RESUMEN
<p><b>OBJECTIVE</b>The effects of lentivirus-mediated suppression of Cyclin Y (CCNY) expression on the proliferation of laryngeal cancer cells were investigated in vitro.</p><p><b>METHODS</b>The lentivirus vectors containing a small hairpin RNA (shRNA) to target CCNY were constructed.Hep-2 cells were divided into the following two experimental groups:the negative control group (control lentivirus infected cells) and CCNY knockdown group (CCNY shRNA-expressing lentivirus infected cells). After Hep-2 cells were infected, Real-time PCR was used to measure CCNY expression. The influence of CCNY on the proliferation of laryngeal cancer cells were assessed using MTT and colony formation experiments.Each experiment was performed in triplicate and repeated three times.</p><p><b>RESULTS</b>Lentiviruses expressing shRNA against CCNY were constructed and Hep-2 cells were infected with above mentioned lentivirus at MOI (Multiplicity of infection) of 120.Real-time PCR analysis showed that the mRNA expression of CCNY in Hep-2 cells in the knockdown group was significantly decreased (P < 0.05); the mRNA level of CCNY was 75.3% lower in the si-CCNY group than in the si-CTRL group. After 5 days of lentiviral infection, the cell viability was significantly lower in cells infected with the CCNY-shRNA lentivirus compared to cells infected with the control lentivirus following a 6-day incubation. The colony number was decreased by 60% in Hep-2 cells infected with the CCNY-shRNA-lentivirus infected cells following a 10-day incubation.</p><p><b>CONCLUSIONS</b>The results suggested that lentivirus-mediated downregulation of CCNY expression decreased the proliferation and growth potency of laryngeal cancer cells.Lentiviruses delivering shRNA against CCNY may be a promising tool for laryngeal cancer therapy.</p>
Asunto(s)
Humanos , Línea Celular Tumoral , Proliferación Celular , Ciclinas , Neoplasias Laríngeas , Metabolismo , Lentivirus , Genética , ARN Interferente Pequeño , GenéticaRESUMEN
<p><b>AIM</b>To investigate the influences of inflammatory mediator on permeability of the blood-brain barrier(BBB) at high altitude environmental exposure as well as relationship and water content in brain.</p><p><b>METHODS</b>Wistar rats were exposed to different altitude gradients, then brain tissue homogenate was prepared, and the activities of tumor necrosis factor-alpha (TNFalpha) and endothelin (ET) in brain tissue homogenate were measured by radioimmunoassay methods. The activities of nitric oxide (NO) in brain tissue homogenate were measured by chemical methods. The evans blue (EB) content in brain tissue was determined by colorimetry and the wet weight/dry weight ratio (W/D) was used to express the water content in brain.</p><p><b>RESULTS</b>With altitude going up, the activities of TNFalpha, NO and ET in the brain of rats rose, and they also rose gradually with time prolonging under high altitude hypoxia exposure. Their most obvious rise was seen during 9 days after ascending 5 000 m high altitude regions. At the same time, the EB and water content in the brain of mice showed the same change trends. Therefore, the inflammatory mediator activity and EB content as well as water content in brain showed an evidently linear relationship.</p><p><b>CONCLUSION</b>The inflammatory mediator plays an important role in the change of permeability of BBB. It's a critical inducing factor in the change of permeability of BBB under high altitude exposure.</p>
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Animales , Femenino , Masculino , Ratas , Altitud , Mal de Altura , Metabolismo , Barrera Hematoencefálica , Fisiología , Edema Encefálico , Metabolismo , Permeabilidad Capilar , Circulación Cerebrovascular , Endotelinas , Metabolismo , Hipoxia , Metabolismo , Mediadores de Inflamación , Fisiología , Óxido Nítrico , Metabolismo , Permeabilidad , Ratas Wistar , Factor de Necrosis Tumoral alfa , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To explore the mechanisms of differentiation and development of pancreatic endocrine cells as well as pancreatic regeneration.</p><p><b>METHODS</b>Human embryonic pancreatic tissue at 7-14 weeks of gestation was collected. Diabetes mellitus rat model was induced with 65 mg/kg of streptozotocin. Insulin, glucagon, somatostatin, nestin, and cytokeratin 19 (CK19) of pancreatic tissues were observed by immunohistochemistry.</p><p><b>RESULTS</b>At 9 weeks of gestation, pancreatic epithelial cells began to co-express insulin, glucagon, somatostatin, and CK19 before migration. Islet cells gradually congregated along with the increase of aging, and at 14 weeks of gestation histological examination showed islet formation. At 12 weeks of gestation, nestin-positive cells could be seen in the pancreatic mesenchyme. During early embryogenesis, islet cells of pancreatic ducts co-expressed insulin, glucagon, and somatostatin. During pancreatic regeneration after damage, nestin expression of islet cells increased.</p><p><b>CONCLUSION</b>In the early stage of embryogenesis, islet cells of primary pancreatic ducts can be differentiated to multipotential endocrine cells before migration. During tissue regeneration, pancreatic stem cells may differentiate and proliferate to form pancreatic islet.</p>
