Expression and function of Delta-like ligand 4 in a rat model of retinopathy of prematurity

The Delta-like ligand 4/Notch signaling pathway was shown to participate in the process of retinal development and angiogenesis. However, the function of the Delta-like ligand 4/Notch signaling pathway in retinopathy of prematurity requires further study. Retinopathy of prematurity was induced in 5-day-old Sprague-Dawley rats exposed to hyperoxia for 7 days, and then returned to room air. Reverse transcription-PCR and western blot revealed that Delta-like ligand 4 levels decreased at postnatal day 12 and increased at postnatal day 17 in retinopathy of prematurity rats. Flat-mounted adenosine diphosphatase stained retina and hematoxylin-eosin stained retinal tissue slices showed that the clock hour scores and the nuclei counts in retinopathy of prematurity rats were significantly different compared to normal control rats. After retinopathy of prematurity rats were intravitreally injected with Delta-like ligand 4 monoclonal antibody to inhibit the Delta-like ligand 4/Notch signaling pathway, there was a significant increase in the severity of retinal neovascularization （clock hours） in the intravitreally injected eyes. The nuclei count was highly correlated with the clock hour score. These results suggest that Delta-like ligand 4/Notch signaling plays an essential role in the process of physiological and pathological angiogenesis in the retina.

Delta-like ligand 4在脑星形细胞瘤中的表达及临床意义

目的探讨Delta-like ligand4(DLL4)在脑星形细胞瘤中的表达及与微血管密度(MVD)的关系。方法脑星形细胞瘤标本40例,分Ⅰ级,Ⅱ级,Ⅲ～Ⅳ级三组,分别使用DLL4单克隆抗体、抗人CD31多克隆抗体行免疫组织化学染色,Image-Pro Plus6.0软件分析DLL4光密度(optical density,OD)半定量分析DLL4表达,统计微血管数。结果各组DLL4蛋白OD值为Ⅰ级514.71±179.56,Ⅱ级961.97±88.53,Ⅲ～Ⅳ级1125.31±225.35,P<0.01;MVD值Ⅰ级24.75±10.33,Ⅱ级57.62±12.45,Ⅲ～Ⅳ级114.60±25.28,P<0.01。DLL4的OD值与MVD的相关系数为γ=0.68(P<0.01)。结论 DLL4表达随星型细胞瘤恶性程度的升高逐步增加,与微血管密度密切相关,对判断星型细胞瘤的恶性程度及血供有指导意义。

Delta-like ligand 4 in hepatocellular carcinoma intrinsically promotes tumour growth and suppresses hepatitis B virus replication

AIM To investigate the role of Delta-like ligand 4(DLL4) on tumour growth in hepatitis B virus(HBV)-associated hepatocellular carcinoma(HCC) in vivo.METHODS We suppressed DLL4 expression in an HBV expressing HCC cell line, HepG2.2.15 and analysed the growth ability of cells as subcutaneous tumours in nude mice. The expression of tumour angiogenesis regulators, VEGF-A and VEGF-R2 in tumour xenografts were examined by western blotting. The tumour proliferation and neovasculature were examined by immunohistochemistry. The viral replication and viral protein expression were measured by quantitative PCR and western blotting, respectively.RESULTS Eighteen days after implantation, tumour volume in mice implanted with sh DLL4 HepG2.2.15 was significantly smaller than in mice implanted with control HepG2.2.15(P < 0.0001). The levels of angiogenesis regulators, VEGF-A and VEGF-R2 were significantly decreased in implanted tumours with suppressed DLL4 compared with the control group(P < 0.001 and P < 0.05, respectively). Furthermore, the suppression of DLL4 expression in tumour cells reduced cell proliferation and the formation of new blood vessels in tumours. Unexpectedly, increased viral replication was observed after suppression of DLL4 in the tumours.CONCLUSION This study demonstrates that DLL4 is important in regulating the tumour growth of HBV-associated HCC as well as the neovascularization and suppression of HBV replication.

