射干制剂对失眠大鼠自主活动及TLR7/MyD88信号通路的影响

目的 探研射干制剂对PCPA致失眠大鼠自主活动、脑电波以及脑组织TLR7、MyD88基因表达的影响。方法 随机将70只健康SD大鼠分为空白对照组、模型对照组、射干制剂低剂量组(26 mg·kg^(-1))、射干制剂中剂量组(52 mg·kg^(-1))、射干制剂高剂量组(104 mg·kg^(-1))、朱砂安神丸组(1.62 g·kg^(-1))、地西泮片组(0.9 mg·kg^(-1))。荧光定量PCR检测各组TLR7、My D88基因的相对表达量;采用Etho Vision大小鼠行为学系统记录大鼠自主活动情况;通过无线遥测系统记录各组睡眠时脑电图。结果 与空白组对照比较,模型对照组脑组织TLR7、MyD88相对表达量显著升高(P<0.01);同模型对照组相比,射干制剂低剂量组、射干制剂中剂量组TLR7、MyD88的mRNA表达量均降低(P<0.05),射干制剂高剂量组TLR7、MyD88 mRNA表达量显著降低(P<0.01);与空白对照组相比,PCPA造模后大鼠运动量明显增加,不同剂量射干制剂组、朱砂安神丸组及地西泮片组运动有不同程度的减少,射干制剂高剂量组运动减少较为明显;给药120 min后,与空白对照组相比,模型对照组脑电波中的δ波百分比降低(P<0.05);与模型对照组相比,射干低中剂量组、朱砂安神丸组、地西泮片组δ波百分比均升高(P<0.05)。结论 射干制剂可有效抑制大鼠兴奋性,增加脑电图δ波百分比,改善睡眠,这些作用可能是通过调节脑组织TLR7和MyD88基因的表达来实现。

Analysis of the autophagy gene expression profile of pancreatic cancer based on autophagy-related protein microtubule-associated protein 1A/1B-light chain 3

BACKGROUND Pancreatic cancer is a highly invasive malignant tumor. Expression levels of the autophagy-related protein microtubule-associated protein 1 A/1 B-light chain 3(LC3) and perineural invasion(PNI) are closely related to its occurrence and development. Our previous results showed that the high expression of LC3 was positively correlated with PNI in the patients with pancreatic cancer. In this study, we further searched for differential genes involved in autophagy of pancreatic cancer by gene expression profiling and analyzed their biological functions in pancreatic cancer, which provides a theoretical basis for elucidating the pathophysiological mechanism of autophagy in pancreatic cancer and PNI.AIM To identify differentially expressed genes involved in pancreatic cancer autophagy and explore the pathogenesis at the molecular level.METHODS Two sets of gene expression profiles of pancreatic cancer/normal tissue(GSE16515 and GSE15471) were collected from the Gene Expression Omnibus.Significance analysis of microarrays algorithm was used to screen differentially expressed genes related to pancreatic cancer. Gene Ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis were used to analyze the functional enrichment of the differentially expressed genes. Protein interaction data containing only differentially expressed genes was downloaded from String database and screened. Module mining was carried out by Cytoscape software and ClusterOne plug-in. The interaction relationship between the modules was analyzed and the pivot nodes between the functional modules were determined according to the information of the functional modules and the data of reliable protein interaction network.RESULTS Based on the above two data sets of pancreatic tissue total gene expression, 6098 and 12928 differentially expressed genes were obtained by analysis of genes with higher phenotypic correlation. After extracting the intersection of the two differential gene sets, 4870 genes were determined. GO analysis showed that 14 significant functional items including negative regulation of protein ubiquitination were closely related to autophagy. A total of 986 differentially expressed genes were enriched in these functional items. After eliminating the autophagy related genes of human cancer cells which had been defined, 347 differentially expressed genes were obtained. KEGG pathway analysis showed that the pathways hsa04144 and hsa04020 were related to autophagy. In addition,65 clustering modules were screened after the protein interaction network was constructed based on String database, and module 32 contains the LC3 gene,which interacts with multiple autophagy-related genes. Moreover, ubiquitin C acts as a pivot node in functional modules to connect multiple modules related to pancreatic cancer and autophagy.CONCLUSION Three hundred and forty-seven genes associated with autophagy in human pancreatic cancer were concentrated, and a key gene ubiquitin C which is closely related to the occurrence of PNI was determined, suggesting that LC3 may influence the PNI and prognosis of pancreatic cancer through ubiquitin C.

