食管癌组织中Krüppel-like Factor4表达与术后复发转移的相关性

目的探究食管癌组织中Krüppel-like Factor4表达与术后复发转移的相关性。方法选取食管癌患者160例并收集其临床资料。采取免疫组化法检测Krüppel-like Factor4表达,根据患者术后是否复发转移将其分为复发转移组和无复发转移组,对比Krüppel-like Factor 4表达水平,并采用多因素logistic回归分析食管癌切除术患者术后复发转移的危险因素。结果Krüppel-like Factor4蛋白表达与分化程度、临床分期和淋巴结转移等临床病理参数显著相关(P<0.05),与患者的年龄、性别、肿瘤直径均无相关性(P>0.05)。复发转移组术后Krüppel-like Factor4阳性表达率显著低于未复发转移组,差异有统计学意义(P<0.05)。多因素分析显示,Krüppel-like Factor4(OR=2.012,P<0.001)是食管癌术后发生复发转移的独立影响因素。结论食管癌组织中Krüppel-like Factor4表达与患者术后复发转移具有相关性,其对患者术后复发转移具有重要预测价值。

Krüppel-like factor 4慢病毒表达载体构建及其对胃癌细胞BGC-823生物学行为的影响

目的构建Krüppel-like factor4(KLF4)过表达慢病毒载体,探讨其对胃癌细胞株BGC-823生物学行为的影响。方法检测BGC-823中KLF4 mRNA的表达水平;用真核表达质粒pcDNA3.1IE-KLF4-EGFP,将KLF4基因连入慢病毒载体pLv-UbC-IRES2-EGFP中,构建pLv-KLF4-IRES2-EGFP重组慢病毒表达载体。将酶切和测序鉴定后的重组质粒转染至BGC-823中,观察转染情况,RT-PCR检测KLF4 mRNA。慢病毒包装后转染BGC-823细胞,Western印迹检测KLF4蛋白。结果重组质粒经酶切和DNA测序证实目的基因插入正确;pcDNA3.1IE-KLF4-EGFP转染BGC-823细胞后KLF4 mRNA升高。慢病毒包装后转染BGC-823检测到目的蛋白KLF4。KLF4能够将细胞阻滞于G1/S,抑制其生长、促进细胞凋亡,减少细胞侵袭能力。结论转染BGC-823后KLF4蛋白检测证实慢病毒载体成功构建,KLF4能抑制胃癌细胞的恶性转化。

Krüppel-like factor 8 overexpression is correlated with angiogenesis and poor prognosis in gastric cancer

AIM:To investigate Krüppel-like factor 8 (KLF8) expression in gastric cancer and its relationship with angiogenesis and prognosis of gastric cancer. METHODS:One hundred and fifty-four patients with gastric cancer who underwent successful curative resection were retrospectively enrolled in the study. Fifty tumor-adjacent healthy gastric tissues (≥ 5 cm from the tumor margin) obtained during the original resection were randomly selected for comparative analysis. In situ expression of KLF8 and CD34 proteins were examined by immunohistochemistry. The intratumoral microvessel density (MVD) was determined by manually counting the immunostained CD34-positive endothelial cells in three consecutive high-magnification fields (× 200). The relationship between differential KLF8 expression and MVD was assessed using Spearman's correlation coefficient test. χ2 test was performed to evaluate the effects of differential KLF8 expression on clinicopathologic factors. Kaplan-Meier and multivariate Cox survival analyses were used to assess the prognostic value of differential KLF8 expression in gastric cancer. RESULTS:Significantly higher levels of KLF8 protein were detected in gastric cancer tissues than in the adjacent non-cancerous tissues (54.5% vs 34.0%, P < 0.05). KLF8 expression was associated with tumor size (P < 0.001), local invasion (P = 0.005), regional lymph node metastasis (P = 0.029), distant metastasis (P = 0.023), and tumor node metastasis (TNM) stage (P = 0.002), as well as the MVD (r = 0.392, P < 0.001). Patients with KLF8 positive expression had poorer overall survival (P < 0.001) and cancer-specific survival (P < 0.001) than those with negative expression. Multivariate analysis demonstrated that KLF8 expression independently affected both overall and cancer-specific survival of gastric cancer patients (P = 0.035 and 0.042, respectively). CONCLUSION:KLF8 is closely associated with gastric tumor progression, angiogenesis and poor prognosis, suggesting it may represent a novel prognostic biomarker and therapeutic target for gastric cancer.

