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We aimed to investigate the target and signal pathway of Smilacis Glabrae Rhixoma (SGR) in the treatment of myasthenia gravis (MG) based on network pharmacology, and to explore its potential molecular mechanism. The main active components of SGR were searched in the pharmacology database of traditional Chinese medicine systems, and analysis platform. The related targets of SGR were obtained by Genecards, connective tissue disease, therapeutic target database, Drugbank, and Online Mendelian Inheritance in Man database. Moreover, the target information was corrected through UniProtKB and also, this data integrated to draw the "Ingredients-targets" network of SGR. Protein interaction analysis was performed in data platform, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways as well as enrichment analysis on disease-drug target was carried out through metascape online platform. A total of 15 active components were collected from SGR, which correspond to 159 targets; There were 1758 MG-related targets; there are 81 targets related to both drug components and diseases, including 12 key targets. In GO bioaccumulation analysis, 1933 GO items were gathered, which were mainly related to the metabolism of active oxygen species and the active factors of postsynaptic neurotransmitter receptor. According to KEGG analysis, SGR may play a role in the treatment of MG through phosphatidylinositol-3-kinase-protein kinase B signaling pathway, T-cell receptor, cAMP, tumor necrosis factor (TNF), and interleukin-17 (IL-17) signaling pathway, Th17 cell differentiation, endocrine resistance, hepatitis, and some cancer pathways. This study shows that SGR mainly treat myasthenia gravis through the regulation of TNF, MAPK1, JUN, TP53 and other targets, T-cell receptor, TNF, and IL-17 signaling pathway, Th17 cell differentiation and other pathways, which reflects the characteristics of multicomponent, multitarget, and multichannel of traditional Chinese medicine, and providing a certain pharmacological basis for the follow-up study.
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Medicamentos Herbarios Chinos , Miastenia Gravis , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico , Estudios de Seguimiento , Humanos , Medicina Tradicional China , Simulación del Acoplamiento Molecular , Miastenia Gravis/tratamiento farmacológico , Miastenia Gravis/genéticaRESUMEN
Objective:To evaluate carotid plaque neovascularization and vessel stability using superb microvascular imaging.Methods:Seventy-two patients with carotid atherosclerotic plaques received treatment in The Seventh People's Hospital of Wenzhou from June 2018 to June 2020 and were included in this study. A total of 100 carotid plaques were surgically removed from these patients. These patients were subject to superb microvascular imaging and contrast-enhanced ultrasonography before carotid plaques were removed. Taking pathological results of carotid plaque as a gold standard, we investigated the efficacy of superb microvascular imaging versus contrast-enhanced ultrasonography in detecting carotid plaque neovascularization and vessel stability and evaluated the detection consistency of each imaging method with the gold standard. Results:The sensitivity, specificity, and accuracy of superb microvascular imaging in detecting carotid plaque neovascularization were 93.24%, 92.31%, and 93.00%, and they were 95.96%, 96.15%, and 96.00% for contrast-enhanced ultrasonography. The Kappa values of consistency of agreement on carotid plaque neovascularization identification were 0.825 and 0.923 for superb microvascular imaging and contrast-enhanced ultrasonography, respectively. The sensitivity, specificity, and accuracy of superb microvascular imaging in detecting vessel stability were 94.74%, 95.35%, and 95.00%, respectively and they were 96.49%, 97.67%, and 97.00%, respectively for contrast-enhanced ultrasonography. The Kappa values of consistency of agreement on vessel stability evaluation were 0.898 and 0.939 for superb microvascular imaging and contrast-enhanced ultrasonography, respectively.Conclusion:Superb microvascular imaging has equivalent efficacy in detecting carotid plaque neovascularization and vessel stability to contrast-enhanced ultrasonography. Superb microvascular imaging is non-invasive, provides ease in operation, and is worthy of clinical promotion.
