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UNLABELLED: Our objective was to investigate the associations between polymorphisms in Wnt pathway genes and peak bone mineral density (BMD) and body composition in young Chinese men. Our study identified that WNT5B and CTNNBL1 for both BMD and body composition, and WNT4 and CTNNB1 gene polymorphisms contribute to the variation in BMD and body composition in young Chinese men, respectively. INTRODUCTION: Our objective was to investigate the associations between polymorphisms in WNT4, WNT5B, WNT10B, WNT16, CTNNB1, and CTNNBL1 genes and peak bone mineral density (BMD), lean mass (LM), and fat mass (FM) in young Chinese men. METHODS: Using SNPscan(TM) kits, 51 single-nucleotide polymorphisms (SNPs) located in the 6 genes were genotyped in a total of 1214 subjects from 399 Chinese nuclear families. BMD, total lean mass (TLM), and total fat mass (TFM) were measured using dual energy X-ray absorptiometry (DXA). The associations between the 51 SNPs and peak BMD and body composition [including the TLM, percentage lean mass (PLM), TFM, percentage fat mass (PFM), and the body mass index (BMI)] were analyzed through quantitative transmission disequilibrium tests (QTDTs). RESULTS: For peak BMD, we found significant within-family associations of rs2240506, rs7308793, and rs4765830 in the WNT5B gene and rs10917157 in the WNT4 gene with the lumbar spine BMD (all P < 0.05). We detected an association of rs11830202, rs3809269, rs1029628, and rs6489301 in the WNT5B gene and rs2293303 in the CTNNB1 gene with body composition (all P < 0.05). For the CTNNBL1 gene, six SNPs (rs6126098, rs6091103, rs238303, rs6067647, rs8126174, and rs4811144) were associated with peak BMD of the lumbar spine, femoral neck, or total hip (all P < 0.05). Furthermore, two of the six SNPs (rs8126174 and rs4811144) were associated with body composition. CONCLUSIONS: This study identified WNT5B and CTNNBL1 for peak BMD and body composition in males from the Han Chinese ethnic group, and the results suggest a site-specific gene regulation. The WNT4 and CTNNB1 gene polymorphisms contribute to the variation in peak BMD and body composition, respectively.
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Pueblo Asiatico/genética , Composición Corporal/genética , Densidad Ósea/genética , Polimorfismo de Nucleótido Simple , Vía de Señalización Wnt/genética , Absorciometría de Fotón/métodos , Adulto , Anciano , Composición Corporal/fisiología , Índice de Masa Corporal , Densidad Ósea/fisiología , Femenino , Cuello Femoral/fisiología , Estudios de Asociación Genética , Genotipo , Haplotipos , Humanos , Desequilibrio de Ligamiento , Vértebras Lumbares/fisiología , Masculino , Persona de Mediana Edad , Núcleo Familiar , Vía de Señalización Wnt/fisiologíaRESUMEN
OBJECTIVE: Dementia and cognitive impairment (CI) are common in Parkinson's disease (PD) and have important clinical consequences. We explored the prognostic factors for CI in patients with PD. METHODS: A total of 102 patients with PD in Xuan wu hospital and Qian dongnan People's Hospital from 2005 to 2010 were included in this study. All patients underwent clinical and neurological assessments. Relevant demographic and performance parameters were analysed to determine variables that may be independently associated with the progression of CI. RESULTS: In the 6-month follow-up group, CI progressed in three out of 58 cases (5%): two cases progressed from mild CI (MiCI) to moderate CI (MoCI), and one case from MoCI to dementia. In the six-month-to-two-year follow-up group, seven out of 46 cases (15%) worsened: one case developed MiCI, three cases progressed from MiCI to MoCI and three other cases from MoCI to dementia. In the two-to-five-year group, 20 out of 44 cases (45%) worsened with one case developing MiCI, 14 cases progressing from MiCi to MoCI and five cases from MoCI to dementia. Compared with other patients, those with worsening of CI symptoms were significantly older in the two-to-five-year group. Progression of CI was also associated with age at onset and initial staging of PD. CONCLUSIONS: Advanced age, late onset of disease and severity of PD are the predictive factors for the progression of CI in PD. The highest probability of progression of CI is in patients with initial severe impairments of visuospatial function.
