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OBJECTIVE@#To analysis clinical phenotype and potential genetic cause of a family affected with hereditary coagulation factor Ⅻ deficiency.@*METHODS@#The prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), D-Dimer (D-D), coagulation factor Ⅻ activity (FⅫ:C) and coagulation factor Ⅻ antigen (FⅫ:Ag) were determined for phenotype diagnosis of the proband and his family members(3 generations and 5 people). Targeted capture and whole exome sequencing were performed in peripheral blood sample of the proband. Possible disease-causing mutations of F12 gene were obtained and further confirmed by Sanger sequencing. The corresponding mutation sites of the family members were analyzed afterwards. The online bioinformatics software AutoPVS1 and Mutation Taster was used to predict the effects of mutation sites on protein function.@*RESULTS@#The APTT of the proband was significantly prolonged, reaching 180.9s. FⅫ:C and FⅫ:Ag of the proband was significantly reduced to 0.8% and 4.17%, respectively. The results of whole exome sequencing displayed that there were compound heterozygous mutations in F12 gene of the proband, including the c.1261G>T heterozygous nonsense mutation in exon 11 (causing p.Glu421*) and the c.251dupG heterozygous frameshift mutation in exon 4 (causing p.Trp85Metfs*53). Both mutations are loss of function mutations with very strong pathogenicity, leading to premature termination of the protein. AutoPVS1 and Mutation Taster software predicted both mutations as pathogenic mutations. The results of Sanger sequencing revealed that c.1261G>T heterozygous mutation of the proband was inherited from his mother, for which his brother and his daughter were c.1261G>T heterozygous carriers. Genotype-phenotype cosegregation was observed in this family.@*CONCLUSION@#The c.1261G>T heterozygous nonsense mutation in exon 11 and the c.251dupG heterozygous frameshift mutation in exon 4 of the F12 gene probably account for coagulation factor Ⅻ deficiency in this family. This study reports two novel pathogenic F12 mutations for the first time worldwide.
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Femenino , Humanos , Masculino , Trastornos de la Coagulación Sanguínea , Codón sin Sentido , Factor XII/genética , Heterocigoto , Mutación , LinajeRESUMEN
Objective: To select the preferred flaps for the reconstruction of different maxillary defects and to propose a new classification of maxillary defects. Methods: A total of 219 patients (136 males and 83 females) underwent the simultaneous reconstruction of maxillary defects in the Beijing Tongren Hospital, Capital Medical University, between January 2005 and December 2018 were reviewed. Age ranged from 16 to 78 years. Based on the proposed new classification of the maxillary defects, 22 patients with class Ⅰ defects (inferior maxillectomy), 44 patients with class Ⅱ defects (supperior maxillectomy), 132 patients with class Ⅲ defects (total maxillectomy) and 21 patients with class Ⅳ defects (extensive maxillectomy) were enrolled. Survival rate, functional and aesthetic outcomes of flaps were evaluated. Survival analysis was performed in 169 patients with malignant tumor, Kaplan-Meier method was used to calculate the survival rate, and Log-rank method was used to compare the difference of survival rate in each group. Results: A total of 234 repairs for maxillary defects were performed in 219 patients. Fibula flaps were used in 4/13 of class Ⅰ defects; temporal muscle flaps (11/24, 45.8%) and anterolateral thigh flaps (6/24, 25.0%) used in class Ⅱ defects; temporal muscle flaps (71/128, 55.5%), anterolateral thigh flaps (6/24, 25.0%) and fibula flaps (12/128, 9.4%) used in class Ⅲ defects; and anterolateral thigh flaps (8/20, 40.0%) and rectus abdominis flaps (8/20, 40.0%) used in class Ⅳ defects. The success rate of local pedicled flaps was 95.6% (109/114) and that of free flaps was 95.8% (115/120). Thrombosis(10/234,4.3%) was a main reason for repair failure. Among the followed-up 88 patients, swallowing and speech functions recovered, 82 (93.2%) of them were satisfied with appearance, and 75 (85.2%) were satisfied with visual field. The 3-year and 5-year overall survival rates were 66.5% and 63.6%, and the 3-year and 5-year disease-free survival rates were 57.1% and 46.2%, respectively, in the 169 patients with malignant tumors. Conclusion: A new classification of maxillary defects is proposed, on which suitable flaps are selected to offer patients good functional and aesthetic outcomes and high quality of life.