桃红四物汤对大鼠血管吻合模型吻合区Delta-like4表达及血管新生的影响

目的:观察桃红四物汤对大鼠血管吻合模型吻合区Delta-like4表达及血管新生的影响。方法:将40只3月龄SPF级SD大鼠随机分为空白对照组、假手术组、生理盐水组、桃红四物汤组,每组10只。桃红四物汤组、生理盐水组行尾部动脉血管离断后吻合术;假手术组仅在相应区域作切口,不进行血管离断吻合;空白对照组不做任何处理。造模术后桃红四物汤组以20 mL·kg^(-1)剂量的桃红四物汤灌胃,生理盐水组给予等量生理盐水灌胃,每日灌胃2次;假手术组和空白对照组不给予任何药物干预。分别在药物干预开始后2 d、5 d从各组随机抽取5只大鼠,采用颈椎脱臼法处死后,桃红四物汤组和生理盐水组切取尾部的吻合处血管,假手术组和空白对照组切取尾部相应部位动脉血管。将所取血管组织制成石蜡切片,分成2份,1份采用免疫组化法检测血管吻合区Delta-like4的蛋白表达(光镜下灰度值与Delta-like4的蛋白表达量呈反比);1份经HE染色后测量吻合区血管内膜面积。结果:(1)血管吻合区Delta-like4的蛋白表达。药物干预开始后2 d,4组大鼠血管吻合区Delta-like4蛋白表达的灰度值比较,差异无统计学意义(126.27±20.56,111.73±7.99,119.30±16.71,124.53±7.35,F=0.624,P=0.062)。药物干预开始后5 d,4组大鼠血管吻合区Delta-like4蛋白表达的灰度值比较,差异有统计学意义(85.60±5.58,99.07±3.84,103.13±11.61,111.60±4.49,F=7.041,P=0.012);进一步两两比较,桃红四物汤组血管吻合区Delta-like4蛋白表达的灰度值低于生理盐水组(LSD-t=-2.334,P=0.048)、假手术组(LSD-t=-3.035,P=0.016)、空白对照组(LSD-t=-4.450,P=0.002);生理盐水组血管吻合区Delta-like4蛋白表达的灰度值与假手术组、空白对照组比较,差异均无统计学意义(LSD-t=-0.703,P=0.502;LSD-t=-2.166,P=0.062);假手术组血管吻合区Delta-like4蛋白表达的灰度值与空白对照组比较,差异无统计学意义(LSD-t=-1.464,P=0.181)。(2)吻合区血管内膜面积。药物干预开始后2 d,4组大鼠吻合区血管内膜面积比较,差异无统计学意义[(0.20±0.01)mm^2,(0.18±0.02)mm^2,(0.16±0.01)mm^2,(0.17±0.02)mm^2,F=2.625,P=0.122)。药物干预开始后5 d,4组大鼠吻合区血管内膜面积比较,差异有统计学意义[(0.30±0.02)mm2,(0.20±0.01)mm^2,(0.21±0.01)mm^2,(0.21±0.01)mm^2,F=17.063,P=0.000];进一步两两比较,桃红四物汤组吻合区血管内膜面积大于生理盐水组(LSD-t=-46.275,P=0.020)、假手术组(LSD-t=-68.142,P=0.000)、空白对照组(LSD-t=-66.154,P=0.000);生理盐水组吻合区血管内膜面积与假手术组、空白对照组比较,差异均无统计学意义(LSD-t=-2.187,P=0.060;LSD-t=-1.988,P=0.082);假手术组吻合区血管内膜面积与空白对照组比较,差异无统计学意义(LSD-t=0.199,P=0.847)。结论:桃红四物汤能上调血管吻合大鼠模型吻合区Delta-like4的表达,促进血管新生。

斑马鱼Notch信号通路配体基因delta-like4对smad基因的调控作用

为研究斑马鱼(Danio rerio)的Notch信号通路配体delta-like 4(dll4)对smad基因(smad1和smad7)的调控作用,利用qRT-PCR方法检测受精后2 d(day post fertilization,dpf)的斑马鱼dll4纯合突变体smad家族中smad1和smad7的表达变化,利用生物信息学软件预测smad1和smad7启动子序列的转录结合位点和CpG岛;采用同源重组的方法,构建p3×Flag-CMV-dll4真核表达载体及pGL3-smad1-Luc和pGL3-smad7-Luc报告基因载体,并通过双荧光素酶试验检测dll4对smad1和smad7的调控活性;分别转染两种启动子报告基因载体到HEK293T细胞,共转染启动子报告基因载体与真核表达载体到HEK293T细胞。结果表明:dll4纯合突变体斑马鱼中smad1和smad7表达量均显著下调(P<0.0001);两种基因启动子均包含HNF-3、GATA-1和Oct-1等转录因子结合位点,只有smad7启动子存在CpG岛;报告基因pGL3-smad1-Luc和pGL3-smad7-Luc的活性分别为对照组的7.3倍和142.7倍,两种报告基因与p3×Flag-CMV-dll4表达载体共转后,转录活性均升高,分别为对照组的2.8倍和2.2倍,说明p3×Flag-CMV-dll4可以促进pGL3-smad1-Luc和pGL3-smad7-Luc的转录表达活性。研究表明,斑马鱼Notch信号通路配体基因dll4可通过smad基因家族在血管生成中发挥调控作用,为损伤血管的修复和肿瘤血管生成提供特异的生物学靶点。

