Predictive value of angiopoietin-like protein 8 in metabolic dysfunction-associated fatty liver disease and its progression:A case-control study

BACKGROUND The prevalence of metabolic dysfunction-associated fatty liver disease(MAFLD)is rapidly increasing,currently affecting approximately 25%of the global population.Liver fibrosis represents a crucial stage in the development of MAFLD,with advanced liver fibrosis elevating the risks of cirrhosis and hepatocellular carcinoma.Simple serum markers are less effective in diagnosing liver fibrosis compared to more complex markers.However,imaging techniques like transient elastography face limitations in clinical application due to equipment and technical constraints.Consequently,it is imperative to identify a straightforward yet effective method for assessing MAFLD-associated liver fibrosis.AIM To investigate the predictive value of angiopoietin-like protein 8(ANGPTL8)in MAFLD and its progression.METHODS We analyzed 160 patients who underwent abdominal ultrasonography in the Endocrinology Department,Xiaogan Central Hospital affiliated to Wuhan University of Science and Technology,during September 2021-July 2022.Using abdominal ultrasonography and MAFLD diagnostic criteria,among the 160 patients,80 patients(50%)were diagnosed with MAFLD.The MAFLD group was divided into the liver fibrosis group(n=23)and non-liver fibrosis group(n=57)by using a cut-off fibrosis-4 index≥1.45.Logistical regression was used to analyze the risk of MAFLD and the risk factors for its progression.Receiver operating characteristic curves were used to evaluate the predictive value of serum ANGPTL8 in MAFLD and its progression.RESULTS Compared with non-MAFLD patients,MAFLD patients had higher serum ANGPTL8 and triglyceride-glucose(TyG)index(both P<0.05).Serum ANGPTL8(r=0.576,P<0.001)and TyG index(r=0.473,P<0.001)were positively correlated with MAFLD.Serum ANGPTL8 was a risk factor for MAFLD[odds ratio(OR):1.123,95%confidence interval(CI):1.066-1.184,P<0.001).Serum ANGPTL8 and ANGPTL8+TyG index predicted MAFLD[area under the curve(AUC):0.832 and 0.886,respectively;both P<0.05].Compared with MAFLD patients without fibrosis,those with fibrosis had higher serum ANGPTL8 and TyG index(both P<0.05),and both parameters were positively correlated with MAFLD-associated fibrosis.Elevated serum ANGPTL8(OR:1.093,95%CI:1.044-1.144,P<0.001)and TyG index(OR:2.383,95%CI:1.199-4.736,P<0.013)were risk factors for MAFLD-associated fibrosis.Serum ANGPTL8 and ANGPTL8+TyG index predicted MAFLD-associated fibrosis(AUC:0.812 and 0.835,respectively;both P<0.05).CONCLUSION The serum levels of ANGPTL8 are elevated and positively correlated with MAFLD.They can serve as predictors for the risk of MAFLD and liver fibrosis,with the ANGPTL8+TyG index potentially exhibiting even higher predictive value.

Neuroprotective effects of acteoside in a glaucoma mouse model by targeting Serta domain-containing protein 4

AIM:To explore the therapeutic effect and main molecular mechanisms of acteoside in a glaucoma model in DBA/2J mice.METHODS:Proteomics was used to compare the differentially expressed proteins of C57 and DBA/2J mice.After acteoside administration in DBA/2J mice,anterior segment observation,intraocular pressure(IOP)monitoring,electrophysiology examination,and hematoxylin and eosin staining were used to analyze any potential effects.Immunohistochemistry(IHC)assays were used to verify the proteomics results.Furthermore,retinal ganglion cell 5(RGC5)cell proliferation was assessed with cell counting kit-8(CCK-8)assays.Serta domain-containing protein 4(Sertad4)mRNA and protein expression levels were measured by qRT-PCR and Western blot analysis,respectively.RESULTS:Proteomics analysis suggested that Sertad4 was the most significantly differentially expressed protein.Compared with the saline group,the acteoside treatment group showed decreased IOP,improved N1-P1 wave amplitudes,thicker retina,and larger numbers of cells in the ganglion cell layer(GCL).The IHC results showed that Sertad4 expression levels in DBA/2J mice treated with acteoside were significantly lower than in the saline group.Acteoside treatment could improve RGC5 cell survival and reduce the Sertad4 mRNA and protein expression levels after glutamate injury.CONCLUSION:Sertad4 is differentially expressed in DBA/2J mice.Acteoside can protect RGCs from damage,possibly through the downregulation of Sertad4,and has a potential use in glaucoma treatment.

