S100 calcium binding protein A6 and associated long noncoding ribonucleic acids as biomarkers in the diagnosis and staging of primary biliary cholangitis

BACKGROUND Primary biliary cholangitis(PBC)is a chronic and slowly progressing cholestatic disease,which causes damage to the small intrahepatic bile duct by immunoregulation,and may lead to cholestasis,liver fibrosis,cirrhosis and,eventually,liver failure.AIM To explore the potential diagnosis and staging value of plasma S100 calcium binding protein A6(S100A6)messenger ribonucleic acid(mRNA),LINC00312,LINC00472,and LINC01257 in primary biliary cholangitis.METHODS A total of 145 PBC patients and 110 healthy controls(HCs)were enrolled.Among them,80 PBC patients and 60 HCs were used as the training set,and 65 PBC patients and 50 HCs were used as the validation set.The relative expression levels of plasma S100A6 mRNA,long noncoding ribonucleic acids LINC00312,LINC00472 and LINC01257 were analyzed using quantitative reverse transcription-polymerase chain reaction.The bile duct ligation(BDL)mouse model was used to simulate PBC.Then double immunofluorescence was conducted to verify the overexpression of S100A6 protein in intrahepatic bile duct cells of BDL mice.Human intrahepatic biliary epithelial cells were treated with glycochenodeoxycholate to simulate the cholestatic environment of intrahepatic biliary epithelial cells in PBC.RESULTS The expression of S100A6 protein in intrahepatic bile duct cells was up-regulated in the BDL mouse model compared with sham mice.The relative expression levels of plasma S100A6 mRNA,log10 LINC00472 and LINC01257 were upregulated while LINC00312 was down-regulated in plasma of PBC patients compared with HCs(3.01±1.04 vs 2.09±0.87,P<0.0001;2.46±1.03 vs 1.77±0.84,P<0.0001;3.49±1.64 vs 2.37±0.96,P<0.0001;1.70±0.33 vs 2.07±0.53,P<0.0001,respectively).The relative expression levels of S100A6 mRNA,LINC00472 and LINC01257 were up-regulated and LINC00312 was down-regulated in human intrahepatic biliary epithelial cells treated with glycochenodeoxycholate compared with control(2.97±0.43 vs 1.09±0.08,P=0.0018;2.70±0.26 vs 1.10±0.10,P=0.0006;2.23±0.21 vs 1.10±0.10,P=0.0011;1.20±0.04 vs 3.03±0.15,P<0.0001,respectively).The mean expression of S100A6 in the advanced stage(III and IV)of PBC was up-regulated compared to that in HCs and the early stage(II)(3.38±0.71 vs 2.09±0.87,P<0.0001;3.38±0.71 vs 2.57±1.21,P=0.0003,respectively);and in the early stage(II),it was higher than that in HCs(2.57±1.21 vs 2.09±0.87,P=0.03).The mean expression of LINC00312 in the advanced stage was lower than that in the early stage and HCs(1.39±0.29 vs 1.56±0.33,P=0.01;1.39±0.29 vs 2.07±0.53,P<0.0001,respectively);in addition,the mean expression of LINC00312 in the early stage was lower than that in HCs(1.56±0.33 vs 2.07±0.53,P<0.0001).The mean expression of log10 LINC00472 in the advanced stage was higher than those in the early stage and HCs(2.99±0.87 vs 1.81±0.83,P<0.0001;2.99±0.87 vs 1.77±0.84,P<0.0001,respectively).The mean expression of LINC01257 in both the early stage and advanced stage were up-regulated compared with HCs(3.88±1.55 vs 2.37±0.96,P<0.0001;3.57±1.79 vs 2.37±0.96,P<0.0001,respectively).The areas under the curves(AUC)for S100A6,LINC00312,log10 LINC00472 and LINC01257 in PBC diagnosis were 0.759,0.7292,0.6942 and 0.7158,respectively.Furthermore,the AUC for these four genes in PBC staging were 0.666,0.661,0.839 and 0.5549,respectively.The expression levels of S100A6 mRNA,log10 LINC00472,and LINC01257 in plasma of PBC patients were decreased(2.35±1.02 vs 3.06±1.04,P=0.0018;1.99±0.83 vs 2.33±0.96,P=0.036;2.84±0.92 vs 3.69±1.54,P=0.0006),and the expression level of LINC00312 was increased(1.95±0.35 vs 1.73±0.32,P=0.0007)after treatment compared with before treatment using the paired t-test.Relative expression of S100A6 mRNA was positively correlated with log10 LINC00472(r=0.683,P<0.0001);serum level of collagen type IV was positively correlated with the relative expression of log10 LINC00472(r=0.482,P<0.0001);relative expression of S100A6 mRNA was positively correlated with the serum level of collagen type IV(r=0.732,P<0.0001).The AUC for the four biomarkers obtained in the validation set were close to the training set.CONCLUSION These four genes may potentially act as novel biomarkers for the diagnosis of PBC.Moreover,LINC00472 acts as a potential biomarker for staging in PBC.