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Trastornos del Conocimiento/etiología , Enfermedad de Parkinson/complicaciones , Adulto , Edad de Inicio , Anciano , China , Trastornos del Conocimiento/fisiopatología , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Enfermedad de Parkinson/fisiopatología , Índice de Severidad de la Enfermedad , Percepción EspacialRESUMEN
In this work, ZnO nanofibers (ZNFs) were successfully prepared via a simple electrospinning technique using polyvinylpyrrolidone (PVP) and zinc acetate dihydrate (Zn(CH3COO)2 2H2O) as precursors. The obtained ZNFs have an average diameter of ca. 95 nm and are composed of crystalline wurtzite phase. Methylene blue (MB) dye was used to investigate the photocatalytic performance of pure ZNFs. The study confirms that ZNFs have favorable catalytic activity, and the best degradation efficiency of MB can exceed 90% under UV light irradiation for 3 hours. In addition, we propose a possible photodegradation mechanism.
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OBJECTIVE: To analyze the epidemiological characteristics of reported imported malaria cases in Zhengzhou City from 2016 to 2020, so as to provide insights into the management of imported malaria in the city. METHODS: All data pertaining to cases with definitive diagnosis of malaria in Zhengzhou City from 2016 to 2020 were captured from the National Notifiable Disease Report System and the Information Management System for Parasitic Disease Control in China, including individual demographic data, and malaria onset, initial diagnosis and definitive diagnosis data. All data were descriptively analyzed. The duration from malaria onset to initial diagnosis, from initial diagnosis to definitive diagnosis and from onset to definitive diagnosis was compared among cases. In addition, the diagnoses of imported malaria cases in which definitive diagnosis was made were compared with the reexaminations by Zhengzhou Municipal Malaria Diagnosis Reference Laboratory. RESULTS: A total of 302 cases with definitive diagnosis of malaria were reported in Zhengzhou City from 2016 to 2020, and all were imported cases, with Plasmodium falciparum malaria as the predominant type (230 cases, 76.2%). There were 293 malaria cases imported from Africa (293 cases, 97.0%), which mainly included Nigeria (48 cases, 15.9%), Angola (40 cases, 13.2%), and the Democratic Republic of the Congo (29 cases, 9.6%). There was no obvious seasonality found in the date of malaria onset and time of reporting malaria. The ratio of male to female malaria cases was 49.3:1, and there were 103 cases (34.1%) with the current residency address in Zhengzhou City, 193 cases (63.9%) with the current residency address in other cities of Henan Province and 6 cases (2.0%) in other provinces of China. There were 271 cases (89.7%) seeking initial diagnosis in medical institutions, and the diagnostic accuracy of malaria was 56.6% (171/302) at initial diagnosis institutions. A total of 122 cases (40.4%) sought medical care on the day of malaria onset, and 252 cases (86.4%) within 3 days; however, only 22 cases (7.3%) were definitively diagnosed on the day of onset, and 162 cases (53.6%) diagnosed within 3 days. There were no significant differences between malaria cases seeking initial diagnosis at medical institutions and disease control and prevention institutions in terms of the duration from malaria onset to initial diagnosis (Z = -1.663, P > 0.05), from initial diagnosis to definitive diagnosis (Z = -0.413, P > 0.05) or from malaria onset to definitive diagnosis (Z = -0.838, P > 0.05). The median duration (interquartile range) from initial diagnosis to definitive diagnosis of malaria was 3.00 (2.00), 3.00 (6.00), 2.00 (4.00) d and 1.00 (1.00) d among cases seeking medical care at township-level and lower, county-, city- and province-level medical institutions, and the median duration from initial diagnosis to definitive diagnosis of malaria was significantly longer among cases seeking medical care at township-level and lower medical institutions than at city (Z = -3.286, P < 0.008 33) and province-level medical institutions (Z = -9.119, P < 0.008 33), while the median duration from initial diagnosis to definitive diagnosis [1.00 (3.00) d vs. 2.00 (4.00) d; Z = -4.099, P < 0.016] and from malaria onset to definitive diagnosis [3.00 (4.00) d vs. 4.00 (5.00) d; Z = -2.868, P < 0.016] among malaria cases with the current residency address in Zhengzhou City was both shorter than in other cities of Henan Province. The diagnostic accuracy was 89.1% (269/302) among malaria cases in which definitive diagnosis was made, and the accuracy of malaria reexaminations was 94.0% (284/302) in Zhengzhou Municipal Malaria Diagnosis Reference Laboratory. CONCLUSIONS: P. falciparum malaria was predominant among reported imported malaria cases in Zhengzhou City from 2016 to 2020, and these imported malaria cases were predominantly diagnosed at medical institutions; however, the diagnostic capability of malaria is poor in township-level and lower medical institutions. Strengthening the collaboration between medical institutions and disease control and prevention institutions and improving the diagnostic capability building at medical institutions are recommended to consolidate malaria elimination achivements.