Quantitative trait loci identification reveals zinc finger protein CONSTANS-LIKE 4 as the key candidate gene of stigma color in watermelon(Citrullus lanatus)

Stigma color is a critical agronomic trait in watermelon that plays an important role in pollination.However,there are few reports on the regulation of stigma color in watermelon.In this study,a genetic analysis of the F2 population derived from ZXG1553(P1,with orange stigma)and W1-17(P2,with yellow stigma)indicated that stigma color is a quantitative trait and the orange stigma is recessive compared with the yellow stigma.Bulk segregant analysis sequencing(BSA-seq)revealed a 3.75 Mb segment on chromosome 6 that is related to stigma color.Also,a major stable effective QTL Clqsc6.1(QTL stigma color)was detected in two years between cleaved amplified polymorphic sequencing(CAPS)markers Chr06_8338913 and Chr06_9344593 spanning a~1.01 Mb interval that harbors 51 annotated genes.Cla97C06G117020(annotated as zinc finger protein CONSTANS-LIKE 4)was identified as the best candidate gene for the stigma color trait through RNA-seq,quantitative real-time PCR(qRT-PCR),and gene structure alignment analysis among the natural watermelon panel.The expression level of Cla97C06G117020 in the orange stigma accession was lower than in the yellow stigma accessions with a significant difference.A nonsynonymous SNP site of the Cla97C06G117020 coding region that causes amino acid variation was related to the stigma color variation among nine watermelon accessions according to their re-sequencing data.Stigma color formation is often related to carotenoids,and we also found that the expression trend of ClCHYB(annotated asβ-carotene hydroxylase)in the carotenoid metabolic pathway was consistent with Cla97C06G117020,and it was expressed in low amounts in the orange stigma accession.These data indicated that Cla97C06G117020 and ClCHYB may interact to form the stigma color.This study provides a theoretical basis for gene fine mapping and mechanisms for the regulation of stigma color in watermelon.

Prognostic and immunological roles of heat shock protein A4 in lung adenocarcinoma

BACKGROUND Heat shock protein A4(HSPA4)belongs to molecular chaperone protein family which plays important roles within variable cellular activities,including cancer initiation and progression.However,the prognostic and immunological significance of HSPA4 in lung adenocarcinoma(LUAD)has not been revealed yet.AIM To explore the prognostic and immunological roles of HSPA4 to identify a novel prognostic biomarker and therapeutic target for LUAD.METHODS We assessed the prognostic and immunological significance of HSPA4 in LUAD using data from The Cancer Genome Atlas database.The association between HSPA4 expression and clinical-pathological features was assessed through Kruskal-Wallis and Wilcoxon signed-rank test.Univariate/multivariate Cox regression analyses and Kaplan-Meier curves were employed to evaluate prognostic factors,including HSPA4,in LUAD.Gene set enrichment analysis(GSEA)was conducted to identify the key signaling pathways associated with HSPA4.The correlation between HSPA4 expression and cancer immune infiltration was evaluated using single-sample gene set enrichment analysis(ssGSEA).RESULTS Overexpressing HSPA4 was significantly related to advanced pathologic TNM stage,advanced pathologic stage,progression disease status of primary therapy outcome and female subgroups with LUAD.In addition,increased HSPA4 expression was found to be related to worse disease-specific survival and overall survival.GSEA analysis indicated a significant correlation between HSPA4 and cell cycle regulation and immune response,particularly through diminishing the function of cytotoxicity cells and CD8 T cells.The ssGSEA algorithm showed a positive correlation between HSPA4 expression and infiltrating levels of Th2 cells,while a negative correlation was observed with cytotoxic cell infiltration levels.CONCLUSION Our findings indicate HSPA4 is related to prognosis and immune cell infiltrates and may act as a novel prognostic biomarker and therapeutic target for LUAD.

AAV-mediated expression of p65shRNA and bone morphogenetic protein 4 synergistically enhances chondrocyte regeneration

BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.

Recombinant angiopoietin-like protein 4 attenuates intestinal barrier structure and function injury after ischemia/reperfusion

BACKGROUND Intestinal barrier breakdown,a frequent complication of intestinal ischemiareperfusion(I/R)including dysfunction and the structure changes of the intestine,is characterized by a loss of tight junction and enhanced permeability of the intestinal barrier and increased mortality.To develop effective and novel therapeutics is important for the improvement of outcome of patients with intestinal barrier deterioration.Recombinant human angiopoietin-like protein 4(rhANGPTL4)is reported to protect the blood-brain barrier when administered exogenously,and endogenous ANGPTL4 deficiency deteriorates radiationinduced intestinal injury.AIM To identify whether rhANGPTL4 may protect intestinal barrier breakdown induced by I/R.METHODS Intestinal I/R injury was elicited through clamping the superior mesenteric artery for 60 min followed by 240 min reperfusion.Intestinal epithelial(Caco-2)cells and human umbilical vein endothelial cells were challenged by hypoxia/reoxygenation to mimic I/R in vitro.RESULTS Indicators including fluorescein isothiocyanate-conjugated dextran(4 kilodaltons;FD-4)clearance,ratio of phosphorylated myosin light chain/total myosin light chain,myosin light chain kinase and loss of zonula occludens-1,claudin-2 and VE-cadherin were significantly increased after intestinal I/R or cell hypoxia/reoxygenation.rhANGPTL4 treatment significantly reversed these indicators,which were associated with inhibiting the inflammatory and oxidative cascade,excessive activation of cellular autophagy and apoptosis and improvement of survival rate.Similar results were observed in vitro when cells were challenged by hypoxia/reoxygenation,whereas rhANGPTL4 reversed the indicators close to normal level in Caco-2 cells and human umbilical vein endothelial cells significantly.CONCLUSION rhANGPTL4 can function as a protective agent against intestinal injury induced by intestinal I/R and improve survival via maintenance of intestinal barrier structure and functions.

