VitD3致大鼠主动脉及心肌损害的超微结构观察

本实验对Vit D_3致大鼠主动脉、心肌损害分别作了SEM、EM观察。结果表明:大剂量VitD_3使主动脉膜内皮细胞萎缩、细胞间隙增宽、局部形成虫蚀状、火山喷口状外观;心肌细胞、主动脉平滑肌细胞浆、内质网、线粒体及主动脉弹力板内出现高电子密度钙盐颗粒,部分细胞变性、坏死。本实验提示:大量VitD_3引起的心肌、主动脉损害主要Ca~(++)超负荷有关,内质网、线粒体一般最先受累。     

Identification of abnormally methylated–differentially expressed genes and pathways in osteoarthritis: a comprehensive bioinformatic study

Clinical Rheumatology - To investigate abnormally methylated–differentially expressed genes (DEGs) and their related pathways in osteoarthritis (OA) by comprehensive bioinformatic analysis....     

抑制miR-30a/HMGA2介导的骨肉瘤细胞自噬对化学药物治疗诱导的细胞凋亡的作用

目的：探讨微小RNA(microRNA,miR)-30a/高迁移率族蛋白A2(high mobility group protein A2,HMGA2)介导的骨肉瘤细胞自噬对化学药物治疗(以下简称化疗)药物诱导的细胞凋亡的影响。方法：随机选取30例经化疗药物治疗后表现为化疗敏感和化疗抵抗的骨肉瘤患者组织,分为化疗敏感组(n=15)和化疗抵抗组(n=15),采用real-time PCR检测两组中miR-30a和HMGA2的mRNA表达水平,以及骨肉瘤细胞U2-OS经不同浓度的化疗药物(顺铂、阿霉素、氨甲蝶呤)处理后miR-30a的mRNA表达水平;采用蛋白质印迹法检测细胞内自噬相关因子Beclin 1,自噬微管相关蛋白1轻链3B(microtubule associated protein 1 light chain 3B,LC3B)、自噬抑制因子P62的表达情况。在骨肉瘤细胞U2-OS中转染miR-30a模拟物和抑制剂,构建miR-30a高表达组、低表达组和对照组,采用蛋白质印迹法检测经过顺铂和阿霉素处理后上述3组细胞内Beclin 1,LC3B和P62的表达情况;采用单丹磺酰尸胺(monodansylcada,MDC)染色法检测细胞内自噬水平,ROS荧光探针-二氢乙啶(dihydroethidium,DHE)检测细胞内ROS水平,细胞计数试剂盒8(cell counting kit-8,CCK-8)检测细胞存活率,流式细胞术检测细胞凋亡程度,线粒体膜电位荧光探针JC-1检测细胞线粒体氧化损伤程度;采用双荧光素酶法检测miR-30a与HMGA2的相互作用,同时通过转染HMGA2模拟物和HMGA2-shRNA干扰质粒载体,构建HMGA2高表达组、低表达组和对照组,采用蛋白质印迹法检测经过顺铂和阿霉素处理后上述3组细胞内Beclin 1,LC3B和P62的表达情况。结果：化疗抵抗组中miR-30a的mRNA水平显著低于化疗敏感组(P<0.05),HMGA2的表达与miR-30a相反(均P<0.05)。在相对低浓度(5 μml/L)的化疗药物刺激下,骨肉瘤细胞U2-OS内的miR-30a mRNA表达下调,Beclin 1和LC3B均显著上调(均P<0.01),P62显著下调(P<0.01)。在miR-30a高表达组中,与对照组相比,Beclin 1和LC3B的表达水平与miR-30a明显下降(P<0.05),P62的表达水平明显升高(P<0.05),在miR-30a低表达组中则相反;在经过化疗药物处理后的miR-30a高表达组中,细胞自噬水平更低,细胞存活率更低,ROS水平更高,线粒体氧化损伤程度更高,细胞凋亡水平更高(均P<0.05),miR-30a低表达组则相反(均P<0.05)。双荧光素酶报告基因检测验证了miR-30a与HMGA2的靶向互补配对关系;与对照组相比,HMGA2高表达组中Beclin 1和LC3B的表达水平与HMGA2明显升高(P<0.05),P62的表达水平明显下降(P<0.05),在HMGA2低表达组中则相反。结论：积极发挥miR-30a/HMGA2抑制骨肉瘤细胞自噬的功能,能够破坏细胞应对ROS介导的自噬与凋亡的平衡,增强化疗药物对细胞的杀伤作用。     

NIR-triggered upconversion nanoparticles@thermo-sensitive liposome hybrid theranostic nanoplatform for controlled drug delivery

