1.Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice
LIU LING ; HUANG ZI-LE ; WANG JIAN-GANG
Chinese Journal of Pharmacology and Toxicology 2017;31(10):964-965
OBJECTIVE To investigate the regulation of {O2 (2,4-dinitrophenyl)1-〔(4-ethoxycarbonyl) piperazin-1-yl〕diazen-1-ium-1,2-diolate}(JS-K), anitric oxide donor, on tumor energy metabolism in H22 tumor- bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The JS-K group and model group were received JS-K (0.75 and 1.5 mg?kg-1) and saline via tail intravenous once every 3 d for 14 d, received 5 injections, respectively. The positive group was received 5-FU 20 mg·kg- 1 by intraperitoneal injection once a day for 14 d. On the 15th day mice were sacrificed. The tumor growth inhibition rate were calculated. The activities of hexokinase (HK), phospho?fructo kinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosine triphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were de?termined by colorimetric method. RESULTS Compared with model group, the tumor mass of JS- K 0.75 and 1.5 mg·kg- 1group was significantly reduced (P<0.01),and the tumor growth inhibition rate was 23.9% and 50.3%, respectively. The activity of HK, PFK, PK, SDH and ATPase of tumor tissue in model group was (22.6±3.7, 14.4±2.6, 12.9±3.2 and 10.5±2.6)U·g-1 protein and (0.70±0.10)μmolPi·mg-1 protein per hour, respectively; which in JS-K 1.5 mg?kg-1 group was dropped by 42.0%, 26.6%, 22.7%, 23.3% and 21.7% (P<0.01, P<0.05). Compared with the model group, the level of ATP and LD in JS-K group was dropped (P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism with inhibition of glycolysis and aerobic oxidation.
2.Nitric oxide donor PABA/NO induced mitochondrial-mediated apoptosis in HepG2 cells
Ling LIU ; Zi-Le HUANG ; Jian-Gang WANG
Chinese Journal of Pharmacology and Toxicology 2017;31(8):786-792
OBJECTIVE To investigate the effects of nitric oxide (NO) donor, O2-{2,4-dinitro-5-[4-(N-methylamino) benzoyloxy]phenyl}1-(N,N-dimethylamino)diazen-1-ium-1,2-diolate (PABA/NO) on apop-tosis in human hepatocarcinoma cells. METHODS Proliferation of HepG2 cells treated with PABA/NO 7.5, 15.0 and 30.0μmol · L-1 was measured with Cell Counting Kit-8 (CCK-8) assay, the morphological features were observed under fluorescence microscopy, the level of NO was measured by DAF-FM DA staining, the apoptosis rate was determined by Annexin Ⅴ-FITC staining, mitochondrial membrane potential was determined by Rhodamine 123 staining, and protein expressions of Bcl-2, Bax, cleaved caspase 3, cytochrome c (Cyt c) and apoptosis inducing factor (AIF) were measured by Western blotting analysis. RESULTS Compared with cell control group, PABA/NO could obviously inhibit the proliferation of HepG2 cells (P<0.01). IC50 value of HepG2 cells treated with PABA/NO for 24 h was (10.8±0.6)μmol·L-1. The cells became round, deformed and appeared shrunken.The level of NO was increased and the fluores-cence intensity was 121 ± 9 (P<0.05), 174 ± 31 (P<0.05) and 230 ± 43 (P<0.01). The apoptosis rate was increased from (2.9 ± 0.5)% to (17.0 ± 4.5)% (P<0.01), (39.8 ± 5.4)% (P<0.01) and (74.3 ± 45.2)% (P<0.01). The fluorescence intensity of Rh123 was reduced from 668±69 to 605±73, 420±65 (P<0.05) and 242±47 (P<0.01). Compared with cell control group, PABA/NO down-regulated Bcl-2, up-regulated Bax and activated cleaved caspase 3. Meanwhile, the expression of Cyt c in the cytoplasm was increased from 0.15±0.04 to 0.27±0.06 (P<0.05), 0.38±0.07 (P<0.01) and 0.82±0.16 (P<0.01). The expression of AIF in the nucleus was increased from 0.183±0.032 to 0.231±0.011, 0.682±0.020 (P<0.01) and 0.966± 0.090 (P<0.01). Addition of carboxy-PTIO (NO scavenger) 50 μmol · L- 1 blocked PABA/NO-induced down-regulation of Bcl-2, up-regulation of Bax and cleaved caspase 3 activation. Additionally, up-regu-lation of Cyt c in the cytoplasm and up-regulation of AIF in the nucleus were also blocked by carboxy-PTIO in PABA/NO-treated HepG2 cells (P<0.01). CONCLUSION PABA/NO may induce HepG2 cell apoptosis through a mitochondrial pathway.