基于心功能及IGFBP7、sST2、CGRP、ET分析沙库巴曲缬沙坦在治疗冠心病合并慢性心力衰竭中的应用效果

目的 分析冠心病(CHD)合并慢性心力衰竭(CHF)患者应用沙库巴曲缬沙坦治疗的效果。方法 选择2020年1月至2023年1月邯郸市第四医院收治的86例CHD合并CHF患者,以随机数字表法将其分为对照组和试验组各43例。两组CHD治疗均应用硝酸酯类、他汀类及抗血小板药物,对照组CHF治疗应用坎地沙坦酯片、醛固酮受体拮抗剂及β受体阻滞剂,试验组治疗则将对照组中的坎地沙坦酯片替换为沙库巴曲缬沙坦钠片。比较两组疗效、不良反应、心功能指标[左室短轴缩短率(LVFS)、左室射血分数(LVEF)、6min步行距离(6 MWD)]、心室重构指标[Ⅲ型胶原前肽(PⅢP)、层粘蛋白(LN)、基质金属蛋白酶-9(MMP-9)]、心肌损伤和血管内皮功能相关指标[胰岛素样生长因子结合蛋白7(IGFBP7)、可溶性生长刺激表达基因2(sST2)、降钙素基因相关肽(CGRP)、内皮素(ET)]。结果与对照组比,试验组治疗3个月后的总有效率更高,差异有统计学意义(P<0.05)。两组治疗3个月后的LVFS、LVEF、6 MWD、IGFBP7、CGRP与治疗前比升高,且试验组与对照组比更高,差异有统计学意义(P<0.05);PⅢP、LN、MMP-9、sST2、ET降低,试验组与对照组比更低,差异有统计学意义(P<0.05)。两组不良反应总发生率对比差异无统计学意义(P>0.05)。结论 沙库巴曲缬沙坦可有效调节CHD合并CHF患者IGFBP7、sST2、CGRP、ET,改善血管内皮功能、心肌损伤、心室重构及心功能,进而可提高疗效,且具有良好的安全性。

ADAMTS-7基因rs3825807及rs1994016位点多态性与KD发病及CAL的关联性分析

目的分析ADAMTS-7基因rs3825807、rs1994016位点多态性与川崎病(KD)发病及其冠状动脉损伤(CAL)是否存在关联性。方法选取KD患儿100例为KD组,健康体检儿童100例为健康组。提取两组外周血DNA进行基因测序,比较两组ADAMTS-7基因rs3825807位点及rs1994016位点的多态性差异。KD组根据是否存在冠状动脉损伤分为冠状动脉损伤组(KD-CAL组)和无冠状动脉损伤组(KD-NCAL组),比较两组ADAMTS-7基因rs3825807位点及rs1994016位点的多态性差异。结果KD组与健康组比较,KD-NCAL组与KD-CAL组比较,ADAMTS-7基因rs3825807位点A、G等位基因频率和AA、AG、GG基因型频率以及rs1994016位点C、T等位基因频率和CC、CT、TT基因型频率差异均无统计学意义(P>0.05)。结论ADAMTS-7基因rs3825807、rs1994016位点多态性与KD的发病及CAL不存在明显关联性。

猪miR-192靶向调控Wnt7a基因表达的研究

【目的】鉴定猪miR-192与Wnt家族成员7A(Wnt7a)基因之间的靶向关系,为进一步构建miR-192介导的胚胎通讯提供依据。【方法】使用miRNA pull-down鉴定miR-192和Wnt7a基因间的互作关系;利用生物信息学软件RNAhybrid 2.2预测miR-192和Wnt7a基因3′-UTR的结合位点,构建含有miR-192结合位点的Wnt7a基因3′-UTR野生型和突变型基因片段,克隆至双荧光素酶报告基因质粒,通过NotⅠ、XhoⅠ酶切和测序进行鉴定;鉴定成功的质粒分别与miR-192 mimics和mimics NC共转染猪子宫内膜上皮细胞,检测双荧光素酶活性;将miR-192 mimics、mimics NC、miR-192 inhibitor、inhibitor NC分别与构建的Wnt7a基因3′-UTR野生型和突变型双荧光素酶报告基因质粒共转染至猪子宫内膜上皮细胞,通过实时荧光定量PCR和Western blotting检测Wnt7a基因mRNA和蛋白的表达水平。【结果】pull-down结果显示,miR-192 mimics组与其阴性对照组相比,Wnt7a基因相对表达量极显著升高(P<0.01),miR-192 input组相对于其阴性对照组极显著降低(P<0.01);RNAhybrid 2.2软件预测到miR-192种子区与Wnt7a基因3′-UTR存在结合位点(UGACCUA);转染Wnt7a野生型质粒的miR-192 mimics和miR-192 mimics NC组相比双荧光素酶活性显著降低(P<0.05),转染Wnt7a突变型质粒的miR-192 mimics和miR-192 mimics NC组之间双荧光素酶活性无显著差异(P>0.05)。实时荧光定量PCR结果表明,miR-192 mimics组Wnt7a基因mRNA相对表达量显著低于其阴性对照组(P<0.05),miR-192 inhibitor组显著高于其阴性对照组(P<0.05)。Western blotting结果表明,过表达miR-192极显著抑制了Wnt7a蛋白的表达(P<0.01),抑制miR-192后Wnt7a蛋白的表达量极显著上升(P<0.01)。【结论】miR-192能够与Wnt7a基因3′-UTR发生靶向作用并抑制其表达。研究结果可为今后深入研究miR-192在妊娠着床过程中的调控机制提供理论依据。

