Purpose:The objective of this study was to identify the risk factors associated with Carbapenem-resistant Enterobacteriaceae (CRE) colonization in intensive care unit (ICU) patients and to develop a predictive risk model for CRE colonization. Patients and Methods:In this study, 121 ICU patients from Fujian Provincial Hospital were enrolled between January 2021 and July 2022. Based on bacterial culture results from rectal and throat swabs, patients were categorized into two groups: CRE-colonized (n = 18) and non-CRE-colonized (n = 103). To address class imbalance, Synthetic Minority Over-sampling Technique (SMOTE) was applied. Statistical analyses including T-tests, Chi-square tests, and Mann-Whitney U-tests were employed to compare differences between the groups. Feature selection was performed using Lasso regression and Random Forest algorithms. A Logistic regression model was then developed to predict CRE colonization risk, and the results were presented in a nomogram. Results:After applying SMOTE, the dataset included 198 CRE-colonized patients and 180 non-CRE-colonized patients, ensuring balanced groups. The two groups were comparable in most clinical characteristics except for diabetes, previous emergency department admission, and abdominal infection. Eight independent risk factors for CRE colonization were identified through Random Forest, Lasso regression, and Logistic regression, including Acute Physiology and Chronic Health Evaluation (APACHE) II score > 16, length of hospital stay > 31 days, female gender, previous carbapenem antibiotic exposure, skin infection, multi-site infection, immunosuppressant exposure, and tracheal intubation. The risk prediction model for CRE colonization demonstrated high accuracy (87.83%), recall rate (89.9%), precision (85.6%), and an AUC value of 0.877. Patients were categorized into low-risk (0-90 points), medium-risk (91-160 points), and high-risk (161-381 points) groups, with corresponding CRE colonization rates of 1.82%, 7.14%, and 58.33%, respectively. Conclusion:This study identified independent risk factors for CRE colonization and developed a predictive model for assessing the risk of CRE colonization.

Contrast-induced nephropathy (CIN) is a complication of patients undergoing percutaneous coronary intervention (PCI). Promising biomarkers for the early prediction of CIN can significantly improve outcomes of these patients. We searched PubMed, EMBASE, Web of Science, and Cochrane Library for studies. Trials reporting an area under the curve (AUC) for the utility of novel biomarkers in the early prediction of CIN in adults after PCI were included. In total, 42 studies comprising 11,984 adult patients undergoing PCI met the criteria. Four urinary biomarkers and four blood biomarkers were included. For urine biomarkers, the pooled AUCs for neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), liver-type fatty acid-binding protein (L-FABP), and kidney injury molecule-1 (KIM-1) were 0.91 (95% CI 0.89–0.94), 0.79 (0.75–0.82), 0.78 (0.74–0.82), and 0.79 (0.76–0.83), respectively. The blood biomarkers NGAL, cystatin C, brain natriuretic peptide (BNP), and C-reactive protein (CRP) had pooled AUCs of 0.93 (0.91–0.95), 0.92 (0.89–0.94), 0.78 (0.74–0.81), and 0.75 (0.71–0.79), respectively. Subgroup analysis showed that blood NGAL in early CIN predictive time (<6 h) was more effective in predicting CIN. The efficiency of cystatin C in predicting CIN was reduced, whereas that of L-FABP was increased among chronic kidney disease (CKD) patients.

ABSTRACT Purpose: Abnormal activation of NOD-like receptor protein 3 (NLRP3) inflammasome can lead to the occurrence and progression of acute pancreatitis. This study investigated the protective effect of MCC950 on pancreatitis mice. Methods: Eighteen mice were randomly divided into control group, severe acute pancreatitis (SAP) group and SAP+MCC950 group. Serum interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) were measured by ELISA. Hematoxylin and eosin (HE) staining was used to evaluate the pathological damage. Western blotting was used to detect the expression of NLRP3 inflammasome and tight junction proteins in the small intestine and pancreas. Results: MCC950 could reduce the levels of IL-6 and IL-1β in SAP mice. After treatment with MCC950, the expression levels of NLRP3 inflammasome in the pancreas of SAP mice were significantly reduced and the pathological damage to the pancreas and intestine was alleviated. Compared with the control group, the expression of tight junction protein (ZO-1,occludin and claudin-4) in the intestinal mucosa of SAP mice was decreased, and the expression of claudin-4 and occludin were upregulated after MCC950 treatment. Conclusions: MCC950 can inhibit NLRP3 inflammasome activation and significantly reduce the inflammatory response and delay the process of pancreatitis. It has therapeutic potential in the treatment of acute pancreatitis.

