Ligand Binding and Polymerization Characteristics of Human Milk Folate Binding Protein Depend on Concentration of Purified Protein and Presence of Amphiphatic Substances

Two folate binding proteins are present in human milk; one of 27 kDa is a cleavage product of the other one (100 kDa) which possesses a hydrophobic membrane anchor. A drastic change of radioligand binding characteristics and appearance of aggregated weak-radioligand affinity forms on gel filtration occurred at low concentrations of both proteins in the absence of Triton X-100 or other amphiphatic substances, e.g. cetyltrimethylammonium and phospholipids. These findings are consistent with a model predicting association between unliganded and liganded monomers resulting in weak-ligand affinity dimers. Amphiphatic substances form micelles and lipid bilayers which could separate hydrophobic unliganded monomers from hydrophilic liganded monomers (monomers become hydrophilic in the liganded state) thereby preventing association between these monomeric forms prevailing at low concentrations of the protein. Bio-Gel P-300 chromatography of the 27 kDa protein revealed a pronounced polymerization tendency, which diminished with decreasing protein concentrations, however, not in the presence of cetyltrimethylammonium. The data could have some bearings on observations indicating that naturally occurring amphiphatic substances, cholesterol and phospholipids, are necessary for the important clustering of membrane folate receptors.     

Stopped-Flow Kinetic Studies of the Interaction of Bovine Folate Binding Protein (FBP) and Folate

The kinetics of the interaction of bovine folate binding protein and folate at pH 7.4 and 5.0 were followed by measuring the changes of the intrinsic protein fluorescence intensity using the stopped-flow technique, which enables the study of reactions from the millisecond time-range. Our results immediately reject a simple one-step binding model, which requires a linear dependence of the observed rate constant on the concentration of the ligand. Thus, we are able to conclude that at pH 5.0 the interaction occurs in two steps and at pH 7.4 in three steps. Changes of fluorescence spectra at equilibrium were used to estimate the overall binding constants. Comparative studies on the binding of folate to human albumin are also reported.     

Ligand Binding Characteristics of a Glycosylphosphatidyl Inositol Membrane-Anchored HeLa Cell Folate Receptor Epitope-Related to Human Milk Folate Binding Protein

The folate receptor (FR) in HeLa cells was characterized as to ligandbinding mechanism, antigenic properties and membrane anchor in order toobtain information to be used for the design of biological agentstargeting FR in malignant tumors. The receptor displayed the followingbinding characteristics in equilibrium dialysis experiments(37°C, pH 7.4) with [³H] folate: a high-affinity type of bindingthat exhibited positive cooperativity with a Hill coefficient >1.0and an upward convex Scatchard plot, a slow radioligand dissociation atpH 7.4 becoming rapid at pH 3.5 and inhibition in the presence of otherfolates. The molecular size of the receptor was 100 kDa on gel filtrationwith Triton X-100, or similar to that of high molecular weight human milkfolate binding protein (FBP). The latter protein represents a 25 kDamolecule which equipped with a hydrophobic glycosylphosphatidylinositol (GPI) membrane anchor susceptible to cleavage byphosphatidylinositol specific phospholipase C (PI-PLC) formsmicelles of 100 kDa size with Triton X-100. The HeLa cell FRimmunoreacted with antibodies against purified human milk FBP inELISA, and in a fluorescence activated cell sorting system, whereHeLa cells exposed to increasing concentrations of antibody showed adose-dependent response. Exposure to PI-PLC decreased the fraction ofimmunolabeled cells indicating a linkage of FR to cell membranes by aGPI anchor. HeLa cells incubated with radiofolate showed a continuousuptake with time, however, with a complete suppression of uptake in thepresence of an excess of cold folate. Prewash of cells at acidic pH toremove endogenous folate increased the uptake. Binding and uptake of [³H]folate was increased in cells grown in a folate-deprived medium. The HeLaFR seems to be epitope related to human milk FBP.     

Ionic Charge, Hydrophobicity and Tryptophan Fluorescence of the Folate Binding Protein Isolated from Cow's Milk

A high-affinity folate binding protein was isolated and purified from cow's milk by a combination of cation exchange chromatography and methotrexate affinity chromatography. Chromatofocusing studies revealed that the protein possessed isoelectric points in the pH-interval 8–7. Polymers of the protein prevailing at pH values close to the isoelectric points seemed to be more hydrophobic than monomers present at pH 5.0 as evidenced by hydrophobic interaction chromatography and turbidity (absorbance at 340 nm) in aqueous buffer solutions (pH 5–8). Ligand binding seemed to induce a conformation change that decreased the hydrophobicity of the protein. In addition, Ligand binding quenched the tryptophan fluorescence of folate binding protein suggesting that tryptophan is present at the binding site and/or ligand binding induces a conformation change that affects tryptophan environment in the protein. There was a noticeable discordance between the ability of individual folate analogues to compete with folate for binding and the quenching effect.     