3.Regulation of nitric oxide donor JS-K on tumor energy metabolism in H22 tumor-bearing mice
Ling LIU ; le Zi HUANG ; na Hui SHEN ; Jing WANG ; chen Mu LIU ; lin Chang HUA
Chinese Journal of Pharmacology and Toxicology 2017;31(7):730-735
OBJECTIVE To investigate the regulation of{O2 (2,4-dinitrophenyl)1-[(4-ethoxycarbonyl) piperazin-1-yl] diazen-1-ium-1, 2-diolate} (JS-K), a nitric oxide donor, on tumor energy metabolism in H22 tumor-bearing mice. METHODS The hepatoma animal model in BALB/c mice was established with H22 cell line. The inoculated mice were randomly divided into four groups. The JS-K group and model group received JS-K (0.75 and 1.50 mg?kg-1) and saline via tail the vein intravenously once every 3 d for 14 d, and 5 injections, respectively. The fluorouracil (5-FU) group received 5-FU 20 mg·kg-1 by intra-peritoneal injection once a day for 14 d. On the 15th day after the first administration, mice were sacri-ficed and the tumor, thymus and spleen were isolated and weighed immediately. The tumor growth inhibitory rate and organ index were calculated. The activities of hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), succinate dehydrogenase (SDH), adenosinetriphosphatase (ATPase), and the levels of lactic acid (LD) and adenosine triphosphate (ATP) in tumor tissues were determined by colorimetric method. The expression of hypoxia-inducible factor 1 alpha (HIF-1α) and hexokinaseⅡ(HKⅡ) in the tumor tissue was analyzed by Western blotting. RESULTS Compared with model group, the tumor mass of JS-K 0.75 and 1.50 mg · kg-1 groups was significantly reduced (P<0.01), and the tumor growth inhibitory rate was 23.9%and 50.3%, respectively. There was no diffrence in thymic and splenic indexes between JS-K group and model group. The activity of HK, PFK, SDH, PK and ATPase of tumor tissue in model group was 22.6±3.7, 14.4±2.6, (10.5±2.6) U·g-1protein, (12.9±3.2) kU·g-1 protein and (0.70 ± 0.10) mmolPi · g-1protein · h-1, respectively, which dropped by 42.0%, 26.6%, 22.7%, 23.3%and 21.7%respectively (P<0.01, P<0.05) in JS-K 1.50 mg?kg-1 group. Compared with the model group, the level of ATP of tumor tissue in JS-K 1.50 mg?kg-1 groups dropped by 16.6%(P<0.01) and the level of LD in JS-K 0.75 and 1.50 mg?kg-1 groups dropped by 38.7%and 59.4%(P<0.01), respectively. In addi-tion, the expression of HIF-1αof tumor tissue in JS-K 1.50 mg?kg-1 group was decreased (P<0.01), and the expression of HKⅡ of tumor tissue in JS-K 0.75 and 1.50 mg?kg-1 groups was decreased signifi-cantly (P<0.05, P<0.01). CONCLUSION JS-K can inhibit the growth of tumor in H22 tumor-bearing mice and its mechanism may be related to regulating the tumor energy metabolism by inhibiting glycolysis and aerobic oxidation.
4.Comparison of lutein, zeaxanthin and β-carotene level in raw and cooked foods consumed in Beijing.
Zi-xin WANG ; Peng-cheng DONG ; Ting-ting SUN ; Xian-rong XU ; Le MA ; Yang-mu HUANG ; Xiao-ming LIN
Chinese Journal of Preventive Medicine 2011;45(1):64-67
OBJECTIVETo measure lutein, zeaxanthin and β-carotene level in foods commonly consumed in Beijing, and compare the content difference between raw and cooked food.
