针刺对脑梗死大鼠缺血脑组织血管新生相关因子表达的影响

目的:动态研究脑梗死后血管新生相关因子的变化规律和针刺干预效应,从促进血管新生角度深入揭示针刺治疗脑梗死的分子机制。方法:以MCAO模型大鼠为研究对象,分为针刺组、模型组和空白组。观察脑梗死后神经功能缺损程度改变,同时采用免疫荧光双标记法观察缺血中心及边缘区的血管新生情况,应用RT-PCR和Western blot法检测脑梗死后缺穴区域血管新生相关因子Ang-1、Ang-2、PDGF-B和bFGF mRNA及蛋白变化规律和电针干预效应。结果:电针组各时相NSS评分均小于同时相模型组。MCAO后9h~7d时电针组血管内皮细胞增殖高于同时相模型组。电针组Ang-1、Ang-2 mRNA及蛋白表达水平明显高于模型组。电针组PDGF-B mRNA在3h~6h、3d~12d时明显高于模型组,蛋白水平在6h、3d~12d时明显高于模型组。针刺组bFGF mRNA在24h~12d、蛋白水平在3d~12d时明显高于模型组。结论:电针水沟穴能明显改善MCAO大鼠神经功能症状;能够促进MCAO模型大鼠梗死区周围血管内皮细胞增殖,并将血管内皮细胞增殖出现时间显著提前,其可能机制为电针水沟穴可上调血管新生相关因子的表达,并使表达时相前移,从而促进血管新生,对改善脑梗死的预后起到了明显的积极作用。

Understanding the interplay between cell force and cell adhesion processes

Cells,wrapped among their neighbors and surrounding extracellular matrix(ECM),form cell-cell adhesions and cell-ECM adhesions.Extracellular biophysical cues exert a far-reaching influence on a sweeping of cell behaviors,including signal transduction,gene expression,and fate determination.Cell-cell adhesions mediated by inter-cellular adhesion molecules bridge the membranes of adjacent cells through either heterophilic or homophilic adhesive interactions,playing a critical part in multicellular structural maintenance and,therefore,a foundation for multicellular organisms.Cell-ECM adhesions are derived from the interaction between cell adhesion receptors and multi-adhesive matrix proteins to ensure cell and tissue cohesion.Whereas cells not only unilaterally respond to certain cues from extracellular environment but can also alter the physicochemical profiles of the externalities and hence hold important implications for clinical applications.The essential function of cell adhesions has cre-ated tremendous interests in developing methods for measuring and studying cell adhesion properties,namely,cellular force.Here,we describe the collection of cell adhesive inputs on cellular signaling cascades and the“crosstalk”between cell-cell adhesions and cell-ECM adhesions.Furthermore,we provide the summary of the current methods to measure such cell adhesive forces.

Engineering of Reversible Luminescent Probes for Real-Time Intravital Imaging of Liver Injury and Repair

As the major organ for drug metabolism and detoxification,the liver is prone to damage and severely impaired functionality.The treatment of liver diseases is based on a clear understanding of the process underlying liver injury and repair.However,intravital real-time imaging of liver injury and repair is still limited due to the lack of in vivo reversible visualization methods.To this end,we proposed a rational design strategy for the development of a reversible upconversion luminescence nanoprobe that allows real-time and in vivo imaging of liver injury and repair processes.As a proof of concept,we first developed a small molecule probe NB3 which can reversibly respond to related analytes of early liver injury[peroxynitrite(ONOO−)]and liver repair[glutathione(GSH)].The small molecule probe was then integrated with a core–shell upconversion nanoparticle to form a sophisticated nanoprobe.Compared with traditional small molecule probes,this nanoprobe exhibited a higher selectivity to ONOO−,longer retention time in liver,and wider dynamic response range to GSH after oxidation by ONOO−.The novel nanoprobe facilitated the successful monitoring and discrimination among the different degrees of liver injury and repair in a mouse model.

Molecular Mechanism of Electroacupuncture Regulating Cerebral Arterial Contractile Protein in Rats with Cerebral Infarction Based on MLCK Pathway

Objective To explore the effect of electroacupuncture(EA)intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction.Methods Male Wistar rats were randomly divided into 3 groups by a random number table:the model group(n=24),the EA group(n=24),and the normal group(n=6).The model and the EA groups were divided into different time subgroups at 0.5,1,3,and 6 h after middle cerebral artery occlusion(MCAO),with 6 rats in each subgroup.MCAO model was established using intraluminal suture occlusion method.The EA group was given EA treatment at acupoint Shuigou(GV 26)instantly after MCAO for 20 min.The contents of cerebrovascular smooth muscle MLCK,the 3 subunits of myosin light chain phosphatase(MLCP)MYPT1,PP1c-δand M20,as well as myosin-ATPase activity were detected using immunohistochemistry and Western blotting.Results The overall expression level of the MYPT1 and PP1c-δin the model group was significantly higher(P<0.01).After EA intervention,the 0.5 h group expression level was close to that of the normal group(P>0.05),and the other subgroups were still significantly higher than the normal group(P<0.01).After EA intervention,the expression level of each subgroup was significantly lower than the corresponding model group.There was a significant difference between the 0.5 and 1 h subgroups(P<0.01),while a difference was also observed between the 3 and 6 h subgroups(P<0.05).The dynamic change rule gradually increased with the prolongation of infarction time within 6 h after infarction.Conclusion EA intervention can inhibit contraction of cerebral vascular smooth muscle cells and regulate smooth muscle relaxation by regulating MLCK pathway.