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Animales , Humanos , Masculino , Ratas , Diferenciación Celular , Diabetes Mellitus Experimental , Metabolismo , Patología , Desarrollo Embrionario , Fisiología , Células Epiteliales , Biología Celular , Fisiología , Células Secretoras de Insulina , Biología Celular , Fisiología , Islotes Pancreáticos , Biología Celular , Fisiología , Páncreas , Biología Celular , Embriología , Fisiología , Conductos Pancreáticos , Biología Celular , Embriología , Fisiología , Ratas Sprague-Dawley , Regeneración , Fisiología , Células Madre , Biología Celular , Metabolismo , FisiologíaRESUMEN
<p><b>OBJECTIVES</b>To study the expression of CD34, CD31, CD14, CD10 and factor VIII on blood island of yolk sac (YS), PAS/aorta-germen-mesonephros (AGM) region and hepatic hematopoietic foci.</p><p><b>METHODS</b>Thirty-two cases of 3rd-12th weeks human embryo were obtained by drug abortion. Paraffin embedded sections with H.E staining and immunohistochemistry reaction (SABC) were performed.</p><p><b>RESULTS</b>YS blood island of 3rd-4th weeks of gestation was consisted of two types of cells. One was vascular endothelial cells located outside and the other hematopoietic cells inside the blood island. Both the two types of cells were CD10, CD14, CD31 and factor VIII positive. Hematopoietic cells were CD34 negative, and vascular endothelial cells were CD34 positive. On 32nd days of gestation, the hematopoietic cells migrated out of YS. On 4th week of gestation, CD34, CD14, CD10, CD31 and factor VIII positive cells appeared in the aorta, mesonephros and hepatic hematopoietic foci. By the 7th week, the number of positive hematopoietic cells reached the peak. In 11th-12th weeks, most cells in these regions were matured red blood cells and were negative for all the antibodies mentioned above excepting for CD34. During 4th-12th weeks, all endothelial cells in embryo were CD34 positive.</p><p><b>CONCLUSIONS</b>The hematopoietic cells and endothelial cells of YS blood island co-expressed CD10, CD14, CD31 and factor VIII. Endothelial cells were CD34 positive but hematopoietic cells were negative in YS blood island. The hematopoietic cells of aorta, mesonephros and hepatic hematopoietic foci expressed CD34, CD10, CD14 and factor VIII from 4th week to 7th week. Anti-CD34 antibody could label endothelial cells of every kinds vessels of embryo from 3rd to 12th weeks.</p>
Asunto(s)
Humanos , Antígenos CD34 , Metabolismo , Factor VIII , Metabolismo , Feto , Metabolismo , Técnicas In Vitro , Receptores de Lipopolisacáridos , Metabolismo , Hígado , Metabolismo , Neprilisina , Metabolismo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta , Metabolismo , Saco Vitelino , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues.</p><p><b>METHODS</b>The tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed.</p><p><b>RESULTS</b>The expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments were found to relate with metastasis, differentiation, inflammation and so on.</p><p><b>CONCLUSION</b>RFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.</p>
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Femenino , Humanos , Adenocarcinoma , Genética , Neoplasias de la Mama , Genética , Carcinoma Ductal de Mama , Genética , Biología Computacional , Electroforesis , Métodos , Perfilación de la Expresión Génica , Métodos , Reacción en Cadena de la Polimerasa , Métodos , Análisis de Secuencia de ADN , MétodosRESUMEN
<p><b>OBJECTIVE</b>To discuss the clinicopathological features and prognostic factors of gastric lymphoma.</p><p><b>METHODS</b>83 gastric lymphoma cases were analyzed retrospectively in accordance to the criteria of the new World Health Organization classification for neoplastic diseases of the hematopoietic and lymphoid tissues. The correlations between clinicopathological features, therapeutic measures and survival were discussed.</p><p><b>RESULTS</b>The age of patients ranged from 25 to 77, with a median of 52. The number of males were similar to that of females. There were no specific symptoms. The most common symptoms were stomach ache (60 cases, 72%) or discomfort. The duration of symptoms was often long and with a history of chronic gastric diseases (21 cases, 25%). 13 cases had multiple lesions in the gastrointestinal mucosa. 51 cases (61%) were accompanied by lymph node involvement. According to the new World Health Organization classification for neoplastic diseases of the hematopoietic and lymphoid tissues, 57 cases were extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT)-type (MALT lymphoma), 23 were diffuse large B cell lymphoma accompanying MALT lymphoma, 2 were diffuse large B cell lymphoma and 1 was follicular lymphoma. Of all the cases, 31 were stage I E, 38 stage II E, 8 stage III E and 6 stage IV by the Ann Arbor staging system (1972). The total 5-year and 10-year survival rates were 77.8% and 70.1% respectively, with the mean survival time of 146 months. The 5-year and 10-year survival rates of MALT lymphoma were 77.4% and 72.3%, the 5-year and 10-year survival rates of diffuse large B cell lymphoma accompanying MALT lymphoma were 81.8% and 68.2%, the 5-year survival rate of diffuse large B cell lymphoma was 50.0%.