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Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Colgajos Tisulares Libres , Maxilar/cirugía , Calidad de Vida , Procedimientos de Cirugía PlásticaRESUMEN
Objective: To study the efficacy and patient comfort of absorbable hemostatic powder after endoscopic sinus surgery (ESS). Methods: A total of 21 (17 males, 4 females) patients with an average age of 42(ranging from 18 to 65) underwent bilateral ESS for chronic rhinosinusitis(CRS) in Beijing Tongren Hospital, Capital Medical University between October 2015 and July 2019 were enrolled to compare the effect of absorbable hemostasis powder with Nasopore using an intrapatient control design. A randomized controlled trial was conducted in the left and right nasal cavities of the same patient. If hemostatic powder was applied in the experiment nasal cavity, the Nasopore was applied in the control nasal cavity. The mean preoperative sinus computed tomography (CT) score was 6.25. All patients competed for symptom diaries using a visual analog scale (VAS, score out of 10) at baseline, through 1, 7, 14 and 30 days. Outcomes including bleeding, facial pain, nasal obstruction, nasal discharges using VAS were recorded separately for both sides. Postoperative endoscopic scores were also investigated. SPSS 22 and Graphpad prism 8.0 statistical softwares were used for the analysis. Paired t-test or nonparametric test was used between the test side and the control side. The difference was statistically significant (P<0.05). Results: The bleeding score and total nasal symptom VAS scores at postoperative days (POD) 1, 7, 14 and 30 were not significantly different(t=1.341, 0.552, 0.631, 0.158, all P>0.05;t=0.944, 1.471, 1.612, 2.251, all P>0.05). There was no significant difference between absorbable hemostasis powder and Nasopore side on POD 1, 7, 14 and 30 in terms of each nasal symptom VAS scores(all P>0.05). On POD 1, 7 and 14, the packing material degeneration scores of the absorbable hemostasis powder side were significantly lower than those of the Nasopore side [(1.33±0.21)vs(2.00±0.00),(0.38±0.18) vs (1.95±0.22), 0 vs (1.80±0.13), all P<0.01]. There were significant differences between absorbable hemostasis powder and Nasopore side on POD 1, 7, 14 and 30 in terms of endoscopic scores (edema, crusting, discharges, scar, polyps and material degeneration, t=3.07, 7.00, 6.41, 2.69, all P<0.05). Conclusions: The absorbable hemostasis powder and Nasopore has similar postoperative hemostasis effect. The absorbable hemostasis powder is rapidly cleared and without negative effects on mucosal wound healing 14 days postoperatively.
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Emodin is a common Chinese medicine compound with anti-inflammatory, antibacterial, anti-oxidant and lipid-lowering effects. Modern studies have found that emodin activates adenylate-activated protein kinase (AMPK) signaling molecules and regulates transcriptional factors and biological functions of relevant pathways. Nonalcoholic fatty liver disease is a chronic liver disease with a high incidence in China. With the global prevalence of obesity and metabolic syndrome, the development of nonalcoholic fatty liver disease (NAFLD) is closely related to the expression of the metabolism-related signal molecule AMPK. AMPK is a key enzyme in glycolipid metabolism that can involve different stages of NAFLD development to non-alcoholic steatohepatitis (NASH) by regulating energy metabolism in the body. In recent years, many studies have suggested that the activation of AMPK signaling molecules is related to the function realization of emodin, and lipid synthesis, fatty acid oxidation, insulin sensitivity and mitochondrial function-related transcription factors affected by AMPK downstream signaling molecules and other biological effects can be interacted with each other. The detailed mechanism of action associated with AMPK activation provides new thought about the treatment of NAFLD by emodin. This paper mainly summarizes the research progress of emodin by participating in the various stages of NAFLD by AMPK-related signaling pathways through literature retrieval and comprehensive analysis. It lays a foundation for further research on the therapeutic effect and mechanism of emodin on NAFLD.