Contrast-induced nephropathy (CIN) is a complication of patients undergoing percutaneous coronary intervention (PCI). Promising biomarkers for the early prediction of CIN can significantly improve outcomes of these patients. We searched PubMed, EMBASE, Web of Science, and Cochrane Library for studies. Trials reporting an area under the curve (AUC) for the utility of novel biomarkers in the early prediction of CIN in adults after PCI were included. In total, 42 studies comprising 11,984 adult patients undergoing PCI met the criteria. Four urinary biomarkers and four blood biomarkers were included. For urine biomarkers, the pooled AUCs for neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), liver-type fatty acid-binding protein (L-FABP), and kidney injury molecule-1 (KIM-1) were 0.91 (95% CI 0.89-0.94), 0.79 (0.75-0.82), 0.78 (0.74-0.82), and 0.79 (0.76-0.83), respectively. The blood biomarkers NGAL, cystatin C, brain natriuretic peptide (BNP), and C-reactive protein (CRP) had pooled AUCs of 0.93 (0.91-0.95), 0.92 (0.89-0.94), 0.78 (0.74-0.81), and 0.75 (0.71-0.79), respectively. Subgroup analysis showed that blood NGAL in early CIN predictive time (<6 h) was more effective in predicting CIN. The efficiency of cystatin C in predicting CIN was reduced, whereas that of L-FABP was increased among chronic kidney disease (CKD) patients.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults, associated with poor prognosis and easy relapse of disease. Circular RNAs (circRNAs) were detected to be m6A modified and the role of m6A circRNAs has been reported in other diseases including cancers, however, their role has not been elucidated in AML yet. In the present study, we aimed to investigate the expression profiling of m6A circRNAs in AML. We performed m6A circRNAs microarray analysis to identify differentially expressed m6A circRNAs in bone marrow samples from AML patients and healthy individuals (control). Furthermore, bioinformatics analysis predicted the potential functions and relevant pathways that may be associated with the m6A circRNAs. The circRNA m6A methylation levels were found to be positively associated with the circRNAs expression, suggesting circRNA m6A modification could contribute to circRNA regulation in AML. Further analysis demonstrated that circRNA m6A modification might influence the circRNA-miRNA-mRNA co-expression network that may contribute to the circRNA regulatory network in AML. Our findings provide evidence of the differential expression profile of m6A circRNAs in AML, and circRNA m6A modification may contribute to circRNA regulatory function in AML.

Cytokines interact closely with each other and play a crucial role in the progression of sepsis. We focused on the associations of a cytokine network with IL-35 in sepsis. First, the retrospective study included 42 patients with sepsis and 23 healthy controls. Blood samples were collected from patients on days 1, 2, 4. Levels of IL-35, IL-1β, IL-4, IL-6, IL-10, IL-17A, TNF-α and IFN-γ were measured. They all increased to various extend on days 1, 2, 4, and strongly associated with markers of disease severity. Network analysis revealed a network formed by IL-35, with IL-6, IL-10, IL-17A, TNF-α and IFN-γ throughout the acute phase of sepsis(days 1, 2 and4). Then, the CLP-induced septic rats were used. The recombinant human IL-35(rIL-35) upregulated the levels of IL-10, but downregulated IL-4, IL-6, IL-17A, TNF-α and IFN-γ, while it had no significant effect on IL-1β, and upregulated the percentages of CD4+CD25+Tregs, and iTR35, but downregulated Teff cells in the peripheral blood. The rIL-35 reduced inflammation damage and improved prognosis of the septic rats. IL-35 forms a network with other cytokines and plays a major role in the immunopathogenesis of sepsis.