Effect of human angiopoietin-like protein 4 overexpression on the growth of esophageal carcinoma EC9706 cells

Objective： The aim of the study was to construct the eukaryotic expression vector of human angiopoietin-like protein 4 （ANGPTL4） and observe the effect of ANGPTL4 overexpression on the growth of esophageal carcinoma EC9706 cells. Methods： Total RNA was extracted from normal hepatic tissue, and ANGPTL4 cDNA was amplified by RT-PCR. The PCR product was doubly digested by Xbal and Sail, and then recombined into eukaryotic expression vector. Then, plRES-GFP-ANGPTL4 was obtained by G418 selection, then plRES-GFP-ANGPTL4 and plRES-GFP were transfected into EC9706 cells with lipidosome-packaged method. Meanwhile, the transfected cells were selected by G418, and then stable transfected cell lines were obtained. ANGPTL4 mRNA levels, the celt cycles and growth curves of EC9706 cells in experiment group （transfected with plRES-GFP-ANGPTL4）, empty vector group （transfected with plRES-GFP） and blank control group （EC9706 cells without transfection） were detected with RT-PCR, flow cytometry and MTT methods, respectively. Results： Eukaryotic ANGPTL4 expression vector plRES-GFP-ANGPTL4 was successfully constructed. The ANGPTL4 mRNA level （0.21 ± 0.03） in experiment group was significantly higher than that of the empty vector group （0.04 ± 0.008） and the blank control group （0.05 ± 0.007）, with significant differences （P 〈 0.01）. The proportion of cells in S phase in experiment group was significantly different with those of the other two groups （P 〈 0.05）. The cell growth of EC9706 cells in experiment group was slower than those of the other two groups. From the third day, the differences began to be significant. Conclusion： ANGPTL4 overexpression in esophageal carcinoma EC9706 cells could inhibit the growth of EC9706 cells.

血清PCT、CRP及IL-4水平预测小儿支原体肺炎病情严重程度的价值

目的:探讨血清降钙素原(PCT)、C反应蛋白(CRP)及白细胞介素-4(IL-4)水平预测支原体肺炎患儿病情严重程度的价值。方法:选取2019年1月—2023年1月淮安市第二人民医院儿科收治的102例支原体肺炎患儿作为研究对象,根据病情将患儿分为轻症组59例和重症组43例。比较两组临床资料及基质细胞衍生因子(CXCL12)、γ干扰素(IFN-γ)、硫化氢(H_(2)S)、超氧化物歧化酶(SOD)、基质金属蛋白酶-9(MMP-9)、PCT、CRP及IL-4水平,多因素分析采取非条件logistic逐步回归分析,采用ROC曲线分析PCT、CRP及IL-4水平对重症支原体肺炎的预测价值。结果:两组性别、年龄、病程及CXCL12、IFN-γ、H_(2)S、SOD、MMP-9水平比较,差异无统计学意义(P>0.05);重症组PCT、CRP、IL-4水平显著高于轻症组,差异有统计学意义(P<0.05)。logistic逐步回归分析结果显示,PCT、CRP及IL-4为重症支原体肺炎独立危险因素(P<0.05)。ROC分析显示,PCT、CRP及IL-4预测重症支原体肺炎的曲线下面积分别为0.896、0.851、0.787。结论:血清PCT、CRP及IL-4水平均参与支气管肺炎患儿的病情进展,且可作为重症支气管肺炎的诊断指标。

老年重症肺炎患者血清4-HNE、APC、sCD163预测预后不良的价值

目的探讨老年重症肺炎(SP)患者血清4-羟基壬烯醛(4-HNE)、活化蛋白C(APC)、可溶性血红蛋白清道夫受体163(sCD163)对预后不良的预测价值。方法选取2020年8月至2022年8月新乡医学院第一附属医院收治的200例老年SP患者纳入SP组,另选取同期、同年龄段200例老年普通肺炎患者纳入普通肺炎组。比较两组患者和SP组不同预后患者的血清4-HNE、APC、sCD163水平,并采用Pearson法分析SP组患者血清4-HNE、APC、sCD163水平与肺部感染评分(CPIS评分)的相关性,采用Logistic回归分析老年SP患者死亡的影响因素,采用受试者工作特性曲线(ROC)分析各指标对预后情况的预测效能。结果SP组患者的血清4-HNE、sCD163水平分别为(21.27±4.02)mg/L、(154.27±56.34)pg/mL,明显高于普通肺炎组的(15.63±3.49)mg/L、(112.17±37.59)pg/mL,APC水平为(25.47±5.06)pmol/L,明显低于普通肺炎组的(30.12±6.14)pmol/L,差异均具有统计学意义(P<0.05);经Pearson法分析结果显示,入院时SP患者的血清4-HNE、sCD163水平与CPIS评分呈正相关(r=0.754、0.723,P<0.05),APC水平与之呈负相关(r=-0.695,P<0.05);入院3 d、7 d后,死亡组患者的血清4-HNE分别为(23.89±6.12)mg/L、(26.01±8.27)mg/L,明显高于生存组的(19.03±4.11)mg/L、(17.25±3.56)mg/L,sCD163水平分别为(182.34±60.33)pg/mL、(219.46±70.41)pg/mL,明显高于生存组的(137.83±30.24)pg/mL、(120.74±25.17)pg/mL,APC水平分别为(23.04±4.89)pmol/L、(20.73±4.25)pmol/L,明显低于生存组的(27.42±4.09)pmol/L、(29.76±4.14)pmol/L,差异均具有统计学意义(P<0.05);Logistic回归分析结果显示,入院3 d、7 d后,血清4-HNE(>20.32 mg/L、>19.57 mg/L)、sCD163(>149.63 pg/mL、>146.90 pg/mL)是老年SP患者治疗28 d后死亡的危险因素,APC(>26.26 pmol/L、>27.37 pmol/L)是其保护因素(P<0.05);ROC分析结果显示,入院3 d后血清各指标水平联合预测死亡的曲线下面积(AUC)为0.910(95%CI:0.861~0.946),最佳预测敏感度、特异度分别为81.13%、86.39%,入院7 d后联合预测死亡的AUC为0.922(95%CI:0.876~0.955),最佳敏感度、特异度分别为90.57%、84.35%。结论血清4-HNE、APC、sCD163水平与老年SP发生、发展相关,各指标水平与CPIS评分均具有一定相关性,联合检测对老年SP患者预后情况具有一定预测价值,可作为临床评估肺部感染程度及预后的辅助指标。