</p><p><b>CONCLUSIONS</b>There are no specific symptoms in gastric lymphoma patients. Extranodal marginal zone lymphoma of MALT-type is the main histopathological type of gastric lymphoma, often accompanied by multiple mucosa involvement and also often accompanied by a history of chronic gastric disease. The lesion is usually localized for a long time, with a very good prognosis. Survival rate has a significant correlation with lymph node involvement and clinical stage. No correlations were found between the survival rates with age, gender, B symptoms, invasive depth of the wall of stomach, the size and range of the tumors or different therapeutic measures.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Quimioterapia Adyuvante , Estudios de Seguimiento , Gastrectomía , Métodos , Metástasis Linfática , Linfoma , Patología , Cirugía General , Terapéutica , Linfoma de Células B , Patología , Cirugía General , Terapéutica , Linfoma de Células B de la Zona Marginal , Patología , Cirugía General , Terapéutica , Linfoma de Células B Grandes Difuso , Patología , Cirugía General , Terapéutica , Estadificación de Neoplasias , Radioterapia Adyuvante , Estudios Retrospectivos , Neoplasias Gástricas , Patología , Cirugía General , Terapéutica , Tasa de SupervivenciaRESUMEN
<p><b>OBJECTIVE</b>Establishing the retinal gene expression profiles of non-diabetic rat and diabetic rat and comparing the profiles in order to analyze the possible genes related with diabetic retinopathy.</p><p><b>METHODS</b>The whole retinal transcriptional fragments of non-diabetic rat and 8-week diabetic rat were obtained by restriction fragments differential display-PCR (RFDD-PCR). Bioinformatic analysis of retinal gene expression was performed using soft wares, including Fragment Analysis. After comparison of the expression profiles, the related gene fragments of diabetic retinopathy were initially selected as the target gene of further approach.</p><p><b>RESULTS</b>A total of 3639 significant fragments were obtained. By means of more than 3-fold contrast of fluorescent intensity as the differential expression standard, the authors got 840 differential fragments, accounting for 23.08% of the expressed numbers and including 5 visual related genes, 13 excitatory neruotransmitter genes and 3 inhibitory neurotransmitter genes. At the 8th week, the expression of Rhodopsin kinase, beta-arrestin, Phosducinìrod photoreceptor cGMP-gated channel and Rpe65 as well as iGlu R1-4 were down-regulated. mGluRs and GABA-Rs were all up-regulated, whereas the expression of GlyR was unchanged.</p><p><b>CONCLUSION</b>These results prompt again that the changes in retinal nervous layer of rat have occurred at an early stage of diabetes. The genes expression pattern of visual related genes and excitatory and inhibitory neurotransmitters in rat diabetic retina have been involved in neuro-dysfunctions of diabetic retina.</p>
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Animales , Femenino , Ratas , Diabetes Mellitus Experimental , Genética , Retinopatía Diabética , Genética , Perfilación de la Expresión Génica , Métodos , Ratas Sprague-Dawley , Retina , Metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , MétodosRESUMEN
The aim was to study the expression of VEGF-A, VEGF-C, angiopoietin-1, angiopoietin-2 and their receptors on development liver during gestation of weeks 3-12 of human embryo. Human embryo contingently aborted at 3-12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 microm serial sections were made. HE staining, immunohistochemistry method and light-microscope were employed. The results showed that at 4-5 weeks of development, liver was constituted by a few hepatic cords. Hematopoietic cell or blood cells were undetectable in the 4 week of gestation. A few cells which were larger, rounded and nucleared cells appeared and expressed VEGFA, flt-4 and Tie-2 proteins strongly in liver at 5 weeks of gestation. The number of these immuno-positive cells was highest in the 7th week and decreased at 11-12 weeks of gestation. These cells expressed flk-1 transiently in the 6th week. VEGF-C and flt-1 were expressed by hepatic cells from weeks 7 to 12 of gestation. The immuno-positive products were deposited in plasma of hepatic cells. Angiopoietin-1, angiopoietin-2 and Tie-2 were detectable on those cells which expressed VEGFA, flt-4 and Tie-2 from weeks 5 to 12 of gestation. The expression of angiopoietin-1 and angiopoietin-2 were weakly and Tie-2 was strongly. They were expressed weakly too by hepatic cells at 5 to 12 weeks of gestation. All factors and their receptors were undetectable on vascular endothelial cells at 4-12 weeks of gestation. It is concluded that the expression patterns of VEGF family on cells of liver are different before and after 7 weeks of gestation. The hematopoiesis in fetal liver may be related to development of hepatic cell.