Levels of soluble delta-like ligand 1 in the serum and cerebrospinal fluid of tuberculous meningitis patients

In this study, the levels of soluble delta-like ligand 1 in cerebmspinal fluid and serum of 50 patients with tuberculous meningitis, 30 patients with viral meningitis, 20 patients with purulent meningitis and 40 subjects without central nervous system disease were determined using an enzyme-linked immunosorbent assay. The mean levels of soluble delta-like ligand 1 in both cerebrospinal fluid and serum from patients with tuberculous meningitis were significantly higher compared with those from patients with viral meningitis or purulent meningitis or from subjects without central nervous system disease. Meanwhile, the level of soluble delta-like ligand 1 gradually decreased as tuberculous meningitis patients recovered. If patients deteriorated after treatment, the level of soluble delta-like ligand 1 in cerebrospinal fluid gradually increased. There was no correlation between the level of soluble delta-like ligand 1 and the protein level/cell number in cerebrospinal fluid. Our findings indicate that the levels of soluble delta-like ligand 1 in cerebrospinal fluid and serum are reliable markers for the diagnosis of tuberculous meningitis and for monitoring treatment progress. At the same time, this index is not influenced by protein levels or cell numbers in cerebrospinal fluid.

Influence of up-regulation of Notch ligand DLL4 on biological behaviors of human gastric cancer cells

AIM: To investigate the potential roles of Delta-like ligand 4 (DLL4) on the biological behavior of gastric cancer cells and its molecular mechanisms. METHODS: A recombinant eukaryotic expression vector containing human DLL4 gene was constructed and transfected into the human gastric cancer cell line SGC7901. Clones with up-regulated DLL4 were selected and amplified. The effect of DLL4 up-regulation on gastric cancer cell growth was assessed using cell growth assay. The migration and invasion were assessed using a transwell migration assay and matrigel invasion assay. Matrix metalloproteinases were detected using the zymogram technique. Cells were implanted subcutaneously into male BALB/c nu/nu mice. Tumor volumes were then calculated and compared. DLL4 staining in the implanted tumor was performed using immunohistochemistry technique. RESULTS: Growth curves over a six-day time course showed significantly promoted cell proliferation of SGC7901 cells with up-regulated DLL4. DLL4 up-regulation in SGC7901 cells promoted the migration (205.4 ± 15.2 vs 22.3 ± 12.1, P < 0.05) and invasion (68.8 ± 5.3 vs 18.2 ± 6.0, P < 0.05) in vitro and tumorigenicity in vivo (2640.5 ± 923.6 mm 3 vs 1115.1 ± 223.8 mm 3 , P < 0.05). Furthermore, significantly increased mRNA level and increased secretion of matrix metalloproteinase-2 (MMP-2) proenzyme were observed in SGC7901 cells with up-regulated DLL4. However, increased MMP-9 mRNA level but decreased extracellular MMP-9 proenzyme level was observed. CONCLUSION: Our observations indicated a mechanism by which activation of DLL4-mediated Notch signaling promotes the expression and secretion of MMP-2 proenzyme and influences the progress of gastric cancer.

利用受体配体亲和技术筛选黄芩中血管紧张素转换酶2和二肽基肽酶4抑制剂

目的筛选中药黄芩中血管紧张素转换酶2(ACE2)和二肽基肽酶4(DPP-4)抑制剂。方法以ACE2和DPP-4作为靶蛋白,利用受体配体亲和超滤和液质联用技术,对黄芩中与ACE2和DPP-4亲和的化合物进行筛选和分析,以筛选具有同时抑制ACE2和DPP-4的化合物。结果从黄芩中筛选出了5,7,2′,6′-4羟基黄酮、5,7,2′,5′-4羟基-8,6′-二甲氧基黄酮、白杨素-7-O-葡萄糖醛酸苷、黄芩素-6-O-葡萄糖醛酸苷、千层纸素A-7-O-葡萄糖醛酸苷、汉黄芩苷、白杨素、千层纸素A,可以同时亲和抑制ACE2和DPP-4,具有潜在的抗新型冠状病毒的作用。结论从黄芩中筛选出与新型冠状病毒肺炎有关的靶蛋白抑制剂,具备潜在开发价值。

