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Objective@#To examine the correlation between the promoter methylation of Sterol regulatory-element binding protein-2 (SREBP-2) and miR-33a expression as well as serum markers in patients with coronary artery disease (CAD).@*Methods@#The case-control study. 100 participants who underwent coronary angiography from August 2017 to April 2018 in TaiheHospital, Hubei University of Medicine, were recruited in this study.The methylation level of two fragments, including 12 CpG sites in the promoter region of SREBP-2, have been detected by pyrosequencing in 50 patients with coronary artery disease (CAD) and 50 non-CAD controls. Serum miR-33a level and a panel of 15 CAD related biomarkers were examined by qPCR and routine biochemistry methods.@*Results@#Methylation level of one CpG site (F1-4 loci) in SREBP-2 promoter region were significant higher in CAD patients than in controls(4.56%±0.70% vs 3.54%±0.72%, t=-3.864, P<0.001); methylation level of F1-4 site was negatively correlates with the serum miR-33a levels and high-density lipoprotein cholesterol (HDL-C) levels(r=-0.318, P=0.001; r=-0.225, P=0.024, respectively). Furthermore, F1-4 hypermethylation was an independent risk factor of CAD, independent of age, gender, histories of hypertension, hyperlipidemia, and diabetes(OR=2.452, 95%CI=1.398-4.299, P=0.002).@*Conclusion@#These results suggest that DNA methylation and miRNA might cooperate to regulate the lipid metabolism in CAD.
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Objective:To examine the correlation between the promoter methylation of Sterol regulatory-element binding protein-2 (SREBP-2) and miR-33a expression as well as serum markers in patients with coronary artery disease (CAD).Methods:The case-control study. 100 participants who underwent coronary angiography from August 2017 to April 2018 in TaiheHospital, Hubei University of Medicine, were recruited in this study.The methylation level of two fragments, including 12 CpG sites in the promoter region of SREBP-2, have been detected by pyrosequencing in 50 patients with coronary artery disease (CAD) and 50 non-CAD controls. Serum miR-33a level and a panel of 15 CAD related biomarkers were examined by qPCR and routine biochemistry methods.Results:Methylation level of one CpG site (F1-4 loci) in SREBP-2 promoter region were significant higher in CAD patients than in controls(4.56%±0.70% vs 3.54%±0.72%, t=-3.864, P<0.001); methylation level of F1-4 site was negatively correlates with the serum miR-33a levels and high-density lipoprotein cholesterol (HDL-C) levels( r=-0.318, P=0.001; r=-0.225, P=0.024, respectively). Furthermore, F1-4 hypermethylation was an independent risk factor of CAD, independent of age, gender, histories of hypertension, hyperlipidemia, and diabetes( OR=2.452, 95 %CI=1.398-4.299, P=0.002). Conclusion:These results suggest that DNA methylation and miRNA might cooperate to regulate the lipid metabolism in CAD.
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Objective To investigate the characteristics of nosocomial infection of carbapenem resistant En-terobacteriaceae ,then to study the risk factors and prognosis of patients ,as to provide evidence for effective control of CRE hospital infection .Methods A retrospective epidemiology study was conducted of CRE infec-ted patients from January 2016 to June 2017 .This was a 1∶2 matched case-control study of patients with in-fection due to carbapenem resistant Enterobacteriaceae and randomly select 218 patients with non CRE infec-tion in the same hospital as control group ,then the risk factors of CRE infection were investigate .Results A-mong the 109 cases CRE infection patients ,the most common bacteria were Klebsiella pneumoniae ,Enter-obacter cloacae and Escherichia coli .The CRE strains were more sensitive to minocycline than other common antibiotics ,the resistance rate to other antibiotics was more than 40% .Univariate analysis showed that ICU staying time more than 7 d ,using beta lactamase inhibitors and carbapenem antibiotics ,combination therapy and mechanical ventilation were the potential risk factors of CRE nosocomial infection .The non conditional multivariate Logistic regression analysis showed that Check in ICU more than 7 d (OR＝ 4 .981 ,95% CI 2 .451-10 .122 ) ,the use of containing beta lactamase inhibitor antibiotics (OR＝ 3 .718 ,95% CI 2 .162-6 .394) ,use of carbapenem antibiotics (OR＝3 .232 ,95% CI 1 .879-5 .561) ,and mechanical ventilation (OR＝5 .26 ,95% CI 2 .576-10 .742) were independent risk factors of CRE nosocomial infection .The nosocomial in- fection CRE strain was with highly antibiotic resistance rate ,and the average hospitalization time and mortality were significantly higher than those of the control group .Conclusion The carbapenem resistant Enterobacte-riaceae infection had multiple independent risk factors ,strengthening of these independent risk factor control can effectively prevent the spread of CRE isolates infection .