Serum Y-Box Binding Protein 1 (YBX-1) and Interleukin 6 (IL-6) Are Associated with Metastasis in Breast Cancer Patients

Objectives: The aim of this study was to assess the levels of Y-box binding protein 1 (YBX-1) and interleukin 6 (IL-6) in the sera of metastatic and non-metastatic breast cancer patients (BC), investigate their clinicopathological significance and to analyze their potential use as biomarkers of breast cancer metastasis. Methods: The study included ninety subjects sub-grouped equally into metastatic BC, non-metastatic BC and healthy volunteers. Serum YBX-1 and IL-6 were quantified using ELISA technique while CA 15-3 was quantified using IRMA kit. Clinical data were collected from patients’ records. Results: YBX-1 (p < 0.001), IL-6 (p < 0.001) and CA15-3 (p = 0.017, 0.001) were significantly elevated in metastatic and non-metastatic BC patients compared to healthy controls, however, only YBX-1 (p 0.001) showed a significant difference with cancer metastasis. Generally, YBX-1 and IL-6 were correlated with worse histological grade and late clinical stage in breast cancer patients and they were also associated with axillary lymph nodes involvement and positive vascular invasion in metastatic BC patients. Serum YBX-1 and IL-6 levels were positively correlated to each other (rs = 0.615, p < 0.001) and they showed high sensitivity and specificity compared to CA 15-3 (p < 0.001 and p = 0.004 for YBX-1 and IL-6 respectively) for predicting cancer metastasis. Conclusions: Serum YBX-1 and IL-6 are potential biomarkers of breast cancer patients with significant correlation with bad clinicopathological characteristics. Serum YBX-1 and IL-6 have superior sensitivity and specificity compared to CA15-3 and can serve as potential follow up and prognostic markers.

Role of the PEST sequence in the long-type GATA-6 DNA-binding protein expressed in human cancer cell lines

GATA-6 mRNA utilizes two Met-codons in frame as translational initiation codons in cultured mammalian cells. Deletion of the nucleotide sequence encoding the PEST sequence between the two initiation codons unusually reduced the protein molecular size on SDS-polyacrylamide gel-electrophoresis. The reduced molecular size is ascribed to the molecular property of GATA-6, since both amino-and carboxy-lterminal tags introduced into GATA-6 were detected on the gel. This PEST sequence seems to contribute to expansion of the long-type GATA-6 molecule. The long-type GATA-6 containing the PEST sequence exhibits more activation potential than that without this sequence, the latter’s activity being similar to that of the short-type GATA-6. We further demonstrated that human colon and lung cancer cell lines express both the long-type GATA-6 and the short-type GATA-6 in their nuclei.

MicroRNA-185-5p mediates regulation of SREBP2 expression by hepatitis C virus core protein

AIM: To investigate the molecular mechanism for regulation of cholesterol metabolism by hepatitis C virus(HCV) core protein in Hep G2 cells.METHODS: HCV genotype 1b core protein was cloned and expressed in Hep G2 cells. The cholesterol content was determined after transfection. The expression of sterol regulatory element binding protein 2(SREBP2) and the rate-limiting enzyme in cholesterol synthesis(HMGCR) was measured by quantitative real-time PCR and immunoblotting after transfection. The effects of core protein on the SREBP2 promoter and 3'-untranslated region were analyzed by luciferase assay. We used different target predictive algorithms, micro RNA(mi RNA) mimics/inhibitors, and site-directed mutation to identify a putative target of a particular mi RNA.RESULTS: HCV core protein expression in Hep G2 cells increased the total intracellular cholesterol level(4.05 ± 0.17 vs 6.47 ± 0.68, P = 0.001), and this increase corresponded to an increase in SREBP2 and HMGCR m RNA levels(P = 0.009 and 0.037, respectively) and protein expression. The molecular mechanism studyrevealed that the HCV core protein increased the expression of SREBP2 by enhancing its promoter activity(P = 0.004). In addition, mi R-185-5p expression was tightly regulated by the HCV core protein(P = 0.041). Moreover, overexpression of mi R-185-5p repressed the SREBP2 m RNA level(P = 0.022) and protein expression. In contrast, inhibition of mi R-185-5p caused upregulation of SREBP2 protein expression. mi R-185-5p was involved in the regulation of SREBP2 expression by HCV core protein. CONCLUSION: HCV core protein disturbs the cholesterol homeostasis in Hep G2 cells via the SREBP2 pathway; mi R-185-5p is involved in the regulation of SREBP2 by the core protein.