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Objective:To explore the application value of case three-dimensional teaching method combined with standardized patient (CTTM+SP) in the clinical clerkship teaching reform of gastrointestinal surgery.Methods:A total of 120 students of clinical medicine having clerkship in Binzhou Medical University Hospital were recruited in this study, and they were randomized to the experimental group and the control group, with 60 students in each group. The experimental group took the CTTM+SP teaching method, and the control group adopted the lecture-based learning teaching method. The comprehensive ability of the two groups was evaluated by department graduation examination and satisfaction survey.Results:The examination performance and the satisfaction rate of students in the experimental group were significantly higher than those of the control group ( P < 0.05). Conclusion:CTTM+SP teaching method is helpful for the probationers to master the theoretical knowledge of gastrointestinal surgery systematically, improve the clinical thinking ability and the ability to solve practical problems, mobilize the students' learning motivation and interest, and improve the teaching effect of gastrointestinal surgery clinical clerkship.
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Objective To explore the clinical features and risk factors of poor prognosis in neonatal necrotizing enterocolitis(NEC).Methods A retrospective study was carried out in the infants with NEC admitted to 6 cooperative hospitals in Guangdong Province between January 2005 and December 2014.The clinical features and risk factors of poor prognosis in preterm and full-term infants diagnosed NEC,early onset and late onset NEC were analyzed.Results A total of 449 cases who met the criteria were admitted during the study time.The mortality was 23.6% (106/449 cases),of which the preterm group was 24.6% (58/238 cases) while the full-term group was 22.7% (48/211 cases),the early onset group was 22.1% (45/204 cases) while the late onset group was 24.3% (57/235 cases).The median number of NEC onset in preterm group was 11 d after birth while the number of the full-term group was 6 d.Full-term infants who diagnosed NEC were more likely to manifest themselves as abdominal distension (52.1% vs.42.0%,x2 =4.597,P =0.032),vomiting(36.5% vs.17.2%,x2 =21.428,P =0.000) and bloody stool(30.3% vs.21.4%,x2 =4.653,P =0.031);but in the onset of NEC,preterm infants more likely to have feeding intolerance (21.0% vs.12.8%,x2=5.309,P =0.021).The early onset group of full-term NEC was much common in twins or multiplets(9.4% vs.1.1%,x2 =6.226,P =0.013),which rate of surgical therapy was much higher (41.0% vs.27.0%,P =0.036) and the breast-feeding rate before NEC was lower than the late onset group(14.5% vs.32.6%,x2 =9.500,P =0.002),the differences were statistically significant.The gestational age and birth weight were bigger in the early onset group of preterm NEC[(33.8 ±2.5) weeks vs.(32.2 ±2.8) weeks,t =4.261,P =0.000;(2.1 ±0.5) kg vs.(1.7 ± 0.5) kg,t =4.735,P =0.000)],but length of stay was shorter than the late onset group (18.0 d vs.26.5 d,P =0.000).Logistic regression analysis showed that the risk factors of poor prognosis of full-term NEC were shock,peritonitis and sepsis;while risk factors of poor prognosis of preterm NEC were small for gestational age infant,pulmonary hemorrhage,shock,intestinal perforation and sepsis;the risk factors of poor prognosis of the early onset group of full-term NEC was shock;while those of the late onset group were shock and peritonitis;the risk factors of poor prognosis in the early onset group of preterm NEC were shock and sepsis,while those in the late onset group were pulmonary hemorrhage,shock,intestinal perforation and sepsis.Conclusions Compared to the preterm NEC,the onset time of full-term NEC was earlier and the clinical manifestations were more typical.Early identification and management of shock,peritonitis,intestinal perforation,sepsis and pulmonary hemorrhage can reduce the risk of poor prognosis of neonate NEC.