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Malaria Falciparum , Malaria , África , China/epidemiología , Ciudades , Femenino , Humanos , Malaria/diagnóstico , Malaria/epidemiología , MasculinoRESUMEN
OBJECTIVE: The aim of this study was to analyze the role of LINC01554 in the pathogenesis of hepatocellular carcinoma (HCC) and explore the potential mechanism through which LINC01554 affects the migration and proliferation of HCC cells. PATIENTS AND METHODS: LINC01554 expression in HCC tissues and its link to the prognosis of patients were analyzed by The Cancer Genome Atlas (TCGA) database. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was carried out to examine LINC01554 levels in 60 cases of HCC clinical tissues and HCC cell lines. Then, LINC01554 overexpression model was constructed using lentivirus in HCC cell lines. HCC proliferation and invasive ability were evaluated through Cell Counting Kit (CCK-8) and transwell tests, respectively. Furthermore, the potential action mechanism of LINC01554 was explored using bioinformatics analysis and in vitro cell experiments. RESULTS: Analysis of the TCGA database revealed that LINC01554 was remarkably under-expressed in HCC tissues. Decreased expression of LINC01554 predicted a poor prognosis for patients. Besides, LINC01554 overexpression markedly blunted the proliferation and migratory capacities of HCC cells. LINC01554 competed with NGFR to bind to microRNA-3681-3p, thereby providing possible mechanisms by which LINC01554 could participate in the progression of HCC. CONCLUSIONS: This study shows for the first time that LINC01554 modulates NGFR expression by binding to microRNA-3681-3p, thereby participating in the progression of HCC.
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Carcinoma Hepatocelular/metabolismo , Neoplasias Hepáticas/metabolismo , MicroARNs/metabolismo , Proteínas del Tejido Nervioso/genética , ARN Largo no Codificante/metabolismo , Receptores de Factor de Crecimiento Nervioso/genética , Sitios de Unión , Carcinoma Hepatocelular/patología , Movimiento Celular , Proliferación Celular , Células Cultivadas , Humanos , Neoplasias Hepáticas/patología , MicroARNs/genética , Proteínas del Tejido Nervioso/metabolismo , ARN Largo no Codificante/genética , Receptores de Factor de Crecimiento Nervioso/metabolismoRESUMEN
AIM: We sought to discuss vascular anastomosis and gut reconstruction in a living-related small bowel transplantation recipient. METHODS: Living-related small bowel transplantation was performed successfully on a boy with short gut syndrome in two stages. In the first stage, 120 cm, of his mother's ileum was implanted into the recipient with the artery and vein anastomosed to the recipient's sigmoid artery and inferior mesenteric vein, respectively. The two ends of the implanted intestine were constructed as stomas. In the second stage, reconstruction of the continuity of the digestive tract was performed at 188 days after the initial transplantation. The residual small bowel was transected and both ends were anastomosed to the proximal and distal end of the graft in end-to-side fashion. The stomas were closed 30 and 43 days later. RESULTS: Both procedures were successful. Postoperative cytomegalovirus infection and acute rejection occurred successively and were controlled. No leakage of the reconstructed gut or other complications developed after the second procedure. The recipient is alive at 15 months with 8 kg an increase in weight. He is caring for himself independently and has a half-liquid diet, sometimes supplied with auxiliary enteral nutrition. A d-xylose test increased from 4.25% to 25% after the small bowel transplantation. CONCLUSIONS: Vascular anastomoses should be performed according to the state of graft and the recipient. The portal route is the first choice when possible. A two-stage gut reconstruction could decrease the incidence of complications, and offer a useful method in living-related small bowel transplantation.
