Graph Regularized L_p Smooth Non-negative Matrix Factorization for Data Representation

This paper proposes a Graph regularized Lpsmooth non-negative matrix factorization(GSNMF) method by incorporating graph regularization and L_p smoothing constraint, which considers the intrinsic geometric information of a data set and produces smooth and stable solutions. The main contributions are as follows: first, graph regularization is added into NMF to discover the hidden semantics and simultaneously respect the intrinsic geometric structure information of a data set. Second,the Lpsmoothing constraint is incorporated into NMF to combine the merits of isotropic(L_2-norm) and anisotropic(L_1-norm)diffusion smoothing, and produces a smooth and more accurate solution to the optimization problem. Finally, the update rules and proof of convergence of GSNMF are given. Experiments on several data sets show that the proposed method outperforms related state-of-the-art methods.

Homocysteine-induced Enhanced Expression of Tissue Factor in Human Vascular Smooth Muscle Cells

The homocysteine （Hcy）-induced tissue factor （TF） expression in human vascular smooth muscle cells （VSMCs） and the effect of Hcy on the activity of nuclear factor-kappaB （NF-кB） and the expression of inducible nitric oxide synthase （iNOS） were investigated. Human umbilical artery VSMCs were cultured by tissue explanting method, identified by α-actin immunohistochemistry, and incubated with different concentrations of Hcy/PTDC （NF-кB inhibitor）. Semi-quantitative RT-PCR was performed to detect the expression of TF mRNA in VSMCs. Flow cytometry was used to assay the expression of TF protein on the surface of VSMCs and the expression of iNOS in VSMCs. Western blot was carried out to detect the expression of NF-кB protein in nuclei. The results showed that Hcy could induce VSMCs expressing TF mRNA significantly after the VSMCs were incubated with Hcy at concentrations of 10, 100, 500 μmol/L respectively. There was low expression level of TF protein on the surface of the resting VSMCs and Hcy could also induce VSMCs expressing TF pro- tein on the cell surface in different concentrations. Additionally, Hcy could rapidly induce the activation of NF-кB and this effect could be significantly inhibited by PDTC. Hcy alone could not induce the expression of iNOS in VSMCs. It was concluded that Hcy could significantly induce the expression of TF in VSMCs and enhance the activation of NF-ΚB, subsequently mediate TF gene expression and protein synthesis. NF-кB-mediated expression of TF in VSMCs might be the important mechanism of atherosclerosis and thrombosis induced by Hcy.

Construction of human eukaryotic expression plasmid vascular endothelial growth factor 165 and its expression in transfected vascular smooth muscles

BACKGROUND: The highly specific vascular endothelialgrowth factor (VEGF) induces the growth of vascular en-dothelial cell. This study was to construct the eukaryoticexpression plasmid of vascular endothelial growth factorl65(VEGF165) and observe its expression in vascular smoothmuscles (VSMCs).METHODS: The primers were designed and synthesizedaccording to the gene sequences of human VEGF165. TheVEGF165 gene was obtained from umbilic artery tissue bythe method of RT-PCR, then it was cloned to eukaryoticexpression plasmid pBudCE4.1 by recombination strategy.The eukaryotic expression plasmid named pBudCE4.1/VEGF165 was identified by restriction enzyme digestion,and was sequenced. The pBudCE4.1/VEGF165 was trans-fected into VSMCs by using lipofection. The VEGF165 ex-pression of mRNA and protein was detected by RT-PCRand Western blot respectively.RESULTS: VEGF165 was shown about 576bp by RT-PCR.Sequencing revealed the amplified VEGF165 gene was iden-tical with that in the GeneBank. Restrictive enzyme (HindBam HI) digestion analysis showed that recombinantexpression plasmid pBudCE4. l/tVEGF165 had been con-structed successfully. The expression of VEGF165 at mRNAand protein levels in the transformed VSMCs had beendemonstrated by RT-PCR and Western blot.CONCLUSIONS: The recombinant eukaryotic expressionplasmid pBudCE4.1/VEGF165 has been successfully con-structed and expressed in transformed VSMCs. The presentstudy has laid a foundation for VEGF165 gene therapy ofvascular stenosis in the transplant organ.