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Objective To investigate the clinical values of detection of plasma miRNA-544a (miR-544a) expression level in the diagnosis and treatment of lung cancer.Methods A total of 110 patients diagnosed with lung cancer in Ⅰ-Ⅱ stage from June 2017 to March 2018 in Shiyan Taihe Hospital were collected,and 35 patients with benign pulmonary nodules and 64 healthy people were also collected as the controls.Realtime quantitative polymerase chain reaction (RT-qPCR) was used to detect the level of miR-S44a in plasma of 64 healthy people,110 patients with lung cancer (50 newly diagnosed patients without anti-tumor treatment,33 patients one week after radical resection,27 patients after one week chemotherapy with the same dose) and 35 patients with benign pulmonary nodules.Of the 50 newly diagnosed patients,42 cases were non-small cell lung cancer and 8 cases were small cell lung cancer.The plasma expression level of miR-544a in each group was compared by using Mann-Whitney U test,and the efficacy of miR-544a in the diagnosis of lung cancer was evaluated by the receiver operationg characteristic (ROC) curve and the area under the curve (AUC).Results The plasma expression levels of miR-544a in the newly diagnosed untreated lung cancer group,one week after operation group,one week chemotherapy group,healthy control group and benign pulmonary nodule group were 1.40 nmol/L (0.55 nmol/L,8.76 nmol/L),33.52 nmol/L (3.64 nmol/L,250.47 nmo/L),8.87 nmol/L (0.68 nmol/L,125.43 nmol/L),0.31 nmol/L (0.17 nmol/L,1.19 nmol/L),1.04 nmol/L (0.31 nmol/L,4.62 nmol/L),respectively,and the differences between untreated lung cancer group and the other 4 groups were statistically significant (Z =-4.483,P ＜ 0.001;Z =-4.274,P ＜ 0.001;Z =-2.562,P =0.01;Z =-2.152,P =0.031).The expression levels of miR-544a in non-small cell lung cancer group and small cell lung cancer group were 1.40 nmol/L (0.66 nmol/L,8.76 nmol/L) and 1.37 nmol/L (0.26 nmol/L,36.97 nmol/L),respectively.The differences between non-small cell lung cancer group and healthy control group and benign pulmonary nodule group were statistically significant (Z =-4.463,P ＜ 0.001;Z =-2.026,P =0.043).Compared with the healthy people,the AUC of miR-544a for diagnosing the lung cancer was 0.841,the sensitivity was 87.5 ％,and the specificity was 68.0 ％.Compared with the benign pulmonary nodule,the AUC for diagnosing lung cancer was 0.638,the sensitive was 45.7 ％,and the specificity was 80.0 ％.Conclusions The plasma expression level of miR-544a has certain significances in the differential diagnosis of early stage lung cancer and benign pulmonary nodules and healthy people,and it can be used as a potential biomarker for diagnosing early stage lung cancer,especially for the non-small cell lung cancer.The plasma expression of miR-544a is increased after surgery or chemotherapy,suggesting that its expression may be related to the occurrence and development of lung cancer,and miR-544a may become a new target for cancer treatment.
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<p><b>OBJECTIVE</b>To assess the association of methylenetetrahydrofolate reductase (MTHFR) gene 677C to T polymorphism with blood homocysteine (Hcy) level among women of childbearing age from Shiyan area.</p><p><b>METHODS</b>PCR-chip hybridization was used to determine the genotype of MTHFR 677C to T, and a biochemical assay was used to determine the total Hcy level among 428 healthy women of childbearing age. Association of MTHFR 677C to T with total Hcy level was assessed.</p><p><b>RESULTS</b>Heterozygous CT mutation was most common form for the MTHFR 677C to T polymorphisms and amounted for 49.77% among the group, while the CC wild type and homozygous TT mutation respectively accounted for 30.61% and 19.63%. These gave a frequency of 44.51% for the 677T allele. The dominant genotype among different age groups were the CT type. Of note, the proportion of MTHFR 677CC is higher in women above 30 years of age. The distribution of MTHFR 677C to T genotypes has differed significantly among different age groups (P<0.05). Compared with those with wild type alleles, carriers of MTHFR mutations had a higher plasma Hcy level. The genotypic frequencies of MTHFR C677T in Shiyan region differed significantly from those of Sichuan, Hebei, Henan and Shandong (P<0.05) but were similar to those of Jiangsu, Guangdong, Ningxia and Xinjiang.</p><p><b>CONCLUSION</b>The distribution of MTHFR C677T polymorphism among women of childbearing age in Shiyan area is influenced by age and is geographically specific and associated with plasma Hcy level. Nearly 50% of women have carried the high risk alleles, for whom folic acid supplementation is crucial for the reduction of birth defect rate.</p>