帕金森病患者血清NPASDP-4,MBP水平表达与认知功能障碍及严重程度的诊断价值研究

目的探讨帕金森病患者血清神经元PAS结构域蛋白4(neuronal Per-Arnt-Sim domain protein 4,NPASDP-4)、髓鞘碱性蛋白(myelin basic protein,MBP)水平表达与认知功能障碍(cognitive impairment,CI)及严重程度的诊断价值研究。方法选取中国人民解放军联勤保障部队第九〇九医院收治的138例帕金森病患者为帕金森病组,同期该院体检中心的健康体检者69例为健康对照组,并根据是否发生CI以及其严重程度进一步将帕金森病组患者分为认知功能正常组(n=55)、轻度CI组(n=51)和痴呆组(n=32)。收集受试者一般资料;ELISA法检测血清NPASDP-4和MBP水平;相关性分析采用Spearman等级相关或Pearson线性相关;诊断价值分析采用ROC曲线;影响因素分析采用多因素Logistic回归。结果与健康对照组比较,帕金森病组血清NPASDP-4(6.75±0.48ng/ml vs2.38±0.31ng/ml),MBP(8.34±0.65μg/L vs 3.54±0.42μg/L)水平升高,差异具有统计学意义(t=68.751,55.761,均P<0.05)。认知功能正常组、轻度CI组、痴呆组H-Y分期比较,差异有统计学意义(χ2=7.788,P<0.05)。UPDRS-Ⅲ评分与认知功能正常组(41.95±10.36分)比较,轻度CI组(47.92±11.63分)、痴呆组(50.78±13.69分)评分升高,差异具有统计学意义(H=6.672,均P<0.05)。认知功能正常组、轻度CI组、痴呆组病程(4.28±0.54,4.71±0.58和5.16±0.63年)及血清NPASDP-4(5.89±0.40,6.83±0.55和8.12±0.54ng/ml),MBP(6.65±0.56,8.94±0.69和10.27±0.70μg/L)水平依次显著升高(H=24.114,207.950,355.594,均P<0.05),MMSE评分(28.47±0.94,24.51±1.35和17.09±2.57分)、MoCA评分(27.45±1.03,20.18±1.92和11.75±2.53分)、GPCOG总分(13.47±0.69,10.25±1.04和8.97±0.82分)依次显著降低(H=515.005,775.933,327.584,均P<0.05),差异具有统计学意义。帕金森病患者血清NPASDP-4,MBP水平均与病程(r=0.316,0.358)、H-Y分期(r=0.345,0.384)、UPDRS-Ⅲ评分(r=0.371,0.396)呈显著正相关(P<0.05),与MMSE评分(r=-0.468,-0.517)、MoCA评分(r=-0.504,-0.569)、GPCOG总分(r=-0.527,-0.538)呈显著负相关(均P<0.05)。血清NPASDP-4,MBP水平及二者联合诊断帕金森病患者CI的曲线下面积(AUC)分别为0.850,0.930和0.960,诊断帕金森病患者CI严重程度的AUC分别为0.866,0.803和0.933。H-Y分期中期[OR(95%CI):4.725(1.742~12.814)],H-Y分期晚期[OR(95%CI):5.083(1.919~13.464)]、UPDRS-Ⅲ评分[OR(95%CI):3.257(1.464~7.246)]、NPASDP-4[OR(95%CI):5.324(1.516~18.701)]和MBP[OR(95%CI):5.769(2.459~13.533)]是帕金森病患者CI的影响因素(均P<0.05);NPASDP-4[OR(95%CI):4.768(2.382~9.543)],MBP[OR(95%CI):5.846(3.141~10.882)]是帕金森病患者CI严重程度的影响因素(均P<0.05)。结论帕金森病患者血清NPASDP-4和MBP呈高水平,且均与CI及其严重程度密切相关,可能具有一定的临床诊断价值。