Transforming growth factor-β and smooth muscle differentiation

Transforming growth factor(TGF)-β family members are multifunctional cytokines regulating diverse cel- lular functions such as growth,adhesion,migration, apoptosis,and differentiation.TGF-βs elicit their effects via specific typeⅠand typeⅡserine/threonine kinase receptors and intracellular Smad transcription factors. Knockout mouse models for the different components of the TGF-β signaling pathway have revealed their critical roles in smooth muscle cell(SMC)differentia- tion.Genetic studies in humans have linked mutations in these signaling components to specific cardiovascular disorders such as aorta aneurysm and congenital heart diseases due to SMC defects.In this review,the current understanding of TGF-β function in SMC differentiation is highlighted,and the role of TGF-βsignaling in SMC- related diseases is discussed.

Partial Purification of Smooth Muscle Cell Derived Growth Factor

The serum free medium conditioned by cultured rabbit aortic smooth muscle cells was partially purified using ultrafiltration and heparin affinity chromatography. Incorporation of [￣3H]-thymidine (￣3H-TdR) into cell DNA was used to measure the mitogenic activity of the fractions from chromatography for NIH 3T3 fibroblasts. The molecular weight and the iso-electric point of these fractions were determined by NaDodSO_4-polyacrylamide gel electrophoresis (SDS-PAGE)and iso-electric focusing, respectively. The results showed that the protein eluted in 1.0-1. 6 mol/L NaCl from the heparin-Sepharose was mitogenic for 3T3 cells,and this protein had a molecular weight of 22. 8-26. 7 ku and an iso-electric point of about 4. 6. The fact that the above-mentioned biochemical properties differed from that of PDGF, IGF and FGF suggests that this mitogenic protein may be a separate growth factor.

Different distributions of interstitial cells of Cajal and platelet-derived growth factor receptor-α positive cells in colonic smooth muscle cell/interstitial cell of Cajal/plateletderived growth factor receptor-α positive cell syncytium in mice

AIM To investigate the distribution and function of interstitialcells of Cajal(ICCs) and platelet-derived growth factor receptor-α positive(PDGFRα+) cells in the proximal and distal colon.METHODS The comparison of colonic transit in the proximal and distal ends was performed by colonic migrating motor complexes(CMMCs). The tension of the colonic smooth muscle was examined by smooth muscle spontaneous contractile experiments with both ends of the smooth muscle strip tied with a silk thread. Intracellular recordings were used to assess electrical field stimulation(EFS)-induced inhibitory junction potentials(IJP) on the colonic smooth muscle. Western blot analysis was used to examine the expression levels of ICCs and PDGFRα in the colonic smooth muscle.RESULTS Treatment with NG-nitro-L-arginine methyl ester hydrochloride(L-NAME) significantly increased the CMMC frequency and spontaneous contractions, especially in the proximal colon, while treatment with MRS2500 increased only distal CMMC activity and smooth muscle contractions. Both CMMCs and spontaneous contractions were markedly inhibited by NPPB, especially in the proximal colon. Accordingly, CyPPA sharply inhibited the distal contraction of both CMMCs and spontaneous contractions. Additionally, the amplitude of stimulationinduced nitric oxide(NO)/ICC-dependent slow IJPs(sIJPs) by intracellular recordings from the smooth muscles in the proximal colon was larger than that in the distal colon, while the amplitude of electric field stimulationinduced purinergic/PDGFRα-dependent fast IJPs(fIJPs) in the distal colon was larger than that in the proximal colon. Consistently, protein expression levels of c-Kit and anoctamin-1(ANO1) in the proximal colon were much higher, while protein expression levels of PDGFRα and small conductance calcium-activated potassium channel 3(SK3) in the distal colon were much higher.CONCLUSION The ICCs are mainly distributed in the proximal colon and there are more PDGFRα+ cells are in the distal colon, which generates a pressure gradient between the two ends of the colon to propel the feces to the anus.

