1.Blood vitamin characteristics and their correlation with severity in patients with metabolic-related fatty liver disease
XIONG Bo ; ZHENG Jinxin ; XIE Yunqi ; RAO Liying ; LIU Xiaojun ; YU Zhijian ; DENG Qiwen
China Tropical Medicine 2024;24(1):60-
Objective To explore the characteristics of blood vitamins A, B2, B6, B12, D, E, K1, K2 and folic acid and their correlation with severity in patients with metabolic-related fatty liver disease (MAFLD). Methods From September to December 2022, a total of 473 cases of residents were recruited through community MAFLD screening activities and their health information was obtained through questionnaire survey and physical examination. The severity of hepatic steatosis was determined with FibroScan, and vitamin concentrations were determined with liquid chromatography-tandem mass spectrometry. Two independent samples' t-tests were used to assess the differences between the two groups, and univariate chi-square tests and multivariate logistic regression analysis were used to explore the related factors of MAFLD. Results Of the 473 inhabitants, 195 (41.23%, 195/473) met the diagnostic criteria for MAFLD, including mild 43 (22.05%, 43/195) cases of fatty liver, 88 (45.13%, 88/195) cases of moderate fatty liver, and 64 (32.82%, 64/195) cases of severe fatty liver. Using healthy residents collected during the same period as controls, the overall mean of vitamins A, E, K1, and K2 in the MAFLD group was higher than that of the healthy group, with a statistical difference (P<0.05). Furthermore, the concentrations of vitamins A, E, K1 and K2 increased with the severity of fatty liver [R=0.149, P=0.004; R=0.245, P<0.001; R=0.110, P=0.032; R=0.129,P=0.012]. There were statistically significant differences (P<0.05) in the blood levels of vitamin A and E between patients with moderate or severe fatty liver and the healthy population. The blood vitamins K1 and K2 in severe fatty liver patients were also different from those of healthy people (P<0.05). However, there was no significance between folic acid, vitamin D, B2, B6, B12, and MAFLD (P>0.05). Through univariate chi-square analysis and multivariate logistic regression analysis, it was found that male [Wald=5.789, P=0.034,OR=1.598(1.037-2.463)] and vitamin E≥8.13 μg/mL[Wald=14.632,P<0.001,OR=2.378(1.522-3.674)] were risk factors for moderate and severe MAFLD. Conclusions The concentrations of vitamin A, E, and K in the blood are increased in patients with MAFLD compared to the healthy population, and they are positively correlated with the severity of MAFLD. ale gender and high levels of vitamin E may be related to moderate to severe MAFLD.
2.Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA
Jiayan XIONG ; Wei LEI ; Bo YOU ; Zhenxin ZHANG ; Haijing XIE ; Ying SHAN ; Tian XIA ; Yong ZHOU
China Oncology 2024;34(5):451-459
Background and purpose:DDX is an adenosine triphosphate(ATP)-dependent RNA helicase closely related to mRNA regulation,tumor proliferation and invasion.This article aimed to explore the effect of DDX6,a member of the DDX family,on the stability of CKMT1A mRNA,as well as the effect of the DDX6 CKMT1A axis on the proliferation and migration ability of human nasopharyngeal carcinoma cell CNE2 and its molecular mechanism.Methods:We retrieved the data of expressions of DDX6 and CKMT1A in human head and neck squamous cell carcinoma from The Cancer Genome Atlas(TCGA)database and performed a correlation analysis.Western blot was performed to detect the expressions of CKMT1A and DDX6 in human nasopharyngeal carcinoma tissues and normal nasopharyngeal tissues preserved by Affiliated Hospital of Nantong University.This study was approved by the Ethics Committee of Affiliated Hospital of Nantong University(Number:2022-L114).We used transwell assay to detect cell migration ability,EdU assay to detect cell proliferation ability,and colony formation assay to detect clone formation ability.We transfect with lentivirus and plasmids to construct sh-DDX6,sh-CKMT1A,sh-CKMT1A+sh-DDX6 and oe-CKMT1A cell models derived from the human nasopharyngeal carcinoma cell line CNE2,preserved by Affiliated Hospital of Nantong University,to clarify the impact of DDX6 and CKMT1A expression levels on the malignant biological phenotypes of nasopharyngeal carcinoma cells.BALB/c nude mice subcutaneous xenograft tumor model was constructed to detect the effects of DDX6 and CKMT1A on nasopharyngeal carcinoma cells in