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Objective To study the composition of gut microbiome in neonates with severe bilinebinemia (serum total bilirubin > 342 μmol/L),and to explore the relationship between gut microbiome and bilirubin brain injury.Methods A prospective study was conducted.The neonates with serum total bilirubin > 342 μmol/L from September 2016 to March 2017 in Guangzhou Women and Children's Medical Center,Guangzhou Medical University,were enrolled in the study and 16S rDNA sequence analysis technology was used to detect the composition of gut microbiome in all subjects.According to the results of brain magnetic resonance imaging (MRI),brain stem auditory evoked potential (BAEP) and clinical manifestations,the subjects were divided into the brain injury group (26 cases) and no brain injury group (28 cases).The differences of the composition of gut microbiome between the 2 groups were compared,and the levels of unconjugated bilirubin in serum and cerebrospinal fluid were also compared.Results The level of unconjugated bilirubin in serum of the brain injury group was (463.51 ± 110.62) μmol/L,but in no brain injury group was(364.18 ±63.13) μmol/L,and there was significant difference between the 2 groups(t =4.090,P =0.000 1).The level of unconjugated bilirubin in the cerebrospinal fluid of the brain injury group was (9.53 ± 2.68) μmol/L,but in no brain injury group was (6.94 ± 2.31) μmol/L,and there was significant difference between the 2 groups (t =3.812,P =0.000 3).There was no correlation between the level of unconjugated bilirubin in the cerebrospinal fluid and serum between the 2 groups(r =0.137,0.081,all P >0.05).The abundance of gut microbiome in the brain injury group was lower than that in no brain injury group in genus level,among which Fusobacterium,Catabacter,Succinivibrio,Clostridium and Bacteroides were significantly different (all P < 0.05).Conclusions The occurrence of bilirubin brain injury depends on the level of unconjugated bilirubin in serum cerebrospinal fluid,but it may be more directly dependent on the level of bilirubin in the cerebrospinal fluid.The diversity of gut microbiome in neonates with bilirubin brain injury was significantly lower than that in no brain injury group.The level of unconjugated bilirubin in cerebrospinal fluid may be related to the different blood-brain barrier permeability caused by different composition of gut microbiome.
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Recently,An increasing number of studies have suggested that interaction between gut microbiome and central nervous system is bidirectional.It is known that altered emotional state and chronic stress can change the composition of gut microbiome,while gut bacteria can impact neurological development,alter behavior and potentially affect the onset and/or severity of nervous system disorders.The review will expound the potential underlying mechanisms from the following several aspects:the effect of gut microbiota on Intestinal permeability,the effect of bacterial metabolites on the central,how the microbiome Influence blood brain barrier integrity,how the gut microbiota regulate neurotransmitter levels,how the microbial control neurological function by the immune system and whether the vagus nerve can act as a mediator of the effect of gut.
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ObjectiveTo analyze the differential expression of microRNA (miRNA) and its significance in patients with neonatal necrotizing enterocolitis (NEC).MethodsTwenty-five patients diagnosed with NEC with Bell stage≥Ⅱ, and 25 non-NEC patients as control group admitted to Guangzhou Women and Children's Medical Center between October 2014 and November 2015 were collected. White blood cells were extracted from the peripheral blood. Five samples were selected randomly each from NEC group and control group, and sequenced by second-generation Illumina high-throughput sequencing, screened for differentially expressed miRNA and analyzed for target genes prediction and biological function. The rest samples of the two groups were detected by real-time fluorescent quantitative polymerase chain reaction technology (RT-qPCR), the results were used to validate the results of high-throughput sequencing. Differentially expressed miRNA in the two groups of data was analyzed using DEGseq software.P<0.05 was considered statistically significant.P<0.01,q<0.001 and丨Log2 Ratio丨≥1 were taken as criteria for screening the differential expression. The differential expressions of miRNA in NEC group and control group were analyzed by cluster analysis using MeV4.6 software.ResultsA total of 482 miRNAs were differentially expressed in the two groups, with significant difference (P<0.05). Among them, 126 were known miRNAs with significantly differential expression in the two groups, with 58 being up-regulated and 68 being down-regulated. The results of up-regulated miRNAs (hsa-miR-223-5p,-183-3p,-222-5p) and down-regulated miRNAs (hsa-miR-23b-5p,-150-5p,-146a-3p,-1298-5p) were confirmed to be consistent with the results of sequencing. Bioinformatics analysis showed that the target genes with differential miRNA expression mainly involved Toll-like receptor signal transduction pathway, mitogen-activated protein kinase pathway, JAK-STAT and other signal transduction pathways.ConclusionsThere are significantly differential expressions of miRNAs in peripheral white blood cells of NEC neonates. These miRNs may be involved in the occurrence and development of NEC via adjusting different target genes to regulate the signal pathway.