An Investigation of the Effects of B7-H4 Gene rs10754339 and miR-125a Gene rs12976445 on Cancer Susceptibility

Objective To investigate the effects of the B7-H4 gene rs10754339 and miR-125a gene rs12976445 on cancer susceptibility through a case-control study and meta-analysis.Methods A total of 1,490 cancer patients(lung/gastric/liver/:550/460/480)and 800 controls were recruited in this case-control study.The meta-analysis was performed by pooling the data from previous related studies and the present study.Results The results of this study showed that in the Hubei Han Chinese population,the rs10754339gene was significantly associated with the risk of lung and gastric cancer but not liver cancer,and the rs12976445 gene was significantly associated with the risk of lung cancer but not liver or gastric cancer.The meta-analysis results indicated that rs10754339 and rs12976445 contributed to cancer susceptibility in the Chinese population and also revealed a significant association between rs10754339and breast cancer risk,as well as between rs12976445 and lung cancer risk.Conclusion The B7-H4 gene rs10754339 and miR-125a gene rs12976445 may be the potential genetic markers for cancer susceptibility in the Chinese population,which should be validated in future studies with larger sample sizes in other ethnic populations.

Codominance Functional Marker of Bacterial Blight Resistance Gene Xa7 in Rice

[Objectives]A codominance functional marker of the broad-spectrum bacterial blight resistance gene,Xa7,of rice was identified for accurate detection,generation tracking,and differentiation between homozygous and hemizygous genotypes of the gene.[Methods]A potential functional marker containing four primers was designed using Premier 5 software and based on the differences on the sequences of Xa7,xa7,and allele-free genomes.The molecular distinctness of the marker in different materials was verified by PCR.Three crossbreed lines of Xa7 and their parents were inoculated with seven bacterial blight strains at the booting stage to examine the affected agronomic traits at maturation.[Results]The homozygous R084 of Xa7 could be amplified into a 91 bp band and the Nip free of allele with a 153 bp band,while the heterozygote Nip/R084,91 bp and 153 bp bands.The candidate codominance marker,Xa7fun,amplified fragments that matched the predicted target bands.No 91 bp fragment was amplified from 18 germplasms of varied types,indicating a lack of Xa7 in them.Whereas Ry1,Ry2 and Ry3 had a 91 bp band,suggesting the inclusion of homozygous Xa7.Under an elevated temperature,Huazhan responded to the seven bacterial blight pathogens as highly susceptible(HS),intermediate susceptible(MS),or susceptible(S);R084 to six of the seven pathogens(HNA1-4,FuJ,GDA2,GD1358,PX086,and YN24)as highly resistant(HR),intermediate resistant(MR)or resistant(R);Ry-1 to five pathogens(GDA2,HNA1-4,FuJ,GD1358,and YN24)as HR or MR;Ry-2 to five pathogens(GDA2,GD1358,HNA1-4,PXO86,and YN24)as HR or R;and Ry-3 to 6 pathogens(HNA1-4,FuJ,GDA2,GD1358,PXO86,and YN24)as HR or MR.Therefore,the infiltration of Xa7 in the improved crossbred lines RY-1,RY-2,and RY-3 significantly accentuated the blight resistance of Huazhan.[Conclusions]Homozygous or hemizygous Xa7 could be accurately differentiated by the currently identified codominance functional marker Xa7 fun.The Xa7 introgression did not significantly alter the critical agronomic traits in the hybridization from generation to generation and could be safely applied in breeding rice varieties with bacterial blight resistance.