胶质瘤组织中TRIP4和DDIT4水平表达及其与临床病理特征和预后的关系

目的研究胶质瘤组织中甲状腺激素受体结合蛋白4(thyroid hormone receptor interacting protein 4,TRIP4)和DNA损伤诱导转录因子4(DNA damage inducing transcription factor 4,DDIT4)水平表达及其与临床病理特征和预后的关系。方法选取2018年2月~2019年2月河北医科大学第一医院收治的94例胶质瘤患者为研究对象。应用免疫组织化学法检测组织中TRIP4和DDIT4蛋白表达。比较不同临床病理特征脑胶质瘤组织中TRIP4和DDIT4蛋白表达。采用Kaplan-Meier生存曲线分析不同TRIP4和DDIT4蛋白表达胶质瘤患者生存预后的差异。单因素和多因素COX回归分析影响胶质瘤患者生存预后的因素。结果胶质瘤组织中TRIP4(68.09%)和DDIT4(65.96%)蛋白阳性率高于瘤旁组织(13.83%,10.64%),差异具有统计学意义(χ^(2)=57.212,60.866,均P<0.05)。胶质瘤组织中TRIP4与DDIT4蛋白表达呈显著正相关性(r=0.722,P<0.05)。WHO分级Ⅲ级、肿瘤直径≥3cm胶质瘤组织中TRIP4和DDIT4蛋白阳性率均高于WHO分级Ⅰ~Ⅱ级、肿瘤直径<3cm胶质瘤组织,差异具有统计学意义(χ^(2)=6.393~14.754,均P<0.05)。TRIP4阳性表达组和阴性表达组三年总体生存率分别为37.50%(24/64),66.67%(20/30),TRIP4阳性表达组三年累积生存率低于阴性表达组,差异具有统计学意义(Log-rankχ^(2)=5.949,P=0.015)。DDIT4阳性表达组和阴性表达组三年总体生存率分别为37.10%(23/62),70.00%(21/30),DDIT4阳性表达组三年累积生存率低于阴性表达组,差异具有统计学意义(Log-rankχ^(2)=7.642,P=0.006)。肿瘤直径≥3cm(HR=1.614,P=0.000),WHO分级Ⅲ级(HR=1.790,P=0.000),TRIP4阳性表达(HR=1.665,P=0.000),DDIT4阳性表达(HR=1.476,P=0.000)是影响胶质瘤患者生存预后的独立危险因素。结论胶质瘤组织中TRIP4和DDIT4蛋白表达升高,两者与肿瘤直径及WHO分级相关,是潜在的评估胶质瘤预后的肿瘤标志物。

血清成纤维细胞生长因子21和脂肪酸结合蛋白4检测对急性ST段抬高型心肌梗死患者经皮冠状动脉介入治疗术后心力衰竭的预测价值

目的探究血清成纤维细胞生长因子21(FGF21)和脂肪酸结合蛋白4(FABP4)检测对急性ST段抬高型心肌梗死(STEMI)患者经皮冠状动脉介入治疗(PCI)术后心力衰竭的预测价值。方法选取2020年9月至2022年9月内蒙古医科大学附属医院接诊的113例STEMI患者为研究对象,依据PCI术后1年是否发生心力衰竭(心衰),将其分为心衰组(n=32)和非心衰组(n=81)。应用ELISA法测定血清FGF21、FABP4表达水平,比较两组血清FGF21、FABP4水平,多因素logistic回归分析影响STEMI患者PCI术后发生心力衰竭的相关因素,ROC曲线评估血清FGF21、FABP4水平对STEMI患者PCI术后心力衰竭发生的预测价值。结果心衰组心率次数、C反应蛋白(CRP)、心肌肌钙蛋白(cTnI)、N末端B型利钠肽原(BNP)、利尿剂使用比例均显著高于非心衰组,左心室射血分数(LVEF)显著低于非心衰组(P<0.05)。心衰组血清FGF21、FABP4表达水平均明显高于非心衰组[(228.37±33.07)ng/L比(185.68±25.52)ng/L、(34.26±5.51)ng/ml比(26.87±4.67)ng/ml,t=7.345、7.195,P<0.05]。血清FGF21(95%CI 1.371~8.191)、FABP4(95%CI 1.176~4.090)及发病到至导丝通过时间(95%CI 1.058~8.157)是影响STEMI患者PCI术后发生心力衰竭的危险因素(OR>1,P<0.05),LVEF(95%CI 0.473~0.913)是保护因素(OR<1,P<0.05)。血清FGF21、FABP4单独及二者联合预测STEMI患者PCI术后发生心力衰竭的曲线下面积(AUC)分别为0.828、0.856、0.934,二者联合优于单一(Z二者联合-FGF21=1.971、Z二者联合-FABP4=2.417,P=0.048、P=0.015)。结论STEMI患者PCI术后发生心力衰竭血清FGF21、FABP4水平均明显升高,二者联合对STEMI患者PCI术后发生心力衰竭的风险具有更高的预测价值。

基于Penton蛋白禽腺病毒血清4型间接ELISA抗体检测方法的建立

Penton蛋白是构成禽腺病毒血清4型(Fowl adenovirus serotype 4,FAdV-4)衣壳的主要结构蛋白,具备高度保守性与良好的免疫原性。以纯化的Penton蛋白为包被抗原,建立一种基于禽腺病毒血清4型Penton蛋白的间接ELISA抗体检测方法。结果显示,Penton蛋白最佳包被质量浓度为2μg/mL,血清稀释倍数为200倍,酶标抗体稀释倍数为1∶5000,阳性临界值为0.222,敏感性可达到1∶6400,与鸡传染性支气管炎病毒、鸡传染性法氏囊病病毒、鸡传染性喉气管炎病毒均无交叉反应,特异性良好,批间批内重复性变异系数均<10%,在100份临床血清中阳性检出率为77%,与商品化试剂盒检出符合率可达98%。综上,建立的ELISA抗体检测方法可以用于禽腺病毒4型临床抗体检测。