mice.RNA stability assay was used to detect the effect of DDX6 knockout on CKMT1A mRNA and further clarify the molecular mechanism of DDX6.Results:DDX6 was highly expressed,CKMT1A level was low in human nasopharyngeal carcinoma tissue,and DDX6 was negatively correlated with CKMT1A expression.DDX6 inhibited protein translation of CKMT1A by disrupting its mRNA stability.Low expression of CKMT1A in CNE2 cells enhanced cell migration and proliferation ability,while high expression inhibited migration and proliferation ability.Knocking out DDX6 reversed the progression of malignant behavior caused by downregulation of CKMT1A.Low expression of CKMT1A promoted tumor cell growth in BALB/c nude mice subcutaneous xenograft tumor model,while low expression of DDX6 inhibited tumor cell growth.Knocking out DDX6 and CKMT1A simultaneously restored the inhibitory effect caused by knocking down DDX6 alone.Conclusion:DDX6 in nasopharyngeal carcinoma cells disrupts the stability of CKMT1A mRNA,negatively regulates CKMT1A protein translation,upregulates the proliferation and migration ability of nasopharyngeal carcinoma cells,and promotes malignant progression of nasopharyngeal carcinoma.
3.Predilection site and risk factor of second primary cancer: A pan-cancer analysis based on the SEER database.
Shan XIONG ; Hengrui LIANG ; Peng LIANG ; Xiuyu CAI ; Caichen LI ; Ran ZHONG ; Jianfu LI ; Bo CHENG ; Feng ZHU ; Limin OU ; Zisheng CHEN ; Yi ZHAO ; Hongsheng DENG ; Zhuxing CHEN ; Zhichao LIU ; Zhanhong XIE ; Feng LI ; Jianxing HE ; Wenhua LIANG
Chinese Medical Journal 2023;136(12):1500-1502
4.Chinese expert consensus on the technical standard of direct anterior hip arthroplasty for elderly femoral neck fracture (version 2023)
Zhonghua XU ; Lun TAO ; Zaiyang LIU ; Yang LI ; Jie LI ; Jun ZHANG ; Xia ZHANG ; Min WANG ; Changqing LI ; Guangxing CHEN ; Liu YANG ; Dawei ZHANG ; Xiaorui CAO ; Guoqiang ZHANG ; Pingyue LI ; Nirong BAO ; Chuan LI ; Shenghu ZHOU ; Zhengqi CHANG ; Bo WU ; Wenwei QIAN ; Weiguo WANG ; Ming LYU ; Hao TANG ; Hu LI ; Chuan HE ; Yunsu CHEN ; Huiwu LI ; Ning HU ; Mao NIE ; Feng XIE ; Zhidong CAO ; Pengde KANG ; Yan SI ; Chen ZHU ; Weihua XU ; Xianzhe LIU ; Xinzhan MAO ; Jie XIE ; Xiaogang ZHANG ; Boyong XU ; Pei YANG ; Wei WANG ; Xiaofeng LI ; Eryou FENG ; Zhen ZHANG ; Baoyi LIU ; Jianbing MA ; Hui LI ; Yuanchen MA ; Li SUN ; Zhifeng ZHANG ; Shuo GENG ; Guanbao LI ; Yuji WANG ; Erhu LI ; Zongke ZHOU ; Wei HUANG ; Yixin ZHOU ; Li CAO ; Wei CHAI ; Yan XIONG ; Yuan ZHANG
Chinese Journal of Trauma 2023;39(11):961-973
Femoral neck fracture (FNF) in the elderly patients is currently a major health challenge worldwide, with excessive consumption of medical resources, high incidence of complications as well as suboptimal outcome and prognosis. Hip joint arthroplasty (HJA) has been the mainstream treatment for FNF in the elderly, but the conventional surgical approaches and techniques are still confronted with a series of bottlenecks such as dislocation, limp and limb length discrepancy. In recent years, direct anterior approach (DAA) for HJA (DAA-HJA) has been a major new choice in the field of joint replacement, which achieves improved clinical effectiveness of HJA in the treatment of elderly FNF, due to the fact that DAA approach involves the neuromuscular interface and accords with the idea of soft tissue retention and enhanced recovery after surgery. However, there is still a lack of unified understanding of standard technique and procedure of DAA-HJA in the treatment of elderly FNF. Therefore, relevant experts from the Hip Joint Group of Chinese Orthopedics Association of Chinese Medical Association, Youth Arthrology Group of Orthopedic Committee of PLA, Orthopedic Committee of Chongqing Medical Association, Branch of Orthopedic Surgeons of Chongqing Medical Doctor Association and Sport Medicine Committee of Chongqing Medical Association were organized to formulate the " Chinese expert consensus on the technical standard of direct anterior hip arthroplasty for elderly femoral neck fracture ( version 2023)" based on evidence-based medicine. This consensus mainly proposed 13 recommendations covering indications, surgical plans, prosthesis selections, surgical techniques and processes, and postoperative management of DAA-HJA in elderly patients with FNF, aiming to promote standardized, systematic and patient-specific diagnosis and treatment to improve the functional prognosis of the patients.