Folate Receptors in Malignant and Benign Tissues of Human Female Genital Tract

We have characterized the folate receptor in malignant and benign tissues of human female genital tract (Fallopian tube and benign and malignant tissues of uterus). Radioligand binding displayed characteristics similar to those of other folate binding proteins. Those include a high-affinity type of binding (K=10¹⁰M^–1), apparent positive cooperativity, a slow dissociation at pH 7.4 becoming rapid at pH 3.5, and inhibition of binding by folate analogues. The gel filtration profile of Triton X-100 solubilized tissue contained two large peaks of ³H-folate labelled protein (>=130 and 100kDa) as well as a 25 kDa peak. Only a single band of 70 kDa was seen on SDS-PAGE immunoblotting. The large molecular size forms on gel filtration appear to represent folate receptors having a hydrophobic membrane anchor inserted into Triton X-100 micelles. The folate receptor of female genital tract showed cross-reactivity in ELISA and positive immunostaining with rabbit antibodies against human milk folate binding protein. Variations in the ratio of immunoresponse to total high affinity folic acid binding suggests the presence of multiple isoforms of the receptor in different types of malignant and benign tissues.     

Preparation and Characterization of Monoclonal Antibodies to Serotonin Binding Protein

Serotonin binding protein (SBP) is a constituent of the synaptic vesicles of serotonergic neurons. Two types of SBP, with molecular masses of 45 kDa and 56 kDa, have been purified. To determine whether there are shared epitopes between the two forms of SBP, we raised and tested for cross-reactivity monoclonal antibodies (MAbs) against each form of SBP. We obtained 12 MAbs, all of which recognize both forms of SBP. Hybridoma clones were produced by fusing P3 X 63Ag8.653 mouse myeloma cells with spleen cells from a mouse that had been immunized with 45-kDa or 56-kDa SBP. Culture supernatants were screened for the presence of anti-SBP antibodies. MAb isotypes were determined by immunodiffusion, using immunoglobulin type-specific antisera. Each antibody to SBP consisted of only a single subclass of immunoglobulin (IgM). We obtained 12 MAbs, each of which interacted with both forms of SBP, as judged by enzyme-linked immunosorbent assay and immunoblot analysis. Ascites fluid to one clone (44-10) was obtained and affinity-purified. In the presence of goat anti-mouse IgM, the partially purified 44-10 antibodies quantitatively immunoprecipitated SBP from crude brain extracts. Immunoblotting revealed two major bands corresponding to 45 kDa and 56 kDa and a minor band corresponding to 68 kDa. MAb 44-10 blocked the binding of [3H]serotonin ([3H]5-HT) to 45-kDa and 56-kDa SBP in a concentration-dependent manner. The 68-kDa protein was found to bind [3H]5-HT. Sites reacting with MAB 44-10 were located immunocytochemically in sections of rat brain.(ABSTRACT TRUNCATED AT 250 WORDS)     

人脂多糖结合蛋白基因的克隆及序列测定

采用PCR技术,从人肝cDNA文库中扩增获得了1.5 kb的脂多糖结合蛋白（LBP）的全长基因.序列分析表明,克隆的LBP基因编码的氨基酸序列与文献报道相同.     

MicroRNA-141 在上皮性卵巢癌患者组织和血清中的表达 及临床意义的探讨

目的:探讨微小RNA-141(miR-141)在卵巢癌患者组织和血清中的表达情况并初步探讨其作为肿瘤标记物早期诊断上皮性卵巢癌的可行性。方法:采用实时荧光定量逆转录聚合酶链反应(real-time RT-PCR)检测16例上皮性卵巢癌患者和4例正常人卵巢组织及血清标本中miR-141的表达;检测4例良性卵巢肿瘤血清中miR-141的表达。结果:miR-141在卵巢癌患者组织中相对表达量分别为(72.846±76.671)显著高于正常人(2.869±3.201)(P<0.05);miR-141在卵巢癌患者血清中相对表达量(31.581±52.885)显著高于良性卵巢肿瘤患者(0.668±1.196)和正常人(1.690±1.697)(P<0.05),后两组间表达无差异(P>0.05);miR-141表达随卵巢癌临床分期的进展呈上升趋势(P<0.01),在无淋巴结转移组明显高于有淋巴结转移组(P<0.05),与组织分级和CA125的升高无关(P>0.05)。在组织中miR-141表达水平与上皮性卵巢癌临床病理特征均未见明显差异(P>0.05)。结论:miR-141可能在上皮性卵巢癌的发生发展中发挥癌基因的作用;miR-141用于检测上皮性卵巢癌敏感性和特异度较高,有望成为上皮性卵巢癌早期诊断的新指标。     