Genetic Predisposition for Type 2 Diabetes Mellitus in a Cameroonian Population: Contribution of rs4731702 (C/T) Polymorphism of Krüppel-Like Factor 14 Gene

Introduction: Krüppel Like Factor 14 (KLF14) gene has recently been identified as a master gene for multiple metabolic phenotypes. The aim of the research study was to investigate the relationship between KLF14 rs4731702 (C/T) gene polymorphism with Type 2 Diabetes Mellitus (T2DM) in a Cameroonian population. Patients and Methods: This case-control study was conducted in 85 patients with T2DM and 95 healthy normoglycemic controls. All were nonrelated, of Cameroonian origin, and were adults aged 24 years old and above. Demographic, clinical and biological data were collected, and biochemical explorations were performed using enzymatic colorimetric methods. The genotyping of KLF14 rs4731702 (CT) gene polymorphism was done by the Polymerase Chain Reaction and Restriction Fragment Length Polymorphism. Results: In comparing the Cameroonian population that consisted of 85 patients with T2DM and 95 healthy controls, the minor or risk allele of the rs4731702 (C/T) polymorphism of the KLF14 gene was T (63.53% diabetic patients vs. 26.32% healthy controls, OR = 4.877 and p < 0.0001) while the protective allele was C (36.47% diabetic patients vs. 73.68% healthy controls, OR = 0.205 and p < 0.0001). The susceptibility to T2DM was higher among subjects having the CT and TT genotypes with OR = 2.721 and p = 0.0145) and OR = 3.907 and p < 0.0001) respectively. This gene polymorphism was not preferentially associated with a specific diabetes phenotype. Conclusion: This study has demonstrated for the first time the relationship between the KLF14 rs4731702 (C/T) gene polymorphism and T2DM in this Cameroonian population. This gene polymorphism could be a promising target for personalized medicine through the development of clinical genetic testing.

Correlation of Krüppel-like factor 9 expression in pancreatic cancer tissue with serum tumor markers and focal cell invasion

Objective:To study the correlation of Krüppel-like factor 9 (KLF9) expressions in pancreatic cancer tissue with serum tumor markers and focal cell invasion.Methods: A total of 58 patients with pancreatic cancer treated in our hospital between June 2012 and May 2016 were collected, the expression of KLF9 in pancreatic cancer tissues and paracancerous tissues were measured and then patients were further divided into high KLF9 expression group and low KLF9 expression group, 29 cases in each group. Serum tumor marker levels as well as invasion gene and tumor suppressor gene expression in tumor tissue were compared between patients with different KLF9 expression.Results: KLF9 expression in pancreatic cancer tissue was significantly lower than that in paracancerous tissue;serum tumor markers CA19-9, CA242, CA50 and CEA levels of low KLF9 expression group were higher than those of high KLF9 expression group;focal invasion genes DKK-1, GSK3β and HOXB7 mRNA expression of low KLF9 expression group were higher than those of high KLF9 expression group while tumor suppressor genes Bach2, SIRT3, DPC4 and Kiss-1 mRNA expression were lower than those of high KLF9 expression group.Conclusion: The expression of KLF9 decreases in pancreatic cancer tissues, and the expression of KLF9 is negatively correlated with the malignant degree of tumor.

首钢水钢含钒钛铁水KR脱硫预处理实践

KR铁水脱硫工艺由于其优秀的搅拌脱硫效果使其得到不断地推广应用.首钢水钢建立两座KR脱硫站进行含钒钛铁水脱硫预处理,铁水脱硫站设备设计处理产能为316.8万t/年.为了探索高效脱硫的工艺过程,制定低成本运行工艺制度.在分析KR脱硫过程机理的基础上,结合对钒钛铁水性质、脱硫剂粒度及成分、搅拌转速、搅拌时间等分析影响脱硫的主要因素,有针对性地提出设计方案和控制措施,为低成本KR脱硫生产提供指导和借鉴.