5.Application of bronchoscopic lavage in children with severe adenovirus pneumonia
Jie HE ; Xinping ZHANG ; Meiyu YANG ; Xiong ZHOU ; Jianshe CAO ; Zili CAI ; Xiayan KANG ; Bo XIE ; Ying LIU ; Zhenghui XIAO
Chinese Pediatric Emergency Medicine 2021;28(6):472-476
Objective:To investigate the efficacy and safety of bronchoscopic lavage in children with severe adenoviral pneumonia.Methods:Patients with severe adenovirus pneumonia who were admitted to ICU department of Hunan Children′s Hospital for bronchoscopy were collected from February to June 2019 and divided into lavage group( n=36) and non-lavage group( n=15) in line with whether lavage was performed.Their results, namely, bronchoscopic diagnosis, blood gas analysis before and 2 hours, 24 hours and 48 hours after bronchoscopy, improvement time of clinical symptoms(fever and pulmonary moist rales), the positive rate of pathogen detection and mortality rate, main vital signs such as heart rate, respiratory rate, mean arterial pressure and bronchoscopy-related complications were recorded before and 1 hour, 2 hours and 24 hours after bronchoscopy. Results:A total of 51 children were collected, all of whom suffered from endobronchitis.More secretions were observed in the airways of 36 patients in the lavage group, and only a little or no secretions were observed in 15 patients in the non-lavage group.P/F value and PCO 2 at 2 hours, 24 hours and 48 hours after treatment in the lavage group were improved comparing to those before treatment and were superior to those in the non-lavage group( P<0.05). P/F values at 24 hours and 48 hours after treatment in the non-lavage group increased and PCO 2 decreased at 48 hours after treatment( P<0.05). The thermal duration, time to resolution of moist rales in the lungs in the lavage group were shorter than those in the non-lavage group( P<0.05). The mortality rate in the lavage group was lower than that in the non-lavage group[2.8%(1/36) vs.26.7%(4/15), P<0.05]. The positive rate of pathogen detection in lavage group was higher than that in non-lavage group[55.6%(20/36) vs. 20.0%(3/15), P<0.05]. There was no significant difference in heart rate, respiratory rate, and mean arterial pressure at each time point before and after bronchoscopic treatment( P>0.05). Associated complications were 11 cases of intraoperative transient hypoxemia, four cases of bronchial mucosal bleeding, and one case each of postoperative hypoxemia, intraoperative hypertension and hypotension.There was no significant difference in the incidence of complications between the two groups( P>0.05). Conclusion:Bronchoscopic lavage, in treating children with severe adenovirus pneumonia, may improve clinical symptoms, respiratory function, and rate of pathogen detection, reduce mortality, and is effective and safe.