Association of gene and protein expression and genetic polymorphism of CC chemokine ligand 4 in colorectal cancer

BACKGROUND Leukocytes,such as T cells and macrophages,play an important role in tumorigenesis.CC chemokine ligand(CCL)4,which is produced by lymphocytes and macrophages,has been found to be expressed in the mucosa of the gastrointestinal tract and is a potent chemoattractant for various leukocytes.AIM To examine CCL4 expression and its genetic polymorphism rs10491121 in patients with colorectal cancer(CRC)and evaluate their prognostic significance.METHODS Luminex technology was used to determine CCL4 Levels in CRC tissue(n=98),compared with paired normal tissue,and in plasma from patients with CRC(n=103),compared with healthy controls(n=97).Included patients had undergone surgical resection for primary colorectal adenocarcinomas between 1996 and 2019 at the Department of Surgery,Ryhov County Hospital,Jönköping,Sweden.Reverse transcription quantitative PCR was used to investigate the CCL4 gene expression in CRC tissue(n=101).Paired normal tissue and TaqMan single nucleotide polymorphism assays were used for the CCL4 rs10491121 polymorphism in 610 CRC patients and 409 healthy controls.RESULTS The CCL4 protein and messenger RNA expression levels were higher in CRC tissue than in normal paired tissue(90%,P<0.001 and 45%,P<0.05,respectively).CRC tissue from patients with localized disease had 2.8-fold higher protein expression levels than that from patients with disseminated disease.Low CCL4 protein expression levels in CRC tissue were associated with a 30%lower cancer-specific survival rate in patients(P<0.01).The level of plasma CCL4 was 11%higher in CRC patients than in healthy controls(P<0.05)and was positively correlated(r=0.56,P<0.01)with the CCL4 protein level in CRC tissue.The analysis of CCL4 gene polymorphism rs10491121 showed a difference(P<0.05)between localized disease and disseminated disease in the right colon,with a dominance of allele A in localized disease.Moreover,the rate of the A allele was higher among CRC patients with mucinous cancer than among those with nonmucinous cancer.CONCLUSION The present study indicates that the CRC tissue levels of CCL4 and CCL4 gene polymorphism rs10491121,particularly in the right colon,are associated with clinical outcome in CRC patients.

基于DLL4/Notch1信号通路探讨人参皂苷Rg-3对AOM/DSS诱导的炎症相关性结直肠癌小鼠血管形成机制

目的基于Delta样配体4(delta-like ligand 4,DLL4)/Notch1信号通路探究人参皂苷Rg-3对氧化偶氮甲烷/葡聚糖硫酸钠(azoxymethane/dextran sulphate sodium,AOM/DSS)诱导的炎症相关性结直肠癌小鼠血管形成机制。方法48只C57BL/6J APCMin/+小鼠随机分为对照组、模型组、人参皂苷Rg-3组(10 mg/kg),每组16只。建立AOM/DSS诱导的结直肠癌模型,采集腹主动脉血与完整结直肠组织,检测血常规与炎症指标,HE染色观察肠组织形态,免疫组化、Western blotting检测肠组织DLL4、Notch1表达。肿瘤细胞、人脐静脉内皮细胞(human umbilical vascular endothelial cells,HUVECs)分为对照组、人参皂苷Rg-3组、人参皂苷Rg-3+DLL4组,通过MTT、克隆形成、划痕、Transwell与血管形成实验检测细胞增殖、迁移、血管形成能力,Western blotting检测DLL4/Notch信号通路、血管形成相关分子表达情况。结果与对照组相比,模型组、人参皂苷Rg-3组RBC、Hct、Hb水平降低(P<0.05),IL-1β、IL-6、TNF-α、DLL4、Notch1表达水平升高(P<0.05);与模型组相比,人参皂苷Rg-3组RBC、Hct、Hb水平提高(P<0.05),IL-1β、IL-6、TNF-α、DLL4、Notch1表达水平下降(P<0.05),肿瘤数量、2~3.5 mm及>3.5 mm的肿瘤数量减少(P<0.05)。细胞实验结果显示,与对照组相比,人参皂苷Rg-3组肿瘤细胞、HUVECs增殖、克隆形成、迁移、血管形成能力下降(P<0.05),DLL4、Notch1、VEGF、VEGFR2、转录因子重组信号结合蛋白Jκ(recombination signal binding protein-Jκ,RBP-Jκ)、Hes1表达水平降低(P<0.05)。结论人参皂苷Rg-3通过抑制DLL4/Notch1信号通路促进血管形成,抑制AOM/DSS诱导结直肠癌进展。