Posterior segment ocular diseases are highly prevalent worldwide due to the lack of suitable noninvasive diagnostic and therapeutic tactics. Herein, concerning this predicament, we designed a hybrid retina-targeted photothermal theranostic nanoplatform (UCNPs@Bi@SiO₂@GE HP-lips), based on the unique upconversion luminescence (UCL) imaging of upconversion nanoparticles (UCNPs), efficient photothermal conversion ability of Bi nanoparticles, and thermal-induced phase transition properties of the liposomes (lips). The nanoplatform was functionalized with penetratin (PNT) and hyaluronic acid (HA), to obtain retina-targeted liposomes (HP-lips). Lipophilic genistein (GE) was entrapped into the liposomes (GE HP-lips). An in vitro release study showed NIR irradiation could photothermally trigger controlled release of GE from the liposomal platform. Moreover, cellular uptake evaluation via UCL imaging demonstrated UCNPs@Bi@SiO₂@GE HP-lips represented the brightest UCL, compared with other formulations, which is beneficial for the accurate evaluation of the prognosis and severity of angiogenesis-related posterior segment disorders. Therefore, UCNPs@Bi@SiO₂@GE HP-lips exhibit promising potential as a theranostic nanoplatform for posterior segment ocular diseases.

A novel hybrid photothermal theranostic nanoplatform UCNPs@Bi@SiO₂@GE HP-lips is developed. Upon NIR irradiation, the nanoplatform could photothermally trigger controlled drug release and present bright upconversion luminescence.

