Evaluating new biomarkers for diabetic nephropathy:Role ofα2-macroglobulin,podocalyxin,α-L-fucosidase,retinol-binding protein-4,and cystatin C

BACKGROUND The intricate relationship between type 2 diabetes mellitus(T2DM)and diabetic nephropathy(DN)presents a challenge in understanding the significance of various biomarkers in diagnosis.AIM To elucidate the roles and diagnostic values ofα2-macroglobulin(α2-MG),podocalyxin(PCX),α-L-fucosidase(AFU),retinol-binding protein-4(RBP-4),and cystatin C(CysC)in DN.METHODS From December 2018 to December 2020,203 T2DM patients were enrolled in the study.Of these,115 were diagnosed with DN(115 patients),while the remaining 88 patients were classified as non-DN.The urinary levels ofα2-MG,PCX,and AFU and the serum concentrations RBP-4 and CysC were measured in conjunction with other relevant clinical indicators to evaluate their potential correlations and diagnostic utility.RESULTS After adjustments for age and gender,significant positive correlations were observed between the biomarkers CysC,RBP-4,α2-MG/urinary creatinine(UCr),PCX/UCr,and AFU/UCr,and clinical indicators such as urinary albumin-to-creatinine ratio(UACR),serum creatinine,urea,24-h total urine protein,and neutrophil-to-lymphocyte ratio(NLR).Conversely,these biomarkers exhibited negative correlations with the estimated glomerular filtration rate(P<0.05).Receiver operating characteristic(ROC)curve analysis further demonstrated the diagnostic performance of these biomarkers,with UACR showcasing the highest area under the ROC curve(AUC^(ROC))at 0.97.CONCLUSION This study underscores the diagnostic significance ofα2-MG,PCX,and AFU in the development of DN.The biomarkers RBP-4,CysC,PCX,AFU,andα2-MG provide promising diagnostic insights,while UACR is the most potent diagnostic biomarker in assessing DN.

Retinol binding protein 4 correlates with and is an early predictor of carotid atherosclerosis in type 2 diabetes mellitus patients

The association of retinol binding protein 4 （RBP4） with atherosclerosis of the carotid artery in type 2 diabetes mellitus （T2DM） remains undefined. We aimed to investigate the correlation of RBP4 expression with atherosclerosis of the carotid artery in T2DM. A total of 1,076 subjects were investigated for intima-media thickness of the bilateral common carotid arteries, and they were divided into three groups： in group Ⅰ, patients had normal neck vascular ultra- sound, in group Ⅱ, intimal carotid artery media thickness was equal to or more than 1 mm, and in group Ⅲ, carotid artery plaque was present. Height, weight, blood pressure （BP）, fasting plasma glucose （FPG）, hemoglobin Alc （HbA1c）, total cholesterol （TC）, triglyceride （TG）, low-density lipoprotein cholesterol （LDL-C）, high-density lipopro- tein cholesterol （HDL-C）, apolipoprotein A-1 （apoA-1）, apolipoprotein B （apoB） and lipoprotein （a） [Lp（a）] were determined by routine laboratory methods. RBP4 and high sensitivity C reactive protein （HsCRP） were measured by an enzyme-linked immuno-sorbent assay, and insulin concentration was measured by an electrochemiluminescence sandwich immunoassay. Duration of diabetes, waist and BP, FPG, HbAlc, TG, TC, LDL-C, APOB, Lp（a）, HsCRP, RBP4 and homeostasis model assessment insulin resistance index （HOMA-IR） were significantly lower in group I than in the other two groups （P〈0.01, P〈0.01）. Plasma levels of HbAlc, RBP4, LDL-C, TC, HOMA-IR, HsCRP and Lp（a）, waist and BP were significantly increased in group III than in group II （P〈0.01）. Multivariate logistic regression analysis showed that there were seven factors associated with the occurrence of carotid artery atherosclero- sis and its risks in descending order were： high LDL-C, high waist, high HsCRP, duration of diabetes, high HOMA-IR, HbAlc and high RBP4. Our finding supported that RBP4 was positively correlated with carotid atherosclerosis in patients with T2DM and could be used as an early predictor of cardiovascular disease.