Prediction of Tumor Microenvironment Characteristics and Treatment Response in Lung Squamous Cell Carcinoma by Pseudogene OR7E47P-related Immune Genes

Objective Pseudogenes are initially regarded as nonfunctional genomic sequences,but some pseudogenes regulate tumor initiation and progression by interacting with other genes to modulate their transcriptional activities.Olfactory receptor family 7 subfamily E member 47 pseudogene(OR7E47P)is expressed broadly in lung tissues and has been identified as a positive regulator in the tumor microenvironment(TME)of lung adenocarcinoma(LUAD).This study aimed to elucidate the correlation between OR7E47P and tumor immunity in lung squamous cell carcinoma(LUSC).Methods Clinical and molecular information from The Cancer Genome Atlas(TCGA)LUSC cohort was used to identify OR7E47P-related immune genes(ORIGs)by weighted gene correlation network analysis(WGCNA).Based on the ORIGs,2 OR7E47P clusters were identified using non-negative matrix factorization(NMF)clustering,and the stability of the clustering was tested by an extreme gradient boosting classifier(XGBoost).LASSO-Cox and stepwise regressions were applied to further select prognostic ORIGs and to construct a predictive model(ORPScore)for immunotherapy.The Botling cohorts and 8 immunotherapy cohorts(the Samstein,Braun,Jung,Gide,IMvigor210,Lauss,Van Allen,and Cho cohorts)were included as independent validation cohorts.Results OR7E47P expression was positively correlated with immune cell infiltration and enrichment of immune-related pathways in LUSC.A total of 57 ORIGs were identified to classify the patients into 2 OR7E47P clusters(Cluster 1 and Cluster 2)with distinct immune,mutation,and stromal programs.Compared to Cluster 1,Cluster 2 had more infiltration by immune and stromal cells,lower mutation rates of driver genes,and higher expression of immune-related proteins.The clustering performed well in the internal and 5 external validation cohorts.Based on the 7 ORIGs(HOPX,STX2,WFS,DUSP22,SLFN13,GGCT,and CCSER2),the ORPScore was constructed to predict the prognosis and the treatment response.In addition,the ORPScore was a better prognostic factor and correlated positively with the immunotherapeutic response in cancer patients.The area under the curve values ranged from 0.584 to 0.805 in the 6 independent immunotherapy cohorts.Conclusion Our study suggests a significant correlation between OR7E47P and TME modulation in LUSC.ORIGs can be applied to molecularly stratify patients,and the ORPScore may serve as a biomarker for clinical decision-making regarding individualized prognostication and immunotherapy.

老年重症肺炎患者血清FOXO1、ATG7水平及与短期预后的关系

目的 探讨老年重症肺炎患者血清叉头状转录因子1(FOXO1)、自噬相关基因7(ATG7)的表达水平及与短期预后的关系。方法 选取2020年8月至2022年8月该院收治的122例老年重症肺炎患者作为病例组,另选取同期来该院进行健康体检的105例老年人作为对照组。采用酶联免疫吸附试验(ELISA)检测血清FOXO1、ATG7的表达水平。根据患者重症肺炎确诊后28 d内是否死亡分为存活组(n=91)和死亡组(n=31)。采用受试者工作特征(ROC)曲线评估血清FOXO1、ATG7表达水平对老年重症肺炎短期死亡的预测价值,采用多因素Logistic回归分析探讨老年重症肺炎短期死亡的影响因素。结果 病例组患者血清FOXO1、ATG7表达水平明显高于对照组,差异有统计学意义(t=7.615、29.591,P<0.05);老年重症肺炎死亡组患者血清FOXO1、ATG7表达水平均高于生存组,差异有统计学意义(t=10.029、9.399,P<0.05)。血清FOXO1、ATG7预测老年重症肺炎患者短期死亡的曲线下面积(AUC)分别为0.733(95%CI:0.673~0.799)、0.826(95%CI:0.756~0.893),两者联合预测的AUC为0.908(95%CI:0.862~0.949)。FOXO1、ATG7与急性生理与慢性健康评估(APACHEⅡ)评分呈正相关(r=0.693、0.627,均P<0.05)。多因素Logistic回归分析显示,APACHEⅡ评分≥22.56分(OR=3.589,95%CI:1.714~7.515),FOXO1≥10.67 ng/mL(OR=2.529,95%CI:1.232~5.193),ATG7≥16.35 pg/mL(OR=2.875,95%CI:1.414~5.845)是老年重症肺炎患者死亡的危险因素(P<0.05)。结论 老年重症肺炎患者血清FOXO1、ATG7表达水平明显升高,二者可用于评估老年重症肺炎患者的短期预后。