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Objective To establish an appropriate neonatal rat model with necrotizing enterocolitis (NEC) and to investigate the protective effects of interleukin-1 receptor associated kinase (IRAK) 1/4 inhibitor on intestinal tissue of neonatal rats with NEC.Methods Thirty-six newborn SD rats were randomly divided into normal control group,NEC model group and IRAK1/4 inhibitor group,12 rats in each group.The rats in normal control group were raised by their mother and they did not receive any intervention.The rats of NEC model group were given artificial feeding,under hypoxia and cold stress.The IRAK1/4 inhibitor group were given IRAK1/4 inhibitor intervention,and also given artificial feeding,under hypoxia and cold stress.Three days later,all rats were sacrificed and intestinal tissues were obtained.The histopathological changes in ileocecal tissues were evaluated by pathological score after hematoxylin-eosin staining.The levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α in intestinal tissues were detected by enzyme-linked immunosorbent assay.Results The rats of NEC model group presented abdominal distention,diarrhea (partly with mucous bloody stool),decreased activity,poor response,lethargy and other symptoms,and the extent was gradually worsened;the rats in IRAK1/4 inhibitor group also had abdominal distention,diarrhea,decreased activity,poor response and other symptoms,but the symptoms emerged later and milder.The histopathological score of intestinal tissues of normal control group was (0.33 ± 0.49) scores,NEC model group was (3.08 ± 0.99) scores,and IRAK1/4 inhibitor group was (1.75 ±0.96) scores.The histopathological scores of NEC model group and IRAK1/4 inhibitors group were significantly higher than those in normal control group (all P < 0.01),and the histopathological scores of IRAK1/4 inhibitor group were significantly lower than those in the NEC model group (P < 0.01).The levels of IL-1β,IL-6 and TNF-α in normal control group separately were (128.76 ± 27.25) ng/L,(0.41 ± 0.10) ng/L,(6.93 ± 1.79) ng/L,respectively;the levels of NEC model group separately were (410.99 ± 44.16) ng/L,(1.79 ± 0.18) ng/L,(44.39 ± 6.00) ng/L;the levels of IRAK1/4 inhibitor group separately were (256.23 ±41.29) ng/L,(1.05 ±0.19) ng/L,(21.45 ± 6.36) ng/L,respectively.The levels of IL-1β,IL-6 and TNF-α of NEC model group had significant differences compared with those of normal control group,respectively (all P <0.01);the levels of IL-1β3,IL-6 and TNF-α of IRAK1/4 inhibitor group had significant differences compared with those of normal control group,respectively (all P < 0.01);the levels of IL-1β,IL-6 and TNF-α of IRAK1/4 inhibitor group had significant differences compared with those of NEC model group (all P < 0.01),respectively.Conclusions IRAK1/4 inhibitor has some protective effects on the intestinal tissues of neonatal rats with NEC,which can reduce the damage to the intestinal tissues,and decrease the secretion of inflammatory cytokines like IL-1β,IL-6 and TNF-α.
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Objective To observe the efficacy of Huoxue Tongmai decoction in the treatment of coronary heart disease with angina pectoris.Methods 60 coronary heart disease patients with angina pectoris were randomly divided into two groups,30 cases in each group.The observation group was treated with Huoxue Tongmai decoction concomitant with routine chemical medicine therapy,and the control group was treated with only routine chemical medicine therapy.The total effective rate,reduced angina occurrences after therapy and ECG were observed in both two groups.Results After treatment,the total effective rate of the observation group was 93.3%,which was signifi-cantly higher than that of the control group(χ2 =4.32,P <0.05).Compared with the two groups of ECG efficacy,the total effective rate of the observation group was 83.3%,which was significantly higher than 66.7% of the control group (χ2 =4.22,P <0.05).After one,three treatment courses,the number of angina attack in the observation group was significantly lower than that in the control group,and the differences were statistically significant(t =6.99, 10.82,all P <0.05).Conclusion Houxue Tongmai decoction combined with routine chemistry medicine is more effective than routine chemistry medicine in treating angina.