Expression of Serum Amyloid A in Human Ovarian Epithelial Tumors: Implication for a Role in Ovarian Tumorigenesis

Serum amyloid A (SAA) is an acute phase protein which is expressed primarily in the liver as a part of the systemic response to various injuries and inflammatory stimuli; its expression in ovarian tumors has not been described. Here, we investigated the expression of SAA in human benign and malignant ovarian epithelial tumors. Non-radioactive in situ hybridization applied on ovarian paraffin tissue sections revealed mostly negative SAA mRNA expression in normal surface epithelium. Expression was increased gradually as epithelial cells progressed through benign and borderline adenomas to primary and metastatic adenocarcinomas. Similar expression pattern of the SAA protein was observed by immunohistochemical staining. RT-PCR analysis confirmed the overexpression of the SAA1 and SAA4 genes in ovarian carcinomas compared with normal ovarian tissues. In addition, strong expression of SAA mRNA and protein was found in the ovarian carcinoma cell line OVCAR-3. Finally, patients with ovarian carcinoma had high SAA serum levels, which strongly correlated with high levels of CA-125 and C-reactive protein. Enhanced expression of SAA in ovarian carcinomas may play a role in ovarian tumorigenesis and may have therapeutic application. (J Histochem Cytochem 58:1015–1023, 2010)     

MicroRNA-141在上皮性卵巢癌患者组织和血清中的表达及临床意义的探讨

目的：探讨微小RNA-141（miR-141）在卵巢癌患者组织和血清中的表达情况并初步探讨其作为肿瘤标记物早期诊断上皮性卵巢癌的可行性。方法：采用实时荧光定量逆转录聚合酶链反应（real-timeRT—PCR）检测16例上皮性卵巢癌患者和4例正常人卵巢组织及血清标本中miR-141的表达;检测4例良性卵巢肿瘤血清中miR-141的表达。结果：miR-141在卵巢癌患者组织中相对表达量分别为（72．846±76．671）显著高于正常人（2．869±3．201）（P〈0．05）;miR-141在卵巢癌患者血清中相对表达量（31．581±52．885）显著高于良性卵巢肿瘤患者（0．668±1．196）和正常人（1．690±1．697）（P〈0．05）,后两组间表达无差异（P〉0．05）;miR-141表达随卵巢癌临床分期的进展呈上升趋势（P〈0．01）,在无淋巴结转移组明显高于有淋巴结转移组（P〈0．05）,与组织分级和CA125的升高无关（P〉0．05）。在组织中miR-141表达水平与上皮性卵巢癌临床病理特征均未见明显差异（P〉0．05）。结论：miR-141可能在上皮性卵巢癌的发生发展中发挥癌基因的作用;miR-141用于检测上皮性卵巢癌敏感性和特异度较高,有望成为上皮性卵巢癌早期诊断的新指标。     

Human 14-3-3 Protein: Radioimmunoassay, Tissue Distribution, and Cerebrospinal Fluid Levels in Patients with Neurological Disorders

Abstract: An antiserum to human 14-3-3 protein has been produced in rabbits. The protein was a poor antigen and attempts to improve immunogenicity were unsuccessful. A radioimmunoassay was developed using the antiserum, ¹²⁵I- 14-3-3-2, and unlabelled 14-3-3-2 as standards. The assay had a sensitivity limit of 2.5 ng.m1⁻¹. The minor component of human 14-3-3 protein (14-3-3-1 protein) cross-reacted to approximately 10% in the assay. Human tissues were surveyed for 14-3-3 protein by two-dimensional electrophoresis and by radioimmunoassay. Two-dimensional electrophoresis showed a 14-3-3 protein complex in brain, intestine, and testis, but not in other tissues. Radioimmunoassay showed that although brain had the highest concentration of 14-3-3 (13.3 μg. mg⁻¹ soluble protein), immunoreactivity was present in all tissues, with the concentration in intestine and testis approaching 50% of the brain level. Lower levels (less than 1.0 μg. mg⁻¹ soluble protein) were seen in liver, kidney, skeletal muscle, and erythrocytes. The immunoreactivity present in tissues other than brain showed the same molecular weight and charge characteristics as authentic 14-3-3 protein. The radioimmunoassay also detected 14-3-3 protein in serum (50 ng.m1⁻¹) and in CSF (5-130 ng.ml⁻¹). The immunoreactivity present in CSF appeared to be intact 14-3-3 protein. CSF 14-3-3 levels were measured in 82 patients with various neurological disorders. Measurements of this protein did not appear sufficiently discriminating to be o f diagnostic value.     