TLR7基因多态性与儿童手足口病易感性及严重程度的关系

目的探讨Toll样受体7(TLR7)基因多态性与儿童手足口病(HFMD)易感性及严重程度的关系。方法选取304例肠道病毒71型(EV71)感染HFMD患儿纳入HFMD组,根据病情严重程度分为轻症组和重症组;另同期选择300例正常健康儿童纳入对照组。比较各组TLR7基因rs3853839、rs179016、rs179009位点基因型和基因频率。Logistic回归分析基因多态性是否HFMD发生的危险因素,比较与否携带C基因患儿的外周血单个核细胞TLR7 mRNA表达水平。结果HFMD组、重症组女童、男童TLR7基因rs3853839位点、rs179016位点C基因型及基因频率分别高于对照组、轻症组(P<0.05)。TLR7基因rs3853839位点、rs179016位点携带C基因是儿童HFMD发生和重症的危险因素(P<0.05)。HFMD轻症和重症患儿rs3853839位点、rs179016位点携带C基因者外周血单个核细胞TLR7 mRNA表达水平均低于未携带C基因者(P<0.05)。结论TLR7基因rs3853839位点、rs179016位点携带C基因是儿童HFMD发生的独立危险因素,且与其严重性相关。

SNHG7对结肠癌预后影响的生物信息学分析

目的研究长链非编码RNA(lncRNA)核仁小RNA宿主基因7(SNHG7)的表达程度与结肠癌预后的关系,以期寻找一种新型潜在的结肠癌标记物及治疗新靶点。方法从癌症基因组图谱(TCGA)数据库下载所有结肠癌转录数据,提取其中表达SNHG7的数据并整理成矩阵资料,利用R软件对数据进行分析,通过SNHG7差异性表达,生存分析、受试者工作特征曲线(ROC曲线),单因素多因素独立预后分析及结合临床数据特征探讨SNHG7与结肠癌患者预后的相互联系。结果SNHG7在结肠癌标本中高表达,SNHG7高表达组与SNHG7低表达组相比总体生存率明显降低,单因素及多因素回归分析显示SNHG7可作为结肠癌患者预后不佳的独立危险因素(P<0.05)。临床数据相关性研究提示SNHG7的表达量与SNHG7甲基化程度相关,SNHG7甲基化程度与结肠癌的N分期、种族相关(P<0.05)。结论SNHG7为结肠癌患者预后差的危险因素,可以作为结肠癌患者诊断和预后的潜在标志物,并为治疗结肠癌靶点提供新方向。

溴氰菊酯胁迫下家蚕品种辐7解毒酶基因的表达模式

昆虫体内解毒酶的过量表达和活性增强是昆虫对杀虫剂产生抗性的主要原因。辐射诱变家蚕品种辐7对菊酯类农药具有较强的耐受能力,研究溴氰菊酯胁迫下辐7体内解毒酶基因的表达模式,对于解析辐7的农药耐受性机制,培育耐农药污染的家蚕品种具有重要意义。根据不同浓度溴氰菊酯处理后辐7雄蛾的求偶和交配行为观察,最终选择2.5 mg/L溴氰菊酯处理组进行解毒酶基因的表达检测。采用荧光定量PCR检测溴氰菊酯处理后3 h和6 h辐7雄蛾触角解毒酶基因(8个羧酸酯酶基因、2个丝氨酸蛋白酶抑制剂基因、1个糜蛋白酶基因、3个脂肪酶基因、1个细胞色素P450基因和1个谷胱甘肽-S-转移酶基因)的转录水平:7个羧酸酯酶基因的表达先上调再下调,1个羧酸酯酶基因表达先下调再上调;1个细胞色素P450基因、1个丝氨酸蛋白酶抑制剂基因和1个糜蛋白酶基因表达先上调再下调;1个谷胱甘肽-S-转移酶基因、1个丝氨酸蛋白酶抑制剂基因和2个脂肪酶基因表达持续上调;1个脂肪酶基因表达在处理后6 h上调。结果表明,溴氰菊酯处理后诱导辐7雄蛾体内解毒酶基因转录水平上调,从而在一定程度上增强了辐7雄蛾对微量菊酯类农药的耐受性。