Is X-linked methyl-CpG binding protein 2 a new target for the treatment of Parkinson's disease?

X-linked methyl-CpG binding protein 2 mutations can induce symptoms similar to those of Parkinson’s disease and dopamine metabolism disorders, but the specific role of X-linked methyl-CpG binding protein 2 in the pathogenesis of Parkinson’s disease remains unknown. In the present study, we used 6-hydroxydopamine-induced human neuroblastoma cell （SH-SY5Y cells） injury as a cell model of Parkinson’s disease. The 6-hydroxydopamine （50 μmol/L） treatment decreased protein levels for both X-linked methyl-CpG binding protein 2 and tyrosine hydroxylase in these cells, and led to cell death. However, overexpression of X-linked methyl-CpG binding protein 2 was able to ameliorate the effects of 6-hydroxydopamine, it reduced 6-hydroxydopamine-induced apoptosis, and increased the levels of tyrosine hydroxylase in SH-SY5Y cells. These findings suggesting that X-linked methyl-CpG binding protein 2 may be a potential therapeutic target for the treatment of Parkinson’s disease.

Hepatitis C virus non-structural 5A protein can enhance full-length core protein-induced nuclear factor-κB activation

AIM： To study the effects of hepatitis C virus （HCV） core and non-structural 5A （NS5A） proteins on nuclear factor- k B （NF- k B） activity for understanding their biological function on chronic hepatitis caused by HCV infection. METHODS： Luciferase assay was used to measure the activity of NF-kB in three different cell lines cotransfected with a series of deletion mutants of core protein alone or together with NS5A protein using pNF- k B-Luc as a reporter plasmid. Western blot and indirect immunofluorescence assays were used to confirm the expression of proteins and to detect their subcellular localization, respectively. Furthermore, Western blot was also used to detect the expression levels of NF- k B/p65, NF- k B/p50, and inhibitor k B-a（k B-a）. RESULTS： The wild-type core protein （C191） and its mutant segments （C173 and C158） could activate NF- k B in Huh7 cells only and activation caused by （C191） could be enhanced by NS5A protein. Moreover, the full-length core protein and its different deletion mutants alone or together with NS5A protein did not enhance the expression level of NF- k B. The NF- k B activity was augmented due to the dissociation of NF-k： B-I k： B complex and the degradation of Ik B-a. CONCLUSION：NF- k B is the key transcription factor that can activate many genes that are involved in the cellular immune response and inflammation. Coexpression of the full-length core protein along with NS5A can enhance the NF- k B activation, and this activation may play a significant role in chronic liver diseases including hepatocellular carcinoma associated with HCV infection.

Effect of Hepatitis C Virus Core Protein on Interferon-Induced Antiviral Genes Expression and Its Mechanisms

Emerging data indicated that HCV subverts the antiviral activity of interferon(IFN);however,whether HCV core protein contributes to the process remains controversial.In the present study,we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway.Our results showed that,following treatment with IFN-α,the transcription of PKR,MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein.In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased.Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1,whereas the level of STAT1 expression was unchanged.Accordingly,SOCS3,the negative regulator of the Jak/STAT pathway,was induced by HCV core protein.These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.

表达HCV core-Ant的重组慢病毒的构建

目的构建表达HCV核心区的重组慢病毒,以探讨HCV感染后该病毒核心蛋白对肝脏干细胞转分化的影响。方法利用PCR体系延伸互为模板的引物,通过T载体克隆法获得HCV core-Ant基因克隆,酶切后连入pLenti6/V5-D-TOPO质粒,得到pLenti6/V5-D-HCV core-Ant,连同pLP/VSVG、pLP1和pLP2四质粒共转染293ET细胞,获得了表达HCV core-Ant重组慢病毒。收集病毒上清,大量扩增,用免疫组化法检测目的基因在Hela细胞的表达。用类似的方法构建了表达EGFP的重组慢病毒并测定其滴度。结果克隆出HCV core-Ant基因,经酶切及测序证实结果正确;使用表达HCV core-Ant重组慢病毒感染Hela细胞,免疫组化实验证实胞浆有HCV core表达。使用表达EGFP的重组慢病毒感染3T3细胞见到绿色荧光,说明构建慢病毒载体有效。结论通过分子克隆体外重组技术成功制备了表达HCV core的重组慢病毒,为进一步研究丙肝病毒致癌机制奠定了基础。