Krüppel样因子6抑制人晶状体上皮细胞增生的研究

背景 研究表明,Kruppel样因子6（KLF6）与生长发育、细胞分化、增生、凋亡、血管生成及组织修复等生理或病理过程有关.在眼科领域,关于人晶状体上皮细胞（LECs）中KLF6蛋白表达水平及KLF6对LECs增生的影响报道较少. 目的 探讨KLF6对LECs株（HLE-B3）增生的抑制作用. 方法 首先用逆转录PCR（RT-PCR）法构建真核表达质粒pEGFP-C2-KLF6,并用双酶切法和PCR法对表达质粒进行鉴定.用含体积分数10％胎牛血清的低糖DMEM对HLE-B3进行培养和传代,按照干预方法的不同将培养的HLE-B3分成4个组,各组均给予KLE-B3转染试剂,空白对照组不加入任何外源质粒及胰岛素样生长因子-1（IGF-1）;单纯IGF-1刺激组仅给予IGF-1刺激;空质粒转染＋IGF-1组加入pEGFP-C2质粒空载体,同时给予IGF-1刺激;pEGFP-C2-KLF6转染＋IGF-1组加入pEGFP-C2-KLF6真核表达质粒,且给予IGF-1刺激.细胞干预24 h后,采用水溶性四唑盐-1（WST-1）实验检测各组细胞的吸光度（A450）值;采用Western blot法测定各组细胞中KLF6蛋白的表达水平.以1＆#215;104个/片的密度将HLE-B3细胞爬片培养24 h,并分别将0、0.10、0.25 μg pEGFP-C2-KLF6转染到各组铺片细胞中,用终质量浓度为50 μg/L的IGF-1刺激24 h,然后应用免疫细胞化学技术检测各组细胞中Ki-67蛋白的相对表达量;采用荧光半定量PCR法检测各组细胞中Ki-67 mRNA的表达变化.结果 扩增得到的PCR产物反应条带与KLF-6基因长度相符,PCR和EcoR I、Sal I限制性内切酶双酶切法鉴定条带大小与预期相符,成功构建pEGFP-C2-KLF6真核表达质粒.WST-1实验显示空白对照组、单纯IGF-1刺激组、空质粒转染＋IGF-1组和pEGFP-C2-KLF6转染＋IGF-1组细胞A值分别为0.86±0.00、2.10±0.01、2.24±0.12和1.06±0.02,4个组间差异有统计学意义（F=38.322,P＜0.05）,其中pEGFP-C2-KLF6转染＋IGF-1组A值明显低于单纯IGF-1刺激组和空质粒转染＋IGF-1组,差异均有统计学意义（q=6.42、7.31,P＜0.05）.Western blot法检测显示,4个组间细胞中KLF6蛋白相对表达量的差异有统计学意义（F=591.858,P＜0.05）,pEGFP-C2-KLF6转染＋IGF-1组KLF6蛋白相对表达量分别是空白对照组、单纯IGF-1刺激组和空质粒转染＋IGF-1组的1.47、2.04、3.27倍.Ki-67蛋白在pEGFP-C2-KLF6转染＋IGF-1组细胞中的表达强度随质粒转染剂量的增加而逐渐减弱,0.10 μg和0.25 μg pEGFP-C2-KLF6转染后细胞中Ki-67 mRNA相对表达值分别为0.15±0.08和0.11±0.03,明显低于0μg pEGFP-C2-KLF6转染的细胞0.77±0.12,组间的总体差异有统计学意义（F=54.825,P＜0.05）. 结论 KLF-6能够抑制IGF-1诱导的HLE-B3的增生,是HLE-B3细胞增生的调控因子.

Sulforaphane ameliorates non-alcoholic steatohepatitis by KLF4-mediated macrophage M2 polarization

Non-alcoholic fatty liver disease (NAFLD) has become a global issue and a severe threat to public health.However, to date, no approved therapeutic drugs have been developed. Dietary interventions with naturalproducts have shown promise in preventing and treating NAFLD. Sulforaphane (SFN) is a phytocompoundwith antioxidant and anti-inflammatory properties, and previous research has demonstrated that SFN canameliorate hepatic lipid accumulation and inflammation. However, the molecular mechanisms underlying thesebeneficial effects remain unclear. In this study, we confirmed the protective effects of SFN on excessive lipidaccumulation and inflammatory injury in a high-fat, high-fructose diet-induced non-alcoholic steatohepatitis(NASH) mouse model. We found that SFN attenuates the inflammatory injury in a macrophage cell line andthe liver of NASH mice, owing to the promotion of M1-type macrophage polarization toward the M2-type andthe regulation of inflammatory mediators. Further analysis demonstrated that this SFN-induced macrophageM2-type polarization occurs in a Krüppel-like factor 4 (KLF4)-dependent manner. In summary, we uncovereda new mechanism of action underlying SFN activity and provide evidence that dietary intervention with SFNmight be protective against NASH.

