared with the cells expressing full-length HAb18G/CD147. Taken together with the obviously inhibitory effects of HAb18 on the function of full-length HAb18G/CD147, these findings suggest that residues (22)AAGTVFTTVEDLGSKILLTCSLNDSATEV(50) may play a critical role in the functions of HAb18G/CD147 on MMP secretion and tumour invasion. These key residues can be used as potential drug target in cancer therapy.


Assuntos
Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/uso terapêutico , Antineoplásicos/uso terapêutico , Basigina/imunologia , Carcinoma Hepatocelular/terapia , Mapeamento de Epitopos/métodos , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/terapia , Sequência de Aminoácidos , Antineoplásicos/metabolismo , Basigina/metabolismo , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/metabolismo , Epitopos/imunologia , Epitopos/metabolismo , Humanos , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/metabolismo , Metaloproteases/antagonistas & inibidores , Metaloproteases/metabolismo , Dados de Sequência Molecular
16.
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
17.
Retina ; 16(2): 139-44, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8724959

RESUMO

PURPOSE: The authors document an animal model of retinal pigment epithelial (RPE) detachments. METHODS: N-ethylmaleimide (NEM) and sodium iodate (sulfhydryl-alkylating agents) were injected into the vitreous of Dutch pigmented rabbits. Subretinal fluid samples were withdrawn 0.5 and 3 hours after NEM injections and analyzed for albumin content by gel electrophoresis. Selected eyes were examined histologically. RESULTS: Injections of 9.2 micrograms NEM caused wrinkling of the retina within 15 minutes, at which time fluorescein angiography revealed multifocal RPE detachments. At this stage, histologic sections showed moderate choroidal edema and RPE detachment. Elevated retinal detachments developed over the next 15 to 25 minutes. Histologic sections showed massive choroidal edema and RPE fragmentation. The albumin concentration of subretinal fluid rose from 60% of serum level at 30 minutes after NEM to 80% after 3 hours. Sodium iodate did not cause choroidal edema or RPE detachment. CONCLUSION: In rabbits, intravitreal NEM causes the rapid appearance of choroidal edema and RPE detachment, followed by serous retinal detachment. The initial effect probably is caused by an alteration of choroidal vascular permeability. The relationship of these effects to sulfhydryl alkylation is unclear because sodium iodate failed to produce RPE detachments. N-ethylmaleimide effects may model aspects of clinical RPE and serous retinal detachments.


Assuntos
Epitélio Pigmentado Ocular/patologia , Descolamento Retiniano/patologia , Animais , Modelos Animais de Doenças , Etilmaleimida , Angiofluoresceinografia , Coelhos , Descolamento Retiniano/induzido quimicamente
18.
Curr Eye Res ; 14(12): 1109-13, 1995 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8974840

RESUMO

This study asks whether retinal adhesiveness is affected by cytochalasin D, a drug that is known to alter the apical morphology of the retinal pigment epithelium (RPE). Cytochalasin D was injected intravitreally in Dutch rabbits and retinal adhesiveness measured 0.5 to 72 h later by two methods: in vitro peeling of the retina from retinal pigment epithelium to observe the amount of adherent pigment, and in vivo measurement of the pressure needed to achieve retinal separation. Electroretinograms were recorded, and RPE apical morphology was examined by scanning electron microscopy. The injection of 60 microM cytochalasin D caused in vitro retinal adhesiveness to fall within 3 h to 10% of normal although the electroretinogram (a, b, and c-waves) remained normal. Smaller doses of cytochalasin D had a lesser effect. The RPE apical surface at 3 h showed large bullet-like microvilli, swollen cone sheaths, and an absence of filamentous microvilli. The severity of these changes was dose-related. At 72 h after cytochalasin D, retinal adhesiveness had largely recovered, and RPE apical morphology appeared normal again. Thus, cytochalasin D weakens retinal adhesiveness acutely but reversibly, and both the initial effect and recovery correlate with changes in RPE microvillar morphology. This suggests that actin microfilaments may be involved in mechanisms of retinal adhesion.


Assuntos
Citocalasina D/farmacologia , Retina/efeitos dos fármacos , Adesividade/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Eletrorretinografia , Microscopia Eletrônica de Varredura , Microvilosidades/efeitos dos fármacos , Epitélio Pigmentado Ocular/efeitos dos fármacos , Coelhos , Retina/fisiologia , Retina/fisiopatologia , Descolamento Retiniano/fisiopatologia , Fatores de Tempo
19.
Arch Ophthalmol ; 113(5): 655-60, 1995 May.
Artigo em Inglês | MEDLINE | ID: mdl-7748138

RESUMO

OBJECTIVE: To assess the participation of cone matrix sheaths, which are domains of the cone photoreceptor-associated interphotoreceptor matrix that extend from the neural retina to the surface of the retinal pigment epithelium (RPE), in retinal adhesion. METHODS: Monkey and human retinas were partially peeled from the RPE, and the tissues were examined by lectin histochemistry to determine the effects of physical separation on the cone matrix sheath. RESULTS: A firm attachment of cone matrix sheaths to both the RPE and the neural retina that was strong enough to cause detachment of sheets of RPE cells from Bruch's membrane or tearing of the sheaths as a result of retinal peeling was observed. Cone matrix sheaths can stretch considerably and contract following tearing. Their integrity was compromised rapidly after the first postmortem minute. CONCLUSION: Cone matrix sheath glycoconjugates are likely to play a major role in mediating retinal adhesion by forming a molecular bridge between the neural retina and the RPE.


Assuntos
Células Fotorreceptoras/metabolismo , Retina/metabolismo , Adulto , Animais , Adesão Celular , Matriz Extracelular/ultraestrutura , Humanos , Lectinas/metabolismo , Macaca fascicularis , Masculino , Células Fotorreceptoras/ultraestrutura , Epitélio Pigmentado Ocular/metabolismo , Epitélio Pigmentado Ocular/ultraestrutura , Retina/ultraestrutura , Células Fotorreceptoras Retinianas Cones/metabolismo
20.
Arch Ophthalmol ; 113(2): 232-8, 1995 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-7864758

RESUMO

OBJECTIVE: To determine the oxygen and glucose dependency of retinal adhesion in primate and rabbit. METHODS: Experiments were performed on Dutch rabbits and monkeys. Retinal adhesiveness was measured by peeling the retina from the retinal pigment epithelium in vitro, under different conditions of PO2 and glucose supply, and by observing the amount of adherent pigment. In vivo ischemia was produced by raising the intraocular pressure. RESULTS: Retinal adhesion failed quickly at low oxygen tensions, but a well-oxygenated solution preserved strong retinal adhesion in vitro for 15 to 20 minutes in rabbit tissue and up to 50 minutes in primate tissue. Ischemic adhesive failure was reversible on raising the PO2. Glucose levels did not affect adhesiveness. Ischemia in vivo for more than 1 minute caused rabbit retina to lose its adhesiveness. CONCLUSIONS: Retinal adhesion is continually and reversibly dependent on oxygenation, and probably on aerobic metabolism. Primate tissue is more resistant to metabolic adhesive failure than is rabbit tissue, but the metabolic requirements appear qualitatively similar.


Assuntos
Glucose/metabolismo , Oxigênio/metabolismo , Retina/fisiologia , Adesividade/efeitos dos fármacos , Animais , Adesão Celular/fisiologia , Glucose/farmacologia , Hipóxia/fisiopatologia , Isquemia/fisiopatologia , Macaca fascicularis , Oxigênio/farmacologia , Coelhos , Retina/metabolismo , Vasos Retinianos/fisiopatologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...