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Objective To investigate the causal association between hip circumference (HC) and type 2 diabetes mellitus (T2DM) based on Mendelian randomization. Methods The genetic variants data of the HC and T2DM from the Genetic Investigation of Anthropometric Traits (GIANT) and DIAbetes Genetics Replication And Meta-analysis (DIAGRAM) database were matched according to the single nucleotide polymorphism (SNP) rsID. Genetic loci strongly related to the HC were used as instrumental variables; and the inverse-variance weighting, MR-Egger regression model and weighting median method were carried out to analyze the causal effect of HC on T2DM. Results Fifty-two, nine and fifteen SNPs were matched in the total cohort, female cohort and male cohort, respectively. Heterogeneity test suggested the SNPs were homogeneous. We found HC to be positively associated with T2DM risk (OR=1.065, 95% CI: 1.030-1.100, OR=1.103, 95% CI: 1.057-1.150 and OR=1.583, 95% CI: 1.273-1.968, respectively) in above three cohorts, respectively. Sensitivity analysis showed the results were robust. Conclusions There is a relationship between HC and T2DM of people, and HC may be the risk factor of T2DM.
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Objective:To explore the hepatotoxic material basis of Polygoni Multiflori Radix with zebrafish model, in order to provide a theoretical basis for the study on the mechanism of Polygoni Multiflori Radix hepatotoxicity. Method:Emodin, rhein, aloe emodin, emodin-1-O-glucoside, physcion-8-O-glucoside and aloe emodin-8-O-glucoside for three days (at the concentrations of 0.000 73, 0.002 22, 0.015 05, 0.002 36, 0.198 95, 0.072 73 g·L-1) selected from the early stage of the experiment were continuously administered to zebrafish fertilized for 72 hours to establish the liver fluorescence transgenic larvae animal model. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutathione (GSH) and total bilirubin (TBIL) in zebrafish at 1, 2, 3 day after administration were measured respectively, and the pathological changes of zebrafish liver tissue were analyzed. Result:Emodin, rhein, emodin-1-O-glucoside and physcion-8-O-glucoside had no significant effect on the activities of ALT, AST, GSH and content of TBIL (PPO-glucoside could significantly reduce the activities of ALT, GSH, whereas increased the content of TBIL (PPConclusion:Aloe-emodin and aloe-emodin-8-O-glucoside in Polygoni Multiflori Radix have a toxic effect on zebrafish liver, suggesting that aloe-emodin and aloe-emodin-8-O-glucoside might be the hepatotoxic material basis of Polygoni Multiflori Radix.
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OBJECTIVE@#Cancer is a serious threat to human health. Despite extensive research on cancer treatment, there is a growing demand for new therapies. CD147 is widely involved in tumor development, but it is unclear whether cancer cell malignancy is affected by CD147 expression level. The first compound (AC-73) targeting CD147 could only act on advanced tumors and inhibit metastasis. Therefore, new compounds with better anticancer activity should be explored.@*METHODS@#Wst-1 assays were used to confirm the effect of novel compounds on proliferation. Apoptosis tests were used to evaluate their proapoptotic capacity. A nude mouse model was used to demonstrate in vivo anticancer activity and safety of the compounds. Western blots were used to suggest a molecule mechanism.@*RESULTS@#There is a positive correlation between CD147 expression and tumor cell proliferation. A new compound, HA-08, was synthesized and proved to be more active than AC-73. HA-08 could inhibit cancer cell viability and promote cancer cell apoptosis both in vitro and in vivo. HA-08 induces cancer apoptosis, mainly by disrupting the CD147-CD44 interaction and then down-regulating the JAK/STAT3/Bcl-2 signaling pathway.@*CONCLUSION@#Our results have clarified the tumor specificity of CD147 and its drug target characteristics. The biological profile of HA-08 suggests that this compound could be developed as a potential anticancer agent.