Elevated retinol binding protein 4 levels are associated with atherosclerosis in diabetic rats via JAK2/STAT3 signaling pathway

BACKGROUND Atherosclerosis is a major cause of mortality worldwide and is driven by multiple risk factors,including diabetes,which results in an increased atherosclerotic burden,but the precise mechanisms for the occurrence and development of diabetic atheroscerosis have not been fully elucidated.AIM To summarize the potential role of retinol binding protein 4(RBP4) in the pathogenesis of diabetic atheroscerosis,particularly in relation to the RBP4-Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway.METHODS Male Wistar rats were randomly divided into three groups,including a control group(NC group),diabetic rat group(DM group),and diabetic atherosclerotic rat group(DA group).The contents of total cholesterol(TC), high-density lipoprotein cholesterol(HDL-c), triglycerides(TG), low-density lipoprotein cholesterol(LDLc), fasting insulin(FINS),fasting plasma glucose,and hemoglobin A1 c(HbA1 c)were measured.Moreover,the adipose and serum levels of RBP4,along with the expression levels of JAK2, phosphorylated JAK2(p-JAK2), STAT3,phosphorylated STAT3(p-STAT3), B-cell lymphoma-2(Bcl-2), and Cyclin D1 in aortic tissues were also measured.Besides,homeostasis model assessment of insulin resistance(HOMA-IR) and atherogenic indexes(AI) were calculated.RESULTS Compared with the NC and DM groups,the levels LDL-c,TG,TC,FINS,HOMAIR,RBP4,and AI were upregulated,whereas that of HDL-c was downregulated in the DA group(P <0.05);the mRNA levels of JAK2,STAT3,Cyclin D1,and Bcl-2 in the DA group were significantly increased compared with the NC group and the DM group;P-JAK2,p-JAK2/JAK2 ratio,p-STAT3,p-STAT3/STAT3 ratio,Cyclin D1,and Bcl-2 at protein levels were significantly upregulated in the DA group compared with the NC group and DM group.In addition,as shown by Pearson analysis,serum RBP4 had a positive correlation with TG,TC,LDL-c,FINS,HbA1 C,p-JAK2,p-STAT3,Bcl-2,Cyclin D1,AI,and HOMA-IR but a negative correlation with HDL-c.In addition,multivariable logistic regression analysis showed that serum RBP4,p-JAK2,p-STAT3,and LDL-c were predictors of the presence of diabetic atherosclerosis.CONCLUSION RBP4 could be involved in the initiation or progression of diabetic atherosclerosis by regulating the JAK2/STAT3 signaling pathway.

Retinoblastoma binding protein 4 up-regulation is correlated with hepatic metastasis and poor prognosis in colon cancer patients

Background: Retinoblastoma binding protein 4 (RBBP4) plays an essential role in the development of multiple cancers. However, its relationship with prognosis in colon cancer and colon cancer hepatic metastasis has not been elucidated. The aim of this study was to explore the relationship between RBBP4 expression and prognosis of colon cancer patients and to evaluate RBBP4 as a new prognostic marker in these patients. Methods: Eighty colon cancer patients underwent surgical resection of the colon were enrolled. Among them, forty colon cancer patients suffered with hepatic metastasis. The colon cancer tissues, para-colon cancer tissues, and hepatic metastatic cancer tissues were collected from the pathological department for further analysis. The expression of RBBP4 proteins was examined by immunohistochemistry and correlated with clinicopathological parameters. The Cancer Genome Atlas (TCGA) database was used to validate the expression and explore its relationship with clinical characteristics. Results: RBBP4 was up-regulated in the colon cancer tissues compared with the para-colon cancer tissues. The analysis of TCGA database verified the upregulation of RBBP4 in the colon cancer tissues and RBBP4 overexpression was correlated with nerve invasion and poor outcomes of chemotherapy. Moreover, the positive rate of RBBP4 expression in 40 colon cancer patients with hepatic metastasis was higher in the hepatic metastatic cancer tissues (39/40, 97.5%) than in the colon cancer tissues (26/40, 65.0%). Our clinicopathological analysis showed that RBBP4 expression was significantly correlated with vascular invasion, hepatic metastasis, and lymph node involvement (all P < 0.05). Additionally, the survival analysis demonstrated that RBBP4 over-expression was correlated with poor prognosis. Conclusions: RBBP4 was upregulated in the colon cancer. RBBP4 may be a novel predictor for poor prognosis of colon cancer and colon cancer hepatic metastasis.