血清PCT、IL-6、HBP和CRP水平对脑梗死合并肺部感染的预测价值

目的探究降钙素原(PCT)、白细胞介素6(IL-6)、肝素结合蛋白(HBP)和C反应蛋白(CRP)对脑梗死合并肺部感染的预测价值。方法选取2022年7月至2023年6月于南京医科大学附属江宁医院就诊的145例脑梗死患者作为脑梗死组,根据其是否并发肺部感染分为合并感染组(76例)和单纯脑梗死组(69例)。另选取同期于该院进行体检的健康人群61例作为健康人对照组。收集各组一般临床资料,检测并比较血清生化指标。采用Logistic回归分析法进行脑梗死合并肺部感染的危险因素分析。绘制ROC曲线,使用ROC曲线下面积(AUC ROC)评估各个指标对脑梗死合并肺部感染的诊断价值。结果年龄、PCT、IL-6、HBP和CRP是脑梗死并发肺部感染的独立危险因素(P<0.05)。ROC曲线分析表明,PCT、IL-6、HBP和CRP对预测脑梗死合并肺部感染均有一定的临床价值,四项联合预测的敏感性和特异性分别为85.47%和75.36%,均高于PCT、IL-6、HBP和CRP单独预测(P<0.05)。结论血清PCT、IL-6、HBP和CRP在脑梗死并发肺部感染患者血清中呈高表达,四者均可作为脑梗死合并肺部感染的诊断指标,且四项联合检测可有效提高诊断的准确性。

IGFBP6在不稳定颈动脉斑块中的作用:生物信息学分析与实验验证

目的探讨不稳定颈动脉粥样硬化斑块的差异表达基因(DEGs)及其分子相互作用。方法从基因表达数据库(GEO)和欧洲生物信息学研究所数据库下载颈动脉斑块患者的基因表达数据集GSE41571、GSE118481和E-MTAB-2055。采用基因本体生物学过程(GO-BP)富集分析、京都基因与基因组百科全书(KEGG)富集分析、蛋白-蛋白相互作用(PPI)网络、miRNAs/转录因子与靶基因的相互关系及药物-基因相互作用等方法,分析至少两个数据集中不稳定颈动脉斑块的共调控DEGs。采用定量实时PCR(qRT-PCR)和酶联免疫吸附试验(ELISA)检测颈动脉粥样硬化斑块患者58例的颈动脉斑块和血浆中部分DEGs的表达水平。结果GO富集分析显示,不稳定颈动脉斑块的DEGs主要富集在与炎症反应相关的基因和细胞外基质结构基因;KEGG富集分析显示,不稳定颈动脉斑块中上调的DEGs富集于细胞外基质受体相互作用、PI3K-Akt、Hippo信号通路及转化生长因子-β(TGF-β)信号通路,下调的DEGs主要富集于溶酶体、吞噬体及趋化因子过程。PPI网络分析结果显示,COL1A2、COL4A2、胰岛素样生长因子结合蛋白6(IGFBP6)、COL4A5、C1QA、CXCL10、CXCL2、CXCR4和CSF1R等可能在PPI网络中起重要作用。药物-基因相互作用的预测显示,CSF1R的药物相互作用最多,CXCL2受药物拮抗程度最高,IGFBP6受药物激活程度最高。qRT-PCR检测结果显示,与稳定斑块组比较,不稳定斑块组IGFBP6表达水平明显降低(P<0.001)。ELISA法检测结果显示,不稳定斑块组血浆IGFBP6浓度明显低于稳定斑块组(P<0.0001)。受试者工作特征曲线分析结果显示,采用血浆IGFBP6水平鉴别不稳定斑块的曲线下面积为0.894(95%CI 0.810~0.977),截断值为142.08 ng/ml。结论IGFBP6可能成为预测不稳定颈动脉斑块的重要生物标志物。

子宫内膜癌组织LAMB3、FABP6表达及其预后价值

目的探讨层黏连蛋白亚基β3(LAMB3)、脂肪酸结合蛋白6(FABP6)在子宫内膜癌组织中的表达及预后价值。方法以本院接受手术治疗的113例子宫内膜癌患者术中收集的子宫内膜癌组织及其癌旁组织为研究样本。检测LAMB3、FABP6在组织中的表达情况;分析LAMB3和FABP6与患者预后的关系;分析影响子宫内膜癌患者预后的危险因素。结果子宫内膜癌组织中LAMB3、FABP6表达水平高于癌旁组织(P<0.05)。子宫内膜癌组织中LAMB3与FABP6表达具有正相关关系(P<0.05)。LAMB3低表达组的生存率高于高表达组(P<0.05),FABP6低表达组的生存率高于高表达组(P<0.05);FIGO分期为Ⅲ期、肌层浸润深度≥1/2、淋巴结转移、低分化、POLE无突变型、LAMB3高表达及FABP6高表达是子宫内膜癌患者3年内死亡的危险因素(P<0.05)。结论LAMB3、FABP6在子宫内膜癌组织中异常高表达,两者与患者临床病理特征及预后密切相关。