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Background@#Air pollutants and their pathogenic effects differ among regions and seasons. We aimed to explore the relationship between fine particulate matter (PM2.5), sulfur dioxide (SO2), and ozone-8 hours (O3-8h) concentrations in heating and non-heating seasons and the associated death risk due to cardiovascular diseases (CDs), respiratory diseases (RDs), and malignant tumors.@*Methods@#Data were collected in Shenyang, China, from April 2013 to March 2016. We analyzed the correlation or lagged effect of atmospheric pollutant concentration, meteorological conditions, and death risk due to disorders of the circulatory system, respiratory system, and malignant tumor in heating and non-heating seasons. We also used multivariate models to analyze the association of air pollutants during holidays with the death risk due to the evaluated diseases while considering the presence or absence of meteorological factors.@*Results@#An increase in the daily average SO2 concentration by 10 μg/m3 increased the death risk by CDs, which reached a maximum of 2.0% (95% confidence interval [CI]: 1.3%–2.7%) on lagging day 4 during the non-heating season and 0.2% (95% CI: 0.1%-0.4%) on lagging day 3 during the heating season. The risk of death caused by RDs peaked on lagging day 1 by 0.8% (95% CI: 0.4%–1.2%) during the heating season. An increase in O3-8h concentration by 10 μg/m3 increased the risk of RD-related death on lagging day 2 by 1.0% (95% CI: 0.4%–1.7%) during the non-heating season, which was significantly higher than the 0.1% (95% CI: 0–0.9%) increase during the heating season. Further, an increase in the daily average PM2.5 concentration by 10 μg/m3 increased the risk of death caused by RDs by 0.3% and 0.8% during heating and non-heating seasons, respectively, which peaked on lagging day 0. However, air pollution was not significantly associated with the risk of death caused by malignant tumors.@*Conclusion@#Short-term exposure to PM2.5, SO2, and O3 during the non-heating season resulted in higher risks of CD-related death, followed by RD-related death.
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BACKGROUND@#Air pollutants and their pathogenic effects differ among regions and seasons. We aimed to explore the relationship between fine particulate matter (PM2.5), sulfur dioxide (SO2), and ozone-8 hours (O3-8h) concentrations in heating and non-heating seasons and the associated death risk due to cardiovascular diseases (CDs), respiratory diseases (RDs), and malignant tumors.@*METHODS@#Data were collected in Shenyang, China, from April 2013 to March 2016. We analyzed the correlation or lagged effect of atmospheric pollutant concentration, meteorological conditions, and death risk due to disorders of the circulatory system, respiratory system, and malignant tumor in heating and non-heating seasons. We also used multivariate models to analyze the association of air pollutants during holidays with the death risk due to the evaluated diseases while considering the presence or absence of meteorological factors.@*RESULTS@#An increase in the daily average SO2 concentration by 10 μg/m increased the death risk by CDs, which reached a maximum of 2.0% (95% confidence interval [CI]: 1.3%-2.7%) on lagging day 4 during the non-heating season and 0.2% (95% CI: 0.1%-0.4%) on lagging day 3 during the heating season. The risk of death caused by RDs peaked on lagging day 1 by 0.8% (95% CI: 0.4%-1.2%) during the heating season. An increase in O3-8h concentration by 10 μg/m increased the risk of RD-related death on lagging day 2 by 1.0% (95% CI: 0.4%-1.7%) during the non-heating season, which was significantly higher than the 0.1% (95% CI: 0-0.9%) increase during the heating season. Further, an increase in the daily average PM2.5 concentration by 10 μg/m increased the risk of death caused by RDs by 0.3% and 0.8% during heating and non-heating seasons, respectively, which peaked on lagging day 0. However, air pollution was not significantly associated with the risk of death caused by malignant tumors.@*CONCLUSION@#Short-term exposure to PM2.5, SO2, and O3 during the non-heating season resulted in higher risks of CD-related death, followed by RD-related death.