Effect of Nuclear Factor-kappa B on Vascular Endothelial Growth Factor mRNA Expression of Human Pulmonary Artery Smooth Muscle Cells in Hypoxia

In order to investigate the effect of nuclear factor kappa B (NF κB) on vascular endothelial growth factor (VEGF) mRNA expression of human pulmonary artery smooth muscle cells (HPASMCs) in hypoxia, the cultured HPASMCs in vitro were stimulated with pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF κB. The NF κB p65 nuclei positive expression was detected by immunocytochemical technique. The IκBα protein expression was measured by Western blot. RT PCR was used to detect the VEGF mRNA expression of HPASMCs. The results showed that no significant change was observed in the NF κB p65 nuclei positive expression of cultured HPASMCs during 6 h-24 h in normoxia, but the levels of NF κB p65 nuclei positive expression of cultured HPASMCs were significantly increased in hypoxia groups as compared with those in all normoxia groups ( P <0.05). The IκBα protein expression of cultured HPASMCs showed no significant change during 6 h-24 h in normoxia, but significantly decreased in hypoxia as comapred with that in normoxia groups ( P <0.05). PDTC (1 to 100 μmol/L) could inhibit the VEGF mRNA expression of HPASMCs in a concentration dependent manner in hypoxia. In conclusion, NF κB can be partly translocation activated from cytoplasm into nuclei in the cultured HPASMCs under hypoxia. The inhibition of NF κB activation can decrease the VEGF mRNA expression. It is suggested that the activation of NF κB is involved in the VEGF mRNA expression of HPASMCs under hypoxia.

Up and Down Expression of Androgen Receptor, Estrogen Receptor beta and Platelet Derived Growth Factor beta by Testosterone in Aortic Vascular Smooth Muscle Tissues

Objectives To investigate the effects of testosterone enanthate (TE) on serum lip- ids and lipoproteins metabolism and the expression of androgen receptor (AR) , estrogen receptor beta ( ER - β) and platelet derived growth factor beta ( PDGFR - β) in aortic vascular smooth muscle tissues (VSMTs). Methods Forty aged male rats were ran- domly divided into 4 groups, group A (placebo group) , group B (2. 5 mg/kg intramuscular injection of TE once a week ) , group C (5.0 mg/kg intramuscular injection of TE once a week ) , group D ( 10. 0 mg/kg intramus- cular injection of TE once a week). All animals were fed freely during 16 - week treatment periods. The ex- pression of AR ,ER - βand PDGFR - β were studied by Western bolt. Results Average serum LDL - C was lower in group D than that in group A ( p < 0. 01 ). Compared with the other groups, average serum TC was also lower in group D (p <0. 05). AR expression in aortic vascular smooth muscle tissues could be regulated by TE: 99.50 ±21.74, 125.38 ±28.68 and 101.98 ± 15.42 for TE concentrations at 2.5 mg/kg, 5.0 mg/kg and 10.0 mg/kg, respectively , the expression of ER - β could be regulated by TE: 92. 34 ± 18. 68, 47. 72 ± 18.12, 82.13 ±23.50, and the expression of PDGFR - β could be regulated as well by TE: 219.70 ±45. 59, 50.16 ±9. 72, 125.36 ±15. 74(Data for AR ,ER-β and PDGFR - β protein band intensity were expressed with x ± s, with control group taken as 100 ).Conclusions This study indicates that androgens have significant effects on serum lipids and lipoprotein metabolism. Testosterone enanthate at 5. 0 mg/kg can stimulate the expression of AR, but inhibite the expres- sion of PDGFR. Testosterone enanthate at the concen- trations of 5. 0 mg/kg and 10. 0 mg/kg can inhibite the expression of ER - β.

Interleukin-1β,Tumor Necrosis Factor-α and Lipopolysaccharide Induce Expression of Monocyte Chemoattractant Protein-1 in Calf Aortic Smooth Muscle Cells