不稳定型心绞痛患者血清RBP-4、Lp-PLA2水平及依折麦布治疗影响研究

目的探究不稳定型心绞痛(UA)患者血清视黄醇结合蛋白-4(RBP-4)、脂蛋白相关磷脂酶A2(Lp-PLA2)水平及依折麦布治疗影响。方法选取2021年1月至2023年1月于唐山市人民医院心内科确诊的UA患者84例为研究对象,其中男性48例,女性36例;根据冠状动脉病变程度分为:Gensini评分<20分为轻度组(27例)、20分≤Gensini评分<40分为中度组(36例)和Gensini评分≥40分为重度组(21例);按随机数字表法分为观察组和对照组,每组各42例。所有患者均行常规治疗,对照组在此基础上给予瑞舒伐他汀治疗,观察组给予瑞舒伐他汀+依折麦布治疗,两组患者均连续治疗6个月。比较不同冠状动脉病变程度组RBP-4、Lp-PLA2水平;比较治疗后观察组和对照组脂代谢、RBP-4、Lp-PLA2水平以及不良心血管事件的发生情况。结果①血清RBP-4、Lp-PLA2水平为:轻度组<中度组<重度组,差异有统计学意义(P<0.05)。斯皮尔曼等级相关分析显示,血清RBP-4、Lp-PLA2水平与UA患者冠状动脉病变程度呈正相关(r=0.844、r=0.729,P<0.05)。②重复测量方差分析显示,血脂水平时点、组间、交互比较差异有统计学意义(P<0.05)。治疗前,两组总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、三酰甘油(TG)水平的比较,差异无统计学意义(P>0.05);治疗3个月、6个月,两组TC、TG、LDL-C的水平均降低(P<0.05),而HDL-C升高(P<0.05),且观察组TC、TG、LDL-C水平低于对照组,HDL-C水平高于对照组,差异有统计学意义(P<0.05)。③重复测量方差分析显示,RBP-4、Lp-PLA2水平时点、组间、交互比较,差异有统计学意义(P<0.05)。治疗前,两组RBP-4、Lp-PLA2水平比较,差异无统计学意义(P>0.05);治疗3个月、6个月,两组RBP-4、Lp-PLA2水平均降低(P<0.05),且观察组RBP-4、Lp-PLA2水平低于对照组,差异有统计学意义(P<0.05)。④观察组心血管事件发生率(7.14%)低于对照组(23.81%),差异有统计学意义(P<0.05)。结论UA患者血清RBP-4、Lp-PLA2水平与冠状动脉病变程度相关,依折麦布可改善不稳定型心绞痛患者的脂代谢水平,调节血清RBP-4、Lp-PLA2水平,减少心血管事件发生。

血清TyG指数和nesfatin-1及RBP4联合预测糖尿病视网膜病变的价值

目的:探究血清甘油三酯-葡萄糖(TyG)指数、摄食抑制因子-1(nesfatin-1)、视黄醇结合蛋白4(RBP4)联合预测糖尿病视网膜病变(DR)的价值,为DR早期预测提供支持。方法:回顾性分析。收集2022-02/2023-12我院接诊的2型糖尿病(T2DM)患者164例的临床资料,按照眼底检查结果分为DR组43例(其中增殖性DR 19例,非增殖性DR 24例),不合并DR的T2DM组121例。入院后记录患者基本资料,检查血清TyG指数、nesfatin-1、RBP4水平。结果:DR组病程长于T2DM组,空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、低密度脂蛋白及TyG指数、RBP4水平高于T2DM组,高密度脂蛋白、nesfatin-1水平低于T2DM组(均P<0.001)。多因素Logistic回归分析可知,T2DM病程(OR=1.338,95%CI:1.059-1.690)、糖化血红蛋白(OR=5.065,95%CI:1.659-15.470)、低密度脂蛋白(OR=12.715,95%CI:2.385-67.790)、TyG指数(OR=23.057,95%CI:2.936-181.073)、RBP4(OR=1.319,95%CI:1.028-1.692)是T2DM患者发生DR的危险因素,nesfatin-1(OR=0.007,95%CI:0.003-0.016)为保护因素。绘制ROC曲线显示,TyG指数、nesfatin-1、RBP4均对T2DM患者并发DR具有一定预测价值,曲线下面积(areas under curve,AUC)分别为0.804、0.878、0.738,各指标联合预测时AUC为0.946,预测敏感度为83.72%、特异度为92.56%。增殖性DR患者TyG指数、RBP4水平高于非增殖性DR患者,nesfatin-1水平低于非增殖性DR患者(均P<0.05)。Spearman相关性分析显示,TyG指数、RBP4水平与DR病情程度呈正相关,nesfatin-1水平与DR病情程度呈负相关(r_(s)=0.557、0.392、-0.359,均P<0.05)。Pearson相关分析显示,T2DM并发DR患者TyG指数与nesfatin-1水平呈负相关,与RBP4水平呈正相关,nesfatin-1与RBP4水平呈负相关(r=-0.486、0.538、-0.592,均P<0.05)。结论:血清TyG指数、nesfatin-1、RBP4水平与DR发病风险及病情程度有关,可作为DR早期预测的标志物,且联合预测效能更好。

Influence of Glyphaea brevis twig extract on nucleus, tight junctions and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells

Objective: To examine the influence of Glyphaea (G.) brevis twig extract on the mitochondrial dehydrogenase activity, integrity of the tight junctions between adjacent cells, mitochondria, apoptosis, nucleus and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells. Methods: TM4 cell line was used in this study as it exhibited properties similar to the Sertoli cells. TM4 Sertoli cells were exposed to G. brevis twig extract (0.1, 1.0, 10.0, 100.0, or 1000.0μg/mL) for 24, 48 and 72 h. Parameters studied included cell viability [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], mitochondrial membrane potential (tetra methyl rhodamine ethyl ester dye), transepithelial electrical resistance, apoptosis (Annexin V Alexa Fluor?488/propidium iodide assay) and mRNA expression (quantitative reverse transcription polymerase chain reaction). Results: G. brevis twig extract had no cytotoxic impact on cell viability, thus, considerably increasing the activity of mitochondrial dehydrogenase enzyme after 24 and 72 h exposure. Transepithelial electrical resistance values revealed substantial (P<0.05) rise in treated groups, especially after 72 h of treatment. Moreover, there was a significant decrease in mitochondrial depolarization of TM4 Sertoli cells exposed to G. brevis twig extract when compared to controls. In addition, G. brevis twig extract significantly reduced necrosis and apoptosis of TM4 Sertoli cells when compared to control. Nevertheless, fluorescence microscopy revealed that the nuclei were egg-shaped and marked uniformly with consistent cell shape at the middle of the TM4 Sertoli cells. Significant stimulatory effects were observed on mRNA levels of inhibin-β, androgen binding protein and stem cell factor. Conclusions: G. brevis twig extract may increase the secretory roles of TM4 Sertoli cells, cells proliferation, as well as cell-cell tight junction integrity. Thus, G. brevis twig may enhance spermatogenesis.

Overexpression of GATA binding protein 4 and myocyte enhancer factor 2C induces differentiation of mesenchymal stem cells into cardiac-like cells

BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as a new area to support heart regeneration.The transcription factors GATA binding protein 4(GATA-4)and myocyte enhancer factor 2C(MEF2C)are considered prominent factors in the development of the cardiovascular system.AIM To explore the potential of GATA-4 and MEF2C for the cardiac differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs).METHODS hUC-MSCs were characterized morphologically and immunologically by the presence of specific markers of MSCs via immunocytochemistry and flow cytometry,and by their potential to differentiate into osteocytes and adipocytes.hUC-MSCs were transfected with GATA-4,MEF2C,and their combination to direct the differentiation.Cardiac differentiation was confirmed by semiquant itative real-time polymerase chain reaction and immunocytochemistry.RESULTS hUC-MSCs expressed specific cell surface markers CD105,CD90,CD44,and vimentin but lack the expression of CD45.The transcription factors GATA-4 and MEF2C,and their combination induced differentiation in hUC-MSCs with significant expression of cardiac genes i.e.,GATA-4,MEF2C,NK2 homeobox 5(NKX2.5),MHC,and connexin-43,and cardiac proteins GATA-4,NKX2.5,cardiac troponin T,and connexin-43.CONCLUSION Transfection with GATA-4,MEF2C,and their combination effectively induces cardiac differentiation in hUC-MSCs.These genetically modified MSCs could be a promising treatment option for heart diseases in the future.

子宫腺肌病IVF-ET助孕病人血清PIBF、RBP-4水平及其对妊娠结局的预测价值

目的:探讨子宫腺肌病体外受精-胚胎移植(IVF-ET)助孕病人血清孕激素诱导阻断因子(PIBF)、视黄醇结合蛋白-4(RBP-4)的表达及其对妊娠结局的预测价值。方法:选取行IVF-ET助孕的子宫腺肌病病人96例为研究对象,根据临床妊娠情况将研究对象分为临床妊娠组(56例)和未妊娠组(40例)。比较2组病人基本特征及治疗数据、血清PIBF、RBP-4水平。Pearson法分析血清PIBF、RBP-4与基础卵泡刺激素(FSH)、抗缪勒管激素(AMH)、基础促黄体生成素(LH)的相关性。多因素logistic回归分析影响子宫腺肌病IVF-ET助孕病人临床妊娠结局的影响因素。受试者工作特征(ROC)曲线分析血清PIBF、RBP-4水平对子宫腺肌病IVF-ET助孕病人妊娠结局的预测价值。结果:临床妊娠组FSH、AMH、促性腺激素(Gn)用量明显高于未妊娠组(P<0.05~P<0.01)。未妊娠组血清PIBF水平明显低于临床妊娠组(P<0.01),RBP-4水平明显高于临床妊娠组(P<0.01)。相关性分析显示,子宫腺肌病病人血清PIBF与基础FSH、AMH呈正相关(r=0.517、0.669,P<0.01),血清RBP-4与基础FSH、AMH呈负相关(r=-0.558、-0.634,P<0.01)。多因素logistic回归分析显示,基础FSH低水平、AMH低水平、PIBF低水平、RBP-4高水平是影响子宫腺肌病IVF-ET助孕病人临床妊娠结局的危险因素(P<0.05~P<0.01)。血清PIBF、RBP-4联合预测子宫腺肌病IVF-ET助孕病人妊娠结局的ROC曲线下面积(AUC)显著大于PIBF、RBP-4单独预测的AUC(P<0.05)。结论:血清PIBF、RBP-4与子宫腺肌病IVF-ET助孕病人妊娠结局相关,两者联合对子宫腺肌病IVF-ET助孕病人妊娠结局具有一定的预测价值。