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Objective:To improve the immobilization efficiency of antibody molecules on immune microarray,the method of es-tablishment and optimization of agarose self-assembled membrane carrier with three-dimensional hydrogel structure was established. Methods: The agarose self-assembled membrane carrier was prepared by using glass slide as the carrier,using agarose and sodium periodate modification on glass surface. The agarose self-assembled membrane carrier was characterized by TEM, AFM and FTIR. The optimum preparation conditions were obtained. The carrier for two different species of fixed source antibody efficiency were studied. Antibody loading capacity of agarose self-assembled membrane carrier and ordinary aldehyde carrier were investigated and compared by fluorescence microscopy imaging and Image J software. Results: The agarose nano-membrane carrier had uniform and compact surface. This structure could increase the specific surface area and improve the probe fixed rate. The optimal concentration of agarose for preparation of carrier was 1. 0% . When the concentration of IgG was 0. 3-0. 4 mg/ml,the oxidized self-assembled chitosan film substrate had highest antibody loading capacity. And it had a 3. 94 fold higher antibody loading capacity than the ordinary aldehyde carrier. Conclusion: The agarose nano-membrane carrier is an ideal method for surface modification of immobilized antibody molecules, which is more suitable for preparation of immune microarray carrier.
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Study on the integrated pharmacokinetics/pharmacodynamics (PK/PD) model of rhubarb in rats with yang-deficiency constipation based on the principle of traditional Chinese medicine system. The rat model of yang-deficiency constipation was established using vinegar and ice water containing activated carbon. The blood samples with 0.5 mL were collected from orbital venous plexus at 0, 5, 10, 15, 30, 60, 120, 240, 480, 720, 1 440 min time points after oral administration of rhubarb decoction, and the dosage is equivalent to crude drug 2.5 g·kg-1. The concentration of aloe-emodin, rhein, emodin and chrysophanol in rat plasma were determined by HPLC, and ELISA method was used to detect the activities of motilin (MTL), gastrin (GT), endothelin (ET) and vasoactive intestinal peptide (VIP) at different time points in serum. SPSS 21.0 software was used for analysis of component correlation and principal component, and WinNonlin 6.30 software was used to fit PK/PD model. Compared with the pharmacokinetic parameters of normal rats, in addition to emodin in the model rats showed characteristics of good absorption and slow to elimination; the content of MTL in model rats was significantly lower than that in normal rats. The composite values of the concentration and effect obtained by principal component analysis were connected by Sigmoid-Emax model. We established the integrated PK/PD model of rhubarb in treating yang-deficiency constipation to provide a new research direction for the material basis and mechanism of rhubarb treatment of yang deficiency constipation.
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To study 48 h processing time of Polygoni Multiflori Radix on its contents and changes of chemical components. HPLC was used to determine the contents of various components in 22 Polygoni Multiflori Radix samples with different processing time, and then the fingerprint similarity analysis and clustering analysis were used for characteristics analysis. Results showed that the similarity was between 0.9-1.0, with good correlation between the samples. In the clustering analysis, the 22 Polygoni Multiflori Radix and processed Polygoni Multiflori Radix samples were classified into 4 types according to the composition changes. The results demonstrated that 4-5 h was the best processing time, providing references for quality control and further study of Polygoni Multiflori Radix.
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This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
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Animales , Perros , Femenino , Masculino , Aconitina , Química , Farmacocinética , Disponibilidad Biológica , Estabilidad de MedicamentosRESUMEN
<p><b>OBJECTIVE</b>To extract two kinds of phenols 4-hydroxy-3, 5-dimethoxy-4-(2-oxopropyl) cyclohexa-2, 5-dien-l-one and 6-methoxy-5,7-dihydroxy coumarin (named as I and H compounds respectively) from Ajania salicifolia and to investigate their antioxidation and cytotoxicity to tumors and explore their pro-apoptosis mechanism.</p><p><b>METHODS</b>The antioxidant activities of two compounds were assessed by ABTS and DPPH radical-scavenging assays. Two compounds were evaluated for their cytotoxicity against human chronic myelogenous leukemia (K562) cells using the MIT assay. The expression of NF-kappaB P65 mRNA in K562 apoptotic cells was measured by reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR. In addition, protein expression levels of the NF-ICB P65, p-Akt, Fas, P-catenina and E-cadherin were also measured by Western blot.