METHODSForty-six commonly consumed foods of 8 classes were collected in Haidian district of Beijing from September to October in 2009. A high performance liquid chromatography method was used to determine the content of lutein, zeaxanthin and β-carotene in both raw and cooked samples.
RESULTSLutein was abundant in cucurbitaceous and solanaceous, allium and nuts, especially in Chinese chive (18 226.9 µg/100 g) and pumpkin (13 265.2 µg/100 g). Major sources of zeaxanthin included round pumpkin, green garlic shoot, corn and eggs, whose level of zeaxanthin were 444.6, 283.5, 279.7, 118.6 - 377.9 µg/100 g, respectively. Zeaxanthin level of those cooked foods changed to 483.9, 239.3, 279.1, 149.5 - 594.7 µg/100 g, respectively. The zeaxanthin level of cooked Chinese chive reached 1081.2 µg/100 g, while we did not detect any zeaxanthin in raw Chinese chive. β-carotene was present in a wide variety of vegetables and fruits. Carrot (17 234.3 µg/100 g) was a good source of β-carotene, while its level in cooked carrot was 17 013.5 µg/100 g.
CONCLUSIONConsuming the proper kinds of foods and changing the method of food processing were beneficial to increase the intake of lutein, zeaxanthin and β-carotene.
China ; Cooking ; Food ; Food Analysis ; Lutein ; analysis ; Xanthophylls ; analysis ; Zeaxanthins ; beta Carotene ; analysis
5.Infiltration of tumor associated macrophages in multiple myeloma and its clinical significance.
Qian Le GUI ; Yan Sheng WANG ; Shan HUANG ; Yang WAN ; Hui Ping WANG ; Zi Gang ZHU ; Man Man LI ; Hai Yan ZHU ; Qian Shan TAO ; Yuan Yuan SHEN ; Qing ZHANG ; Hui QIN
Chinese Journal of Hematology 2018;39(2):122-127
Objective: To investigate the clinical significance of tumor associated macrophages (TAM) in multiple myeloma (MM) and the relationship with angiogenesis and immunosuppression. Methods: Seventy cases of MM patients diagnosed from August 2015 to June 2017 were enrolled in the study as experimental group, 20 cases of benign hematological diseases (13 with iron deficiency anemia and 7 with megaloblastic anemia) patients as control group. Immunohistochemical method was used to detect the expression of CD163, CD34 and VEGF in bone marrow samples, and flow cytometry was used to detect the proportion of regulatory T cell (Treg cells), ELISA was used to detect the level of IL-10, and the clinical features were analyzed. Results: ①Among the 70 patients, there were 31 males and 39 females with a median age of 65 (50~78) years old. TAM infiltration density, microvascular density (MVD), VEGF expression level, Treg ratio and IL-10 level in bone marrow samples of 70 MM patients were significantly higher than those of benign hematological diseases (P<0.05). ②In the MM group, the above indexes of the patients with disease stabilized (15 cases) were lower than those of the newly diagnosed group (35 cases) and the relapse refractory group (20 cases) (P<0.05), those of relapse refractory group were higher than those of newly diagnosed group (P>0.05). ③Of the 35 newly diagnosed MM patients, 27 completed 4 courses of treatment. In the effective group (15 cases), the TAM infiltration density after treatment was significantly lower than that before treatment, the difference was statistically significant[(20.20±7.66) vs (28.87±11.97), t=2.362, P=0.025]; while in the ineffective group of 12 cases, the difference of the TAM infiltration density before and after treatment was not statistically significant[(42.00±13.76) vs (48.25±13.59), t=1.119, P=0.275]. ④TAM infiltration density in the effective group after bortezomib treatment (21 cases) were lower than those in the non-bortezomib treatment group (18 cases)[(16.52 ±4.26) vs (19.27 ±5.82), t=1.662, P=0.170]. ⑤The TAM infiltration density in MM patients was positively correlated with MVD, VEGF expression level, Treg cell ratio and IL-10 level (P<0.001). Conclusion: The infiltration of TAM in the microenvironment of MM, which may promoting angiogenesis and inhibiting immune response, is related to the occurrence, development, therapeutic effect and drug resistance of MM.