肝特异性Rbp4基因敲除小鼠的建立及糖代谢特征分析

目的 建立肝特异性Rbp4基因敲除小鼠模型,并初步探索肝特异性Rbp4基因缺失对糖代谢的影响。方法 利用Cre-LoxP技术,使用C57/BL6J小鼠和Alb-Cre小鼠构建肝特异性Rbp4基因敲除小鼠模型。利用PCR及琼脂糖凝胶电泳鉴定小鼠基因型。选取10只18周龄C57/BL6J雄性小鼠为野生对照组(WT),10只同周龄flox纯合且Alb-Cre阴性小鼠为实验对照组(Rbp4~(flox/flox):Cre~-),10只同周龄flox纯合且Alb-Cre阳性小鼠为实验组(Rbp4~(flox/flox):Cre~+)。分别利用Western Blot及qRT-PCR验证小鼠肝中RBP4蛋白及Rbp4 mRNA表达水平。利用qRT-PCR检测其他组织中Rbp4 mRNA表达水平。采用苏木素-伊红(HE)染色观察肝组织形态。利用血糖仪检测小鼠尾静脉血液标本血糖值,进行葡萄糖耐量及胰岛素耐量实验。利用qRT-PCR检测肝糖代谢基因磷酸烯醇丙酮酸羧化酶(Pepck)和葡萄糖-6-磷酸酶(G6pase)表达水平。结果 成功繁育并鉴定出肝特异性Rbp4基因敲除小鼠。Rbp4~(flox/flox):Cre~+组小鼠肝中RBP4蛋白表达显著减少(P<0.05),Rbp4 mRNA表达显著减少(P<0.05)。三组小鼠脂肪、肾、胰、脾、心脏和肌肉组织中Rbp4 mRNA的相对表达量差异无显著性(P>0.05)。HE染色、葡萄糖耐量及胰岛素耐量实验结果表明肝特异性Rbp4基因敲除对肝组织形态、葡萄糖耐量及胰岛素耐量无显著影响(P>0.05)。三组小鼠肝中Pepck mRNA表达差异具有显著性(P<0.05),两两比较显示,Rbp4~(flox/flox):Cre~+组小鼠肝中Pepck mRNA相对表达量较Rbp4~(flox/flox):Cre~-组小鼠降低(P<0.05)。三组小鼠肝中G6pase mRNA表达差异无显著性(P>0.05)。结论 成功构建了肝特异性Rbp4基因敲除小鼠模型,基因缺失可抑制小鼠肝Pepck mRNA表达,为进一步探索该基因在小鼠糖代谢中的作用提供依据。