趋化因子受体4及其配体12在下咽鳞状细胞癌中的表达及其临床意义

目的 探究趋化因子受体4(CXCR4)及趋化因子配体12(CXCL12)在下咽鳞状细胞癌(HSCC)中的表达及其临床意义。方法 以2017年1月—2022年11月收治的90例HSCC患者为研究对象,以HSCC患者手术切除的HSCC组织、癌旁组织为实验材料,采用免疫组织化学染色法检测HSCC组织和癌旁组织中CXCR4、CXCL12的表达情况;采用实时荧光定量PCR(qRT-PCR)法检测HSCC组织和癌旁组织中CXCR4 mRNA、CXCL12 mRNA水平;收集并记录HSCC患者临床病理特征,分析CXCR4、CXCL12表达水平与患者临床病理特征的关系;采用Pearson分析CXCR4与CXCL12水平相关性。结果 CXCR4在HSCC组织中的表达率为60.00%,明显高于癌旁组织的17.78%(P<0.05);CXCL12在HSCC组织中的表达率为57.78%,明显高于癌旁组织的24.44%(P<0.05)。HSCC组织中CXCR4、CXCL12水平均与淋巴结是否转移、TNM分期、分化程度有关(P<0.05)。与癌旁组织相比较,HSCC组织CXCR4、CXCL12水平均显著升高(P<0.05)。Pearson分析显示,HSCC患者中CXCR4水平与CXCL12水平呈正相关(r=0.538,P<0.05)。结论 CXCR4、CXCL12在HSCC组织中呈高表达,两者与HSCC淋巴结转移、分化程度、TNM分期有关。

亲水性CIT-NaYF_(4)∶Yb^(3+),Ho^(3+)纳米粒子的制备及其发光性能研究

采用溶剂热法制备了油溶性稀土掺杂上转换纳米粒子NaYF_(4)∶Yb^(3+),Ho^(3+),并利用TEM、XRD、IR、荧光光谱法等考察了反应温度、反应时间、掺杂比等反应条件对纳米粒子形貌、结构、发光性能的影响。结果表明,在反应温度为300℃、反应时间为50 min、稀土离子掺杂比为NaYF_(4)∶20%Yb^(3+),2%Ho^(3+)条件下,制备出的油溶性NaYF_(4)∶Yb^(3+),Ho^(3+)纳米粒子呈标准六方晶型,尺寸均一、单分散性好,平均粒径为31.55 nm。通过表面配体交换法成功将柠檬酸修饰在其表面,转为亲水性CIT-NaYF_(4)∶Yb^(3+),Ho^(3+)纳米粒子。

Syntheses, Characterization and Biological Activity of Coordination Compounds of 3-Hydroxy-2-methyl-4<i>H</i>-pyran-4-one and Its Mixed Ligand Complexes with 1,2-Diaminocyclohexane

Coordination compounds of 3-hydroxy-2-methyl-4H-pyran-4-one with iron(III), cobat(III) and chromium(III) were synthesized with M:L (1:2). Mixed ligand coordination compounds of 3-hydroxy-2-methyl-4H-pyran-4-one and 1,2-diaminocyclohexane using the same metal ions were also synthesized M:L1:L2 (1:1:1) where L1 is 3-hydroxy-2-methyl-4H-pyran-4-one and L2 is 1,2-diaminocyclohexane. The coordination compounds obtained were characterized using electronic and infrared spectral analyses, magnetic susceptibility and percentage metal analysis. They were also evaluated for their cytotoxic and antioxidant activities. The result obtained suggested that octahedral geometry was obtained for all the compounds, as a result of additional two molecules of the solvent coordinated to the metal ions. Both the primary and secondary ligands coordinated in a bidentate fashion. The synthesized compounds exhibited moderate cytotoxicity, although none was as active as the standard. The cobalt(III) mixed ligand complex elicited the highest activity. The synthesized compounds all exhibited good to moderate antioxidant activity.