Posterior segment ocular diseases, such as age-related macular degeneration (AMD) and diabetic retinopathy (DR), have drawn considerable attention worldwide. Both AMD and DR are highly prevalent and have been proven to be the leading causes of blindness in the developed nations, resulting in a heavy social burden.^1,2 It is common sense that neovascularization in the posterior segment, including choroidal neovascularization (CNV) and retinal neovascularization (RNV), is the primary cause of severe visual impairment.³ Research has shown that suitable therapeutic tactics, as well as timely and precise diagnostic avenues, are highly crucial to the prevention and/or treatment of angiogenesis-related ocular disorders. Therefore, it is high time to develop proper platforms with desirable diagnostic and therapeutic abilities for angiogenesis-related ocular disorders.For accurate evaluation of the prognosis and severity of angiogenesis-related posterior segment disorders, convenient and efficient imaging techniques of angiogenesis are the prerequisites. Diverse nanoprobes for bioimaging are one of the best fruits of the quick development of nanotechnology.^4–7 Many organic imaging agents have been employed in the field of bioimaging.^8–10 Besides organic bioimaging agents, inorganic lanthanide (Ln³⁺)-doped upconversion nanoparticles (UCNPs), which can convert low-energy NIR light into higher-energy UV/visible/NIR light, have attractive superiorities over conventional organic dyes and quantum dots, such as high signal-to-noise ratio, low auto-fluorescence background, narrow emission peak width, superb stability, and absence of photodamage to live organisms.^7,11,12 Accordingly, UCNPs have been broadly investigated as emerging luminescent nanoprobes for real-time bioimaging and drug release tracking in recent decades.^13,14 Especially, NIR light shows an adequate penetration depth in deep tissue, which promotes in vivo bioimaging application of UCNPs.¹⁵ Additionally, UCNPs have been widely employed as a building block to construct multimodal imaging nanoplatforms to enhance their imaging precision or equipped with therapeutic functionality by surface functionalization, designing hierarchical nanostructures with other functional nanoparticles and introducing diversiform doped ions.^16,17 Rieffel et al. have designed porphyrin–phospholipid-coated UCNPs, realizing at least six imaging modalities, including upconversion luminescence imaging, fluorescence imaging, positron emission tomography, computed tomography, photoacoustic imaging, and Cerenkov luminescence imaging.¹⁸ To integrate the photothermal function into UCNPs, diverse UCNPs-based nanocomposites with different nanostructure have been developed, such as UCNPs-Au,¹⁹ UCNPs-CuS,²⁰ UCNPs-Cu₂S,²¹ UCNPs-Ag₂S,²² and UCNPs-Bi₂Se₃.¹⁶ These nanocomposites structurally combining UCNPs with photothermal agents would not only achieve real-time monitoring of in vivo agent distribution, but can also endow UCNPs with photothermal therapeutic ability. Numerous studies of UCNPs-based nanoplatforms for diagnosis and therapy of tumors present an undeniable interest in the biomedical application of UCNPs.^5,23–25 Very recently, UCNPs have been utilized in the field of ophthalmic application. Ma et al. designed retinal photoreceptor-binding upconversion nanoparticles (pbUCNPs) and endowed the mice with NIR light image vision while their normal vision and related behavioral responses were not compromised.²⁶ Microenvironment-triggered degradable hydrogels were designed by Li et al. using ultrasmall UCNPs, and exhibited its potential application in non-invasive NIR-II imaging diagnosis and combined therapy in human choroidal melanoma.²⁷Current therapeutic strategies of angiogenesis-related ocular disorders are far from satisfactory. It is universally acknowledged that intraocular injection of anti-vascular endothelial growth factor (VEGF) agents has been broadly accepted as a “gold standard” treatment for angiogenesis-related ocular diseases and in a way, can ameliorate the visual impairment of DR and AMD patients.²⁸ Nonetheless, repeated invasive injections enhance the risk of complications, compromise patient compliance, and aggravate the burden of healthcare.²⁹ Topical instillation of eye drops is the most common and accessible route to treat ocular diseases due to its convenient and painless properties, however, multiple ocular barriers (including cornea, conjunctiva, sclera, blood-retinal barrier, and blood-aqueous barrier) result in a negligible permeated amount of drug to the posterior segment by topical administration.² Recently, photo-driven therapy, such as photodynamic therapy (PDT) and photothermal therapy (PTT), have been widely used in biomedicine and many organic components play key roles.^30,31 Additionally, light has been widely employed as a non-invasive external stimulus for remote spatiotemporal controlled drug release.^7,32 Light-triggered drug delivery platforms exhibit attractive advantages, including low cost, outstanding therapeutic effect and little damage to physiological tissue, and hence are suitable for non-invasive ocular drug delivery, in terms of intrinsic anatomical and biological constraints of eyes. It is reported that cell penetrating peptides (CPPs), which can promote the transport of various cargos across the cell membrane, have evoked considerable interest in pharmaceutical applications.^33,34 Among the peptides, penetratin (PNT), a cationic and amphipathic CPP, was demonstrated to have excellent penetration through multiple physiological barriers in the eyes, promoting drug transport to the posterior segment.^35,36 Angiogenesis-related posterior segment diseases can result in the overexpression of cluster of differentiation 44 (CD44) in the retina.^37,38 As we all know, hyaluronic acid (HA) is a specific ligand of the CD44 receptor. Accordingly, HA can be applied as a retina-targeted ligand for posterior ocular drug delivery. In short, PNT and HA are ideal constituent parts to design retina-targeted theranostic nanoplatforms towards diagnosis and therapy for angiogenesis-related posterior ocular diseases.Herein, we designed a hybrid photothermal stimuli-responsive theranostic nanoplatform, UCNPs@Bi@SiO₂@GE HP-lips with the retina-targeted ability (Fig. 1), to realize the combined diagnosis and therapy for angiogenesis-related posterior diseases. Genistein (GE), an anti-angiogenic agent that could suppress CNV formation, was selected as the model drug.³⁹In vitro release study revealed UCNPs@Bi@SiO₂ in the aqueous core of the liposomal platform could generate mild heat upon NIR irradiation, which facilitated phase transition of thermo-sensitive phospholipid DPPC, enhanced the fluidity of lipid layer, and hence triggered the release of GE from the liposomal platform. The admirable cytocompatibility of the nanocomposites with ARPE-19 cells was demonstrated by CCK-8 assay and live/dead cell staining. Moreover, UCL imaging proved cellular uptake of the nanocomposites by ARPE-19 cells was time-dependent and UCNPs@Bi@SiO₂@HP-lips showed the brightest UCL, compared with other formulations. Therefore, UCNPs@Bi@SiO₂@GE HP-lips could be a potential theranostic nanoplatform for posterior segment diseases.Open in a separate windowFig. 1Schematic diagram of UCNPs@Bi@SiO₂@GE HP-lips for NIR-triggered drug release. (A) The fabrication procedure of UCNPs@Bi@SiO₂@GE HP-lips. As proposed, hydrophilic UCNPs@Bi@SiO₂ and lipophilic drug GE were loaded in the aqueous core and lipid bilayer of the liposome frameworks, respectively. Cell penetrating peptide PNT and retina-targeted ligand HA were conjugated by amidation reaction in the preformed liposome bilayer. (B) Upon NIR irradiation, UCNPs@Bi@SiO₂ in the aqueous core of the liposomal platform could generate mild heat, which realized real-time green UCL monitoring and photothermally triggered drug release.     

手术治疗急性出血性胰腺坏死24例疗效观察

手术治疗急性胰腺炎疗效显著,施术者要尽量保存有活力的胰腺组织,既要清除胰腺及胰腺外的坏死组织,又要尽量减少术中、术后出血,要保证术后腹膜后坏死组织和渗出能最大限度地排出及引流.     