妊娠合并甲状腺功能减退患者血清PAPP-A、RBP-4水平及其对妊娠结局的预测价值

目的探讨血清妊娠相关血浆蛋白A(PAPP-A)和视黄醇结合蛋白4(RBP-4)水平对甲状腺功能减退(简称甲减)患者妊娠结局的预测价值。方法回顾性选取2021年6月至2023年6月在该院产检的149例妊娠合并甲减患者作为观察组,另选取同期在该院产检的50例产检正常的健康孕妇作为对照组。对观察组患者进行随访,根据患者妊娠结局划分为结局良好组和结局不良组;采用酶联免疫吸附试验检测各组甲状腺激素指标和血清PAPP-A、RBP-4水平;采用Pearson相关分析妊娠合并甲减患者血清PAPP-A与RBP-4水平间的相关性;采用多因素Logistic回归分析妊娠合并甲减患者妊娠结局不良的影响因素;采用受试者工作特征(ROC)曲线分析PAPP-A、RBP-4血清水平对妊娠合并甲减患者妊娠结局不良的预测价值。结果与对照组比较,观察组患者PAPP-A、游离三碘甲状腺原氨酸(FT3)和游离甲状腺素(FT4)水平明显降低(P<0.05),RBP-4和促甲状腺素(TSH)水平明显升高(P<0.05)。根据观察组患者妊娠结局分为结局良好组96例,结局不良组53例。与结局良好组比较,结局不良组患者TSH、RBP-4水平明显升高(P<0.05),FT4、PAPP-A水平明显降低(P<0.05)。妊娠合并甲减患者血清PAPP-A水平与RBP-4水平呈负相关(r=-0.380,P<0.001)。多因素Logistic回归分析表明,血清FT4、PAPP-A水平升高是妊娠合并甲减患者妊娠结局不良的保护因素(P<0.05),血清TSH、RBP-4水平升高是妊娠合并甲减患者妊娠结局不良的危险因素(P<0.05)。ROC曲线分析结果显示,血清PAPP-A、RBP-4水平预测妊娠合并甲减患者妊娠结局不良的曲线下面积(AUC)分别为0.857(95%CI:0.799~0.914)、0.876(95%CI:0.823~0.929),低于2项联合预测的AUC[0.958(95%CI:0.930~0.986)],差异均有统计学意义(Z=3.136,P=0.002;Z=2.696,P=0.007)。结论妊娠合并甲减患者的血清PAPP-A、RBP-4水平与妊娠结局密切相关,2项联合对妊娠合并甲减患者的妊娠结局具有一定的临床预测价值。

高纯度RBP4抗体的制备及在水动力转染小鼠肝癌模型中的应用

目的:制备高纯度视黄醇结合蛋白4(RBP4)单克隆抗体并应用其检测高压水动力转染小鼠肝癌模型中RBP4的表达水平。方法:将RBP4重组蛋白表达菌株培养并诱导表达,使用AKTA系统纯化蛋白,通过杂交瘤的方式制备高纯度RBP4单克隆抗体;将7周龄SPF级C57BL/6J小鼠随机分为实验组和对照组,每组6只,雌雄各半,利用高压水动力转染法联合SB11转座子系统与CRISPR/Cas9系统构建实验组小鼠肝癌模型;将RBP4单克隆抗体应用于免疫组化法检测小鼠肝癌组织中RBP4的表达水平。结果:成功获得纯度95%以上的重组RBP4蛋白,制备RBP4单克隆抗体过程中杂交瘤细胞融合率60%,最终获得3株阳性单克隆细胞株,经纯化获得高效价(1∶243 000)、高纯度(96.77%)单克隆抗体。成功建立高压水动力转染小鼠肝癌模型,实验组肝癌发生率100%,肝脏呈现典型肝癌病理特征,对照未出现肝癌。利用制备的RBP4单克隆抗体进行免疫组化检测,发现实验组肝癌组织中RBP4表达水平显著低于对照组(P<0.001)。结论:本研究制备的RBP4单克隆抗体具有高效价和高纯度特点,RBP4蛋白表达水平在高压水动力转染小鼠肝癌组织中明显降低,RBP4与肝癌发生和发展的关系值得进一步研究。

精神分裂症患者并发糖尿病的危险因素分析及RBP4、Vaspin对糖尿病的预测价值

目的分析精神分裂症患者并发糖尿病的危险因素分析及视黄醇结合蛋白4(RBP4)、脂肪特异性丝氨酸蛋白酶抑制剂(Vaspin)对糖尿病的预测价值。方法选取2018年6月至2023年6月于我院治疗的90例精神分裂症患者为研究对象,根据患者糖尿病发生情况分为未并发糖尿病组67例,并发糖尿病组23例,分析精神分裂症患者并发糖尿病的危险因素,采用ROC曲线评价RBP4、Vaspin对糖尿病的预测价值。结果单因素分析显示,精神分裂症患者糖尿病的发生与性别、居住地、职业、文化程度、婚姻状况、具有精神病家族史无关,P>0.05;年龄>60岁、体质量为肥胖、病程>10年、住院时间>6个月、使用氯氮平、具有糖尿病遗传史、存在高血脂症、胆固醇偏高患者糖尿病发生率较高(P<0.05)。多因素Logistics回归分析显示,年龄、体质量、病程、住院时间、抗精神病药物、糖尿病遗传史、高血脂症、胆固醇为影响精神分裂症患者并发糖尿病的危险因素(P<0.05)。与未并发糖尿病组对比,并发糖尿病组患者的RBP4、Vaspin表达水平升高(P<0.05)。ROC曲线显示,与RBP4、Vaspin单项预测对比,两项联合对糖尿病具有较高的预测价值(P<0.05)。结论年龄>60岁、体质量为肥胖、病程>10年、住院时间>6个月、使用氯氮平、具有糖尿病遗传史、存在高血脂症、胆固醇偏高为精神分裂症患者并发糖尿病的危险因素,检测机体内RBP4、Vaspin表达水平,可用于糖尿病的预测。