6.Clinical predictive value of TCD in monitoring cerebral circulation in sepsis patients
Ying FENG ; Ying TAN ; Bo XIE ; Weilv XIONG ; Xiaowei JI ; Qin ZHOU ; Jianhong LU ; Yongbin WANG
Chinese Journal of Emergency Medicine 2020;29(8):1087-1092
Objective:To study the dynamic evaluation value of transcranial Doppler (TCD) in cerebral circulation and brain function of sepsis patients and its predictive value of the prognosis.Methods:A retrospective analysis was performed on 98 sepsis patients admitted to our hospital from January 2017 to June 2018 who were divided into the survival group (56 cases) and death group (42 cases) according to the 28-day prognosis. The general clinical data of the patients, acute physiology and chronic health evaluation II (APACHE II), delirium assessment scale (CAM-ICU), and TCD parameters PSV, Vm, EDV, PI, RI on the first day and the third day of admission were collected, and the differences of TCD parameters at different time points were analyzed between the two groups, the area under the ROC curve (AUC) was calculated and evaluating their predictive value of sepsis outcome was evaluated.Results:APACHE Ⅱscore and the number of patients with sepsis encephalopathy were significantly higher in the death group than those in the survival group ( P < 0.05). There were no significant differences in PSV, Vm and EDV between the two groups on the first day and the third day. The PI and RI of the survival group were significantly lower than those of the death group ( P<0.01). In the survival group, PI and RI on the third day decreased significantly compared with those on the first day ( P < 0.05), while in the death group, PI and RI on the first day and the third day did not change significantly. ROC survival curves showed that Day1PI, Day3PI, Day1PI - Day3PI, Day1RI, Day3RI, Day1RI - Day3RI and APACHEⅡ predicted AUC of the prognosis of sepsis patients were 0.657, 0.835, 0.730, 0.707, 0.826, 0.705, and 0.815 ( P < 0.01). When PI cutoff value on the third day became 1.02, the sensitivity was 85.7% and the specificity was 61.3%. When RI cutoff value on the third day became 0.62, the sensitivity was 78.6% and the specificity was 71.4%. Conclusions:TCD parameters, PI and RI, are of certain value in the dynamic evaluation of cerebral circulation and brain function in sepsis patients. PI and RI on the first day and the third day and the differences between them are all valuable in predicting the prognosis, and PI and RI on the third day have greater predictive value.
7.Clinical features of severe type 7 adenovirus pneumonia: an analysis of 45 cases.
Xin-Ping ZHANG ; Mei-Yu YANG ; Xiong ZHOU ; Jian-She CAO ; Zi-Li CAI ; Xia-Yan KANG ; Bo XIE ; Ying LIU ; Jie HE ; Zheng-Hui XIAO
Chinese Journal of Contemporary Pediatrics 2020;22(5):429-434
OBJECTIVE:
To study the clinical features of severe type 7 adenovirus pneumonia in children.
METHODS:
A retrospective analysis was performed for the clinical data of children who were diagnosed with severe type 7 adenovirus pneumonia from February to June, 2019.
RESULTS:
Among the 45 children, the male/female ratio was 3:2 and the median age was 14 months. All children had repeated fever, cough, and pulmonary moist rales, and the mean duration of fever was 14±4 days. The median time from fever to dyspnea was 8 days, and the time from fever to mechanical ventilation was 11.6±2.5 d. There was no significant increase in white blood cell count, with neutrophils as the main type. There were slight reductions in hemoglobin and albumin, while platelet and fibrinogen remained normal. There were increases in aspartate aminotransferase, lactate dehydrogenase, procalcitonin, and C-reaction protein. The detection rate of mixed pathogens was 84%. Effusion in both lungs was the major change on chest imaging (64%). Bronchoscopic manifestations were endobronchitis, tracheomalacia, and plastic bronchitis. The incidence rate of respiratory complications was 100%, and extrapulmonary complications mainly involved the circulatory system (47%), digestive system (36%), and nervous system (31%). Among the 45 children, 16 were administered with 400 mg/kg intravenous immunoglobulin (IVIG) daily for 5 days, with a mean duration of fever of 16±5 days, and 29 were administered with 1 g/kg IVIG daily for 2 days, with a mean duration of fever of 13±4 days; there was a significant difference in the mean duration of fever between the two groups (P=0.046). The overall mortality rate was 11%.
CONCLUSIONS
Severe type 7 adenovirus pneumonia in children has severe conditions, with a high incidence rate of complications and a high mortality rate, so it should be diagnosed and treated as early as possible.