急性髓系白血病患者血清lncRNA DANCR、lncRNA SNHG7表达及与病理特征、预后的关系

目的探究急性髓系白血病(AML)患者血清长链非编码RNA抗分化非编码RNA(lncRNA DANCR)、长链非编码RNA小核仁RNA宿主基因7(lncRNA SNHG7)表达及与病理特征、预后的关系。方法选取2017年10月至2018年10月期间在本院就诊后出院的120例AML患者记为AML组,另选取120例在本院体检的志愿者记为对照组。观察并记录所有参与者年龄、性别、AML形态学分型等临床指标。实时荧光定量PCR(qRT-PCR)方法检测lncRNA DANCR、lncRNA SNHG7表达量;t检验方法分析lncRNA DANCR、lncRNA SNHG7表达差异及其与病理特征的关系;Pearson相关性分析二者表达水平的相关性;根据二者表达水平均值将AML患者分为lncRNA DANCR高表达组(n=65)和lncRNA DANCR低表达组(n=55)及lncRNA SNHG7高表达组(n=74)和lncRNA SNHG7低表达组(n=46);Kaplan-Meier法进行生存分析。结果AML患者lncRNA DANCR、lncRNA SNHG7水平显著高于对照组(P<0.05),且二者具有正相关性(r=0.414,P<0.001);LncRNA DANCR、lncRNA SNHG7表达水平与危险度分层、白细胞(WBC)、血红蛋白(Hb)及血小板计数(PLT)相关(P<0.05);高表达lncRNA DANCR、lncRNA SNHG7的AML患者5年生存率分别为30.77%、31.08%,低表达lncRNA DANCR、lncRNA SNHG7的AML患者5年生存率分别为50.91%、54.35%(P<0.05),低表达lncRNA DANCR、lncRNA SNHG7的AML患者生存率显著更高(P<0.05)。结论LncRNA DANCR、lncRNA SNHG7在AML患者血清中表达水平较高,两者具有的正相关性,可作为AML患者预后不良的重要指标。

Evaluation of autophagy-related genes and lncRNAs signature for prognositic prediction in thyroid carcinoma via bioinformatics analysis

Autophagy plays a significant role in the pathogenesis and prognosis of thyroid carcinoma.The role of autophagy-related genes and long non-coding RNAs,as well as the risk model of thyroid carcinoma patients were investigated to predict clinical outcome of thyroid carcinoma.Different expression of autophagy-related genes and long non-coding RNAs in thyroid carcinoma patients was identified in The Cancer Genome Atlas database.Functional enrichment analysis and gene set enrichment analysis was used to hint the mechanism that autophagy might act in thyroid carcinoma.Univariate and multivariate Cox regression analyses were performed for screening the prognostic autophagy-related genes and long non-coding RNAs to construct prognostic related risk model.thyroid carcinoma patients were divided into the low-risk and high-risk groups.The overall survival time was both shorter in the high-risk groups than that in the low-risk groups.As for autophagy-related genes prognostic risk model,age and autophagy-related genes risk score are independent prognostic factors that affect the survival of thyroid carcinoma.ATIC and CDKN2A expression was closely related to pathological stage and T status,DNAJB1 expression was closely related to M status,age and gender.While autophagy-associated long non-coding RNA related prognostic risk model consequently demonstrated that the long non-coding RNA risk score could significantly predict the survival rate of thyroid carcinoma patients with areas under the curve of 0.972.gene set enrichment analysis presented that a total of 16 gene sets including 10 up-regulated and 6 down-regulated gene sets were significantly enriched.The autophagy-related genes and long non-coding RNAs based prognostic risk models are a reliable forecasting tool for thyroid carcinoma patients.

TCF7L2基因多态性、HbA1c、血浆致动脉粥样硬化指数与2型糖尿病合并冠心病的关系研究

目的探讨转录因子7类似物2(TCF7L2)基因多态性、糖化血红蛋白(HbA1c)、血浆致动脉粥样硬化指数(AIP)与2型糖尿病合并冠心病的关系。方法回顾性选取2020年11月至2022年6月在重庆大学附属江津医院治疗的2型糖尿病患者200例,其中合并冠心病患者67例(观察组),未合并冠心病患者133例(对照组)。比较观察组和对照组的临床资料、HbA1c、AIP、TCF7L2基因多态性差异,并比较观察组不同狭窄程度患者HbA1c、AIP及TCF7L2基因多态性;分析2型糖尿病合并冠心病的影响因素。结果观察组年龄、吸烟患者比率、空腹血糖、HbA1c、糖尿病病程、AIP和甘油三酯分别为(68.82±8.10)岁、49.25%、(9.28±2.12)mmol/L、(8.89±1.82)%、(12.54±2.11)年、1.01±0.32、(2.10±0.67)mmol/L,明显高于对照组[(61.19±9.40)岁、28.57%、(8.33±2.03)mmol/L、(7.50±1.60)%、(9.93±1.96)年、0.60±0.27、(1.89±0.60)mmol/L],差异均有统计学意义(P<0.05)。观察组患者rs290481位点AG+GG基因比例为67.16%,明显高于对照组(34.59%),差异有统计学意义(P<0.05)。观察组和对照组rs7903146位点多态性比较,差异无统计学意义(P>0.05)。中重度狭窄患者HbA1c和AIP分别为(10.30±1.56)%和1.30±0.26,明显高于轻度狭窄患者[(8.20±1.45)%,0.87±0.24],差异均有统计学意义(P<0.05)。中重度狭窄患者TCF7L2基因rs290481位点AG+GG基因型比例为86.36%,明显高于轻度狭窄患者(13.64%),差异有统计学意义(P<0.05)。Logistic回归分析显示:年龄、吸烟、HbA1c、糖尿病病程、AIP及TCF7L2基因rs290481位点多态性为2型糖尿病合并冠心病的影响因素(P<0.05)。结论TCF7L2基因rs290481位点多态性、HbA1c、AIP是2型糖尿病合并冠心病的影响因素,同时与患者冠状动脉狭窄程度有一定关系。