人附睾蛋白4与血清CA125联合检测在卵巢癌诊断中的应用价值

目的：探讨人附睾蛋白4（HE4）和CA125联合检测在上皮性卵巢癌临床诊断中的应用价值。方法：用化学发光法测定51例卵巢癌患者、64例卵巢良性疾病患者和50例健康对照者血清中的HE4和CA125水平,并对结果进行比较分析。结果：卵巢癌组血清中HE4和CA125水平均显著高于卵巢良性肿瘤组和健康对照组,差异有统计学意义（P〈0．05）,卵巢良性肿瘤组与健康对照组比较无显著性差异;HE4在早期卵巢癌中的阳性率高于CA125,两者联合检测可提高卯巢癌诊断的阳性率;卵巢癌组联合检测ROC曲线下面积大于单项检测曲线下面积。结论：联合检测HE4和CA125有助于卵巢癌的早期诊断。     

血清钙结合蛋白S100A12、脂联素及IL-17与稳定期COPD患者严重程度及肺功能的相关性

摘要 目的：探讨血清钙结合蛋白S100A12、脂联素及白细胞介素-17（Interleukin-17,IL-17）水平与稳定期慢性阻塞性肺疾病（COPD）病情严重程度和肺功能的关系。方法：选择稳定期COPD患者（稳定期COPD组）和健康体检者（对照组）各80例,稳定期COPD患者分为四级：I级轻度（22例）,Ⅱ级中度（24例）,Ⅲ级重度（19例）,Ⅳ级极重度（15例）。使用肺功能仪对所有研究对象的肺功能进行检查,采用血气分析仪检测动脉血中的氧分压 (PaO₂) 和二氧化碳分压 (PaCO₂),采用酶联免疫吸附试验检测血清中钙结合蛋白S100A12、脂联素和IL-17水平,采用 Pearson相关分析各指标之间的相关性。结果：稳定期COPD组肺功能[第一秒用力呼出气量容积占预计值百分比（FEV₁%）和第一秒用力呼出气量容积/用力肺活量（FEV1/FVC%）]和PaO₂显著低于对照组（P<0.05）,并随着COPD严重程度增加而降低（P<0.05）;稳定期COPD组血清钙结合蛋白S100A12、脂联素、IL-17水平和PaCO₂均高于对照组（P<0.05）,并随COPD严重程度增加而升高（P <0.05）。Pearson相关分析显示：稳定期COPD患者血清钙结合蛋白S100A12、脂联素和IL-17水平与FEV₁%、FEV₁/FVC%和PaO₂呈负相关（P<0.05）,与PaCO₂呈正相关（P<0.05）。结论：钙结合蛋白S100A12、脂联素和IL-17可能共同参与了稳定期COPD慢性炎症过程,引起气流受限,影响肺通气功能,可以辅助评估稳定期COPD病情的严重程度。     

人心型脂肪酸结合蛋白(H-FABP)的原核表达、纯化和冻干品的制备

目的：获得重组人心型脂肪酸结合蛋白,分析其活性及制备冻干品。方法：从GenBank中检索人源H-FABP CDs 序列,合成基因后构建原核表达载体pET-28a（+）-H-FABP。将表达载体转入E.coli BL21（DE3）,摸索pET-28a（+）-H-FABP最佳诱导条件,表达并纯化重组蛋白。使用检测试剂检测纯化后的重组H-FABP活性,研究最佳冻干方案并考察冻干品的稳定性。结果：经过优化诱导条件,重组H-FABP在BL21（DE3）中以可溶性蛋白形式表达。诱导条件为OD₆₀₀≈0.6,IPTG终浓度0.4mmol/L,30℃诱导4h。通过Ni²⁺亲和层析纯化可得到纯度大于95%的重组H-FABP蛋白。重组H-FABP冻干品可以在37℃稳定保存12d,25℃、4℃稳定保存至少4个月。结论：本项研究中的重组H-FABP表达体系成熟、蛋白活性高,冻干品稳定性好,为后续研究提供了稳定高效的生物原材料。     