血管生成素样蛋白4通过调节成纤维细胞和内皮细胞功能影响糖尿病足进程

背景:研究表明,血管因素对糖尿病足的发生具有重要影响。目的:探讨血管生成素样蛋白4在糖尿病足形成中的重要作用。方法:①对糖尿病足患者的基因表达谱数据进行生物信息学分析,找到关键基因。收集糖尿病足患者以及无糖尿病健康人的皮肤标本进行苏木精-伊红染色、免疫组化染色以及qRT-PCR实验,检测血管生成素样蛋白4表达情况。②培养人永生化皮肤成纤维细胞系和原代人脐静脉内皮细胞,将2种细胞分别分为对照组和外源性补充血管生成素样蛋白4组,通过划痕实验以及CCK-8实验分别检测成纤维细胞的迁移能力和增殖能力,通过Ki67实验检测内皮细胞的增殖能力。结果与结论:①生信分析发现,血管生成素样蛋白4基因的下调可能是导致糖尿病足形成的关键基因。②苏木精-伊红染色结果显示,与正常皮肤相比,血管生成素样蛋白在糖尿病足皮肤内弱表达,且其mRNA水平相对表达量降低(P<0.01)。③划痕实验结果显示,与对照组相比,血管生成素样蛋白4组成纤维细胞迁移能力明显增强;CCK-8细胞增殖实验显示,血管生成素样蛋白4组成纤维细胞的吸光度值在24,48 h均高于对照组(P<0.01,P<0.001);提示血管生成素样蛋白4可增强高糖处理的成纤维细胞迁移及增殖能力。④Ki67实验结果显示,与对照组相比,血管生成素样蛋白4组内皮细胞Ki67阳性细胞数目明显多于对照组;CCK-8细胞增殖实验显示,血管生成素样蛋白4组内皮细胞的吸光度值在24,48 h均高于对照组(P<0.05,P<0.001)。(5)以上结果均提示血管生成素样蛋白4可增强高糖处理内皮细胞的增殖能力。

Lnc-PCIR通过调控PABPC4的泛素化促进喉癌细胞的增殖和迁移

目的:探究LncRNA RP11-214F16.8(Lnc-PCIR)通过调控多腺苷酸结合蛋白质4[poly(A)-binding protein cytoplasmic 4,PABPC4]的泛素化对喉癌细胞增殖、迁移的影响。方法:利用Human Protein Atlas、GEPIA数据库分析Lnc-PCIR、PABPC4在头颈鳞状细胞癌组织中的水平及患者的总体生存期。选取我院130例喉癌患者的癌及癌旁组织标本,RT-qPCR、Western blot检测组织Lnc-PCIR、PABPC4 mRNA和蛋白水平,并分析Lnc-PCIR水平与患者临床病理参数的关系。将HEP-2、TU212细胞进行不同转染,分为si-Lnc-NC组、si-Lnc-PCIR组、Lnc-NC组、Lnc-PCIR组、si-Lnc-PCIR+NC组、si-Lnc-PCIR+PABPC4组。CCK-8实验、EdU染色和Transwell实验检测细胞增殖和迁移能力;RT-qPCR检测细胞Lnc-PCIR、PABPC4 mRNA水平;HEP-2、TU212细胞分别经放线菌酮(cycloheximide,CHX)、MG132、ML364处理后,Western blot检测细胞PABPC4蛋白水平。20只裸鼠经皮下注射已转染的HEP-2细胞悬液,分为si-Lnc-NC组、si-Lnc-PCIR组;1个月后,测定移植瘤体积、质量和Lnc-PCIR、PABPC4蛋白水平。结果:数据库显示头颈鳞状细胞癌组织中Lnc-PCIR、PABPC4水平高于正常组织,且Lnc-PCIR、PABPC4水平与总体生存期呈负相关(均P<0.05)。与正常组织相比,患者喉癌组织中Lnc-PCIR、PABPC4 mRNA和蛋白水平升高,且Lnc-PCIR水平与患者性别、TNM分期、肿瘤分化程度、淋巴结转移相关(均P<0.05)。敲低Lnc-PCIR后,细胞Lnc-PCIR、PABPC4蛋白水平降低;细胞培养1 d、2 d、3 d后的OD值、EdU阳性率降低,细胞迁移数减少。而过表达PABPC4能部分逆转敲低Lnc-PCIR对细胞增殖和迁移的影响(均P<0.05);过表达Lnc-PCIR能降低细胞PABPC4泛素化水平(P<0.05)。敲低Lnc-PCIR能抑制小鼠移植瘤生长(P<0.05)。结论:Lnc-PCIR通过调控PABPC4的泛素化促进喉癌细胞的增殖和迁移。