副猪嗜血杆菌血清7型crp基因缺失株的部分生物学特性

副猪嗜血杆菌(HPS)是引起猪格拉瑟病(Glasser’s disease)的病原菌,给全球和中国规模化养猪业造成了重大经济损失。副猪嗜血杆菌血清型众多,其中血清7型是近年来分离比例越来越高的血清型,也是危害越来越严重的血清型。crp是最重要的系统调控因子之一,在细菌感染过程中对细菌适应环境变化起着至关重要的作用。为了筛选副猪嗜血杆菌血清7型crp基因缺失弱毒株,采用自然转化法构建了副猪嗜血杆菌血清7型临床分离株HPS7的crp基因缺失株,对HPS7及其crp基因缺失株的生长特性、生物膜形成、抗药性、毒力等进行了研究。结果表明,HPS7的crp基因缺失后,生长显著减慢,自凝速度减慢,生物膜形成能力明显减弱,对大部分抗生素的抗性降低,对豚鼠的毒力降低。以上结果说明crp基因对HPS7的生长、抗药性、毒力均有明显影响。本研究结果为筛选副猪嗜血杆菌血清7型弱毒株提供了参考。

Cloning and expression of MXR7 gene in human HCC tissue

AIM To clone and identify the whole cDNA ofMXR7 gene and to find out its expression inhuman HCC,and normal tissues.METHODS The DNA primers were designed andsynthesized according to the whole cDNAsequence of MXR? gene.The cDNA of humanHCC was taken as the template while the cDNA ofMXR7 gene was synthesized by polymerasechain reaction（PCR）.Recombinant DNAconforming to reading frame was constructed byconnecting purified PCR product of the cDNA ofMXR? gene with expression vector pGEX-5X-1 offusion protein.The plasmid MXRT/pGEX-5X-1was identified by sequencing.Using 32p labeledMXR? cDNA as probe,MXR7 mRNA expressionwas detected by Northern blot analysis in 12different human normal tissues,7 preoperativelyuntreated non-liver tumor tissues,30preoperatively untreated HCC,theparacancerous liver tissues and 12 normal livertissues samples.RESULTS Restriction enzyme and sequenceanalysis confirmed that the insertion sequence invector pGEX-5X-1 was the same as the cDNAsequence of MXR7 gene.Northern blot analysisshowed no expression of MXR? mRNA in 12 kindsof normal human tissues including liver,7 tumortissues in other sites and 12 normal livertissues,the frequencies of MXR7 mRNA expression in HCC and paracancerous livertissues were 76.6% and 13.3%,respectively.The frequency of MXR7 mRNA expression in HCCwithout elevation of serum AFP and in HCC【5cm was 90%（9/10）and 83.3%（5/6）,respectively.CONCLUSION MXR7 mRNA is highly expressedin human HCC,which is specific and occurs at anearly stage of HCC,suggesting MXR7 mRNA canbe a tumor biomarker for HCC.The detection ofMXR7 mRNA expression in the biopsied livertissue is helpful in discovering early subclinicalliver cancer in those with negative serum AFP.

Roles and regulation of bone morphogenetic protein-7 in kidney development and diseases

The gene encoding bone morphogenetic protein-7(BMP7) is expressed in the developing kidney in embryos and also in the mature organ in adults. During kidney development, expression of BMP7 is essential to determine the final number of nephrons in and proper size of the organ. The secreted BMP7 acts on the nephron progenitor cells to exert its dual functions: To maintain and expand the progenitor population and to provide them with competence to respond to differentiation cues, each relying on distinct signaling pathways. Intriguingly, in the adult organ, BMP7 has been implicated in protection against and regeneration from injury. Exogenous administration of recombinant BMP7 to animal models of kidney diseases has shown promising effects in counteracting inflammation, apoptosis and fibrosis evoked upon injury. Although the expression pattern of BMP7 has been well described, the mechanisms by which it is regulated have remained elusive and the processes by which the secretion sites of BMP7 impinge upon its functions in kidney development and diseases have not yet been assessed. Understanding the regulatory mechanisms will pave the way towards gaining better insight into the roles of BMP7, and to achieving desired control of the gene expression as a therapeutic strategy for kidney diseases.