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Objective To investigate the changes of plasma miR-29a level in patients with hepatocellular carcinoma(HCC) and its clinical significance.Methods Case-control study,30 patients with HCC,30 patients with cirrhosis of liver (LC) and 30 healthy controls (HC) were recruited from Shiyan Taihe Hospital,2016 January to June.The level of microRNA-29a (miR-29a) in plasma was detected by real time quantitative PCR,and the sensitivity and specificity of plasma miR-29a expression in the diagnosis of HCC were analyzed by receiver operating characteristic curve (ROC),and to analyze the correlation between the miR-29a and the alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma,the combination of miR-29a and AFP could improve the diagnostic efficiency of HCC.Results The relative expression of miR-29a was significantly different between HCC group (3.38±8.37),LC group (8.79±3.80) and HC group (11.98±6.64),the expression level of miR-29a in HCC group was significantly lower than that in LC group (P=0.046) and HC group (P=0.001),and that in LC group was significantly lower than that in HC group (P=0.026).The area under the ROC curve of miR-29a was 0.816 (95％ CI 0.695 to 0.938) less than AFP 0.918 (95％ CI 0.853 to 0.982),and the diagnostic efficiency of miR-29a was not as good as that of AFP.The levels of miR-29a and AFP in plasma of patients with hepatocellular carcinoma were significantly correlated (P=0.002).Conclusion The level of miR 29a in plasma of patients with HCC decreased,which may become the reference index of HCC diagnosis.
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Objective To investigate the expression and significance of CCR9/CCL25 pathway in acute rejection of mouse skin transplantation.Method BALB/c mice and C57BL/6 mice were selected as allogeneic and syngeneic skin graft donors and C57BL/6 as recipients,then established a murine skin transplantation model of acute rejection.Allogeneic transplant recipient mice were injected with an AntiCCL25(2 g,as experimental group) mAb or IgG(2 g,as control group) every other day via tail vein,a total of 10 injections.The transplanted graft was scored visually daily,and then we collected skin graft and spleen from recipient mice of each group at 3,5,7 days after transplantation.HE staining was done to analyze necrosis of skin tissue Confocal and immunohistochemistry were also used to measure CCR9 and CCL25 expression in recipient skin grafts.Result HE staining indicated that there was a widespread inflammatory cell infiltration in the skin from allo-transplantation mice,and CCR9 expression measured by immunohistochemistry and confocal was significantly elevated in the surface of the infiltrated CD3 + T cells from skin grafts tissue and spleen.Neutralization of CCL25 with Anti-CCL25 mAb significantly prolonged allogra,ft survival and markedly reduced inflammation.Conclusion CCR9 was highly expressed in the spleen and skin grafts tissue of allogeneic transplanted mice Neutralization of CCL25 by intravenous injection of Anti-CCL25 monoclonal antibody significantly prolonged skin allograft survival.Our study indicates that CCR9/CCL25 pathway is involved in acute rejection process of skin transplantation model in mice were used as syngeneic and skin grit skin graft donor mice.
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MicroRNA (miRNA) is an important gene regulatory molecule involved in the occurrence of a variety of diseases and is relat-ed to tumors. MicroRNA has become a new direction of oncology research in recent years. Studies showed that miR-29 plays dual roles, as tumor suppressor and tumor promoter. The expression of miR-29 significantly differs between cancer and normal tissues. miR-29 is predicted to be a biomarker for early diagnosis and prognosis prediction of certain cancer. This paper reviews the role of miR-29 in the pathogenesis of cancer.
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Objective To investigate the effects of 5-Aza-2′-deoxycytidine (5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific pol-ymerase chain reaetioll (MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression of hMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively. Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter region of the hM-LH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.
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Objective To investigate the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the apoptosis of A549/DDP cells and the expression of hMLH1 gene.Methods A549/DDP cells were treated with 5-Aza-CdR at 0.5,5,50 μmol/L.The growth curve of A549/DDP cells was investigated by MTT assay.The methylation status of hMLH1 gene was detected by methylation specific PCR (MSP).The expression of hMLH1 mRNA was evaluated by FQ-PCR.The apoptosis rate of A549/DDP cells was analyzed by flow cytometry.Results A549/DDP cells treated with 5-Aza-CdR showed a slow growth in comparison with the control cells,and the growth rates were decreased with the increasing of 5-Aza-CdR concentration.The apoptosis rate after treatment was higher than that before treatment in A549/DDP cells (P ＜ 0.05),and had a positive correlation with 5-Aza-CdR dose (P ＜ 0.001).hMLH1 mRNA expression level was increased in a 5-Aza-CdR concentration dependent manner (P ＜ 0.05).hMLH1 promoter in A549/DDP cells was methylated and hMLH1 mRNA was negatively expressed before treatment,but the mRNA was positively expressed after treatment with 5-Aza-CdR.Conclusions 5-Aza-2'-CdR can induce apoptosis of A549/DDP cells by inducing demethylation of hMLH1 promoter and thereby enhancing hMLH1 gene expression and its tumor suppressor function.