One of the most common causes of mortality and disability across the world is sepsis, a condition marked by an abnormal immunological reaction in the body. The prognosis of patients with sepsis is much improved when they are diagnosed early and provided with proper therapy. Soluble triggering receptors expressed on myeloid cells (STREM-1), Interleukin-10 (IL-10), neuronal serum enolase (NSE), and so on are possible biomarkers for the diagnosis of sepsis based on the pathophysiology. Blood purification treatment to control the cytokine storm induced by sepsis was regarded to be promising. Recently, the treatment of sepsis is likely to shift toward a multimodal approach. Hence in this paper, we investigated the effect of a continuous blood purification technique (hemofiltration without heparin followed by hemoadsorption) on STREM-1, NSE, and IL-10 levels in patients suffering from sepsis. A sample of hundred patients suffering from sepsis was randomly allocated to one of the two groups (study and control groups). The control group got standard sepsis care, whereas the research group got continuous blood purification treatment. To compare the differences between the two groups, we used t-statistical analysis. Blood STREM-1, IL-10, and NSE concentrations of the study group were significantly lesser than that of the control group after therapy. As a result, continuous blood purification can significantly minimize the dysregulated immune response in patients with sepsis, promote immune function and improve the survival rate.

Canonical epigenetic modifications, which include histone modification, chromatin remodeling and DNA methylation, play key roles in numerous cellular processes. Epigenetics underlies how cells that posses DNA with similar sequences develop into different cell types with different functions in an organism. Earlier epigenetic research has primarily been focused at the chromatin level. However, the number of studies on epigenetic modifications of RNA, such as N-1-methyladenosine, 2 '-O-ribosemethylation, inosine, 5-methylcytidine, N-6-methyladenosine (m(6)A) and pseudouridine, has seen an increase. Circular RNAs (circRNAs), a type of RNA species that lacks a 5 ' cap or 3 ' poly(A) tail, are abundantly expressed in acute myeloid leukemia (AML) and may regulate disease progression. circRNAs possess various functions, including microRNA sponging, gene transcription regulation and RNA-binding protein interaction. Furthermore, circRNAs are m(6)A methylated in other types of cancer, such as colorectal and hypopharyngeal squamous cell cancers. Therefore, the critical roles of circRNA epigenetic modifications, particularly m(6)A, and their possible involvement in AML are discussed in the present review. Epigenetic modification of circRNAs may become a diagnostic and therapeutic target for AML in the future.

Objective This study was designed to uncover the mechanism of miR-34b-5p-mediated aquaporin-2 (AQP2) in sepsis-induced injury using human renal tubular epithelial cells (HK-2).Methods Serum levels of miR-34b-5p, TNF-α, IL-1β, IL-6, serum creatinine (SCr), and blood urea nitrogen (BUN) in septic patients with acute kidney injury (AKI) and healthy controls were detected. Lipopolysaccharide (LPS) was used to induce sepsis in HK-2 cells. LPS-induced HK-2 cells were transfected with miR-34b-5p inhibitor, miR-34b-5p mimic, pcDNA3.1-AQP2, si-AQP2, miR-34b-5p inhibitor + si-NC, or miR-34b-5p inhibitor + si-AQP2. The expressions of miR-34b-5p, AQP2, Bax, Bcl-2, cleaved caspase-3, TNF-α, IL-1β, and IL-6 in HK-2 cells were detected. TUNEL staining revealed the apoptosis of HK-2 cells. Dual-luciferase reporter assay verified the binding between miR-34b-5p and AQP2.Results The expression of miR-34b-5p and the inflammatory responses were augmented in septic AKI patients. miR-34b-5p was up-regulated and AQP2 was down-regulated in LPS-induced HK-2 cells. miR-34b-5p inhibition or AQP2 overexpression ameliorated apoptosis and inflammation in LPS-induced HK-2 cells. In contrast, overexpressing miR-34b-5p deteriorated LPS-induced injury in HK-2 cells. AQP2 was a downstream target of miR-34b-5p. AQP2 silencing abolished the suppressive effects of miR-34b-5p inhibition on LPS-induced apoptosis and inflammatory response in HK-2 cells.Conclusion miR-34b-5p inhibits AQP2 to promote LPS-induced injury in HK-2 cells.