Aged
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Female
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Humans
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Macrophages
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Male
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Middle Aged
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Multiple Myeloma
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Neoplasm Recurrence, Local
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Neovascularization, Pathologic
6.Laboratory diagnosis and molecular characterization of highly pathogenic avian influenza virus (H5N1) in human in Hunan Province in 2005-2006.
Yi-Wei HUANG ; Zi LI ; Hong ZHANG ; Yun-Zhi LIU ; Le-Ying WEN ; Jun-Hua LI ; Li-Dong GAO ; Xin-Sheng ZHAO ; Zhong-Jie LI ; Bi-Yun CHEN ; Yu LAN ; Lei ZHOU ; Yue-Long SHU
Chinese Journal of Virology 2007;23(6):434-439
To determine the etiologic agents of two atypical pneumonia human cases in Hunan Province in 2005-2006 and to study their pathogenic potential, the patients' respiratory tract samples and sera were collected. The respiratory tract samples were tested by real-time RT-PCR and RT-PCR methods, and the sera by hemagglutination-inhibition assay. Virus was isolated from case 2 and its genome was sequenced and analyzed. Results showed the H5 nucleic acid tests of two cases were positive. The H5-specific antibody titer of the convalescence serum of case 1 showed a 4-fold greater rise than that of the acute phase. And case 2's antibody titer of acute phase was negative. The two atypical pneumonia cases were confirmed as the avian influenza A (H5N1) infection cases. Viral strain A/Hunan/1/2006 was isolated from case 2. Phylogenetic and molecular analysis suggested that 8 gene segments of A/Hunan/1/2006 originated from avian viruses. And A/Hunan/1/2006 was similar with viruses isolated from avian in Hunan Province. The isolated virus did not recombine with human influenza viruses and no obvious variation was observed.
Adult
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Antibodies, Viral
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blood
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Child
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China
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Female
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Hemagglutination Inhibition Tests
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Hemagglutinin Glycoproteins, Influenza Virus
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genetics
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Humans
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Influenza A Virus, H5N1 Subtype
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classification
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genetics
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immunology
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isolation & purification
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Influenza, Human
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diagnosis
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virology
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Male
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Phylogeny
7.Characteristic analysis of NA gene of human influenza viruses (H3N2) isolated from 1996 to 2005 in China.
Wei-Juan HUANG ; Li-Bo DONG ; Ye ZHANG ; Le-Ying WEN ; Xiang ZHAO ; Man-Xiang LI ; Jun-Feng GUO ; Yu LAN ; Zi LI ; Min WANG ; Jie DONG ; Yuan-Ji GUO ; Yue-Long SHU
Chinese Journal of Virology 2007;23(5):345-349
The NA genes of 395 strains of human H3N2 influenza virus isolated from 1996 to 2005 in China were sequenced, analyzed with bioinformatics tools. The NA nucleotide sequence of phylogenetic tree showed a main evolution branch with multiple short side branches. The strains in the same year may be divided into several branches. There was an obvious lag between vaccine strains recommended by WHO and the Chinese circulating strains in phylogenetic tree of the NA nucleotide. The result also showed no amino acid deletion and insertion in the NA. In NA antigen sites, where including residues 197-199 aa, 431-434 aa and 339-347aa the mutation was higher, in contrast, the residues including 153 aa, 328-336 aa, 367-370aa and 400-403 aa, the mutation was lower. Besides the antigenic determinant sites, there also had the other amino acid mutated highly, such as 18, 23, 30, 93, 143, 208, 216, 221, 249, 265, 267, 307, 385 and 437 aa, among them 143 and 267 mutation were higher than that in antigenic determinant sites, their biological significance are not clear yet. The neuraminidase active-site residues in NA were highly conservative and the same were the disulphide bond and the glycosylation sites in NA. In conclusion, our analysis provides some information for influenza prevention and control and the NA inhibitor medicine application.
China
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Genes, Viral
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Humans
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Influenza A Virus, H3N2 Subtype
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genetics
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Mutation
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Neuraminidase
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genetics
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Phylogeny
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Time Factors
8.Study on the correlation of human influenza A/H3N2 hemagglutinin gene variation and the epidemic from 1995 to 2005 in China.