Gold （III） Complexes with Thiolactate and bis-（1,4-Sodiumthiolactate） Butane Ligands

The new ligand bis-（1,4-sodium thiolactate） butane （L）-O2CCH3S-（CH2）4SCHCH3CO2- has been prepared from the reaction of disodium salt of thiolactic acid and 1,4-dichlorobutane, while the disodium thiolactate was prepared instanteously through the reaction of thiolactic acid with NaOH. Mono and dinuclear complexes were obtained by direct reaction of the above ligands with H[AuCI4] in 1 ： 1, 2：1, 1：2, 2：2 and 3：1 ligands to metal molar ratio. The prepared complexes were characterized by elemental analysis, spectral studies FTIR （Fourier transform Infrared） and UV-Vis, magnetic measurement, conductivity measurement and IHNMR for the ligand （L） and some of the complexes. The conductance data indicate that the complexes of the formulas Na[Au（SCH3CHCOO）2], Na[Au（SCH3CHCOO）（OOCCHCH3SH）2] and [Au（L）]Cl are 1：1 electrolyte. Electronic spectra and magnetic moment values indicate the presence of square planner geometry around Au（III） ions.

棕矢车菊素调节SDF-1α/CXCR4信号通路对妊娠糖尿病大鼠炎症反应的影响

目的探讨棕矢车菊素对妊娠糖尿病(GDM)大鼠炎症反应的影响及可能机制。方法将妊娠大鼠腹腔注射链脲佐菌素建立GDM模型,将建模成功大鼠分为GDM组,棕矢车菊素-L、M、H组,棕矢车菊素+CTCE-0214[基质细胞衍生因子1α(SDF-1α)/趋化因子受体4(CXCR4)激活剂]组,每组10只;另取10只健康妊娠大鼠作为对照组。棕矢车菊素-L、M、H组分别灌胃2、3、4 mg/kg棕矢车菊素,棕矢车菊素+CTCE-0214组腹腔注射10 mg/kg CTCE-0214并灌胃4 mg/kg棕矢车菊素,对照组和GDM组给予等量生理盐水。比较各组大鼠空腹血糖、空腹胰岛素以及胰岛素抵抗指数差异;酶联免疫吸附测定法检测大鼠胎盘组织中白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α水平;HE染色观察胎盘组织病理变化;TUNEL检测胎盘组织细胞凋亡;Western blot检测大鼠胎盘组织中SDF-1α/CXCR4通路相关蛋白水平。结果不同剂量棕矢车菊素干预GDM大鼠引起大鼠空腹血糖、空腹胰岛素、胰岛素抵抗指数水平降低,促炎细胞因子水平降低,改善GDM大鼠胎盘损伤和细胞凋亡,并抑制SDF-1α/CXCR4信号通路激活,呈剂量依赖性(P<0.05),SDF-1α/CXCR4激活剂和棕矢车菊素联合干预后抑制棕矢车菊素上述作用。结论棕矢车菊素可通过抑制SDF-1α/CXCR4信号通路激活来减轻GDM大鼠炎症反应和胰岛素抵抗。

趋化因子C-C-基元配体4通过激活人单核白血病细胞THP-1源性巨噬细胞NOD样受体热蛋白结构域相关蛋白3炎症小体促进骨关节炎发展实验研究

目的:探讨趋化因子C-C-基元配体4(CCL4)通过激活人单核白血病细胞THP-1源性巨噬细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体促进骨关节炎(OA)发展的潜在作用机制。方法:用CCL4和NLRP3小干扰RNA(siRNA)处理THP-1源性巨噬细胞,然后将预处理的THP-1源性巨噬细胞与软骨细胞共培养。采用流式细胞术检测细胞凋亡和活性氧(ROS);采用酶联免疫吸附测定(ELISA)法检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶1(MMP-1)和MMP-13水平;采用胶原降解试验检测胶原活性;采用Western blot分析NLRP3和凋亡相关微粒蛋白(ASC)蛋白的表达。结果:CCL4不会诱导THP-1源性巨噬细胞凋亡,但可上调炎症因子(IL-1β和TNF-α)和炎症相关蛋白(NLRP3和ASC)的表达。将CCL4预处理的THP-1源性巨噬细胞与软骨细胞共培养可以上调软骨细胞中MMP-1和MMP-13的表达,增强胶原酶活性,促进ROS的产生及软骨细胞的凋亡。抑制NLRP3的表达可以部分逆转CCL4对炎症因子的影响,以及与THP-1源性巨噬细胞共培养的软骨细胞相关功能的影响。结论:CCL4处理的THP-1源性巨噬细胞可通过激活NLRP3加速软骨细胞OA的进展。