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Intestino Delgado/cirugía , Donadores Vivos , Síndrome del Intestino Corto/cirugía , Adolescente , Anastomosis Quirúrgica , Femenino , Humanos , Arteria Ilíaca/cirugía , Masculino , Persona de Mediana Edad , Núcleo Familiar , Recolección de Tejidos y ÓrganosRESUMEN
INTRODUCTION: The aim of this study was to develop a rat model of chronic irreversible rejection, which is a major causes of late graft loss and retransplantation after orthotopic liver allotransplantation. METHODS: Allogeneic liver transplantation was performed in a rat combination of Dark Agouti (DA) to Brown Norway (BN). Group A was left without treatment, group B received cyclosporine' (CsA; 1 mg/kg/d) and group C, CsA (4 mg/kg/d). Animals were followed for 6 months. Liver tissue was harvested to construct a time course of histological changes after liver transplantation using histopathological and morphometric techniques. We compared the total histological score of rejection activity index and survival rates. RESULTS: In untreated animals, irreversible acute rejection developed, all animals died within 15 days. In the low-dose CsA group, all animals that survived more than 30 days developed moderate to severe manifestations of chronic liver rejection, with graft infiltration, ductular damage or proliferation, obliterative arteriopathy, and liver fibrosis. No apparent histological alterations were observed in group C. Survival analysis showed significant differences between the three groups. CONCLUSIONS: In the rat strain combination of DA --> BN with low-dose immunosuppression, early mild inflammation was followed by the development of chronic rejection.
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Rechazo de Injerto/patología , Trasplante de Hígado/patología , Animales , Ciclosporina/uso terapéutico , Modelos Animales de Enfermedad , Rechazo de Injerto/mortalidad , Rechazo de Injerto/prevención & control , Trasplante de Hígado/inmunología , Trasplante de Hígado/mortalidad , Ratas , Ratas Endogámicas BN , Ratas EndogámicasRESUMEN
During our screening of pharmacologically active principles from Indonesian medicinal plants by a hypothermic effect the methanol-extract of Brucea javanica (L.) Merr. has been found to exhibit a lethal toxicity to mice. The toxic components were isolated and identified with bruceoside A and B and yadanzioside F. Correction of 13C-nuclear magnetic resonance assignment was also mentioned.
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Fiebre/inducido químicamente , Glaucarrubina/análogos & derivados , Extractos Vegetales/toxicidad , Plantas Medicinales/análisis , Cuassinas , Animales , Glaucarrubina/aislamiento & purificación , Glaucarrubina/toxicidad , Indonesia , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Extractos Vegetales/farmacologíaRESUMEN
OBJECTIVE: Analysis of the pathogenesis of cytomegalovirus (CMV) infection for rat chronic liver rejection. METHODS: Based on animal models that we previously developed, we divided control and a CMV infection group to analyze survival, histopathology and rejection activity. The level and mRNA expression of transforming growth factor-ß1, platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF) were measured in peripheral blood serum and liver tissue. RESULTS: Obliterative arteriopathy and fibrosis was significantly greater in the CMV infection versus the control group (P < .05). PDGF mRNA expression in rat liver and in peripheral blood were significantly higher in the CMV infection versus the control group (P < .05). bFGF mRNA expression in the CMV infection group was also significantly higher than among the controls (P < .05). CONCLUSIONS: CMV infection aggravated foam cell obliterative arteriopathy and liver fibrosis in chronic liver rejection, processes that accelerate the immune response, which may correlate with early high PDGF and bFGF expression.