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Objective To discuss the risk factors of progressive hemorrhagic injury(PHI)in patients with craniocerebral injury.Methods Clinical data of 149 patients with closed craniocerebral injury were retrospectively analyzed,and the patients were divided into PHI group (42 cases)and non -PHI group (107 cases)according to PHI appeared or not.The patients were immediately given CT scan after admitted,the first CT review was given in the non -PHI group within routine 4 -8h after first CT scan,and due to deterioration of clinical symptoms,the PHI group was given CT review in advance.The intracranial hematoma volume changes between first CT and first CT review in the two groups were observed,then clinical symptoms,signs,biochemical indicators and CT performance in the two groups were compared,and analyzed risk factors of PHI.Results The intracranial hematoma volume showed in CT scan,first CT review and increment volume of the PHI group were significantly higher than the non -PHI group [(14.59 ±4.60)mL vs.(7.28 ±2.94)mL,(25.92 ±8.84)mL vs.(8.35 ±3.41)mL,(10.20 ±3.45)mL vs. (2.10 ±0.65)mL],the differences were significant (t =6.796,11.894,9.367,all P 10mL were major risk factors of PHI (P 50 years old,mydriasis,conscious disturbance,intracranial hematoma volume >10mL in first CT scan.
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Objective To observe the vertebrobasilar artery and its branch courses,diameters and the included angles in the starting part,offering morphological basis for clinic intervention and vascular image.Methods Layer by layer dissecting head,neck and chest of nine adult specimens affused with red latex,and after affused carotid artery with red latex,the origin,course,branches,distribution,anastomosis,and accompanying of the vertebrobasilar artery were observed in three normal adult corroded specimens.Results The degree of the left included angle between subclavian artery and vertebral was (91.70 ± 0.90) ° and that of the right one was (115.80 ± 1.60) ° (P < 0.05).A percentage (14.1%) of them contacted with vagus and glosssopharyngeal nerve.The degree of the left included angle between posterior cerebal lower artery and vertebral artery was (118.60 ± 3.70) ° and that of the right one (77.60 ± 7.90) °.A percentage (27.6 %) of itself contacted or pressed by root of facial nerve.The degree of the included angle on the left side between anterior cerebellar lower artery and brobasilar artery was (91.60 ± 2.40) ° and that of the right one was (27.10 ±2.70)°(P <0.05).The degree of the left included angle between posterior cerebral artery and basilar artery was (93.90 ± 3.40) ° and that of its right one was (130.80 ± 3.20) ° (p < 0.05).Conclusions Both vertebral artery in every part out of skull and the data of the each branch pipe diameter,the starting included angle of vertebrobasilar artery had statistical significance,which should select personalized catheter or operation method in endovascular intervention or craniocervical surgery.
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BACKGROUND:Cerebrovascular stent may destroy the vessel walls,which can lead to vascular restenosis.There are different versions about the safety,pathologic pharmacology reasons and clinical effect.OBJECTIVE:To evaluate the safety and short-term effect of endovascular stent implantation for symptomatic artery stenosis.METHODS:Totally 20 patients with total 22 lesions diagnosed symptomatic artery stenosis were treated with endovascular stenting.The vascular stenotic lesions involved middle cerebral artery in 6 cases,internal carotid artery in 6 cases,vertebral artery in 4 cases,basilar artery in 3 cases and vertebro-basilar artery in 3 cases.The length of vascular stenotic lesions was 3-10 mm with the average of 7 mm.Both balloon and self-expandable stents were used in 12 cases with embolus protection device.RESULTS AND CONCLUSION:All of the 22 stents in 20 patients were placed successfully in one time.After stent implantation,the vascular angiography showed that the vessels were reformed obviously with the degree of stenosis no more than 20%.The perfusion in cerebrum was improved in parenchymal phase and the symptom was also improved clearly.At 6-24 months follow-up,20 patients never had cerebral ischemia.With follow-up for 12-24 months in 7 patients,digital subtraction angiography displayed that intima hyperplasia was occurred in stent in one patient with no symptom.The stenotic vascular were unobstructed and no intima hyperplasia by transcranial Doppler examinations.The results demonstrated that endovascular stent implantation is a safe and effective treatment for intracranial symptomatic artery stenosis,while its long-term effect needs further study.
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Objective To analyze the positive rate of HBsAg in different population at different periods after widely using Hepatitis B Vaccine and provide strategy for Hepatitis B Control.Methods Using ELISA method to detect the HBsAg carrier in polulation under 18 years old in local area.Results After six year investigating,the HBsAg carrier rate in tested population decreased.For 0-4,4-8,8-12,12-15,15-18 age group,the carrier rate were 3.76% vs 0.30%,4.74% vs 1.39%,5.59% vs 2.52%,8.51% vs 3.03%,and 12.58% vs 2.98% in 2000 and 2006.Conclusion The reduction in carrier rate had a direct relationship with Hepatitis B Vaccine inoculation.Hepatitis B Vaccine had important effect to Hepatitis B control.We should gradually intensify Hepatitis B Vaccine inoculation.It is not only for newborn but also for adult.