基于生物信息学分析RBP4在肝细胞癌中的功能作用

为了揭示视黄醇结合蛋白(RBP4)在肝细胞癌(LIHC)中的功能作用,对RBP4的结构及理化性质进行了分析。基于公共数据库明确了RBP4在正常组织和LIHC中的表达,分析了其与LIHC患者的病理状态与临床生存预后的关系。结果显示:RBP4的等电点(pI)为5.76,其β-桶核心是视黄醇的结合中心,亲水性强,具有一个信号肽;RBP4在正常肝组织中的表达高于LIHC,并且高表达RBP4有更好的预后生存率(log rank P<0.001);RBP4的表达可能会缓解LIHC的进展,也可作为潜在生物标志物应用于LIHC的临床诊断与预后中。

连续性血液净化和间歇性血液透析对老年急性肾功能衰竭患者血清RBP4 Tfβ_(2)-MG水平的影响

目的:探讨连续性血液净化(CBP)和间歇性血液透析(IHD)对老年急性肾功能衰竭(ARF)患者血清视黄醇结合蛋白4(RBP4)、转铁蛋白(Tf)、β_(2)微球蛋白(β_(2)-MG)水平的影响。方法:纳入2021年1月至2023年1月在我院收治的114例ARF患者为本次研究的研究对象,采取随机数字表对入组ARF患者进行简单随机分组,各57例(观察组、对照组)。对照组予以IHD,观察组予以CBP,两组患者透析周期均为2周。比较两组患者临床疗效,透析前、透析2周后血清检测水平[RBP4、Tf、β_(2)-MG、Scr和BUN]与电解质指标(K^(+)、Cl^(-)、Na^(+)),记录两组患者并发症发生情况。结果:透析结束后,观察组与对照组ARF患者总有效率分别为94.74%(54/57)、78.95%(45/57),差异有统计学意义(P<0.05)。透析2周后两组患者血清RBP4、Tf、β_(2)-MG、Scr、BUN水平较之透析前明显降低,且观察组血清RBP4、Tf、β_(2)-MG、Scr、BUN水平下降幅度均大于对照组(P<0.05)。透析2周后两组患者血清K^(+)、Cl^(-)和Na^(+)水平较之透析前明显升高,且观察组K^(+)、Cl^(-)和Na^(+)水平升高幅度均大于对照组(P<0.05)。两组ARF患者透析期间并发症发生率(5.26%、7.02%)比较差异不显著(P>0.05)。结论:老年ARF患者应用CBP透析效果较好,可以较大程度清除血液中的大小分子毒素,并维持电解质的稳定。

血尿酸及血清RBP4、Cys-C、Lp-PLA2与糖尿病视网膜病变合并糖尿病肾病的相关性

目的分析血尿酸(SUA)及血清视黄醇结合蛋白4(RBP4)、胱抑素C(Cys-C)、脂蛋白相关磷脂酶A2(Lp-PLA2)与糖尿病视网膜病变(DR)合并糖尿病肾病(DKD)的相关性。方法选取2021年1月至2023年3月在河南省荣军医院诊治的82例DR患者,按照是否合并DKD分为合并DKD组(21例)和未合并DKD组(61例),比较两组SUA及血清RBP4、Cys-C、Lp-PLA2水平,并分析SUA及血清RBP4、Cys-C、Lp-PLA2与DR患者合并DKD的相关性以及对DR患者合并DKD的预测价值。结果合并DKD组SUA、RBP4、Cys-C、Lp-PLA2与未合并DKD组比较,更高(P<0.05)。经Spearman分析,SUA及血清RBP4、Cys-C、Lp-PLA2与DR合并DKD呈正相关(r=0.507、0.440、0.402、0.494,P<0.05)。受试者工作特征曲线分析显示,SUA、RBP4、Cys-C、Lp-PLA2、联合预测DR患者合并DKD的曲线下面积分别为0.835、0.791、0.766、0.827、0.913,在最佳临街值对应的敏感度、特异度分别为:SUA为90.5%、62.3%,RBP4为52.4%、100.0%,Cys-C为52.4%、100.0%,Lp-PLA2为61.9%、100.0%,联合为85.7%、88.5%。结论DR患者合并DKD其SUA及血清RBP4、Cys-C、Lp-PLA2水平升高,且与DR患者合并DKD呈正相关,SUA、RBP4、Cys-C、Lp-PLA2联合对DR患者合并DKD具有较高的预测价值。