血清视黄醇结合蛋白-4、生长停滞特异性蛋白6水平与急性心肌梗死合并心力衰竭患者预后的关系

目的探讨血清视黄醇结合蛋白-4(RBP-4)、生长停滞特异性蛋白6(GAS6)水平与急性心肌梗死(AMI)合并心力衰竭(HF)患者预后的关系。方法选取2020年1月至2022年1月内蒙古自治区通辽市医院收治的186例AMI合并HF患者,根据随访1年的预后将其分为预后不良组(61例)和预后良好组(125例)。采用酶联免疫吸附试验检测两组RBP-4、GAS6水平。采用多因素logistic回归模型分析AMI合并HF患者预后不良的影响因素,采用受试者操作特征曲线分析血清RBP-4、GAS6水平对AMI合并HF患者预后不良的预测价值。结果预后不良组年龄、急性HF占比、N末端前体B型钠尿肽、RBP-4水平高于预后良好组,左室射血分数、GAS6低于预后良好组,差异有统计学意义(P<0.05)。多因素分析显示,年龄(OR=1.091,95%CI:1.016~1.173)、急性HF(OR=2.468,95%CI:1.030~5.913)、N末端前体B型钠尿肽(OR=1.002,95%CI:1.001~1.003)、RBP-4(OR=1.156,95%CI:1.076~1.242)、左室射血分数(OR=0.829,95%CI:0.725~0.948)、GAS6(OR=0.342,95%CI:0.195~0.599)均为AMI合并HF患者预后不良的影响因素(P<0.05)。受试者操作特征曲线分析显示,血清RBP-4、GAS6水平单独及联合预测AMI合并HF患者预后不良的曲线下面积分别为0.786、0.790、0.895(P<0.05)。结论血清RBP-4水平升高和GAS6水平降低与AMI合并HF患者预后不良有关,二者联合对AMI合并HF患者预后不良具有良好预测价值。

重症社区获得性肺炎患者PIV、IL-6、HBP、PAB水平对疗效的评估价值

目的分析重症社区获得性肺炎(CAP)患者的泛免疫炎症值(PIV)、白细胞介素(IL)-6、肝素结合蛋白(HBP)、前白蛋白(PAB)水平变化及其对入院72 h治疗效果的评估价值。方法120例重症CAP患者(重症组)按照初始治疗效果分为有效组87例和失败组33例,另选取同期收治的非重症CAP患者120例为非重症组。比较重症组与非重症组、失败组与有效组入院次日PIV、IL-6、HBP及PAB水平,以受试者工作特征(ROC)曲线分析以上指标单独及联合检测对CAP病情及入院72 h治疗效果的评估价值。结果重症组PIV、IL-6及HBP水平高于非重症组,PAB水平低于非重症组(P<0.05);ROC曲线分析示,单独检测时PIV评估重症CAP的曲线下面积(AUC)最高,为0.830(95%CI:0.780~0.881);PIV联合IL-6、HBP及PAB评估重症CAP的AUC为0.929(95%CI:0.892~0.967),高于各单一指标检测(P<0.05)。失败组PIV、IL-6及HBP水平高于有效组,PAB水平低于有效组(P<0.05);ROC曲线分析示,单独检测时PIV评估重症CAP入院72 h治疗效果的AUC最高,为0.777(95%CI:0.692~0.862);PIV联合IL-6、HBP及PAB评估重症CAP入院72 h治疗效果的AUC为0.916(95%CI:0.846~0.986),高于各单一指标检测(P<0.05)。结论PIV联合IL-6、HBP、PAB检测对CAP患者病情及重症CAP入院72 h治疗效果评估均具有良好的价值。

Mechanisms of Steatosis-Derived Hepatocarcinogenesis:Lessons from HCV Core Gene Transgenic Mice

Hepatitis C virus(HCV)is a major cause of chronic hepatitis,liver cirrhosis,and hepatocellular carcinoma(HCC)worldwide.Among the structural proteins of HCV,the HCV core protein has the ability to regulate gene transcription,lipid metabolism,cell proliferation,apoptosis,and autophagy,all of which are closely related to the development of HCC.Transgenic mice carrying the HCV core gene exhibited age-dependent insulin resistance,hepatic steatosis,and HCC that resembled the clinical characteristics of chronic hepatitis C patients.Several dietary modifications,including calorie restriction and diets rich in saturated fatty acids,trans fatty acids,or cholesterol,were found to influence hepatic steatogenesis and tumorigenesis in HCV core gene transgenic mice.These strategies modulated hepatocellular stress and proliferation,in addition to hepatic fibrotic processes and the microenvironment,thereby corroborating a close interconnection between dietary habits and steatosis-related hepatocarcinogenesis.In this review,we summarize the findings obtained from mouse models transgenic for the HCV genome,with a special focus on HCV core gene transgenic mice,and discuss the mechanisms of steatogenesis and hepatocarcinogenesis induced by the HCV core protein and the impact of dietary habits on steatosis-derived HCC development.