KR法铁水脱硫技术研究

过对影响铁水脱硫的因素进行分析与讨论，发现在脱硫剂中加入适量增铝剂，用硅铝复合脱氧，会生成低熔点硅铝酸盐，有助于提高脱硫效率；铁水温度和CaO／CaF：比应分别控制在1360—1390℃与14～16区间，过高的温度和CaF2含量会在石灰颗粒表面形成较多的液相，造成石灰颗粒团聚，减小铁水与石灰的接触面积，降低脱硫反应速率；铁水硅含量在0．40％-0．50％区间时，脱硫率最高，达到88．68％。

KR脱硫效果影响因素分析

从原材料条件、工况动力学特性及扒渣程度三方面出发,分析不同因素对KR铁水脱硫效果的影响,这些因素包括铁水初始(S)、铁水温度、脱硫剂尺寸、搅拌效果、渣的流动性、扒渣操作等,最后提出改善炼钢厂KR铁水脱硫效果的建议。

Krüppel-like factor 8 is a potential prognostic factor for pancreatic cancer

Background Pancreatic cancer is a lethal disease that is often diagnosed at an advanced stage.There is a lack of information to predict the prognosis of pancreatic cancer.Krüppel-like factor （KLF） 8 has been found to be deregulated in multiple cancers,and its high expression was correlated with poor prognosis.However,so far,no information was reported about the expression of KLF8 in pancreatic cancer.In the present study,we investigated,possibly for the first time,the expression of KLF8 in pancreatic cancer samples and analyzed its correlation with clinical parameters and overall survival （OS） rate.Methods We used immunohistochemical staining to detect KLF8 in 68 samples from patients who underwent surgery and its correlation with the clinicopathological characteristics.We used Kaplan-Meier curve to analyze the relationship between KLF8 expression and the OS time.Univariate analysis was performed in addition to multivariate hazard models with clinicopathological features to assess KLF8 as an independent prognostic factor.Results KLF8 was present in the cytoplasm of pancreatic cancer cells and 52.9％ of the 68 cases had positive expression.KLF8 expression was not associated with sex,age,tumor location,lymph node stage,and metastasis stage,but was associated with tumor stage （P=0.04）.Kaplan-Meier method demonstrated that patients with negative expression of KLF8 had a better prognosis.In univariate and multivariate models,KLF8 was a significant predictor of OS in pancreatic cancer.Conclusion Our results revealed that KLF8 may be a potential prognostic factor for pancreatic cancer.

Current knowledge of Krüppel-like factor 5 and vascular remodeling: providing insights for therapeutic strategies

Vascular remodeling is a pathological basis of various disorders. Therefore, it is necessary to understand the occurrence, prevention, and treatment of vascular remodeling. Krüppel-like factor 5 (KLF5) has been identified as a significant factor in cardiovascular diseases during the last two decades. This review provides a mechanism network of function and regulation of KLF5 in vascular remodeling based on newly published data and gives a summary of its potential therapeutic applications. KLF5 modulates numerous biological processes, which play essential parts in the development of vascular remodeling, such as cell proliferation, phenotype switch, extracellular matrix deposition, inflammation, and angiogenesis by altering downstream genes and signaling pathways. Considering its essential functions, KLF5 could be developed as a potent therapeutic target in vascular disorders.