Summary: To investigate whether interleukin-1β(IL-1β), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) induce expression of monocyte chemoattractant protein-1 (MCP-1 ) mRNA and protein in calf aortic smooth muscle cells(SMCs), calf aortic SMCs were cultured by a substrate-attached explant method. The cultured SMCs were used between the third to the fifth passage. After the cells became confluent, the SMCs were exposed to 2 ng/ml IL-1β, 20ng/ml TNF-1α and 100 ng/ml LPS respectively, and the total RNA of SMCs which were incubated for 4 h at 37℃ were extracted from the cells by using guanidinium isothiocyanate method. The expres- ion of MCP-1 mRNA in SMCs was detected by using dot blotting analysis using a probe of γ-32 P- end-labelled 35-mer oligonucleotide. After a 24-h incubation, the media conditioned by the cul- tured SMCs were collected. The MCP-1 protein content in the conditioned media was determined by using sandwich ELISA. The results were as follows: Dot blotting analysis showed that the cul- tured SMCs could express MCP-1 mRNA. After a 4-h exposure to IL-1β, TNF-α and LPS, the MCP-1 mRNA expression in SMCs was increased (3.6-fold, 2. 3-fold and 1. 6-fold, respectively). ELISA showed that the levels of MCP-1 protein in the conditioned media were also increased (2.9- fold, 1.7-fold and 1.1-fold, respectively). The results suggest that calf aortic SMCs could ex- press MCP-1 mRNA and protein. IL-1β and TNF-α can induce strong expression of MCP-1 mRNA and protein, and the former is more effective than the latter.

CXCL5通过诱导血管钙化参与颈动脉斑块的形成

背景:CXC基序趋化因子5(CXC-motif chemokine 5,CXCL5)为上皮细胞衍生的中性粒细胞激活肽,研究发现其可能参与动脉病变。然而,CXCL5在血管钙化中的作用未见报道。目的:探讨CXCL5在颈动脉粥样硬化的血管钙化中的作用。方法:①细胞实验:将小鼠血管平滑肌细胞分成以下各组:成骨培养基组,Vector组(空白质粒转染到细胞中),CXCL5组(CXCL5质粒转染到细胞中),si-NC组(CXCL5阴性对照siRNA转染到细胞中),si-CXCL5组(CXCL5 siRNA转染到细胞中),Vector+LY2157299组和CXCL5+LY2157299组(细胞转染24 h后,将转化生长因子β受体1激酶抑制剂LY2157299加入细胞中)。进行茜素红染色、碱性磷酸酶染色和钙含量测定以评估血管平滑肌细胞成骨分化水平。②动物实验:48只ApoE-/-小鼠随机分成4组:Con+si-NC组、Con+si-CXCL5组、CAS+si-NC组和CAS+si-CXCL5组,前2组不造模,尾静脉注射si-NC或si-CXCL5慢病毒;后2组制备颈动脉粥样硬化模型,尾静脉注射si-NC或si-CXCL5慢病毒。采用Von Kossa染色和免疫组织化学染色评估小鼠颈动脉血管钙化以及CXCL5、转化生长因子β受体1表达情况。结果与结论:①CXCL5组细胞Runt相关转录因子2蛋白水平上调、α-平滑肌肌动蛋白水平下调,si-CXCL5组中的发现与其相反;CXCL5过表达上调了转化生长因子β受体1水平,而CXCL5敲低抑制了转化生长因子β受体1水平。②与Vector组相比,CXCL5组细胞茜素红染色的强度、碱性磷酸酶活性和钙含量显著增加(P<0.05);与si-NC组相比,si-CXCL5组上述2项指标显著降低(P<0.05);当用LY2157299抑制转化生长因子β受体1表达时,CXCL5对平滑肌细胞的成骨转化作用减弱。③与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中CXCL5蛋白表达和血管钙化面积显著增加(P<0.05);与CAS+si-NC组相比,CAS+si-CXCL5组颈动脉中上述2项指标显著降低(P<0.05)。④与Con+si-NC组相比,CAS+si-NC组大鼠颈动脉中Runt相关转录因子2蛋白表达显著增加(P<0.05)和α-平滑肌肌动蛋白表达显著降低(P<0.05);与CAS+si-NC组相比,CAS+si-CXCL5组颈动脉中上述2项指标呈相反变化(P<0.05)。⑤结果说明,CXCL5通过激活转化生长因子β受体1通路诱导血管平滑肌细胞成骨样转化,抑制CXCL5表达对于改善颈动脉粥样硬化小鼠颈动脉血管钙化是有效的。