感染可能性评分联合围术期HBP和IL-6变化率对泌尿系结石术后尿源性脓毒血症的早期识别价值

目的研究感染可能性评分(IPS)联合围术期肝素结合蛋白(HBP)和白细胞介素-6(IL-6)变化率对泌尿系结石术后尿源性脓毒血症(US)的早期识别价值。方法选取2021年4月至2024年5月新乡医学院第一附属医院收治的198例泌尿系结石手术患者进行前瞻性研究,根据术后是否发生US分为US组41例和非US组157例。比较两组患者的基线资料及IPS评分、HBP和IL-6水平,采用Spearman法分析IPS评分、HBP和IL-6变化率与US相关性,偏相关系数进一步分析其偏相关性,受试者工作特征(ROC)曲线分析各指标变化率对US的预测效能。结果US组患者的结石直径、手术时间明显长于非US组,鹿角形结石形态占比明显高于非US组,差异均有统计学意义(P<0.05);US组患者术前IL-6水平明显高于非US组,术后24 h的IPS评分、HBP、IL-6水平明显高于术前,非US组患者术后24 h的IL-6明显高于术前,US组患者术后24 h的IPS评分、HBP、IL-6及三者变化率明显高于非US组,差异均有统计学意义(P<0.05);Spearman法分析结果显示,IPS评分、HBP和IL-6变化率均与US呈正相关(r=0.816、0.907、0.785,P<0.01);偏相关性分析结果显示,控制结石直径等相关因素后,IPS评分、HBP和IL-6变化率仍与US显著相关(偏相关性系数=0.808、0.901、0.769,P<0.01);ROC分析结果显示,IPS评分变化率+HBP变化率+IL-6变化率预测US的曲线下面积(AUC)为0.931(95%CI:0.887~0.963),大于各指标变化率单独预测AUC 0.755(95%CI:0.689~0.813)、0.765(95%CI:0.699~0.822)、0.838(95%CI:0.779~0.886)。结论IPS评分、HBP、IL-6变化率与泌尿系结石患者术后US发生均具有一定相关性,联合应用对US具有较高预测价值,可作为临床预测US的辅助指标,并指导后续临床诊疗工作。

类风湿关节炎患者血清lncRNA GATA6-AS1和GATA6 mRNA水平与疾病活动度指标、免疫功能及疾病严重程度的关系

目的分析类风湿关节炎患者血清中长链非编码RNA GATA结合蛋白6反义RNA1(lncRNA GATA6-AS1)和GATA结合蛋白6(GATA6)信使RNA(mRNA)水平与疾病活动度指标、免疫功能及疾病严重程度的关系。方法选取2021年4月至2023年3月我院收治的74例类风湿关节炎缓解期患者作为缓解期组,81例类风湿关节炎活动期患者作为活动期组,另选取同期体检健康者80例为对照组。收集受试者一般资料和疾病活动度指标[类风湿因子(RF)、血细胞沉降率(ESR)、C反应蛋白(CRP)、抗环瓜氨酸肽(CCP)抗体、肿瘤坏死因子(TNF)-α、28处关节疾病活动度(DAS28)评分];荧光定量PCR法检测血清中lncRNA GATA6-AS1和GATA6 mRNA水平;并检测受试者免疫功能指标[免疫球蛋白(Ig)A、IgM、IgG、CD3^(+)T、CD4^(+)T、CD8^(+)T细胞,计算CD4^(+)/CD8^(+)比值]。分析类风湿关节炎患者血清lncRNA GATA6-AS1、GATA6 mRNA、疾病活动度指标、免疫功能指标的相关性,影响类风湿关节炎患者活动期发生的危险因素,血清lncRNA GATA6-AS1、GATA6 mRNA对类风湿关节炎患者活动期发生的预测价值。结果对照组、缓解期组、活动期组RF、ESR、CRP、抗CCP抗体、TNF-α、DAS28评分、血清lncRNA GATA6-AS1、GATA6 mRNA、IgA、IgM、IgG、外周血CD3^(+)T、CD4^(+)T、CD8^(+)T细胞比例、CD4^(+)/CD8^(+)比值水平依次升高(P<0.05);风湿关节炎患者血清lncRNA GATA6-AS1与GATA6 mRNA呈正相关(P<0.05);血清lncRNA GATA6-AS1和GATA6 mRNA水平均与RF、ESR、CRP、抗CCP抗体、TNF-α、DAS28评分、IgA、IgM、IgG、CD3^(+)T细胞、CD4^(+)T细胞、CD8^(+)T细胞和CD4^(+)/CD8^(+)比值呈正相关(P<0.05);RF、ESR、CRP、抗CCP抗体、TNF-α、DAS28评分、lncRNA GATA6-AS1、GATA6 mRNA均是影响类风湿关节炎患者活动期发生的独立危险因素(P<0.05);血清lncRNA GATA6-AS1、GATA6 mRNA、二者联合预测类风湿关节炎患者活动期发生的曲线下面积(AUC)分别为0.843、0.861、0.928;二者联合预测的AUC高于血清lncRNA GATA6-AS1、GATA6 mRNA水平各自单独预测的AUC(P<0.05)。结论类风湿关节炎患者血清中lncRNA GATA6-AS1和GATA6 mRNA呈现高水平,且活动期患者二者水平更高,二者与疾病活动度指标、免疫功能以及疾病严重程度密切相关,可以很好地预测类风湿关节炎患者活动期的发生。