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Femenino , Humanos , Embarazo , Angiopoyetina 1 , Angiopoyetina 2 , Proteínas de la Matriz Extracelular , Edad Gestacional , Inmunohistoquímica , Hígado , Química , Embriología , Receptor TIE-2 , Factor A de Crecimiento Endotelial Vascular , Factor C de Crecimiento Endotelial Vascular , Receptor 1 de Factores de Crecimiento Endotelial Vascular , Receptor 3 de Factores de Crecimiento Endotelial VascularRESUMEN
The study was to investigate the expression of VEGFA, VEGFC, angiopoietin-1, angiopoietin-2 and their receptors on yolk sac blood island, AGM region during gestation of 3th-12th weeks of human embryo. Human embryo contingently aborted at 3 - 12 weeks of gestation were collected with signed agreements of the pregnant women suffered from accidental abortions. The specimens were fixed by 4% paraformaldehyde and embedded by paraffin. 5 micro m serial sections were made. HE and immunohistochemistry method (SABC) and light-microscope were employed. The results showed that VEGFA and its receptors flt1/flk-1, VEGFC and its receptor flt-4, angiopoietin-2 and its receptor tie-2 proteins were expressed strongly and angiopoietin-1 was weakly expressed by hematopoietic cells and vascular endothelial cells of blood island at 21 and 25 days of gestation. In the 4th week of gestation, immuno-positive reaction of these factors and their receptors appeared in the aorta and mesonephros deposited in larger, rounded and nucleated cells which represented hematopoietic cells. Up to 7th week, positive hematopoietic cells in the regions were much abundant. The number of positive cells decreased at 8th week. Up to 12th week, almost all blood cells were immuno-negative. VEGFA, flt-1, flt-4, angiopoietin-1, angiopoietin-2 and Tie-2 protein were expressed mainly by gonad at 6 - 8 weeks, but it did not express VEGFC and flk-1. The immuno-reaction of the factors and their receptors could not detected in vascular endothelial cells during 3-12th weeks of gestation. It is concluded that hematopoietic cells and endothelial cells in blood island of yolk sac, mesonephros and dorsal aorta co-expressed some factors and their receptors in relation to vasculogenesis and hematopoiesis. Intraembryonic hematopoiesis began in the 4th week of gestation.
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Humanos , Angiopoyetina 1 , Angiopoyetina 2 , Embrión de Mamíferos , Química , Proteínas de la Matriz Extracelular , Inmunohistoquímica , Receptor TIE-2 , Receptores de Factores de Crecimiento Endotelial Vascular , Factor A de Crecimiento Endotelial Vascular , Factor C de Crecimiento Endotelial Vascular , Receptor 1 de Factores de Crecimiento Endotelial Vascular , Receptor 2 de Factores de Crecimiento Endotelial Vascular , Receptor 3 de Factores de Crecimiento Endotelial Vascular , Saco Vitelino , QuímicaRESUMEN
@#ObjectiveTo observe the effect of comprehensive rehabilitation on shoulder subluxation of patients with stroke hemiplegia.MethodsComprehensive rehabilitation was practiced on 20 hemiplegic post-stroke patients with shoulder subluxation, including putting injured limb at correct position, keeping the range of passive motion, resistance exercise of scapula muscle group, and shoulder sting. The therapeutic effect was evaluated after 6 weeks treatment.ResultsThere was a significant difference in the shoulder subluxation and movement function of the upper extremities between pre-therapy and after therapy ( P<0.05~0.01).ConclusionComprehensive rehabilitation has obvious therapeutic effect on shoulder subluxation and motor dysfunction of patients with stroke hemiplegia.