基于改进蚁群算法的机器人全局路径规划

针对传统蚁群算法存在初始信息素缺乏、收敛速度慢以及无法有效躲避障碍物等问题,文中提出了一种基于改进蚁群算法的全局路径规划。引入正态分布函数改进传统启发函数,提高了算法效率,缩短了算法收敛所需时间。自适应调整信息素挥发系数,限定信息素范围,避免过早收敛。对算法路径平滑处理,缩短路径长度,从而实现机器人的全局路径规划。仿真结果表明,在20×20环境下,文中算法平均迭代次数比传统蚁群算法减少了28代,收敛速度更快。平均拐点减少了33.3%,使路径更为平滑,克服了初始信息素缺乏,加快了收敛速度,减少了拐点数量,能够有效躲避环境中的障碍物,证明了该算法的可行性。

轻度慢性阻塞性肺疾病的病理特征及其与炎性因子的相关性

目的 探索轻度慢性阻塞性肺疾病(COPD)患者肺组织病理特征及其与炎性因子的相关性。方法 前瞻性纳入因肺小结节行手术治疗的患者70例,分为正常组10例、轻度COPD组50例、中重度COPD组10例。肺组织分别经HE、Masson、EVG染色后评估病理改变;免疫组化染色检测肺组织平滑肌肌动蛋白(SMA)、肌动蛋白(Actin)、CD31蛋白表达;免疫组化及q PCR检测肺组织肿瘤坏死因子(TNF)-α、白细胞介素(IL)-10蛋白及mRNA水平。结果 轻度COPD肺组织可见肺泡间隔增宽,小气道扩张,血管壁轻度增厚,多见以淋巴细胞为主的炎症反应。免疫组化染色提示轻度COPD肺组织中SMA、Actin蛋白含量较正常组升高(P<0.05)。此外,轻度COPD肺组织中TNF-α的mRNA水平及TNF-α阳性率明显高于正常组,而IL-10的mRNA水平明显低于正常组(均P<0.05)。SMA、Actin与炎性因子TNF-α阳性表达呈正相关,而与IL-10阳性表达呈负相关(均P<0.05)。结论 轻度COPD肺组织主要病理改变包括以小血管平滑肌层增厚为特征的肺小血管重构和以淋巴细胞浸润为主的炎症反应,同时促炎因子TNF-α升高和抗炎因子IL-10降低与COPD病理改变相关。

抗差Vondrak滤波方法在时间频率传递中的应用

时间频率传递的结果会受到非模型化误差和观测噪声的影响,其噪声常为高频信号,构建低通滤波器可在一定程度上消除观测值序列中的高频噪声信号.本文对Vondrak滤波函数的本质进行剖析,通过IGG3算法对钟差序列进行定权并采用频率响应法选择适合的滤波因子;对不同的链路分别进行卫星双向时间频率传递(two-way satellite time and frequency transfer,TWSTFT)、基于软件接收机的卫星双向时间传递(two-way satellite time and frequency transfer based on software defined receiver,SDR-TWSTFT)和短基线共视时间频率传递实验,并对钟差结果采用抗差Vondrak滤波进行平滑去噪.结果表明:滤波后的钟差序列能够很好地反映原始钟差序列的趋势;平滑后的TWSTFT钟差结果,日波动效应得到了有效的抑制,精度有明显提升;对于共视钟差结果,精度有明显提升,与精密单点定位(precise point positioning,PPP)时间传递结果的差值保持在−1.0~1.0 ns范围内.