冷诱导RNA结合蛋白联合IL-6的检测在妊娠期高血压疾病中的诊断价值

目的探讨冷诱导RNA结合蛋白(CIRBP)联合白细胞介素-6(IL-6)检测在妊娠期高血压疾病中的诊断价值。方法前瞻性选取2022年7月至2023年10月入上海市第四人民医院妇产科病区经剖宫产分娩的高血压产妇30例作为实验组,选取正常产妇30例作为对照组,分别采用免疫组织化学染色、Western blot、qRT-PCR和ELISA法检测两组研究对象胎盘组织及血浆中CIRBP、IL-6的表达并进行对比分析。根据疾病程度将实验组患者分为妊娠期高血压组(12例)和重度子痫前期(18例)。比较两组患者的血浆中CIRBP和IL-6的表达水平。采用ROC曲线分析CIRBP+IL-6联合检测对疾病的诊断效能。结果实验组患者胎盘组织中CIRBP和IL-6阳性表达率明显高于对照组(P<0.05);实验组患者胎盘组织中CIRBP和IL-6 mRNA和蛋白表达明显高于对照组(P<0.05);实验组患者中血浆中CIRBP和IL-6表达明显高于对照组(P<0.05);重度子痫前期组患者血浆中CIRBP和IL-6表达明显高于妊娠期高血压组(P<0.05)。结论妊娠高血压患者胎盘组织及血浆中CIRBP和IL-6均呈高表达,且随着病程的加重不断升高,对诊断高血压产妇有深远的影响。

骆驼刺提取物对脂多糖诱导的IEC-6细胞损伤模型NLRP3炎症小体及相关细胞因子的影响

目的研究骆驼刺提取物(Alhagi pseudalhagi(M.B.)Desv.Extract,APE)对脂多糖诱导的大鼠小肠隐窝上皮细胞(Intestinal epithelial cell,IEC-6)损伤模型NLRP3炎症小体及相关细胞因子的影响。方法培养IEC-6细胞,将其分为空白组、模型组、APE低、中、高浓度组,用1.0μg/mL的脂多糖(Lipopolysaccharide,LPS)诱导建立细胞炎症损伤模型,APE(低、中、高浓度:15、25、35μg/mL)干预后采用CCK-8法检测细胞的存活率,通过ELISA试剂盒检测炎症因子IL-1β、IL-18、TNF-α的分泌水平。蛋白质印迹法(WB)检测核苷酸结合寡聚化结构域样受体蛋白3(Nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体信号通路5个关键蛋白:NLRP3、半胱氨酸天冬氨酸蛋白酶1(Cystein-asparate protease-1,Caspase-l)、凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing a CARD,ASC)及抗凋亡蛋白Bcl-2(Anti-apoptosis Protein Bcl-2)和Bcl-xl(Anti-apoptosis Protein Bcl-xl)表达。结果与空白组比较,模型组IEC-6细胞的存活率降低,NLRP3、Caspase-1、ASC蛋白表达水平升高,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平降低,促炎因子IL-1β、IL-18和TNF-α的分泌水平升高,差异有统计学意义(P<0.05)。与模型组比较,APE低、中、高浓度组细胞存活率升高,35μg/mL APE组IEC-6细胞的NLRP3、Caspase-1、ASC蛋白相对表达水平降低,抗凋亡蛋白Bcl-2、Bcl-xl的表达水平升高,差异有统计学意义(P<0.05)。中、高浓度的APE能够抑制炎症因子分泌,25μg/mL APE对IL-1β、IL-18、TNF-α炎症因子分泌水平抑制率分别为31.60%、31.19%和31.09%(P<0.05)。结论骆驼刺提取物通过提高抗凋亡蛋白Bcl-2、Bcl-xl的表达水平,下调NLRP3炎症小体组成成分以及促炎因子IL-1β、IL-18和TNF-α分泌,从而抑制NLRP3炎症小体组装和激活,实现缓解LPS对IEC-6细胞的损伤。