血清RBP4联合全身免疫炎症指数对急性冠状动脉综合征患者介入术后主要心血管不良事件的预测价值

目的 研究血清视黄醇结合蛋白4(RBP4)联合全身免疫炎症指数(SII)对急性冠状动脉综合征(ACS)患者介入术后主要心血管不良事件(MACE)的预测价值。方法 选取2021年3月至2023年10月在重庆市云阳县人民医院接受介入术的ACS患者152例(ACS组),根据介入术后6个月是否发生MACE分为MACE组(32例)和非MACE组(120例),另选取同期57名健康体检志愿者(对照组)。收集ACS患者临床资料,检测ACS组和对照组血清RBP4水平,并计算SII。分析ACS患者介入术后MACE的影响因素和血清RBP4、SII对其的预测价值。结果 ACS组血清RBP4水平和SII明显高于对照组(P <0.05)。年龄增加、急性心肌梗死、Killip分级≥Ⅱ级、RBP4升高、SII升高为ACS患者介入术后MACE的独立危险因素(P <0.05),左心室射血分数升高为其独立保护因素(P <0.05)。血清RBP4水平联合SII预测ACS患者介入术后MACE的曲线下面积为0.833,大于RBP4、SII单独预测的0.744、0.750(P <0.05)。结论血清RBP4水平联合SII对预测ACS患者介入术后MACE的价值较高,且对预后的评估具有一定指导意义。

小尾寒羊高繁殖力候选基因RBP4的研究

以视黄醇结合蛋白4(retinol binding protein 4,RBP4)基因为候选基因,采用PCR-SSCP技术分析了RBP4基因在高繁殖力绵羊品种(小尾寒羊、湖羊)以及低繁殖力绵羊品种(多赛特羊、萨福克羊)中的单核苷酸多态性,同时研究这个基因对小尾寒羊高繁殖力的影响。结果表明:RBP4基因扩增片段在4个绵羊品种中存在PCR-SSCP多态性。BB基因型只出现在高繁殖力绵羊品种中,而低繁殖力绵羊品种则没有BB基因型;AB基因型频率随着绵羊繁殖力的降低而升高;AA基因型只出现在小尾寒羊、多赛特羊中。BB基因型小尾寒羊产羔数分别比AA和AB基因型多0.52只(P<0.05)和0.67只(P<0.05),AA和AB基因型小尾寒羊产羔数没有显著差异(P>0.05)。本研究检测的绵羊RBP4基因座位与小尾寒羊高繁殖力相关。

血清GA、ADP、RBP4、Hcy与妊娠糖尿病胰岛素抵抗的相关性研究

目的探讨糖化白蛋白(GA)、脂联素(ADP)、视黄醇结合蛋白4(RBP4)、同型半胱氨酸(Hcy)与妊娠糖尿病(GDM)胰岛素抵抗的关系。方法选取2014年7月—2015年7月保定市妇幼保健院GDM患者80例(GDM组)、正常孕妇80例(正常妊娠组)及正常非妊娠期妇女80例(正常非妊娠组),比较3组血清GA、ADP、RBP4、Hcy水平,分析GA、ADP、RBP4、Hcy与GDM孕妇胰岛素抵抗的关系。结果 GDM组血清GA、RBP4、Hcy水平分别为(18.36±0.98)%、(42.98±2.28)mg/L、(22.36±2.58)μg/L,均显著高于正常妊娠组的(14.22±2.05)%、(34.22±3.48)mg/L、(11.25±2.98)μg/L以及正常非妊娠组的(14.36±2.69)%、(30.02±2.96)mg/L、(11.02±2.79)μg/L,而ADP水平为(4.23±1.42)mg/L,显著低于正常妊娠组的(9.23±2.58)mg/L以及正常非妊娠组的(9.78±2.69)mg/L(P<0.05),正常妊娠组RBP4水平高于正常非妊娠组(P<0.05)。GDM组空腹血糖(FPG)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)高于正常妊娠组及正常非妊娠组(P<0.05),正常妊娠组FPG、FINS、HOMA—IR高于正常非妊娠组,但差异无统计学意义(P>0.05)。经Pearson相关因素分析可知,GA、RBP4、Hcy与FPG、FINS、HOMA-IR水平呈正相关(P<0.05),而ADP与FPG、FINS、HOMA-IR水平呈负相关(P<0.05)。GA、ADP、RBP4、Hcy联合诊断GDM的ROC曲线下面积(AUC)为0.902,大于单一指标,灵敏度、特异度分别为87.2%、88.6%,均高于单一指标。结论血清GA、ADP、RBP4、Hcy水平异常与GDM患者胰岛素抵抗有密切的关系,通过检测血清GA、ADP、RBP4、Hcy水平能快速、有效地对GDM作出诊断,联合多指标检测可提高诊断灵敏度和特异度。

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。