Objective:To systematically assess the effect of electrical impedance tomography (EIT) guided positive end-expiratory pressure (PEEP) titration on patients with acute respiratory distress syndrome (ARDS) / acute respiratory failure (ARF).Methods:Systematic searches were performed in the PubMed, Embase, Web of science, Cochrane library, CNKI, VIP and Wanfang databases for articles published up to March 10, 2020 about effects of EIT guided PEEP titration on ARDS / ARF patients. Two reviewers independently screened articles, extracted data and evaluated the quality of included studies. Then meta-analysis was conducted using RevMan 5.3 software.Results:Seven articles with 147 patients were enrolled in this study. Meta-analysis showed that compared with traditional PEEP setting methods (ARDS network, pressure / volume curve, flow rate / volume curve), the PEEP [mean difference (MD) = 2.35, 95% confidence interval (CI) (0.38, 4.32), Z = 2.34, P = 0.02] and oxygenation index [MD = 26.63, 95%CI (7.89, 45.38), Z = 2.78, P = 0.005] were higher, and the lung compliance [MD = 5.06, 95%CI (2.49, 7.62), Z = 3.86, P = 0.000 1] was better in ARDS / ARF patients with EIT guided PEEP titration. However, there was no significant difference in the mean arterial pressure [MD = -0.97, 95%CI (-6.31, 4.36), Z = 0.36, P = 0.72] and in-hospital mortality [odds ratio = 0.40, 95%CI (0.14, 1.12), Z = 1.74, P = 0.08] between the EIT guided PEEP titration and traditional PEEP setting methods.Conclusion:Compared with traditional PEEP setting methods, EIT guided PEEP titration can improve the PEEP, oxygenation index and lung compliance, but has no significant effect on the mean arterial pressure and in-hospital mortality of ARDS / ARF patients.

目的 探索凝血指标抗凝血酶Ⅲ(ATⅢ)在急性肺动脉血栓栓塞症(PTE)中的临床应用价值.方法 采用回顾性研究方法,纳入2012年5月至2019年6月福建省立医院收住的204例已确诊的PTE患者,研究资料包括患者基本情况、基础疾病及入院24 h内实验室检查、评分系统,分析ATⅢ与PTE院内死亡及不良事件之间的关系,并进一步探究ATⅢ优化危险分层的价值.结果 ATⅢ水平预测PTE患者院内死亡率的受试者工作特征曲线下面积(AUC)为0.719,其中截断值为77.7％(敏感性64.71％,特异性80.21％).根据截断值水平分为ATⅢ≤77.7％组(n=48)及ATⅢ＞77.7％组(n=156),慢性心力衰竭、白细胞计数、血小板计数、谷丙转氨酶、白蛋白、肌钙蛋白Ⅰ组间差异有统计学意义(P＜0.05).据院内死亡情况分为死亡组(n=17)与存活组(n=187),比较白细胞计数、ATⅢ、D-二聚体、谷丙转氨酶、白蛋白、肾小球滤过率、APACHE Ⅱ评分组间差异有统计学意义(P＜0.05).Logistics回归分析发现ATⅢ≤77.7％、白细胞计数为院内死亡的独立危险因素.运用受试者工作特征曲线分别对危险分层及危险分层联合ATⅢ进行院内死亡预测,AUC分别为0.705、0.813,差异有统计学意义(P＜0.05).进一步采用列线图展示危险分层联合ATⅢ新评分模型.结论 ATⅢ≤77.7％是院内死亡的独立危险因素,并有利于优化危险分层,其机制可能与血栓形成、右心功能障碍及炎症反应等有关.