Adenoviridae
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Bronchitis
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Female
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Fever
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Humans
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Infant
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Male
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Pneumonia, Viral
;
Retrospective Studies
8.Study on the Optimization of the Extraction Technology of Shengmaiyin Polysaccharide and Its Regulation Effects on Intest- inal Function of Spleen Deficiency Model Rats
Yu YOU ; Lin LUO ; Zhejie CHEN ; Meisi LIN ; Heying XU ; Bo XIONG ; Chaomei FU ; Tian XIE
China Pharmacy 2019;30(4):493-498
OBJECTIVE: To optimize the extraction technology of Shengmaiyin polysaccharide, and to investigate the regulation effects of Shengmaiyin and its polysaccharide on intestinal function of spleen deficiency model rats. METHODS: The contents of polysaccharide were determined by phenol-sulfuric acid method, and the extraction rate of polysaccharide was calculated. Using extraction rate of Shengmaiyin polysaccharide as investigation index, singel factor and orthogonal tests were used to optimize material-liquid ratio, extraction time, extraction temperature and extraction times of Shengmaiyin polysaccharide. Validation test was also conducted. Totally 80 male SD rats were randomly divided into blank group, model group, Shengmaiyin low-dose, medium-dose and high-dose groups (350, 700, 1 400 g/L, by crude drug), Shengmaiyin polysaccharide low-dose, medium-dose and high-dose groups (24.5, 49, 98 g/L, by crude drug), with 10 rats in each group. Except for blank group, other groups were given Rheum palmatum water decoction 10 mL/kg to induce spleen deficiency model, once a day, for consecutive 15 d. Since the 16th day, blank group and model group were given isovolumic water intragastrically, while other groups were given corresponding drugs, once a day, for consecutive 10 d. The general status of rats and body weights were recorded in each group. The serum contents of D-xylose, gastrin (GAS) and vasoactive intestinal peptide (VIP) were detected by phloroglucinol method or ELISA. RESULTS: The optimal extraction technology of Shengmaiyin polysaccharide was material-liquid ratio 1 ∶ 10(g/mL), extraction time 45 min, extraction temperature 80 ℃, extracting for 1 time. Results of validation test showed that extraction rates of the polysaccharide in 3 times were 7.43%, 7.64%, 7.80% (RSD=1.01%, n=3). After modeling, except for blank group, other groups suffered from loose stools, thin body and reduced food intake, and the body weight and serum level of D-xylose were decreased significantly compared with blank group (P<0.01). After last medication, above symptoms of administration groups were improved to different extents. Except for model group, body weight and serum contents of D-xylose in other groups were increased significantly than those before modeling or before medication (P<0.05 or P<0.01). Compared with blank group, body weight and serum content of GAS were decreased significantly in model group, while serum content of VIP was increased significantly (P<0.01). Compared with model group, body weight of Shengmaiyin medium-dose group and Shengmaiyin polysaccharide low-dose and high-dose groups, serum contents of D-xylose and GAS in Shengmaiyin medium-dose and high-dose groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were increased significantly, while serum contents of VIP in Shengmaiyin groups and Shengmaiyin polysaccharide low-dose and medium-dose groups were all decreased significantly (P<0.05 or P<0.01). CONCLUSIONS: The optimized extraction technology of Shengmaiyin polysaccharide is stable and feasible. Shengmaiyin and its polysaccharide contribute to the recovery of intestinal function of spleen deficiency model rat, the effects of which may be associated with the secretion regulation of GAS and VIP.