姜黄素对鼻咽癌细胞RECK基因甲基化以及MMP-9表达与活性的影响

目的探讨姜黄素对鼻咽癌细胞回复引导半胱氨酸丰富蛋白含kazal基元（ RECK）基因甲基化以及基质金属蛋白酶9（MMP-9）表达的影响。方法体外培养鼻咽癌细胞系CNE-1,用1、10、30μmol/L姜黄素处理后,采用Western blot和实时定量PCR分别检测RECK、MMP-9蛋白和mRNA表达;高效液相色谱-电喷雾质谱检测RECK甲基化;MTT法检测CNE-1细胞的增殖。同时采用明胶酶谱实验观察姜黄素处理前后MMP-9酶活性变化。结果 CNE-1细胞未刺激时RECK表达水平较低,而经1--30μmol/L姜黄素处理后,能显著增强RECK蛋白和mRNA的表达。高效液相色谱-电喷雾质谱结果显示,30μmol/L姜黄素处理后,RECK启动子、全基因组以及细胞核内甲基化水平分别降低为（69.04±10.62）%、（61.13±7.08）%、2.80±1.32。同时,姜黄素能显著降低CNE-1细胞中MMP-9蛋白和mRNA表达以及MMP-9的酶活性。结论姜黄素可能通过能上调RECK基因表达,降低细胞内甲基化水平,从而抑制MMP-9的表达与活性而发挥对CNE-1细胞生长抑制作用。     

无创机械通气治疗对老年COPD伴呼吸衰竭患者α-HBDH、β-EP、MMP-9及TIMP-1的影响

 目的 探讨无创机械通气治疗对老年慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)伴呼吸衰竭患者α-羟丁酸脱氢酶(α-hydroxybutyrate dehydrogenase,αHBDH)、β-内啡肽(β-endorphin,β-EP)、金属蛋白酶9(matrix metalloprotein-9,MMP-9)及组织抑制因子-1(tissueinhibitor of metallo proteinase-1,TIMP-1)的影响。方法 选择76例老年COPD伴呼吸衰竭患者为研究对象,随机分为两组。对照组予以噻托溴铵治疗,实验组在对照组基础上予以无创机械通气治疗,对比两组治疗前后α-HBDH、β-EP、MMP-9及TIMP-1 的改善情况。结果 治疗前,两组患者的各项指标差异均无统计学意义;治疗后,实验组的αHBDH、β-EP、MMP-9、TIMP-1 水平均低于对照组,差异有统计学意义(P＜0.05);实验组6 min步行距离较对照组延长,圣乔治呼吸问卷评分较对照组降低,差异有统计学意义(P＜0.05)。结论 无创机械通气治疗老年COPD伴呼吸衰竭可有效降低αHBDH、β-EP、MMP-9及TIMP-1水平,促进病情缓解。     

五味子醇甲药理研究进展

目的 探讨五味子醇甲的药理研究进展.方法 计算机检索CNKI、Pubmed等数据库,收集近几年国内外对五味子醇甲的药理研究进行综述.背景五味子在临床的广泛应用.五味子是一种常用的补益药,为木兰科植物五味子的成熟果实,其作为一味传统中药在临床上得到广泛的应用,其具有补益心肾,宁心安神的作用.而五味子醇甲是五味子的主要成分...     

从“治未病”角度探析儿童骨质疏松的中医防治思路

文章从“治未病”角度探讨儿童骨质疏松的中医防治思路,对“未病先防”“饮食有节”“既病防变”三个方面进行剖析,提出了治未病思想在儿童骨质疏松中运用的建议。     

治疗前CT影像组学结合机器学习预测非小细胞肺癌患者EGFR突变亚型

目的探讨基于治疗前胸部平扫CT影像组学特征和临床特征结合机器学习算法预测非小细胞肺癌(NSCLC)患者表皮生长因子受体(EGFR)突变状态和突变亚型(19Del/21L858R)的可行性和价值。方法回顾性分析南华大学附属第一医院和附属第二医院经活检病理证实和接受EGFR基因检测的280例NSCLC患者的治疗前胸部平扫CT和临床特征数据, 其中EFGR突变患者为136例。由两位高年资影像和肿瘤医师勾画原发肺部大体肿瘤区域(GTV), 然后提取851个影像组学特征, 采用Spearman相关分析和RELIEFF算法筛选具有预测性的特征, 两家医院分别为训练组和验证组。经特征选择的影像组学特征和临床特征构建临床-影像组学模型, 并与单独采用影像组学特征和临床特征模型进行比较。采用序贯建模流程, 使用支持向量机(SVM)建立机器学习模型预测EGFR突变状态和突变亚型。受试者工作曲线下面积(AUC-ROC)评估预测模型的诊断效能。结果经特征筛选各有21个影像组学特征在预测EGFR突变和突变亚型时具有预测效能并用于建立影像组学模型。临床-影像组学模型表现出最好的预测效能, 预测EGFR突变状态的模...