Objective: Pulmonary embolism is a terrible cardiovascular condition with considerable morbidity and mortality. Previous studies have investigated systolic blood pressure (systolic BP) and diastolic blood pressure (diastolic BP) as being related to 30-day and in-hospital mortality. We aimed to determine whether the average mean arterial pressure (aMAP) in the first 24 hours of hospital admission is useful in predicting short-term outcomes of intermediate-risk and high-risk PE patients. Method: We conducted a single-center retrospective study. From May 2012 to April 2019, 122 intermediate-risk and high-risk PE patients were included. The primary outcome was in-hospital mortality. The secondary outcome was adverse events. ROC curves and cut-off values for aMAP predicting in-hospital death were computed. According to cut-off values, we categorized five groups defined as follows: group 1: aMAP<70 mmHg; group 2: 70 mmHg≤aMAP<80 mmHg; group 3: 80 mmHg≤aMAP<90 mmHg; group 4: 90 mmHg≤aMAP<100 mmHg; and group 5: aMAP≥100 mmHg. Cox regression models were calculated to investigate associations between aMAP and in-hospital death. Results: In the study group of 122 patients, 15 patients (12.30%) died in the hospital due to PE. ROC analysis for MAP predicting in-hospital death revealed an AUC of 0.729 with a cut-off value of 79.4 mmHg. Cox regression models showed a significant association between in-hospital death and aMAP group 1 (ref), aMAP group 2 (OR 1.680, 95% CI 0.020-140.335), aMAP group 3 (OR 0.003, 95% CI 0.0001-0.343), aMAP group 4 (OR 0.006, 95% CI 0.0001-1.671), and aMAP group 5 (OR 0.003, 95% CI 0.0001-9.744). In particular, those with an aMAP of 80-90 mmHg suffer from minimum adverse events. Conclusion: The prognostic role of MAP during the first 24 hours of hospital admission should be emphasized in patients with PE. The optimal range of MAP for intermediate-risk and high-risk PE patients may be 80 to 90 mmHg.

目的 探究新型炎症指标单核细胞计数(monocyte)与高密度脂蛋白(high density lipoprotein,HDL)比值(monocyte/HDL ratio,MHR)与急性肺栓塞(acute pulmonary embolism,APE)病情严重程度及院内病死率的关系.方法 采用回顾性队列研究方法,选择2012年5月至2019年3月福建省立医院收治的286例APE患者,收集入组患者年龄、基础疾病等基本信息,进行血白细胞计数(WBC)、单核细胞计数、HDL、肌钙蛋白(cTn)及D-二聚体等实验室检查,计算MHR,分析不同病情严重程度院内生存和死亡患者的MHR水平.结果 根据院内生存情况分为生存组(n=271)及死亡组(n=15),两组患者年龄、高血压、糖尿病及脑梗死等基础疾病比较差异无统计学意义(P＞0.05),血WBC、单核细胞计数、白蛋白、HDL及MHR等差异有统计学意义(P＜0.05).利用上述指标绘制受试者工作特征曲线(ROC曲线)对院内病死率进行预测,曲线下面积(AUG)最大分别为血WBC 0.798,MHR 0.675.根据MHR截断值0.37分为MHR＞ 0.37组及MHR≤0.37组,发现MHR＞ 0.37组患者D-二聚体、cTn水平明显高于MHR≤0.37组,分别为[4.08(1.91,7.54)vs.2.59(0.72,5.41)]、[0.02(0.00,0.09)vs.0.02(0.00,0.02)],差异有统计学意义(P＜0.05).根据《2019年欧洲心脏病协会与呼吸协会共同制定的急性肺栓塞诊断与管理指南》[1]中危险分层将APE患者分为高危、中高危、中低危及低危.中低危、中高危及高危组患者MHR水平明显高于低危组患者,差异均有统计学意义(P值分别为0.003、0.022、0.004).应用二元Logistic回归分析发现,MHR、血WBC、D-二聚体与肌酐(Cr)升高及HDL降低均为APE患者院内死亡的独立危险因素.结论 MHR、血WBC、D-二聚体与血Cr升高及HDL降低均是APE院内死亡的独立危险因素;中危及高危APE患者MHR水平高于低危患者,MHR可以预测APE患者病情严重程度.