</p><p><b>RESULTS</b>(1) We found that compound I displayed significant inoxidizability, while compound II had no obvious antioxidizability. (2) In cytotoxicity experiments, compound I didn't display cytotoxicity while compound H displayed obvious cytotoxicity. (3) Compared with the blank group, the expression of NF-kappaB P65 mRNA in K562 cell after treatment with compound II was obviously up-regulated. (4) Compared with the blank group, the expression levels of NF-kappaB P65, Fas, beta-catenina and E-cadherin were significantly increased in compound II treated groups and it appeared obvious dose-effect relationship between the expression of protein and drug concentration.</p><p><b>CONCLUSION</b>Two phenols have obvious antioxidizability and cytotoxicity respectively. On the one hand, the tumor-suppressing mechanism of compound II maybe act by up-regulation the expression of NF-kappaB P65 and Fas protein; thereby, affecting the classical Fas apoptosis signaling pathways. On the other hand, it can also up-regulate the expression of protein beta-catenin and E-cadherin, which participate in the adhesion between cells, and accordingly, playing an important role in preventing the proliferation and metastasis of cancer cells.</p>
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Humanos , Apoptosis , Asteraceae , Química , Cadherinas , Metabolismo , Células K562 , Proteína Oncogénica v-akt , Metabolismo , Fenoles , Química , Transducción de Señal , Factor de Transcripción ReIA , Metabolismo , Regulación hacia Arriba , beta Catenina , Metabolismo , Receptor fas , MetabolismoRESUMEN
Objective: To optimize the preparation for Xinjianwei Tablet, and to establish a method for the quantitative determination of hesperidin. Methods: The new process used dried tangerine peel raw powder as medicine to replace the traditional water decocting process. The effect of the new process was verified with the content of hesperidin as evaluation index. Results: The new process is superior to the traditional one on the premise of the basic substances and the total yield did not change basically. RP-HPLC was used to determine hesperidin and the linearity was good in the range of 10.1-202.5 μg/mL, r = 0.9999. The average recovery was 98.37% with RSD = 1.0%. Conclusion: The improved process is more rational and simple, and the method based on HPLC is simple, reliable, and reproducible. It could be used effectively for the quality control of Xinjianwei Tablet.
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<p><b>OBJECTIVE</b>To establish a three-dimensional digital dental model through scanning dental impression directly with micro-CT.</p><p><b>METHODS</b>The polyvinyl siloxane (PVS) impression of the plaster model was taken and scanned with micro-CT. VGStudio MAX and Imageware softwares were used to obtain the digital dental model.</p><p><b>RESULTS</b>The three-dimensional digital model was established successfully. The scanning layer was 90 µm.</p><p><b>CONCLUSIONS</b>A new way of establishing the digital dental models could be achieved with micro-CT.</p>
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Humanos , Diseño Asistido por Computadora , Materiales de Impresión Dental , Química , Modelos Dentales , Procesamiento de Imagen Asistido por Computador , Imagenología Tridimensional , Polivinilos , Química , Siloxanos , Química , Programas Informáticos , Microtomografía por Rayos X , MétodosRESUMEN
Objective: To investigate the protective effect of deferoxamine preconditioning against hypoxia injury in astrocytes and the underlying mechanisms. Methods: Astrocytes were cultured under ischemia stress, which was mimicked by oxygen and glucose deprivation (OGD) with deferoxamine. Astrocytes were divided into three groups: normally cultured group; Deferoxamine pretreated group: astrocytes were pretreated with Deferoxamine and then treated with Deferoxamine OGD; and OGD group; astrocytes were treated with Deferoxamine OGD. The cell viability examination, the ratio of condensed nuclei, and the morphological changes were used to assess the protective effect of deferoxamine. The expression of HIF-1a and EPO protein and mRNA in astrocytes was examined by immunofluorescence staining and RT-PCR, respectively. Results: The morphology of AS in the deferoxamine pretreated group kept intact. AS viability in deferoxamine pretreated group was 58% of the normally cultured group, and that in the OGD group was 25% of the normally cultured group (P<0. 05). The ratio of condensed nuclei in deferoxamine pretreated group was 38% and that in OGD group was 30% (P<0. 05). Immunofluorescence staining found that expression HIF-1a, and EPO protein appeared in cultured astrocytes after deferoxamine pretreatment. RT-PCR confirmed that mRNA of HIF-1a and EPO was up-regulated after deferoxamine pretreatment. Conclusion: Deferoxamine preconditioning can protect the astrocytes from hypoxia damage, which is related to deferoxamine-induced increase of HIF-1a and EPO expression.