9. Effect of Shenqi Compound Recipe on Intestinal Microecology of Patients with Qi and Yin Deficiency and Blood Stasis Syndrome Newly Diagnosed Type 2 Diabetes Mellitus
Xin-xia ZHANG ; Wan-fu LIU ; Ran XIONG ; Bo-tong YANG ; Min ZHONG ; Xue-hui LIU ; Chun-guang XIE
Chinese Journal of Experimental Traditional Medical Formulae 2019;25(22):72-77
Objective: To observe the effect of Shenqi compound recipe on glucose and lipid metabolism in patients with Qi and Yin deficiency and blood stasis syndrome newly diagnosed type 2 diabetes mellitus (T2DM), and its intervention effect on intestinal microecology and serum proinflammatory factors. Method: The 106 eligible patients were divided into the observation group (54 cases) and the control group (52 cases) by random number table method. Another 40 healthy volunteers in physical examination center of the hospital during the same period were enrolled as health control group. On the basis of Guidelines for the Prevention and Treatment of Type 2 Diabetes in China(2013 edition), control group was provided lifestyle interventions, such as reasonable diet, weight control, moderate exercise, salt restriction, tobacco control, alcohol restriction and psychological balance. In addition to the therapy of the control group, the observation group was given Shengi compound for oral administration, 2 times/days. Both groups were treated for 8 weeks. The fasting blood glucose (FBG), postprandial 2 h blood glucose (PBG), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index (HOMA-IR), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) before and after treatment were evaluated. The structure and quantity of intestinal flora before and after treatment were detected. The traditional Chinese medicine(TCM)symptom was scored. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), IL-8, and tumor necrosis factor-α (TNF-α) were measured before and after treatment. Result: FBG, PBG, HbA1c and HOMA-IR levels in observation group were lower than those in control group (P<0.05). The operational taxonomy unit (OUT) of observation group was higher than that of control group (P<0.05). The abundances of Bacteroides and Klebsiella in observation group were higher than those in control group, while actinomyces abundance was significantly lower than that in control group (P<0.05). TC, TG and LDL-C levels in observation group were lower than those in control group, while HDL-C was higher than that in control group (P<0.05). IL-1β, IL-6, IL-8 and TNF-α levels in observation group were lower than those in control group (P<0.01). Rank-sum test showed that the efficacy on diseases of observation group was better than that of control group after treatment (Z=2.134, P<0.05). Conclusion: Shenqi compound can regulate blood glucose and blood lipid in patients with newly diagnosed T2DM (Qi and Yin deficiency and blood stasis syndrome), improve IR, intestinal microecology imbalance, and reduce non-specific inflammatory response, with a good clinical efficacy on intestinal microecology of patients with Qi and Yin deficiency and blood stasis syndrome newly diagnosed type 2 diabetes mellitus.
10.Clinical features and prognosis of patients with acute ST-segment elevation myocardial infarction comorbid with diabetes mellitus
Yi LUAN ; Wei LI ; Li-Rong WU ; Xing-De LIU ; Ping LI ; Jin-Feng LIANG ; Bo WEI ; Zheng SHEN ; Deng-Hai XIE ; An-Min LI ; Yun CHEN ; Guo-Bao XIONG ; Hong-Ling WU ; Dong-Jiang LI ; Zong-Gang DUAN
Chinese Journal of Interventional Cardiology 2018;26(2):87-92
Objective To investigate the clinical features of patients with acute ST-segment elevation myocardial infarction (STEMI) comorbid with diabetes mellitus (DM) and to analyze the prognosis within 12 months after primary percutaneous coronary intervention (pre-PCI). Methods A total of 375 STEMI patients were divided into the diabetes group (n=140) and the normal blood glucose group(n=235) according to whether they met the diagnostic criteria of DH. The clinical data,characteristics of coronary artery lesions,type of stent implant,rate of coronary slow flow or no-reflow after pre-PCI, and the prognosis within 12 months after PCI of the two groups were investigated.Results Patient in the diabetes group presented with higher mean age ,higher comorbid rates of hypertension , hyperlipidemia and heart function of Killip class Ш and above than patients in the normal blood glucose group (all P<0.05). patients in the diabetes group had higher rates of slow reflow /no-reflow after PCI(12.9% vs.5.5%,P=0.013),higher percentages of 3-ressel disease(40.7% vs. 28.9%,P=0.019)and lef t main lesions(13.6% vs. 7.2%,P=0.044). The in-hospital mortality rates(6.4% vs.1.7%,P=0.020),revascularization rates within 12 months(7.9% vs.0.9%,P=0.001)and incidence of heart failure(7.9% vs. 2.6%,P=0.017)were all higher in the diabetes group. Conclusions STEMI patients comorbid with DM were relatively older, had higher comorbidities of hypertension,hyperlipidemia, three-vessel disease, left main coronary lesions and higher mortality during hospitalization. No significant increase in cardiac death and recurrent myocardial infarction were deserved during the follow-up period. These patients may benefit more from early intervention.

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