OBJECTIVETo investigate the down-regulation effect of let-7b-5p on the expression of FTO in acute myeloid leukemia cell line THP-1 and inhibitory effect on THP-1 proliferation via m6A/MYC signaling pathway.METHODSThe acute myeloid leukemia cell line THP-1 and the normal human peripheral blood mononuclear cells (PBMNC) were selected as subjects. The expression of let-7b-5p and FTO mRNA in those cells was detected by qPCR, further the expression of FTO protein in those cells was detected by Western blot. And, the luciferase reporter gene assay was used to verify the targeting effect of let-7b-5p on FTO. Finally, THP-1 cells were transfected respectively with let-7b-5p mimic, and PBMNC with let-7b-5p inhibitor, there after the C-MYC mRNA m6A enrichment level in transfected cells was analyzed by dot blot, the expression levels of let-7b-5p, FTO and c-MYC were assayed by RT-PCR, the expressions of FTO and c-MYC protein were verified by Western blot, and the proliferation level of cells after transfection was detected by MTT assay.RESULTSCompared with PBMNC, the expression of let-7b-5p in THP-1 significantly decreased, while the expression of FTO was significantly increased (P<0.05). After transfection with let-7b-5p mimic combined with FTO 3'-UTR, the luciferase activity of transfected THP-1 cells significantly decreased, but the luciferase activity significantly increased after transfection with mutant 3'-UTR, which was significantly different from the negative control group(blank vector) (P<0.05). Let-7b-5p inhibitor down-regulated c-MYC mRNA m6A enrichment, and then up-regulated the expression of FTO in transfected PBMNC cells, the effects of which were significant (P<0.05). However, let-7b-5p mimic up-regulated c-MYC mRNA m6A enrichment level and down-regulated the expression of FTO in the transfected THP-1 cells, and the cell proliferation rate was significantly lower than that in the negative control group (blank vector) (P<0.05).CONCLUSIONHuman acute myeloid leukemia cell line THP-1 low expresses the let-7b-5p, which regulates c-MYC expression through let-7b-5p-/FTO-/m6A axis and promotes the proliferation of leukemia cell line THP-1.

Objective To investigate the expression of interleukin-35 (IL-35) in patients with sepsis and multiple organ dysfunction syndrome (MODS),and to explore its effect on disease prognosis.Methods A prospective study was conducted.Twenty-two patients with sepsis and MODS were selected as study group,and 20 healthy volunteers were selected as controls.Blood samples of the patients and the volunteers were taken within 1 hour of the patient's visit,and cytokines,such as IL-35,IL-4,IL-10,IL-17,INF-γ,and TGF-β,were simultaneously detected by enzyme-linked immunosorbent assay (ELISA).The expression level of CD4+CD25+FOXP3+ Treg cells was detected by flow cytometry,and the acute physiology and chronic health status scoring system Ⅱ (APACHE Ⅱ) was calculated.According to the survival outcome at 28 days after admission,the expression levels of IL-35 in the survival group (12 cases)and the death group (8 cases) were compared.The Spearman method was used to analyze the correlation between IL-35 level and the.above indicators in patients with sepsis and MODS.Results Compared with the control group,the IL-35 levels were significantly increased in the study group (P＜0.05),and the IL-35 levels began to increase gradually on the first day of the disease,and significantly higher on the 7th day (P＜0.01).The number of CD4+CD25+FOXP3+regulatory T cells increased in the study group,especially on the 4th day (P＜0.01),and on the first day of the disease,the IL-35 levels were positively correlated to the number of CD4+CD25+FOXP3+Treg cells (r=0.60,P＜0.05).Compared with the control group,the levels of IL-10,IL-4,IL-17,INF-γ and TGF-β in the study group increased gradually with the course of disease,and were significantly higher on the 7th day than those on the 1st day and the 4th day (P＜0.05).The IL-35 levels in the study group were positively correlated with INF-γ and TGF-β cytokines (P＜0.05),whereas there was no significant relationship between the levels of IL-10,IL-4,and IL-17 (P＞0.05).The IL-35 level in the death group was significantly lower than that in the survival group (P＜0.05).IL-35 levels in the survival and death groups were analyzed by ROC curve with a AUC of 0.78 (P=0.03).The ~ IL-35 level was negatively correlated with the APACHE-Ⅱ score in the study group (r=-0.78,P＜0.01).Conclusions The IL-35 levels in patients with sepsis and MODS is significantly higher than that in healthy controls,and negatively correlated with APACHE Ⅱ score.The level of IL-35 has an important implication for the prognosis for patients with sepsis.