超重/肥胖合并2型糖尿病患者血清ANGPTL4、HSP70、IL-34水平与胰岛素抵抗的相关性

目的 探讨超重/肥胖合并2型糖尿病(T2DM)患者血清血管生成素样蛋白4(ANGPTL4)、热休克蛋白(HSP)70、白细胞介素-34(IL-34)水平与胰岛素抵抗的相关性。方法 选取2020年5月—2022年5月保定市第一中心医院T2DM患者182例(T2DM组)。参考相关诊断标准,将T2DM患者分为超重/肥胖T2DM组(90例)和体重正常T2DM组(92例)。另选取同期健康体检者90名作为正常对照组,其中超重/肥胖者40名(超重/肥胖对照组)、体重正常者50名(体重正常对照组)。检测所有研究对象血清ANGPTL4、HSP70、IL-34、胰岛素和血糖水平,计算稳态模型胰岛素抵抗指数(HOMA-IR)。T2DM患者治疗3个月后,再次检测其血清ANGPTL4、HSP70、IL-34水平和HOMA-IR。采用Pearson相关分析评估血清ANGPTL4、HSP70、IL-34与HOMA-IR的相关性。结果 与正常对照组和体重正常T2DM组比较,超重/肥胖T2DM组血清ANGPTL4和HSP70显著降低(P<0.05),血清IL-34和HOMA-IR显著升高(P<0.05)。与正常对照组比较,体重正常T2DM组血清ANGPTL4和HSP70显著降低(P<0.05),血清IL-34和HOMA-IR显著升高(P<0.05)。Pearson相关分析结果显示,血清ANGPTL4、HSP70与HOMA-IR呈负相关(r值分别为-0.733、-0.758,P<0.001),IL-34和HOMA-IR呈正相关(r=0.705,P<0.001)。治疗后,超重/肥胖T2DM组和体重正常T2DM组血清ANGPTL4和HSP70均明显升高,血清IL-34和HOMA-IR明显降低;且相对于超重/肥胖T2DM组,体重正常T2DM组血清ANGPTL4和HSP70升高更显著(P<0.05),血清IL-34和HOMA-IR降低更显著(P<0.05)。结论 超重/肥胖合并T2DM患者ANGPTL4、HSP70和IL-34与胰岛素抵抗显著相关,或可作为超重/肥胖合并T2DM的疗效监测指标。

视网膜静脉阻塞病人血清胶质纤维酸性蛋白和水通道蛋白4水平变化及其与预后的关系

目的检测视网膜静脉阻塞(RVO)病人血清中胶质纤维酸性蛋白(GFAP)、水通道蛋白4(AQP4)水平,探究二者表达水平与RVO病人预后的关系。方法回顾性选取2019年8月至2021年8月西安医学院第二附属医院收治的94例RVO病人为研究组,另取同期体检健康者85例为对照组。收集病人一般临床资料,对研究组和对照组的血清GFAP、AQP4水平进行检测;根据研究组病人预后情况将其分为预后良好组(39例)和预后不良组(55例);多因素logistic回归分析RVO病人预后的影响因素;绘制血清GFAP、AQP4对RVO病人预后评估的受试者操作特征曲线(ROC曲线)。结果研究组血清GFAP水平(3.56±0.74)μg/L显著高于对照组(1.85±0.41)μg/L,研究组血清AQP4水平(15.97±2.82)μg/L显著高于对照组(10.56±2.54)μg/L(P<0.05)。预后不良组血清GFAP水平(3.91±0.84)μg/L显著高于预后良好组(3.06±0.62)μg/L;预后不良组血清AQP4水平(17.25±3.29)μg/L显著高于预后良好组(14.18±2.18)μg/L(P<0.05)。预后良好组与预后不良组病人高血压史、空腹血糖、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C)差异有统计学意义(P<0.05)。logistic回归分析显示,血清GFAP、AQP4、高血压史、空腹血糖、TC、TG、LDL-C和HDL-C均是RVO病人预后不良的影响因素(P<0.05)。血清中GFAP、AQP4、二者联合预测RVO病人预后的AUC分别是0.73、0.79、0.92,灵敏度分别为47.27%、78.85%、77.08%,特异度分别为89.74%、72.97%、97.14%,约登指数分别为0.370、0.518、0.742;二者联合优于GFAP、AQP4各自单独预测(均P<0.05)。结论RVO病人血清GFAP、AQP4水平显著升高,对病人的预后状况具有较高的预测效能,可为临床的合理干预和改善病人预后提供依据。