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Infecciones por Citomegalovirus/complicaciones , Rechazo de Injerto/etiología , Trasplante de Hígado/efectos adversos , Animales , Arteriopatías Oclusivas/etiología , Biomarcadores/sangre , Enfermedad Crónica , Infecciones por Citomegalovirus/sangre , Infecciones por Citomegalovirus/genética , Infecciones por Citomegalovirus/virología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Factor 2 de Crecimiento de Fibroblastos/sangre , Factor 2 de Crecimiento de Fibroblastos/genética , Rechazo de Injerto/sangre , Rechazo de Injerto/genética , Rechazo de Injerto/patología , Rechazo de Injerto/virología , Cirrosis Hepática/etiología , Masculino , Factor de Crecimiento Derivado de Plaquetas/genética , Factor de Crecimiento Derivado de Plaquetas/metabolismo , ARN Mensajero/sangre , Ratas , Factores de Tiempo , Factor de Crecimiento Transformador beta1/sangre , Factor de Crecimiento Transformador beta1/genéticaRESUMEN
A laser damage test facility delivering pulses from 100 fs to 3 ps and designed to operate at 1030 nm is presented. The different details of its implementation and performances are given. The originality of this system relies the online damage detection system based on Nomarski microscopy and the use of a non-conventional energy detection method based on the utilization of a cooled CCD that offers the possibility to obtain the laser induced damage threshold (LIDT) with high accuracy. Applications of this instrument to study thin films under laser irradiation are presented. Particularly the deterministic behavior of the sub-picosecond damage is investigated in the case of fused silica and oxide films. It is demonstrated that the transition of 0-1 damage probability is very sharp and the LIDT is perfectly deterministic at few hundreds of femtoseconds. The damage process in dielectric materials being the results of electronic processes, specific information such as the material bandgap is needed for the interpretation of results and applications of scaling laws. A review of the different approaches for the estimation of the absorption gap of optical dielectric coatings is conducted and the results given by the different methods are compared and discussed. The LIDT and gap of several oxide materials are then measured with the presented instrument: Al(2)O(3), Nb(2)O(5), HfO(2), SiO(2), Ta(2)O(5), and ZrO(2). The obtained relation between the LIDT and gap at 1030 nm confirms the linear evolution of the threshold with the bandgap that exists at 800 nm, and our work expands the number of tested materials.
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AIM: The study is aim to compare two kinds of cleaning score system for capsule endoscopy, with a view of these two cleaning score system can help to evaluate small bowel cleanliness. METHODS: Three readers evaluated these two cleaning score system by assessing the inter-observer, intra-patient, and intraobserver agreement. RESULTS: The assessment of the reliability and concordance, inter-observer agreement and intra-patient agreement of System1 and System2 was excellent with the intraclass correlation coefficient (ICC) values of 0.873, 0.821, 0.863 and 0.772. The data regarding the assessment on intra-observer agreement and intrapatient agreement of System1 and System2 were available and the results were also excellent with ICC values of 0.887, 0.846, 0.870 and 0.809. The overall adequacy assessment of System1 and System2, there was no significant difference among the three readers of inter-observer agreement (X2 = 0.051, P = 0.822, X2 = 0.085, P = 0.081, X2 = 0.048, P = 0.827) and intra- patient agreement (X2 = 0.196, P = 0.658, X2 = 0.208, P = 0.648, X2 = 0.054, P = 0.817), neither was intra-observer agreement (X2 = 0.208, P = 0.648, X2 = 0.223, P = 0.637, X2 = 0.484, P = 0.487) and intrapatient agreement (X2 = 0.054, P = 0.817, X2 = 0.054, P = 0.817, X2 = 0519, P = 0.471). CONCLUSION: The two system both are simple, operable, and can be used in clinical practice.
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Endoscopía Capsular , Irrigación Terapéutica/métodos , Humanos , Procesamiento de Imagen Asistido por Computador , Intestino Delgado , Polietilenglicoles/administración & dosificación , Reproducibilidad de los Resultados , Estudios RetrospectivosRESUMEN
The analgesic components were isolated from a Bornean medicinal plant, Tabernaemontana pauciflora Blume (syn. Ervatamia blumeana Mark gr.), and the major components were identified as coronaridine and 3-(2-oxopropyl)coronaridine. Four minor components were estimated to be 5R- and 5S-(2-oxopropyl)coronaridine, 3-(2-oxopropyl)voacangine and 3,3'-(oxopropyl)dicoronaridine, which might be produced during the isolation process. Voacangine was also isolated as a major component of T. pandacaqui Poir. Coronaridine and voacangine exhibited significant analgesic and hypothermic effects in mice at a dose of 25 mg/kg, p.o., while 3-(2-oxopropyl)coronaridine was less effective. The former two compounds also revealed a surface anesthesia.