急性肾损伤(acute kidney injury, AKI)作为常见的临床症候,与重症患者的不良预后密切相关.AKI以肾功能迅速恶化为特征,是危重症常见的并发症 [1],也是造成危重症患者死亡的主要原因之一,AKI患者病死率是未发生AKI患者的2～5倍[2].高发病率、高病死率、高医疗费用、预后差等特点,使AKI成为一个备受关注的健康问题 [3].因此,阐明其发病机制,早期诊断,提高治疗效果,降低病死率及终末期肾脏病(end stage renal disease, ESRD)比例,具有重要的临床意义.

目的:探讨分析依赖热稳定解旋酶DNA恒温扩增技术检测烟曲霉;方法:将烟曲霉基因作为目的片段,以此设计特异度引物,并且在恒温条件下建立快速检测烟曲霉依赖热稳定解旋酶DNA恒温扩增技术,进行特异度和灵敏度实验,并与普通聚合酶链式反应(PCR)方法进行比较.结果:热稳定的依赖解旋酶恒温核酸扩增(tHDA)产物序列与EnsemblFungi数据库的序列相似率高达95％;在特异度检测中,黄曲霉的电泳结果为阳性,在100 bp处能够观察到清晰的条带;在灵敏度检测中,不同稀释浓度均能够明显观察到条带;结论:烟曲霉的tHDA检测方法具有普通PCR的特异、灵敏等特点,并且对仪器要求低,普通水浴槽即可进行反应.

The present study was designed to investigate the relationship among epigenetic changes in Wnt antagonists, histone H4K20me1 and the expression of tumor-suppressor genes in acute leukemia (AL) to better understand the pathogenesis of leukemia. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect messenger RNA (mRNA) expression levels of Wnt antagonists (Wnt5a, HDPR1, DKK1 and DKK3) in patients with AL and in normal controls; pyrophosphate sequencing was performed to detect the methylation status of the Wnt5a promoter; and western blotting was performed to detect the overall expression levels of Wnt5a protein and histone H4K20me1 in patients with acute myeloid leukemia (AML) and in normal controls. The relationship between Wnt5a protein expression and histone H4K20me1 was analyzed. Chromatin immunoprecipitation-qPCR (ChIP-qPCR) was performed to investigate the recruitment of H4K20me1 and SET8 to the Wnt5a promoter and coding regions. Our results demonstrated that the expression levels of Wnt antagonists were generally low in AML, but showed differential expression in acute lymphocytic leukemia (ALL). In most cases of AML, methylation of the Wnt5a promoter was observed and Wnt5a protein expression was low. In some cases of AML, the overall level of H4K20me1 protein was higher than that in normal controls. In addition, Wnt5a expression was positively correlated with H4K20me1 expression and was unrelated to the methylation status of its promoter. Moreover, H4K20me1 and SET8 were enriched in the Wnt5a promoter region and coding region. By contrast, Wnt5a expression was unrelated to H4K20me1 expression in normal controls. Moreover, we observed that the methylation of Wnt antagonists was often found in patients with AL, particularly those with AML, whereas the extent of methylation was variable in ALL patients. Wnt5a expression was positively correlated with the enrichment of H4K20me1 and SET8 at the Wnt5a promoter and coding regions. H4K20me1 increased Wnt5a expression by promoting transcription initiation and elongation.

Our purpose was to investigate the effect of 3-deazaneplanocin A (DZNep) on human T-cell acute lymphoid leukemia (T-ALL) cells, and to explore the underlying molecular mechanisms. The human T-ALL cell line Molt4 was treated with DZNep, and cell proliferation was examined. The expression of Mps one binder kinase activator 1 gene (MOB1) mRNA and protein was determined by RT-PCR and Western blotting, respectively. The histone modification effect of DZNep on the lysine 9 of histone 3 associated with MOB1 promoters was examined with chromatin immunoprecipitation and quantitative PCR, and CpG methylation in MOB1 promoters was detected by bisulfite sequencing PCR. DZNep treatment inhibited the growth of Molt4 cells. The expressions of MOB1 genes were upregulated by DZNep treatment, and histone methylations in their promoters were significantly reduced. The results indicate that DZNep is a promising therapeutic compound for the treatment of human T-ALL.