Loss of the Krüppel-like factor 4 tumor suppressor is associated with epithelial-mesenchymal transition in colorectal cancer

Aim: Colorectal cancer (CRC) is the third leading cancer-related cause of death due to its propensity to metastasize. Epithelial-mesenchymal transition (EMT) is a multistep process important for invasion and metastasis of CRC. Krüppel-like factor 4 (KLF4) is a zinc finger transcription factor highly expressed in differentiated cells of the intestinal epithelium. KLF4 has been shown to play a tumor suppressor role during CRC tumorigenesis - its loss accelerates development and progression of cancer. The present study examined the relationship between KLF4 and markers of EMT in CRC. Methods: Immunofluorescence staining for KLF4 and EMT markers was performed on archived patient samples after colorectal cancer resection and on colonic tissues of mice with colitis-associated cancer. Results: We found that KLF4 expression is lost in tumor sections obtained from CRC patients and in those of mouse colon following azoxymethane and dextran sodium sulfate (AOM/DSS) treatment when compared to their respective normal appearing mucosa. Importantly, in CRC patient tumor sections, we observed a negative correlation between KLF4 levels and mesenchymal markers including TWIST, β-catenin, claudin-1, N-cadherin, and ;vimentin. Similarly, in tumor tissues from AOM/DSS-treated mice, KLF4 levels were negatively correlated with mesenchymal markers including SNAI2, β-catenin, and vimentin and positively correlated with the epithelial marker E-cadherin. Conclusion: These findings suggest that the loss of KLF4 expression is a potentially significant indicator of EMT in CRC.

E-cadherin和KLF 4表达对胃癌侵袭转移的作用

观察E-cadherin,Krüppel-like factor 4(KLF4)蛋白在胃癌和正常胃黏膜组织中的表达,分析其与胃癌浸润、转移的关系.应用免疫组织化学SP法检测84例手术切除的胃癌标本及对应正常胃黏膜组织中E-cadherin,KLF4蛋白的表达.各指标之间相关因素的差异性比较采用χ2检验,E-cadherin,KLF4相关性研究采用Spearman相关分析.结果显示,与正常胃组织相比,E-cadherin、KLF4蛋白在胃癌组织中均呈低表达或者缺失(分别42.9%vs.95.24%,8.3%vs 81%,P<0.05).E-cadherin、KLF4蛋白的阳性表达率与组织分级(P<0.05)、肿瘤浸润深度(P<0.05)、淋巴转移(P<0.05)明确相关.Spearman相关分析显示KLF4蛋白与E-cadherin蛋白的表达呈正相关(P<0.05).因此,E-cadherin,KLF4蛋白水平低表达可能与胃癌浸润和转移有关,而联合检测更能有效判断胃癌这一生物学行为.

KLF6与肿瘤研究进展

Krüppel样转录因子6(Krüppel-like factor 6,KLF6)是哺乳动物细胞中普遍表达的核内转录因子,其与肿瘤的发生、发展密切相关,从KLF6的结构及其在肿瘤发生中的作用机制2个方面进行综述。

平面状态下EWK延性断裂准则与K准则对比研究

为研究EWK延性断裂准则对金属起裂及扩展的预测能力,将EWK断裂算法嵌入ABAQUS主程序。针对金属I型裂纹,在平面应变条件下,通过试验与有限元计算相互对比的方法,研究了该准则与应力强度因子K准则对紧凑拉伸CT试样起裂的预测结果,发现两者有近似等效性。在平面应力条件下,以紧凑拉伸CS试样为模型,通过有限元计算,提取由EWK控制断裂的结构KR-曲线,将该曲线与试验资料对比,发现两者吻合良好。

FBW7-mediated ubiquitination and degradation of KLF5

Krüppel-like factor(KLF) family proteins are transcription factors that regulate numerous cellular functions, such as cell proliferation, differentiation, and cell death. Posttranslational modification of KLF proteins is important for their transcriptional activities and biological functions. One KLF family member with important roles in cell proliferation and tumorigenesis is KLF5. The function of KLF5 is tightly controlled by post-translational modifications, including SUMOylation, phosphorylation, and ubiquitination. Recent studies from our lab and others' have demonstrated that the tumor suppressor FBW7 is an essential E3 ubiquitin ligase that targets KLF5 for ubiquitination and degradation. KLF5 contains functional Cdc4 phospho-degrons(CPDs), which are required for its interaction with FBW7. Mutation of CPDs in KLF5 blocks the ubiquitination and degradation of KLF5 by FBW7. The protein kinase Glycogen synthase kinase 3β is involved in the phosphorylation of KLF5 CPDs. In both cancer cell lines and mousemodels, it has been shown that FBW7 regulates the expression of KLF5 target genes through the modulation of KLF5 stability. In this review, we summarize the current progress on delineating FBW7-mediated KLF5 ubiquitination and degradation.