胆汁三烯结合蛋白在大肠杆菌中的表达、复性与纯化

目的：合成胆汁三烯结合蛋白（BBP）基因并在大肠杆菌中表达,获得重组BBP纯化制品。方法：根据天然BBP的基因序列和大肠杆菌偏好密码子设计并合成BBP基因的引物,PCR扩增优化的BBP基因序列,克隆至载体pEasy-T3;测序正确后,将该序列克隆至表达载体pET-32a上,构建表达质粒,转化至大肠杆菌BL21（DE3）pLysS,在IPTG诱导下表达融合蛋白;采用Ni柱纯化融合蛋白。结果：PCR扩增获得了优化后的BBP基因序列,构建了表达载体pET-32a-BBP;SDS-PAGE分析表明表达的融合蛋白相对分子质量为20×10^3,以包涵体形式存在,占全菌蛋白的40%以上;变性、复性后经Ni2＋柱纯化,获得纯度达98%以上的重组蛋白。结论：优化并合成了BBP全基因序列,获得了高纯度重组融合蛋白,为进一步鉴定其生物活性及筛选小分子的研究奠定了基础。     

曲美他嗪人血浆蛋白结合率的测定

目的:建立测定曲美他嗪与人血浆蛋白结合率的高效液相色谱法(HPLC),测定曲美他嗪在人血浆中的蛋白结合率。方法:采用平衡透析法,结合HPLC测定曲美他嗪在人血浆中的蛋白结合率。结果:低、中、高3个浓度下,曲美他嗪的血浆蛋白结合率分别为14.7%,16.2%和15.5%。结论:本方法灵敏度高,重现性好,操作简单,能满足生物样品分析的要求。曲美他嗪与血浆蛋白结合率较低。     

Comparison of Antibodies Hydrolyzing Myelin Basic Protein from the Cerebrospinal Fluid and Serum of Patients with Multiple Sclerosis

It was found that antibodies (Abs) against myelin basic protein (MBP) are the major components of the antibody response in multiple sclerosis (MS) patients. We have recently shown that IgGs from sera of MS patients are active in the hydrolysis of MBP. However, in literature there are no available data concerning possible MBP-hydrolyzing Abs in cerebrospinal fluid (CSF) of MS patients. We have shown that the average content of IgGs in their sera is about 195-fold higher than that in their CSF. Here we have compared, for the first time, the average content of lambda- and kappa-IgGs as well as IgGs of four different subclasses (IgG1-IgG4) in CSF and sera of MS patients. The average relative content of lambda-IgGs and kappa –IgGs in the case of CSFs (8.0 and 92.0%) and sera (12.3 and 87.7%) are comparable, while IgG1, IgG2, IgG3, and IgG4: CSF - 40.4, 49.0, 8.2, and 2.5% of total IgGs, respectively and the sera - 53.6, 36.0, 5.6, and 4.8%, decreased in different order. Electrophoretically and immunologically homogeneous IgGs were obtained by sequential affinity chromatography of the CSF proteins on protein G-Sepharose and FPLC gel filtration. We present first evidence showing that IgGs from CSF efficiently hydrolyze MBP and that their average specific catalytic activity is unpredictably ∼54-fold higher than that of Abs from sera of the same MS patients. Some possible reasons of these findings are discussed. We suggest that anti-MBP abzymes of CSF may promote important neuropathologic mechanisms in this chronic inflammatory disorder and in MS pathogenesis development.     