6E-UDCA通过FABP7信号通路对MAFLD小鼠脂质代谢的影响研究

目的探讨6位乙基取代的熊去氧胆酸(6E-UDCA)通过重组人脂肪酸结合蛋白7(FABP7)信号通路对代谢相关脂肪性肝病(MAFLD)小鼠脂质代谢的影响。方法选取野生型(WT)、WT-1小鼠、脑FABP7特异性敲除(B-FABP7^(-/-))小鼠各6只作为3组,均采用高脂肪饲料喂养16周法构建MAFLD小鼠模型,其中WT-1组、B-FABP7^(-/-)组的饲料中加6E-UDCA(23.50 mg/kg),记录各组小鼠体重和摄食情况。第16周末小鼠在麻醉下剖腹并采集腹主动脉血,检测血清TC、TG、ALT、血糖、尿酸、游离长链脂肪酸(LCFA)水平;小鼠麻醉下失血死亡后取肝脏组织,采用HE染色观察肝脏组织病理学变化,分离小鼠肝脏并称取质量,计算肝脏系数。结果WT组小鼠肝脏组织中存在明显炎性细胞浸润情况,且有大量脂肪堆积,肝细胞排列紊乱;WT-1组、B-FABP7^(-/-)组小鼠肝脏组织中炎性细胞浸润减轻,轻微脂肪堆积,且肝细胞排列紊乱明显减轻。WT-1组和B-FABP7^(-/-)组小鼠体重、摄食量、肝脏质量、肝脏系数以及TC、TG、ALT、血糖、尿酸水平均明显低于WT组(均P<0.05),游离LCFA水平高于WT组(P<0.05);B-FABP7^(-/-)组小鼠体重、摄食量、肝脏质量、肝脏系数以及TC、TG、ALT、血糖、尿酸水平均明显低于WT-1组(均P<0.05),游离LCFA水平高于WT-1组(P<0.05)。结论6E-UDCA可通过调控FABP7信号通路,进而改善MAFLD小鼠脂质代谢,提高游离LCFA水平。

妊娠期糖尿病患者孕6~14周血清CTRP3、FGF19及SHBG的变化及预测价值

目的探讨妊娠期糖尿病(GDM)患者孕6~14周血清补体C1q/肿瘤坏死因子相关蛋白3(CTRP3)、成纤维细胞生长因子19(FGF19)及性激素结合球蛋白(SHBG)的变化及预测价值。方法回顾性分析2019年6月至2022年10月在秦皇岛市第一医院分娩的684例单胎妊娠孕妇的临床资料。统计GDM发生率,根据是否患有GDM分为病例组(n=266)和对照组(n=418)。比较两组CTRP3、FGF19及SHBG水平,并分析其联合预测GDM的价值,采用单因素和多因素Logistic回归分析影响GDM发生的危险因素。结果684例孕妇GDM发生率为38.89%(266/684)。与对照组比较,病例组CTRP3、FGF19及SHBG水平更低(P<0.05)。受试者工作特征(ROC)曲线结果显示,CTRP3、FGF19及SHBG联合预测GDM的曲线下面积(AUC)高于单项预测(P<0.05)。两组年龄、孕前体重指数(BMI)、不良孕产史、糖尿病家族史、妊娠期高血压、月经周期紊乱、CTRP3、FGF19、SHBG比较,差异具有统计学意义(P<0.05);两组孕次、产次比较,差异无统计学意义(P>0.05)。年龄≥35岁、孕前BMI≥24 kg/m^(2)、不良孕产史、糖尿病家族史、CTRP3<417.82 ng/L、FGF19<139.23 pg/mL、SHBG<429.59 mmol/L是影响GDM发生的独立危险因素(P<0.05)。结论GDM患者孕6~14周CTRP3、FGF19及SHBG水平均会下降,通过CTRP3、FGF19及SHBG联合预测GDM具有较高的应用价值。