沉默TUFM通过AMPK/mTOR信号通路调控线粒体自噬对肺源性心脏病模型大鼠肺动脉高压的影响

目的探讨线粒体翻译延伸因子Tu(TUFM)通过线粒体自噬促进肺动脉高压(PAH)血管重塑的作用机制。方法2022年1月—2023年6月于辽宁省人民医院中心实验室进行实验。将36只健康雄性Sprague-Dawley大鼠随机分为空白对照(Ctrl)组、模型(PAH)组、TUFM过表达(OE)组、OE阴性对照(OE-NC)组、短发夹RNA(Sh)敲除TUFM(Sh)组和Sh-NC阴性对照(Sh-NC)组,每组6只。除Ctrl组外,其余大鼠均一次性腹腔注射1%野百合碱(60 mg/kg)诱导心源性肺水肿PAH大鼠模型;大鼠肺动脉平滑肌细胞(PASMC)在低氧(3%O 2)条件下培养24 h模拟体内肺动脉高压微环境,分为常氧(Norm)组、低氧(Hyp)组、小干扰RNA(SiRNA)-1组、SiRNA-2组、Si-NC组、OE-NC组和OE组。右心导管插管和脉冲多普勒超声检测大鼠肺血流动力学;苏木素-伊红染色检测肺小动脉病理结构;免疫荧光共染检测TUFM组织定位;细胞计数法检测细胞增殖;透射电镜观察线粒体结构和自噬小体;蛋白免疫印迹检测TUFM、自噬、凋亡和磷酸腺苷活化蛋白激酶(AMPK)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白表达。结果与Ctrl组比较,PAH组大鼠TUFM蛋白表达升高,且主要与PASMC标志物α平滑肌肌动蛋白(α-SMA)在肺小动脉内膜存在共定位,而与内皮细胞标志物CD31无共定位,肺动脉收缩压(PASP)升高,肺动脉血流加速时间(PAAT)缩短,远端肺小动脉管壁呈向心性增厚,管腔狭窄几乎堵塞,TUFM、苄氯素1重组蛋白(BECN1)、人微管相关蛋白轻链3(LC3)II/I和B淋巴细胞瘤2(Bcl2)蛋白表达升高,P62、Bcl2相关X蛋白(Bax)和凋亡酶激活因子(Apaf)蛋白表达降低(P<0.05);与PAH组比较,OE组PASP升高,PAAT缩短,肺小动脉管壁厚度升高,肺动脉TUFM、BECN1、LC3II/I和Bcl2表达升高,P62、Bax和Apaf表达降低(P<0.05);与PAH组比较,Sh组PASP降低,PAAT延长,肺小动脉管壁厚度和管腔狭窄度有所改善,TUFM、BECN1、LC3II/I和Bcl2表达降低,P62、Bax和Apaf表达升高(P<0.05)。与Norm组比较,Hyp组PASMC细胞TUFM蛋白表达升高;与Si-NC组细胞相比,SiRNA-1和SiRNA-2组P62、Bax蛋白表达升高,BECN1、LC3II/I、Bcl2、TUFM表达降低,线粒体结构完整,PASMC细胞增殖活性降低,细胞p-AMPK表达降低,p-mTOR表达升高(P<0.05);与OE-NC组比较,OE组细胞P62和Bax蛋白表达降低,BECN1、LC3II/I、Bcl2和TUFM表达升高,部分线粒体损伤崩解,嵴断裂消失,PASMC细胞增殖活性明显升高,细胞p-AMPK表达升高,p-mTOR表达降低(P<0.05)。结论沉默TUFM可通过激活AMPK/mTOR信号通路促进线粒体自噬加速PAH肺动脉平滑肌细胞凋亡。

光伏组件输出特性的异常分类研究

光伏组件异常直接影响到发电量与组件的使用寿命,及时发现光伏组件异常并消除异常,将直接提高光伏组件发电效率。为准确检测光伏组件的异常状态,以Matlab平台为基础,模拟6种光伏组件异常状态,并分析输出特性,提出以输出特性曲线的五特征法判断异常类型。用Python语言搭建概率神经网络,并用粒子群优化算法优化平滑因子。用粒子群优化概率神经网络(PSO-PNN)模型训练异常数据,结果表明:概率神经网络对数据集较敏感,在较大的数据集,模型分类准确率高,能有效检测光伏组件异常状态。

基于4阶Bézier曲线的路径平滑方法研究

路径平滑技术旨在采用以曲代直的方法解决路径规划中存在的尖角问题,实现路径的平滑化,以满足外形设计和运动规划等工程应用需求。文中在深入探讨了Bézier曲线在路径平滑领域的应用优势及其在路径拟合中产生的较大偏差后,提出了一种基于内外比例因子的4阶Bézier曲线路径平滑新方法。通过严谨的理论分析,证明了该方法能够有效控制平滑误差,在不考虑路径节点坐标的情况下,误差上限为4.25,从而实现对原始路径的高精度平滑近似。在无人机路径平滑的仿真实验中,该方法耗时0.0061 s,证明了其在平滑处理速度方面的优越性。综合理论分析与实验结果,该方法不仅在平滑效果上表现出色,而且在处理速度上也具有明显优势。同时由于仅涉及内外比例因子两个自由参数,因此,操作简单,易于实现,具有较强的适用性和灵活性。