支原体肺炎患儿血清FABP3、SDC-4水平与心肌损害的关系研究

目的 分析支原体肺炎患儿血清心脏型脂肪酸结合蛋白(fatty acid-binding protein 3,FABP3)、多配体蛋白聚糖-4(Syndecan-4,SDC-4)水平与心肌损害的关系。方法 选取本院2022年6月到2023年6月期间收治的支原体肺炎患儿98例作为此次研究对象,根据患儿的病情严重程度分为轻症组(58例)和重症组(40例),另选取同期健康体检儿童98例作为对照组。根据心肌损害情况将患儿分为心肌损害组(n=43)和非心肌损害组(n=55);血清FABP3、SDC-4水平检测采用酶联免疫吸附法(Enzyme-linked immunosorbent assay,ELISA);使用全自动生化分析仪和电化学发光法检测心肌酶谱指标;采用Pearson法分析血清FABP3、SDC-4水平以及与心肌酶谱相关指标的相关性;采用Logistic回归分析支原体肺炎患儿的影响因素。结果 重症组和轻症组血清FABP3、SDC-4水平显著高于对照组(P<0.05)。重症组血清FABP3、SDC-4水平显著高于轻症组(P<0.05)。心肌损害组和非心肌损害组FABP3、SDC-4、谷草转氨酶(aspartate transferase,AST)、乳酸脱氢酶(lactate dehydrogenase,LDH)、α-羟丁酸脱氢酶(α-hydroxybutyrate dehydrogenase,α-HBDH)、磷酸肌酸激酶(creatine phosphokinase,CK)、肌酸激酶同工酶(creatine kinase isoenzyme,CK-MB)以及超敏肌钙蛋白I(high sensitive troponin I,hs-cTnI)水平显著高于对照组(P<0.05),且心肌损害组FABP3、SDC-4、AST、LDH、α-HBDH、CK、CK-MB以及hs-cTnI水平显著高于非心肌损害组(P<0.05)。根据Pearson相关性分析得知,支原体肺炎患儿血清FABP3、SDC-4水平呈正相关(P<0.05),血清FABP3、SDC-4水平均与AST、LDH、α-HBDH、CK、CK-MB以及hs-cTnI水平呈正相关(P<0.05)。根据Logistic回归分析得知,FABP3、SDC-4、AST、LDH、α-HBDH、CK、CK-MB以及hs-cTnI升高是影响支原体肺炎合并心肌损害的危险因素(P<0.05)。结论 FABP3、SDC-4在支原体肺炎患儿血清中显著升高,二者与心肌损害密切相关。

溃疡性结肠炎患者血清趋化因子配体19、干扰素调节因子4水平及临床意义

目的探讨溃疡性结肠炎(UC)患者血清趋化因子配体19(CCL19)、干扰素调节因子4(IRF4)水平及临床意义。方法选择2020年6月至2022年5月在该院住院治疗的105例UC患者为UC组,另选取83例健康体检者为对照组。根据Mayo评分将UC患者分为缓解期组(n=35)和活动期组(n=70)。根据活动期病情严重程度,将活动期UC患者分为轻度组(n=19)、中度组(n=23)和重度组(n=28)。根据患者预后情况分为预后良好组和预后不良组。采用酶联免疫吸附试验检测血清CCL19、IRF4水平;采用Pearson相关分析血清CCL19、IRF4水平与Mayo评分的相关性;采用多因素Logistic回归分析影响UC患者预后的因素;采用受试者工作特征(ROC)曲线分析血清CCL19、IRF4水平对UC患者预后不良的预测价值。结果UC组血清CCL19、IRF4水平明显高于对照组,差异均有统计学意义(P<0.05)。缓解期组血清CCL19、IRF4水平低于活动期组,差异均有统计学意义(P<0.05)。重度组血清CCL19、IRF4水平明显高于轻度组和中度组,且中度组明显高于轻度组,差异均有统计学意义(P<0.05)。预后良好组CCL19、IRF4、C反应蛋白(CRP)、红细胞沉降率(ESR)水平明显低于预后不良组,差异有统计学意义(P<0.05)。相关分析结果显示,UC患者血清CCL19、IRF4水平均与Mayo评分呈正相关(r=0.426、0.471,P<0.05)。血清CCL19、IRF4水平升高是UC患者预后不良的危险因素(P<0.05)。ROC曲线分析结果显示,血清CCL19、IRF4水平单独及联合预测UC患者预后不良的曲线下面积(AUC)分别为0.798、0.752、0.845。结论UC患者血清CCL19、IRF4水平均升高,且与患者病情和临床预后关系密切,可作为UC临床预后评估的潜在指标。