Near index matching enables solid diffractive optical element fabrication via additive manufacturing

Diffractive optical elements(DOEs)have a wide range of applications in optics and photonics,thanks to their capability to perform complex wavefront shaping in a compact form.However,widespread applicability of DOEs is still limited,because existing fabrication methods are cumbersome and expensive.Here,we present a simple and cost-effective fabrication approach for solid,high-performance DOEs.The method is based on conjugating two nearly refractive index-matched solidifiable transparent materials.The index matching allows for extreme scaling up of the elements in the axial dimension,which enables simple fabrication of a template using commercially available 3D printing at tens-of-micrometer resolution.We demonstrated the approach by fabricating and using DOEs serving as microlens arrays,vortex plates,including for highly sensitive applications such as vector beam generation and super-resolution microscopy using MINSTED,and phase-masks for three-dimensional single-molecule localization microscopy.Beyond the advantage of making DOEs widely accessible by drastically simplifying their production,the method also overcomes difficulties faced by existing methods in fabricating highly complex elements,such as high-order vortex plates,and spectrum-encoding phase masks for microscopy.

Effect of Electro-acupuncture on Vasomotor Symptoms in Rats with Acute Cerebral Infarction Based on Phosphatidylinositol System

Objective:To investigate the effect of electro-acupuncture(EA)on vasomotor symptoms in rats with acute cerebral infarction,by observing the changes in the expression of factors related to the phosphatidylinositol(PI)system.Methods:Forty-two Wistar rats were randomly divided into 3 groups by a random number table:the control group(n=6),the model group(n=18)and the EA group(n=18).The EA group was given EA treatment at Shuigou(GV 26)instantly after modeling with middle cerebral artery occlusion(MCAO)method,while the model and control groups were not given any treatment.The degrees of neurological deficiency were evaluated using neurological severity scores(NSS)and the brain blood flow was evaluated by a laser scanning confocal microscope.Western blot analysis was conducted to detect the expression levels of G-protein subtype(Gq)and calmodulin(CaM).Competition for protein binding was conducted to detect the expression level of inositol triphosphate(IP3).Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol(DAG).The expression level of intracellular concentration of free calcium ion([Ca^(2+)]i)was detected by flow cytometry.Results:The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO(P<0.01),while the EA group was significantly lower than the model group at 6 h(P<0.01).The cerebral blood flow in the model group was significantly lower than the control group at 1,3 and 6 h after MCAO(P<0.01),while for the EA group it was remarkably higher than the model group at the same time points(P<0.01).The expressions of Gq,CaM,IP3,DAG and[Ca^(2+)]i in the model group were significantly higher than the control group(P<0.05 or P<0.01),and those in the EA group were significantly lower than the model group at the same time points(P<0.05 or P<0.01).Conclusion:EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction,improve cerebral autonomy movement,and alleviate cerebral vascular spasm.

A new silicon phthalocyanine dye induces pyroptosis in prostate cancer cells during photoimmunotherapy

Photoimmunotherapy(PIT)combines the specificity of antibodies with the cytotoxicity of light activatable photosensitizers(PS)and is a promising new cancer therapy.We designed and synthesized,in a highly convergent manner,the silicon phthalocyanine dye WB692-CB2,which is novel for being the first light-activatable PS that can be directly conjugated via a maleimide linker to cysteines.In the present study we conjugated WB692-CB2 to a humanized antibody with engineered cysteines in the heavy chains that specifically targets the prostate-specific membrane antigen(PSMA).The resulting antibody dye conjugate revealed high affinity and specificity towards PSMA-expressing prostate cancer cells and induced cell death after irradiation with red light.Treated cells exhibited morphological characteristics associated with pyroptosis.Mechanistic studies revealed the generation of reactive oxygen species,triggering a cascade of intracellular events involving lipid peroxidation,caspase-1 activation,gasdermin D cleavage and membrane rupture followed by release of pro-inflammatory cellular contents.In first in vivo experiments,PIT with our antibody dye conjugate led to a significant reduction of tumor growth and enhanced overall survival in mice bearing subcutaneous prostate tumor xenografts.Our study highlights the future potential of the new phthalocyanine dye WB692-CB2 as PS for the fluorescence-based detection and PIT of cancer,including local prostate tumor lesions,and systemic activation of anti-tumor immune responses by the induction of pyroptosis.