缺氧诱导SD大鼠阴茎海绵体平滑肌细胞内炎症反应及番茄红素的干预作用

目的:探索缺氧对SD大鼠海绵体平滑肌细胞(CCSMCs)内炎症反应的诱导作用及番茄红素调节效应。方法:分离SD大鼠原代CCSMCs并连续观察第4代海绵体平滑肌细胞内的形态稳定性;为了验证所培养的CCSMCs的纯度,进行了α-平滑肌肌动蛋白(α-SMA)和结蛋白(Desmin)的免疫荧光染色;不同浓度的CoCl_(2)和番茄红素单独或联合干预海绵体平滑肌细胞24 h后,使用细胞计数试剂盒-8(CCK8)检测细胞活力;设立3个细胞处理组别:对照组、缺氧组(CoCl_(2)200μmol/L)和番茄红素治疗组(40μmol/L);培养24 h后,应用蛋白质印迹法(Western blotting)和反转录定量聚合酶链式反应(RT-qPCR)检测缺氧诱导因子1α(HIF-1α)、肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6、IL-18、IL-1β水平。结果:提取的SD大鼠海绵体平滑肌细胞在培养至第4代时依旧保持了形态上的均一性,经α-SMA和Desmin染色均显示出纯度较高的阳性反应。200μmol/LCoCl_(2)处理后CCSMCs的活力下降到50%,且Western blotting和RT-qPCR检测细胞内HIF-1α、TNF-α、IL-6、IL-18的水平均呈上升趋势(P<0.05),IL-1β的mRNA表达亦显著升高(P<0.05),但蛋白表达量仅有升高趋势(P>0.05);40μmol/L番茄红素能将CCSMCs的活力得以提升至正常状态的80%,同时显著降低HIF-1α、TNF-α、IL-18的蛋白表达量和TNF-α、IL-18、IL-1β的mRNA表达量(P<0.05),IL-6的蛋白、mRNA及IL-1β的蛋白表达量仅有下降趋势(P>0.05)。结论:缺氧条件下,CCSMCs中炎症因子表达上调,番茄红素能有效抑制炎症因子表达。

基于蚁群算法的物资运送小车路径规划研究

针对路径规划蚁群算法的盲目性、收敛速度慢、路径较长和路径折点多等问题,提出了一种改进蚁群路径规划算法。首先通过改进启发信息的数学模型,限制轮盘赌在8个方向的选择概率,降低迭代次数;然后建立自适应更新影响因子,通过实时监测目标点位置,进一步提高路径的选择方式和算法的鲁棒性;最后通过路径二次寻优,对改进蚁群路径规划算法形成的最优路径进一步消除冗余节点,在已知最优路径进一步寻优,从而提高路径平滑度、减少路径折点,以及缩短路径长度,提高物资运送小车的使用效率。通过栅格环境地图中障碍物不同占比的仿真试验,验证了所提出的改进蚁群算法的迭代速率更快、寻优能力更强、鲁棒性更好和路径更短。