食管鳞癌患者血清中lncRNA-TUSC7、lncRNA-UCA1表达与血管生成拟态的关系及意义

目的探讨食管鳞状细胞癌(ESCC)患者血清长链非编码肿瘤抑制候选基因-7(lncRNA-TUSC7)、长链非编码尿路上皮癌相关基因1(lncRNA-UCA1)表达与血管生成拟态(VM)的关系及意义。方法回顾性分析2020年3月至2021年12月经该院确诊为ESCC的133例患者,按照是否形成VM将其分为有VM组76例及无VM组57例,实时荧光定量聚合酶链反应(qPCR)检测两组患者血清lncRNA-TUSC7、lncRNA-UCA1表达;分析lncRNA-TUSC7、lncRNA-UCA1表达与ESCC患者不同临床参数之间的关系;Spearman相关系数分析lncRNA-TUSC7、lncRNA-UCA1表达与VM生成之间的相关性;受试者工作特征曲线(ROC曲线)分析联合lncRNA-TUSC7、lncRNA-UCA1检测对ESCC患者形成VM的预测效能。结果相比于无VM组,VM组患者lncRNA-TUSC7表达显著降低,而lncRNA-UCA1表达显著增高,差异有统计学意义(P<0.05);lncRNA-TUSC7与ESCC患者肿瘤分化程度、临床分期、淋巴结转移情况及是否发生VM具有密切联系(均P<0.05);lncRNA-UCA1与ESCC患者肿瘤分化程度、临床分期、淋巴结转移情况、浸润程度及是否发生VM具有密切联系(均P<0.05);Spearman相关系数显示,lncRNA-TUSC7与VM生成之间呈负相关(r=-0.782,P<0.001),而lncRNA-UCA1与VM生成之间呈正相关(r=0.766,P<0.001)。ROC曲线显示lncRNA-TUSC7、lncRNA-UCA1联合检测的曲线下面积显著高于单一检测(Z=2.428,P<0.05),灵敏度为89.40%,特异度为83.20%,对ESCC发生VM具有较高的诊断效能。结论通过定时监测ESCC患者lncRNA-TUSC7、lncRNA-UCA1表达能够对其是否合并VM做出有效评估,并对及时采取治疗措施预防奠定了可信的生物标志物基础,同时也为日后靶向治疗ESCC合并VM患者提供了可能的治疗靶点,具有较为深远的临床意义。

Effects of Mitofusin-2 Gene on Cell Proliferation and Chemotherapy Sensitivity of MCF-7

In order to evaluate the effect of mitofusin-2 gene （mfn2） on proliferation and chemotherapy sensitivity of human breast carcinoma cell line MCF-7 in vitro, pEGFPmfn2 plasmid carrying full length of mitofusin-2 gene was transfected, by using sofast, into MCF-7 cells. Mitofusin-2 gene expression in MCF-7 cells transfected by sofast after 48 h was detected by PCR and Western blotting, and the stable expression of GFP protein in MCF-7 cells by Western blot analysis. The proliferation of MCF-7 cells was assayed by MTT and cell counting. By using PI method, the effects of mfn2 on the cell cycle distribution of MCF-7 were measured. Annexin-Ⅴ/PI double labeling method was employed to detect the changes in apoptosis induced by chemotherapeutics before and after transfection. The results showed that the MCF-7 cells transfected with mfn2 gene could stably and highly express GFP protein. MTT assay revealed that after transfection of mfn2 cDNA, the proliferation of MCF-7 cells was significantly inhibited. DNA histogram showed that cells arrested in S phase, and the percentage of S phase cells was 42.7, 17.2 and 19.6 in mfn2 cDNA transfection group, blank plasmid transfection group and blank control group, respectively （P〈0.05）. The apoptosis ratio of the cells transfected with mfn2 gene was increased from 3.56% to 15.95%, that of the cells treated with camptothecin （CAMP） followed by mfn2 gene transfection was 69.6%, and that in blank plasmid transfection group and blank control group was 31.0% and 23.4% respectively （P〈0.05）. It was suggested that transfection of mfn2 gene could significantly inhibit the proliferation of MCF-7 cells and promote their sensitivity to CAMP with a synergic effect.