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Objective To investigate the distribution and clinical significance of D‐Dimer positive patients .Methods 1 003 D‐di‐mer positive patients were enrolled in the study ,which were measured by latex enhanced immune turbidimetry .Results The total positive rate of ICU ,cardiology ,respiratory medicine ,orthopedics ,general surgery ,liver disease ,neurosurgery ,obstetrics and gyne‐cology ,oncology departments was 44 .1% .The numbers of D‐dimer positive patients with diffuse intravascular coagulation ,deep vein thrombosis ,pulmonary embolism ,heart cerebrovascular disease ,liver disease ,malignant tumor were 86 ,34 ,26 ,24 ,18 and 12 , respectively .Conclusion The determination of plasma D‐dimer could be used in thrombotic disease prevention and monitoring .
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Objective To detect the expression of p16 and discuss the significance in genesis and development of cervical cancer. Methods FQ-PCR method was used to detect the expression levels of p16 mRNA in cervical cancer(CC),cervical intraepithelial neoplasia(CIN),and normal tissues.And in situ hybridization was used to detect HR-HPV in these tissues.Results The expres-sion levels of p16 were 0.79±0.34,0.70±0.36,0.26±0.21 in CC,CIN and normal cervical tissues,respectively.The difference was significant (P 0.05).The expression level of p16 in HR-HPV positive cases was significantly higher than that in HR-HPV negative cases (P <0.05).Conclusion p16 shows high expression level in cervical cancer,which is closely related to the infection of HR-HPV infection.It suggests that the high ex-pression of p16 play an important role in genesis and development of cervical cancer.
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Objective To analyze the drug resistance of Escherichia coli isolated from urinary tract infection and risk factors of quinolone resistance strains.Methods A total of 705 cases (strains) with Escherichia coli drug resistance isolated from urine specimens were divided into quinolone sensitive group [474 cases(strains)] and quinolone resistance group [231 cases(strains)].The risk factors of the quinolone resistance strains were analyzed.Results The sensitivity rate of amoxicillin/clavulanic acid,cefalotin,ceftazidime,aztreonam,piperacillin,amikacin,compound sulfamethoxazole,ciprofloxacin,gentamicin,levofloxacin,cefepime in quinolone resistance group was higher than that in quinolone sensitive group [50.2％(238/474) vs.78.8％(182/231),11.6％(55/474) vs.48.5％(112/231),17.9％(85/474) vs.63.2％(146/231),15.0％(71/474) vs.57.6％(133/231),3.2％(15/474) vs.27.7％(64/231),80.8％(383/474)vs.93.1％(215/231),16.0％(76/474) vs.49.8％(115/231),0 vs.100.0％(231/231),32.5％ (154/474)vs.70.6％ (163/231),3.8％ (18/474) vs.98.7％ (228/231),18.6％ (88/474) vs.63.2％ (146/231),P ＜0.05].Logistic regression analysis showed history of using the third generation cephalosporins and quinolones,urinary drainage and bacterium producing extra-broad spectrum beta-lactamase was independent risk factor for quinolone resistance Escherichia coli (P ＜ 0.05).Conclusions The epidemic of quinolone resistance Escherichia coli isolated from urine specimens is extremely serious.The quinolone resistance is strong,and infection patients have a high medical cost and average length of stay.The quinolone resistance Escherichia coli infection has multiple independent risk factors.To strengthen the control of the independent risk factors can effectively prevent quinolone resistance strains infection spread.
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Objective To investigate the status and diagnostic value of aberrant phosphatase and tensin homology deleted on chromosometen(PTEN) gene promoter in tissues, peripheral plasma and BALF of lung cancer patients. Methods We analyzed the methylation of PTEN gene in tissues, peripheral plasma and BALF by methylation specific-PCR. Results The frequency of methylation of promoter of PTEN gene was 26.67%(12/45) in lung cancer tissues, 15.56%(7/45) in peripheral plasma, 22.22%(10/45) in BALF, no methylation product was found in lung tissues without cancer, normal plasma and BALF controls (P
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0.05).Conclusions:Detection of hypermethylation change of BRCA1 promoter promises a definite value in histologic type,malignant metastases and early prognostic in sporadic breast cancer.