2型糖尿病伴干眼症病人血清和泪液分泌型卷曲相关蛋白5、脂肪酸结合蛋白4水平与病情严重程度的相关性

目的分析分泌型卷曲相关蛋白5(SFRP-5)、脂肪酸结合蛋白4(FABP4)在2型糖尿病(T2DM)伴干眼症病人血清和泪液中的表达及其与病情严重程度的相关性。方法选取2020年12月至2021年12月唐山市眼科医院收治的T2DM病人145例,其中单纯T2DM病人84例168眼(T2DM组),伴干眼症病人61例122眼(T2DM伴干眼症组),另选取同期该院体检健康者50例100眼作为对照组。T2DM伴干眼症病人又分为轻度组(29例)、中度组(17例)、重度组(15例)。利用酶联免疫吸附法测定所有受试者血清和泪液中SFRP-5、FABP4水平;相关性分析采用Pearson法或Spearman法;logistic回归分析影响T2DM病人干眼症发生的因素。结果T2DM伴干眼症组、T2DM组血清和泪液SFRP-5水平均低于对照组(106.09±8.37、135.72±9.26比158.34±9.45,28.85±5.13、58.27±6.14比45.18±5.92),T2DM伴干眼症组低于T2DM组(P<0.05);T2DM伴干眼症组、T2DM组血清和泪液FABP4水平均高于对照组(70.63±6.59、58.27±6.14比45.18±5.92,15.91±3.76、10.28±3.58比7.72±3.29),T2DM伴干眼症组高于T2DM组(P<0.05)。重度组、中度组血清和泪液SFRP-5水平(68.29±7.15、95.54±8.34比131.82±9.02,12.83±4.62、24.72±5.49比39.56±5.18)、泪膜破裂时间(BUT)、泪液分泌试验(SIT)低于轻度组,重度组低于中度组(P<0.05);重度组、中度组血清和泪液FABP4水平(84.56±6.83、73.18±6.94比61.93±6.27,25.64±4.19、17.15±3.86比10.16±3.47)及眼表疾病指数量表(OSDI)积分高于轻度组,重度组高于中度组(P<0.05)。T2DM伴干眼症病人血清与泪液SFRP-5水平呈正相关,血清与泪液FABP4水平也呈正相关(P<0.05)。T2DM伴干眼症病人血清和泪液SFRP-5水平与OSDI积分均呈负相关,与BUT、SIT均呈正相关(P<0.05);血清和泪液FABP4水平与OSDI积分均呈正相关,与BUT、SIT均呈负相关(P<0.05)。血清和泪液SFRP-5水平是影响T2DM病人干眼症发生的独立保护因素,而血清和泪液FABP4水平是独立危险因素(P<0.05)。结论SFRP-5在T2DM伴干眼症病人血清和泪液中均低表达,FABP4均高表达,二者与病情严重程度密切相关。

下调PDCD4抑制MAP2K3和p38 MAPK的表达减轻LPS诱导的肾小管上皮细胞炎症和凋亡

目的研究程序性细胞死亡蛋白4(programmed cell death protein 4,PDCD4)在脓毒症诱导的急性肾损伤(acute kidney injury,AKI)中的作用机制,以及调控PDCD4表达通过丝裂原活化蛋白激酶3(mitogen-activated protein kinase 3,MAP2K3)和p38蛋白激酶(p38 mitogen-activated protein kinase,p38 MAPK)对脓毒症AKI起到潜在治疗作用。方法用脂多糖(lipopolysaccharide,LPS)刺激人肾小管上皮细胞(HK-2)构建脓毒症AKI细胞模型。进一步用腺病毒介导siRNA和过表达载体抑制和上调AKI细胞模型中PDCD4的表达;CCK-8法检测细胞增殖;用DCFH-DA及激光共聚焦显微镜检测细胞中ROS水平,用总SOD活性检测试剂和MDA检测试剂盒检测细胞中SOD和MDA水平;免疫共沉淀验证PDCD4和MAP2K3之间的蛋白相互作用;TUNEL染色法检测细胞凋亡;RT-qPCR和Western blot检测PDCD4及相关基因的mRNA和蛋白表达水平;ELISA法检测患者血清中炎症相关因子水平。结果LPS诱导可以促进HK-2细胞中PDCD4表达,下调PDCD4可抑制LPS诱导的HK-2细胞的炎症、氧化应激及细胞凋亡。数据库预测及免疫共沉淀证实PDCD4可以与MAP2K3相互作用,且在LPS诱导的HK-2细胞中,MAP2K3表达水平显著增强。MAP2K3过表达和p38 MAPK激动剂可以减轻PDCD4下调对LPS诱导的细胞炎症和氧化应激的影响并抑制细胞凋亡。结论下调PDCD4可以通过抑制MAP2K3和p38 MAPK从而抑制LPS诱导的肾小管上皮细胞的炎症和凋亡。

中晚期宫颈癌患者血清HE4、TK1、DCLK1水平变化及其与化疗效果的关系

目的 分析中晚期宫颈癌患者血清人附睾蛋白4(HE4)、胸苷激酶1(TK1)、双皮质素样激酶1(DCLK1)变化及其与化疗效果的关系。方法 收集2020年1月至2023年1月于郑州大学第一附属医院接受化疗的215例中晚期宫颈癌患者的病历资料,根据纳入患者的化疗效果将其分为良好组和不良组,其中良好组疗效评估结果为完全缓解(CR)与部分缓解(PR),共173例,不良组疗效评估结果为稳定(SD)与进展(PD),共42例。比较两组血清HE4、TK1、DCLK1水平等临床资料,分析中晚期宫颈癌患者血清HE4、TK1、DCLK1水平与化疗效果的关系。结果 良好组临床分期为Ⅱ期比例、高分化比例、无淋巴结转移比例均高于不良组,肿瘤最大直径以及血清HE4、TK1、DCLK1水平均低于不良组,差异均有统计学意义(P<0.05);经logistic多因素分析显示,肿瘤的临床分期达到Ⅳ期、分化程度为中低分化、淋巴结转移以及血清HE4、TK1、DCLK1水平升高均为影响中晚期宫颈癌患者化疗效果的独立因素(P<0.05);经Spearman相关性分析显示,中晚期宫颈癌患者临床分期、淋巴结转移以及血清HE4、TK1、DCLK1水平与其化疗效果成负相关,分化程度与其化疗效果成正相关(P<0.05)。结论 中晚期宫颈癌患者临床分期越晚、分化程度越差、临床转移以及HE4、TK1、DCLK1水平越高,越不利于患者的化疗,上述指标对其预后均具有一定预测价值。