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BACKGROUND:Antisense gene therapy offers immense promise for the management of human cerebral arteriovenous malformation through inhibiting expression of vascular endothelial growth factor and angiogenesis in endothelial cells.OBJECTIVE: To observe the inhibitory effect of vascular endothelial growth factor-antisense oligonucleotide (VEGF-ASODN) on the proliferation of vascular endothelial cells in human cerebral arteriovenous malformation.DESIGN: Observational contrast study.SETTING: Department of Neurosurgery, General Hospital of Shenyang Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Neuromedical Institute, General Hospital of Shenyang Military Area Command of Chinese PLA from August to December 2006. A total of 18 patients with human cerebral arteriovenous malformation were selected from Department of Neurosurgery, Shenyang General Hospital of Military Area Command of Chinese PLA. There were 12 males and 6 females and their mean age was 40 years. Cerebral arteriovenous malformation was classified based on Spetzler grade: grade Ⅱ (n =10) and grade Ⅲ (n=8). All cases were diagnosed with whole cerebral angiography before operation and they provided the confirmed consent. Main reagents were detailed as follows: endothelial cell growth supplements (ECGS, Sigma, USA), 391 DNA automatic synthetic device (Shanghai Shenggong Liyong Company, PE, USA), anaerobic incubator (DY-1, Zhejiang), human vascular endothelial growth factor enzyme-linked kit (TBD Company, Beijing), 96E enzyme-labeling device (ERMA, INC), cell cycle analytical reagent kit (BD Company), and flow cytometer (FACS Calibur, BD Company).METHODS: ①Experimental procedure: Tissue explants adherent method was used to culture vascular endothelial cells from human cerebral arteriovenous malformation. The third generated cells were used and randomly divided into antisense group, sense group and control group with four bottles of cells in each group. Sense and antisense phosphorothioate oligodeoxynucleotides of artificial vascular endothelial growth factor selected from the antisense group and the sense group were covered with positive liposomes, and then they were used to transfected vascular endothelial cells cultured from human cerebral arteriovenous malformation; however, cells in the control group were not dealt with any treatments. Cells in the three groups were incubated in anaerobic incubator (including 0.95 volume fraction of N2 and 0.05 volume fraction of CO2) at 37 ℃ for 2, 4 and 8 hours, respectively. ② Experimental evaluation: Cell cycles were measured, protein content of vascular endothelial growth factor was measured, and mRNA expression of vascular endothelial growth factor was also detected.MAIN OUTCOME MEASURES: Expression of mRNA and protein of vascular endothelial growth factor and proliferation exponent at different times of hypoxia.RESULTS: ①mRNA expression of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, mRNA expression of vascular endothelial growth factor was higher than that before hypoxia in the control group (P < 0.05);however, mRNA expression was lower in the antisense group than that in the control group (P < 0.05). ② Protein content of vascular endothelial growth factor: At 2, 4 and 8 hours after hypoxia, protein content of vascular endothelial growth factor was higher than that before hypoxia in the control group (P < 0.05); however, protein content was lower in the antisense group than that in the control group (P < 0.05). ③ Proliferation exponent: At 4 and 8 hours after hypoxia,proliferation exponent of endothelial cells cultured from human cerebral arteriovenous malformation was higher than that before hypoxia in the control group (P < 0.05); however, proliferation exponent was lower in the antisense group than that in the control group (P < 0.05).CONCLUSION: Hypoxia may induce gene expression of vascular endothelial growth factor in endothelial cells at the transcriptional level. Antisense vascular endothelial growth factor can obviously inhibit gene expression of vascular endothelial growth factor cultured from human cerebral arteriovenous malformation and proliferation under hypoxic conditions.
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The working principle and application characteristics of ECG machine were introduced in detail from such aspects as the development history,structure,classification,new usage and procurement principle.The objective is to direct new equipment procurement and facilitate clinical application.