补骨脂提取物对骨质疏松模型大鼠趋化因子配体炎4、症反应及骨代谢的影响及机制研究

目的 探讨补骨脂提取物对骨质疏松模型大鼠趋化因子配体4 (CCL4)、炎症反应及骨代谢的影响及机制。方法 选择无特定病原体(SPF)级SD雌性大鼠55只,选取10只大鼠作为空白组,其余45只建立骨质疏松模型,将40只造模成功大鼠分为模型组及补骨脂提取物低、中、高剂量组,各10只。造模第9周,补骨脂提取物低、中、高剂量组分别给予补骨脂提取物药液50、100、200 mg/(kg·d)灌胃,空白组、模型组给予生理盐水10 mL/(kg·d)灌胃。均连续干预8周。干预8周后,处死大鼠,获取股骨组织,HE染色光镜下观察股骨病理形态改变;酶联免疫吸附法检测股骨组织中CCL4、炎症因子[肿瘤坏死因子-ɑ(TNF-ɑ)、白细胞介素6 (IL-6)];用电化学发光法检测骨代谢指标[骨钙素(OC)、Ⅰ型原胶原N-端前肽(P1NP)];用Western blotting法检测股骨组织中转化生长因子β1 (TGF-β1)/Smad4信号通路相关蛋白表达。结果 空白组股骨骨髓腔面积最小,模型组骨髓腔面积最大,补骨脂提取物低、中、高剂量组骨髓腔面积依次缩小,但仍大于空白组。与模型组比较,补骨脂提取物低、中、高剂量组CCL4水平低(P<0.05),且呈剂量依赖性(P<0.05)。与空白组比较,模型组OC、PINP水平低(P<0.05);与模型组比较,补骨脂提取物低、中、高剂量组OC、PINP水平高(P<0.05),且呈剂量依赖性(P<0.05)。与空白组比较,模型组TNF-ɑ、IL-6水平高(P<0.05);与模型组比较,补骨脂提取物低、中、高剂量组TNF-ɑ、IL-6水平低(P<0.05),且呈剂量依赖性(P<0.05)。与空白组比较,模型组TGF-β1、Smad4蛋白相对表达量低(P<0.05);与模型组比较,补骨脂提取物低、中、高剂量组TGF-β1、Smad4蛋白相对表达量高(P<0.05),且呈剂量依赖性(P<0.05)。结论 补骨脂提取物可使骨质疏松大鼠CCL4水平下降,抑制炎症反应,改善骨代谢,其机制可能与TGF-β1/Smad4信号通路被抑制有关。

糖尿病视网膜病变患者血清Hcy、DLL4和SUA水平与糖脂代谢、胰岛素抵抗的关系

目的:探讨糖尿病视网膜病变(DR)患者血清同型半胱氨酸(Hcy)、Delta样配体4(DLL4)和血尿酸(SUA)与糖脂代谢、胰岛素抵抗水平(HOMA-IR)的关系。方法:选取2021-01-2024-01在本院治疗的DR患者68例作为DR组,同时选取单纯糖尿病患者130例(糖尿病组)和健康体检者100例(健康组),比较各组Hcy、DLL4、SUA、空腹血糖(FBG)、糖化血红蛋白(HbA1c)、HOMA-IR差异,以及不同视网膜病变程度患者间上述指标的差异。结果:DR组Hcy、DLL4、SUA、FBG、HbA1c和HOMA-IR水平明显高于糖尿病组和健康组(P<0.05);DRⅢ-Ⅳ期患者Hcy、DLL4和SUA水平明显高于Ⅰ-Ⅱ期患者(P<0.05);DR患者Hcy、DLL4和SUA与FBG、HbA1c和HOMA-IR水平均呈正相关(P<0.05)。结论:DR患者血清Hcy、DLL4和SUA水平升高,可能影响患者血糖、血脂代谢,并参与疾病进展。