骨形态发生蛋白2介导同型半胱氨酸促进血管钙化

背景:高同型半胱氨酸血症与血管钙化之间存在一定的内部联系。但是,目前有关高同型半胱氨酸血症促进血管钙化的发病机制还不清楚。目的:探讨骨形态发生蛋白2在高同型半胱氨酸血症诱导血管钙化中的作用。方法:人颈动脉蜡块标本根据有无钙化斑块分为钙化组(n=29)与非钙化组(n=13)。16只ApoE-/-小鼠随机分为对照组和高同型半胱氨酸血症组,每组8只。骨形态发生蛋白2转染大鼠胸大动脉平滑肌A7r5细胞,然后应用梯度浓度同型半胱氨酸(50,100,200,400μmol/L)干预A7r5细胞。茜素红染色和苏木精-伊红染色法检测钙化反应,免疫荧光法检测骨形态发生蛋白2与Runt相关转录因子2相互作用关系,免疫组化法和Western blot检测骨形态发生蛋白2、Runt相关转录因子2、α-平滑肌肌动蛋白的表达。结果与结论:①人体颈动脉组织染色显示,与非钙化组比较,钙化组炎性细胞增多,钙化阳性率增加(P<0.05);与非钙化组比较,钙化组骨形态发生蛋白2和Runt相关转录因子2表达量均上调,α-平滑肌肌动蛋白表达量下降(P均<0.05);②小鼠动脉标本染色显示,高同型半胱氨酸血症组钙化面积阳性率高于对照组(P<0.05);高同型半胱氨酸血症组血清同型半胱氨酸水平明显高于对照组(P<0.05);与对照组比较,高同型半胱氨酸血症组骨形态发生蛋白2和Runt相关转录因子2表达量均上调,α-平滑肌肌动蛋白表达量下降(P均<0.05);③A7r5细胞培养分析,随着同型半胱氨酸浓度梯度增加,A7r5细胞钙化程度、骨形态发生蛋白2和Runt相关转录因子2的蛋白表达增加(P<0.05),α-平滑肌肌动蛋白的蛋白表达降低(P<0.05);④过表达骨形态发生蛋白2的A7r5细胞培养分析,骨形态发生蛋白2过表达组较相对应的对照组、β-甘油磷酸钠组和同型半胱氨酸组钙化程度增加;Runt相关转录因子2表达上调(P<0.05),α-平滑肌肌动蛋白表达下调(P<0.05);⑤同型半胱氨酸钙化培养基培养的骨形态发生蛋白2过表达A7r5细胞中骨形态发生蛋白2表达量增加,且Runt相关转录因子2表达量随骨形态发生蛋白2表达量增加而增加;⑥结果表明,骨形态发生蛋白2是高同型半胱氨酸血症调控平滑肌细胞表型转化导致血管钙化的关键靶基因。靶向调控骨形态发生蛋白2可降低高同型半胱氨酸诱导的血管钙化。

Nrf2调控的氧化应激在Adropin抑制低氧肺动脉平滑肌细胞增殖中的作用研究

目的 观察Adropin对低氧诱导的SD大鼠肺动脉平滑肌细胞(PASMCs)增殖及氧化应激的影响,并探讨其可能机制。方法 以1%的O_(2)作为诱导剂,建立PASMCs增殖及氧化应激的细胞模型,通过活性氧(ROS)激活剂H_(2)O_(2),抗氧化剂N-乙酰半胱氨酸(NAC)及不同浓度Adropin(100、300、1000 nmol·L^(-1))干预24 h后,检测细胞增殖及ROS,筛选Adropin抑制增殖及氧化应激的最适浓度。选择1000 nmol·L^(-1) Adropin和/或核因子E2相关因子2(Nfr2)抑制剂ML385在低氧条件下孵育PASMCs 24 h,并设置Nrf2激活剂富马酸二甲酯(DMF)作为阳性对照组,通过CCK-8试剂盒检测细胞增殖;DCFH-DA探针联合荧光酶标仪测定细胞内ROS的水平;试剂盒检测细胞内超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)、过氧化氢酶(CAT)、丙二醛(MDA)的水平;流式细胞仪检测细胞周期;Western blot检测Nrf2、细胞周期蛋白(Cyclin)D1、Cyclin E的表达。结果 与对照组相比较,H_(2)O_(2)及低氧均能够诱导PASMCs增殖及ROS生成,而不同浓度的Adropin(100、300、1000nmol·L^(-1))和NAC均可抑制低氧诱导的PASMCs增殖及ROS产生,且Adropin抑制增殖及ROS产生具有浓度依赖性。与低氧组相比较,DMF或Adropin(1000 nmol·L^(-1))干预后,PASMCs增殖及细胞内ROS水平下降,SOD、GPx、CAT活性增加,MDA水平下降,Nrf2表达上调(P<0.05),而ML385能够逆转Adropin的上述作用(P<0.05);DMF或Adropin能够通过抑制Cyclin D1与Cyclin E的表达,阻滞细胞周期于G_(0)/G_(1)期,从而抑制PASMCs增殖(P<0.05),而ML385能够逆转Adropin上述作用(P<0.05)。结论 Adropin可通过抑制氧化应激抑制低氧诱导的PASMCs增殖,其机制可能与激活Nrf2有关。