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Analgésicos/síntesis química , Plantas Medicinales/química , Analgésicos/farmacología , Animales , Temperatura Corporal/efectos de los fármacos , Borneo , Cobayas , Masculino , Ratones , Ratones EndogámicosRESUMEN
Comparison studies of recombinant prourokinase(pro-UK) in various host cells, and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro-UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200, 12.5 and 50 IU/(10(6) cells 24 h). pro-UK purities separated by immunoaffinity chromatograph are above 90%. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production.
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Precursores Enzimáticos/biosíntesis , Proteínas Recombinantes/biosíntesis , Activador de Plasminógeno de Tipo Uroquinasa/biosíntesis , Animales , Células CHO , Chlorocebus aethiops , Cricetinae , Peso Molecular , Células VeroRESUMEN
Two preparations of glutamate receptors were purified from the synaptic junctions of pig brain by a combination of detergent solubilization, anion-exchange chromatography, wheat-germ agglutinin affinity chromatography and sedimentation through sucrose gradients. These preparations were enriched in specific L-[3H]glutamate binding activity (> 5000 pmol of glutamate binding sites/mg of protein), and the rank order of ligand affinity for binding to these preparations was: quisqualate > 6-cyano-7- nitroquinoxaline-2,3-dione > alpha-amino-3-hydroxy-5-methyl-4- isoxazolepropionic acid (AMPA) > L-glutamate > kainate > > N-methyl-D-aspartate approximately L-2-amino-4-phosphonobutyrate. SDS/PAGE analysis revealed that more than 80% of the protein in either of these preparations appeared as a single protein band of 106 kDa. Two-dimensional gel electrophoresis further revealed that these 106 kDa proteins consisted of a series of acidic proteins which were recognized by antibodies against rat AMPA receptor subunits. These 106 kDa proteins were also recognized by wheatgerm agglutinin and concanavalin A; in addition, peptide N-glycosidase F treatment of these preparations decreased their size to 99 kDa. Our results suggest that the putative glutamate receptors isolated here are likely to belong to the AMPA subtype of glutamate receptors in pig brain. Using the purification procedure reported here, 5 micrograms of AMPA receptor proteins can be isolated from 250 g of pig brain tissue.
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Química Encefálica , Receptores AMPA/aislamiento & purificación , Animales , Centrifugación por Gradiente de Densidad , Cromatografía de Afinidad , Cromatografía en Gel , Electroforesis en Gel Bidimensional , Ácido Glutámico/metabolismo , Ácido Kaínico/metabolismo , Peso Molecular , Octoxinol , Polietilenglicoles/farmacología , Cloruro de Potasio/farmacología , Ratas , Receptores AMPA/metabolismo , Receptores de Ácido Kaínico/metabolismo , Solubilidad , Porcinos , Membranas Sinápticas/química , Ácido alfa-Amino-3-hidroxi-5-metil-4-isoxazol Propiónico/metabolismoRESUMEN
A novel technique was developed to deal with apoptosis in large-scale animal cell culture. By means of replacing part of Cytopore porous microcarriers at regular intervals, a rCHO cell line, which produces urokinase-type plasminogen activitor(u-PA), was cultivated continuously with serum-free medium in a 30 L stirred tank for 91 days. The cell density was maintained at (1.3-2.6) x 10(7)/mL, and > 90% of cells was viable. In order to reduce the effect of cell density on cell growth and expression, a cyclic pressure oscillation was exerted on a 7.5 L reactor headspace to enhance cell expression at high cell density to a certain extent. During the 67 days of medium-replacement culture, the maximal cell density reached 2.64 x 10(7)/mL, and cell viability was always kept above 95% when combined with microcarrier-replacement. Compare to control culture, culture with cyclic pressure oscillation could enhance cell expression level and reduce the ratio of glucose metabolized anaerobically to produce lactate. With four-step purification process, about 80 g u-PA(approximately 90% scu-PA) was recovered from approximately 2100 liters supernatant which contained approximately 135 g u-PA.