Li-Bo DONG ; Ye ZHANG ; Le-Ying WEN ; Xiang ZHAO ; Wei-Juan HUANG ; Yu LAN ; Jun-Feng GUO ; Zi LI ; Min WANG ; Jie DONG ; Yuan-Ji GUO ; Yue-Long SHU
Chinese Journal of Virology 2007;23(5):339-344
To study the correlation of human influenza A/H3N2 hemagglutinin gene variation and the epidemic from 1995 to 2005 in China, 550 HA1 sequences of H3N2 viruses isolated in China were analyzed with phylogenetic tree. The results showed that the evolution of HA1 represented a long trunk with short side branches. The animo acid changes of HA1 mostly located at the antigenic sites or aside of them, but also may occur at the other sites simultaneously. The analysis also showed three possibilities of HA1 variation to cause H3N2 epidemic, the first is multiple site mutations happened simultaneously; the second is mutation sites happened gradually and then accumulated to multiple sites; the third is a single antigenic site mutation occurred simultaneously with the receptor binding site variation.
China
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Disease Outbreaks
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Genes, Viral
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Genetic Variation
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Hemagglutinin Glycoproteins, Influenza Virus
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genetics
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Humans
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Influenza A Virus, H3N2 Subtype
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genetics
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Influenza, Human
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epidemiology
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Mutation
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Time Factors
9.Application of Cardiopulmonary Exercise Testing in Enhanced Recovery after Surgery.
Liang Yan ZHANG ; Zi Jia LIU ; Le SHEN ; Yu Guang HUANG
Acta Academiae Medicinae Sinicae 2017;39(6):831-835
Enhanced recovery after surgery (ERAS) is a new perioperative concept that aims to reduce perioperative stress response and accelerate rehabilitation of patients through a variety of optimized management. With the wider application of this concept,the effective implementation of ERAS program has become a new challenge. Cardiopulmonary exercise testing (CPET) has shown promising value in the preoperative assessment,perioperative optimization,and postoperative rehabilitation of ERAS. This article reviews the application of CPET in ERAS,with an attempt to provide evidence for more detailed and comprehensive ERAS program.
10.Clinical Utility of Revised Cardiac Risk Index to Predict Perioperative Cardiac Events in Elderly Patients with Coronary Heart Disease Undergoing Non-cardiac Surgery.
Le Yi LIU ; Zi Jia LIU ; Guang Yan XU ; Feng Yan ZHANG ; Li XU ; Yu Guang HUANG
Acta Academiae Medicinae Sinicae 2020;42(6):732-739
Objective To explore the predictive ability of the revised cardiac risk index(RCRI)in elderly patients with coronary heart disease(CHD)undergoing non-cardiac surgery. Methods We performed a retrospective study including a total of 2100 patients,aged≥65 with a history of CHD who underwent non-cardiac surgery form January 2013 to September 2019.The preoperative,intraoperative and postoperative clinical data were extracted from an electronic database.The RCRI and reconstructed-RCRI(R-RCRI)score of each patient were calculated.The primary end point was defined as an occurrence of perioperative MACE.Multivariate logistic regression analysis was performed to evaluate the risk factors of perioperative MACE.The area under the receiver operating characteristic(ROC)curve was used to compare the predictive value of RCRI,R-RCRI,and the new risk scoring system of the study for perioperative MACE. Results The incidence of perioperative MACE in elderly patients with CHD was 5.4%.Six independent risk factors of perioperative MACE for this population were identified:age≥80 years;female;history of heart failure;insulin-depended diabetes mellitus;preoperative ST segment abnormality;American Society of Anesthesiologists grade≥Ⅲ,and the risk index was 2,2,2,2,2 and 3 respectively.The area under ROC curve of RCRI,R-RCRI and risk scoring system in this study were 0.586,0.552 and 0.741. Conclusion The correlation between RCRI score and perioperative MACE was poor in elderly patients with CHD undergoing non-cardiac surgery,and a better cardiac risk assessment method should be established for this population.
Aged
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Coronary Disease/complications*
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Female
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Humans
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Logistic Models
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Male
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Middle Aged
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Multivariate Analysis
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Postoperative Complications/epidemiology*
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Retrospective Studies
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Risk Assessment
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Risk Factors
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Surgical Procedures, Operative