Salidroside Ameliorates Vascular Endothelial Cell Senescence through Downregulation of KLF4

Salidroside is extensively used as a herbal medicine worldwide, and it has been shown to protect against disruption of endothelial homeostasis and act as an anti-aging agent. The present study aimed to investigate the ameliorative effects of salidroside on homocysteine (Hcy)-induced cell senescence in human umbilical vein endothelial cells (HUVECs) that were mediated via inhibition of Krüppel-like factor 4 (KLF4). An endothelial cell senescence model was induced by Hcy. The cell viability, activities of telomerase and lactate dehydrogenase (LDH), and the level of reactive oxygen species were determined using commercial kits. The expression levels of KLF4, p53 and p21 were determined via western blot analysis, whereas the mRNA expression levels of KLF4 were detected by reverse transcription-quantitative PCR. Small interfering RNA-mediated knockdown of KLF4 was found to reverse Hcy-induced cell senescence. Hcy treatment led to an accelerated cell senescence, as evidenced by decreases in both cell viability and telomerase activity, whereas increases were noted in the leakage of LDH and the level of reactive oxygen species, in addition to an up-regulation of the protein levels of p53 and p21, and up-regulation of KLF4 at both the mRNA and protein level. Treatment with salidroside ameliorated Hcy-induced cell senescence in a dose-dependent manner. Taken together, these results suggested that Hcy may induce cell senescence through upregulation of KLF4, and this may be reversed by treatment with salidroside. Therefore, salidroside was shown to inhibit Hcy-induced cell senescence through KLF4 inhibition.

首钢京唐KR高效脱硫技术

首钢京唐采用KR进行100%全量铁水脱硫预处理,从生产布局上可以同时满足脱磷炉和脱碳炉的生产需要。为了实现KR的高效脱硫,对影响脱硫的因素进行了分析和讨论,认为脱硫剂中添加一定量的CaF2可生成一定量的共熔晶体,提高了铁水中硫元素的传输和反应速率;铁水中加入一定量的铝渣可以降低铁水中的氧活度,提高脱硫反应速度;铁水温度应控制在1300~1380℃之间,温度太高会在石灰颗粒表面形成较多的液相,造成石灰颗粒聚团,减少铁水与脱硫剂的接触面积,降低了反应速率;良好的石灰质量和搅拌头形状也有利于KR脱硫。通过以上措施,铁水经过KR脱硫预处理后w[S]≤0.002%比例达到98%以上,转炉终点平均硫质量分数为0.005%。

KLF2对 ox-LDL 诱导内皮细胞 microRNA-146 a及促炎细胞因子表达的影响

目的：探讨Kr＆#252;ppel样因子2（KLF2）对氧化型低密度脂蛋白（oxidized low density lipo-protein， ox-LDL）诱导人脐静脉内皮细胞（HUVECs）微小RNA 146a（microRNA-146a， miR-146a）及细胞因子单核细胞趋化蛋白（MCP-1）和白细胞介素-6（IL-6）表达的影响。方法构建 KLF2过表达腺病毒载体（ rAAV-KLF2）及KLF2-siRNA，分别转染HUVECs及ox-LDL诱导的HUVECs。于0 h、3 h、6 h、12 h、24 h、48 h收集HUVECs，采用实时荧光定量PCR，检测HUVECs miR-146a的表达；设计合成锁核酸（the locked nucleic acid，LNA））修饰的anti-miR-146a的反义核苷酸（LNA-anti-miR-146a）转染HUVECs。收集各组细胞上清，采用双抗体夹心ABC-ELISA方法检测MCP-1及IL-6的表达。结果KLF2明显抑制静息及ox-LDL诱导的HUVECs miR-146a的表达，并显著减少ox-LDL诱导HUVECs MCP-1、IL-6的表达，且具有时间依赖性；miR-146 a 沉默可削弱ox-LDL诱导的内皮细胞炎症反应，而且部分逆转了KLF2对内皮细胞炎症反应的抑制作用。结论 KLF2通过下调miR-146 a的表达抑制ox-LDL活化的HUVECs MCP-1、IL-6的分泌。