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<p><b>OBJECTIVE</b>To investigate whether desferoxamine (DFO) preconditioning can induce tolerance against cerebral ischemia and its effect on the expression of hypoxia inducible factor 1alpha (HIF-1alpha) and erythropoietin (EPO) in vivo and in vitro.</p><p><b>METHODS</b>Rat model of cerebral ischemia was established by middle cerebral artery occlusion with or without DFO administration. Infarct size was examined by TTC staining, and the neurological severity score was evaluated according to published method. Cortical neurons were cultured under ischemia stress which was mimicked by oxygen-glucose deprivation (OGD), and the neuron damage was assessed by MTT assay. Immunofluorescent staining was employed to detect the expressions of HIF-1alpha and EPO.</p><p><b>RESULTS</b>The protective effect induced by DFO (decreasing the infarction volume and ameliorating the neurological function) appeared at 2 d after administration of DFO (post-DFO), lasted until 7 d and disappeared at 14 d (P < 0.05); the most effective action was observed at 3 d post-DFO. DFO induced tolerance of cultured neurons against OGD: neuronal viability was increased 23%, 34%, 40%, 48% and 56% at 8 h, 12 h, 24 h, 36 h, and 48 h, respectively, post-DFO (P < 0.05). Immunofluorescent staining found that HIF-1alpha and EPO were upregulated in the neurons of rat brain at 3 d and 7 d post-DFO; increase of HIF-1alpha and EPO appeared in cultured cortex neurons at 36 h and 48 h post-DFO.</p><p><b>CONCLUSION</b>DFO induced tolerance against focal cerebral ischemia in rats, and exerted protective effect on OGD cultured cortical neurons. DFO significant induced the expression of HIF-1alpha and EPO both in vivo and in vitro. DFO preconditioning can protect against cerebral ischemia, which may be associated with the synthesis of HIF-1alpha and EPO.</p>
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Animales , Ratas , Isquemia Encefálica , Quimioterapia , Metabolismo , Células Cultivadas , Infarto Cerebral , Quimioterapia , Metabolismo , Deferoxamina , Farmacología , Usos Terapéuticos , Modelos Animales de Enfermedad , Eritropoyetina , Metabolismo , Técnica del Anticuerpo Fluorescente , Subunidad alfa del Factor 1 Inducible por Hipoxia , Metabolismo , Hipoxia-Isquemia Encefálica , Quimioterapia , Metabolismo , Infarto de la Arteria Cerebral Media , Quimioterapia , Metabolismo , Hierro , Metabolismo , Precondicionamiento Isquémico , Métodos , Degeneración Nerviosa , Quimioterapia , Metabolismo , Neuronas , Metabolismo , Patología , Ratas Sprague-Dawley , Sideróforos , Farmacología , Usos Terapéuticos , Factores de Tiempo , Resultado del Tratamiento , Regulación hacia Arriba , FisiologíaRESUMEN
Objective To investigate the activity change pattern and corresponding significance of gly- colytic enzymes including alsolase A(ALDA)and lactate dehydrogenase M(LDH-M)regulated by hy- poxia-inducible factor 1?(HIF-1?)in spinal cord injury(SCI)of rats.Methods SD rats were ran- domly divided into control group and groups at 12 hours,1,2 and 3 days,1 and 2 weeks after compres- sive SCI,in which the activity changes of ALDA and LDH-M in the injured spinal cord were observed at different time points by means of enzyme histochemistry.Results Opitical density(A)value of AL- DA continued significant increase from two days to one week after SCI(P<0.05)and decreased gradual- ly at 2 weeks after SCI.A value of LDH-M began significant increase at day 1 after SCI and recovered to normal level at 2 weeks after SCI(P<0.05).Conclusion Activities of ALDA and LDH-M regulated by HIF-1?in spinal cord injury is significantly increased.