特发性肺纤维化患者血清血管生成素-2、潜在转化生长因子结合蛋白2、趋化因子配体14水平与预后的关系研究

摘要 目的：研究特发性肺纤维化（IPF）患者血清血管生成素-2（Ang-2）、潜在转化生长因子结合蛋白2（LTBP2）、趋化因子配体14（CXCL14）水平与预后的关系。方法：选取2019年3月～2020年3月河北医科大学第二医院东院区呼吸内科收治的100例IPF患者作为研究对象。将其按照随访结局分成预后不良组59例及预后良好组41例。比较两组Ang-2、LTBP2、CXCL14水平、肺功能指标、实验室指标以及一般资料,Pearson相关分析血清Ang-2、LTBP2、CXCL14水平与肺功能指标的相关性,多因素Logistic回归分析IPF患者预后的影响因素,受试者工作特征（ROC）曲线分析Ang-2、LTBP2、CXCL14对IPF患者预后的预测价值。结果：预后不良组血清Ang-2、LTBP2、CXCL14水平均高于预后良好组（P＜0.05）。预后不良组第1s用力呼气容积（FVC₁）、一氧化碳弥散量占预计值百分比（DLCO%）、动脉血氧分压（PaO₂）/氧体积分数（FiO₂）均低于预后良好组,而吸烟史、肺动脉高压人数占比高于预后不良组（P＜0.05）。Pearson相关性分析显示：IPF患者Ang-2、LTBP2、CXCL14水平均和FVC₁、DLCO%呈负相关（P＜0.05）。多因素Logistic回归分析结果显示：FVC₁、DLCO%、PaO₂/FiO₂、吸烟史、肺动脉高压以及血清Ang-2、LTBP2、CXCL14水平是IPF患者预后不良的影响因素（P＜0.05）。ROC曲线分析结果显示：血清Ang-2、LTBP2、CXCL14三项联合检测预测IPF患者预后的曲线下面积高于上述三项指标单独检测。结论：IPF患者血清Ang-2、LTBP2、CXCL14水平与其预后密切相关,可作为评估其预后的辅助性指标。     

兔抗人热激蛋白70样蛋白1多克隆抗体的制备与初步鉴定

目的：制备兔抗人热激蛋白70样蛋白1（HSP70L1）的多克隆抗体并进行初步鉴定。方法：在大肠杆菌中重组表达融合蛋白GST-HSP70L1和His-HSP70L1并纯化;将GST-HSP70L1融合蛋白用于免疫新西兰大耳白兔获得多克隆抗体,用His-HSP70L1对抗血清进行分离纯化,得到抗HSP70L1多克隆抗体,用Western印迹、免疫沉淀对其进行初步鉴定。结果：获得了高表达的GST-HSP70L1和His-HSP70L1重组融合蛋白;纯化获得抗HSP70L1抗体,此抗体可用于Western印迹和免疫沉淀实验。结论：获得了兔抗人HSP70L1的多克隆抗体,为进一步研究HSP70L1的生物功能提供了有用的工具。     

2型糖尿病患者血清三叶因子3、磷酸酪氨酸衔接蛋白1水平与肾功能及炎症反应的关系研究

目的:探讨2型糖尿病(T2DM)患者血清三叶因子3(TFF3)、磷酸酪氨酸衔接蛋白1(APPL1)水平,并分析其与肾功能及炎症反应之间的关系。方法:选择2017年1月至2019年1月期间我院收治的T2DM患者168例,根据尿白蛋白/肌酐比率(UACR)将患者分为正常白蛋白尿组(UACR30 mg/g,n=79)、微量白蛋白尿组(30 mg/g≤UACR≤300 mg/g,n=65)和大量白蛋白尿组(UACR300 mg/g,n=24),另选择同期行体检的健康志愿者57例作为对照组。对比四组受试者血清TFF3、APPL1水平、肾功能指标[血尿素(UREA)、血肌酐(Scr)、胱抑素C(Cys C)和肾小球滤过率(GFR)]及炎症指标[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)及单核细胞趋化因子1(MCP-1)]变化,Pearson相关分析血清TFF3、APPL1水平与肾功能指标及炎症因子的相关性。结果:四组受试者血清TFF3、APPL1、肾功能指标、TNF-α、IL-6及MCP-1之间的差异具有统计学意义(P0.05);与对照组相比,T2DM患者血清TFF3、APPL1、UREA、Scr、Cys C、TNF-α、IL-6及MCP-1水平均明显升高,而GFR显著下降,且差异均具有统计学意义(PP0.05);不同UACR水平的T2DM患者血清TFF3、APPL1、肾功能指标、TNF-α、IL-6及MCP-1之间的差异具有统计学意义(P0.05);Pearson相关分析结果显示,T2DM患者血清TFF3、APPL1分别与UREA、Scr、Cys C、TNF-α、IL-6、MCP-1呈正相关(P0.05),与GFR呈负相关(P0.05),但是与IL-10无相关性(P0.05)。结论:血清TFF3和APPL1可能通过影响肾功能及炎症反应而影响T2DM的发生及发展,可能作为T2DM临床诊断的生物学指标,为T2DM的治疗提供新靶点。