CHIP相关基因与MPN患者心脑血管事件的风险分析

目的:分析骨髓增殖性肿瘤(MPN)患者不确定潜能的克隆性造血(CHIP)相关基因突变谱和临床特征,探讨CHIP相关基因与其心脑血管事件(CCE)的相关性及可能作用机制。方法:回顾性分析2019年8月-2022年7月首都医科大学附属北京安贞医院血液科收治的73例MPN患者的临床资料和二代测序结果,采用Logistic回归分析CHIP相关基因、炎症细胞因子对MPN患者CCE的影响。结果:55例(75.3%)MPN患者检出CHIP相关基因,原发性血小板增多症(ET)和真性红细胞增多症(PV)患者CHIP相关基因各突变频率差异无统计学意义。CHIP相关基因突变以单基因形式为主,检出率从高至低依次为JAK2V617F(63.0%,46/73)、ASXL1(16.4%,12/73)、TET2(11.0%,8/73)、DNMT3A(9.6%,7/73)、SRSF2(6.9%,5/73)、SF3B1(4.1%,3/73)、TP53(1.4%,1/73)和PPMID(1.4%,1/73)。年龄>60岁患者CHIP相关基因检出率明显高于≤60岁者[91.7%(33/36)vs 59.5%(22/37)]。27例(37.0%)MPN患者伴CCE(MPN/CCE),2次CCE者5例,均为动脉事件。CCE组患者年龄(62.8±12.8 vs 53.9±15.8岁,P=0.015)、IL-1β水平(17.7±26.0vs 4.3±8.6,P=0.012)、IL-8水平(360.7±598.6 vs 108.3±317.0,P=0.045)、血栓形成史(29.6%vs 2.2%,P=0.020)和CHIP相关基因检出率(88.9%vs 67.4%,P=0.040)高于无CCE组。多因素Logistic回归分析结果显示,年龄(OR=0.917,95%CI:0.843-0.999,P=0.047)、血栓形成史(OR=34.148,95%CI:2.392-487.535,P=0.009)、任何1个CHIP相关基因突变(OR=16.065,95%CI:1.217-212.024,P=0.035)和IL-1β水平升高(OR=0.929,95%CI:0.870-0.992,P=0.027)均是MPN/CCE的独立危险因素;CHIP相关单基因突变与MPN/CCE无关,但DNMT3A(OR=88.717,95%CI:2.690-292.482,P=0.012)、ASXL1(OR=7.941,95%CI:1.045-60.353,P=0.045)突变是PV/CCE的独立危险因素。结论:MPN患者CHIP相关基因突变率高,尤其是60岁以上患者;高龄、血栓形成史、CHIP相关基因突变和IL-1β水平升高是MPN发生CCE的独立危险因素。DNMT3A、ASXL1单基因突变是PV患者CCE的独立危险因素。CHIP相关基因突变及炎症细胞因子IL-1β升高是MPN新的CCE危险因素。

替普瑞酮通过E3泛素连接酶CHIP减轻LPS引起的心肌炎症反应和心功能障碍

目的:探究替普瑞酮又称香叶基香叶基丙酮,GGA)对脂多糖(LPS)诱导的心功能障碍的治疗作用及机制。方法:(1)取8周龄C57BL/6雄性野生型小鼠和热休克蛋白70(HSP70)羧基末端相互作用蛋白(CHIP)基因敲除小鼠,随机分为对照组、LPS组、LPS+GGA组和GGA组,每组8只。用腹腔注射LPS(25 mg/kg)的方法建立模型,于LPS刺激后1 h给予小鼠腹腔注射GGA(100 mg/kg)。利用小动物超声系统评估小鼠心脏功能;采集各组小鼠血清,检测血清肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)水平;HE染色观察病理学改变;ELISA检测心脏组织中炎症因子肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的水平;Western blot检测各组心脏组织HSP70、CHIP、核转运蛋白α2(KPNA2)、髓过氧化物酶(MPO)、血管细胞黏附分子(VCAM)和细胞间黏附分子(ICAM)的蛋白表达以及细胞核NF-κB的水平。(2)利用小鼠心肌细胞HL-1,建立LPS刺激的离体细胞炎症模型。ELISA检测细胞上清中TNF-α和IL-6的水平;Western blot检测心肌细胞中HSP70、CHIP和KPNA2蛋白表达;免疫荧光染色观察细胞核NF-κB的表达。结果:(1)GGA有效改善LPS刺激小鼠的心脏功能,显著提高射血分数和左室短轴缩短率(P<0.01),减少血清CK-MB和LDH含量(P<0.01),减轻心肌损伤。(2)GGA显著减少LPS引起的TNF-α和IL-6炎症因子的释放(P<0.01),以及NF-κB的入核,降低心肌组织中KPNA2、MPO、VCAM和ICAM蛋白表达,增加心肌组织和细胞HSP70的水平(P<0.01)。(3)在CHIP基因敲除的心肌细胞和小鼠中,GGA不能抑制LPS引起的炎症反应,失去了改善LPS刺激小鼠心脏功能的作用。结论:GGA能够减轻LPS引起的心功能障碍,其作用机制与升高HSP70的表达,促进CHIP的活化,减少NF-κB的入核,抑制炎症因子的释放有关。CHIP的敲除使GGA丧失了减轻LPS诱导的炎症反应和心肌损伤的作用。

Simulation and fabrication of in vitro microfluidic microelectrode array chip for patterned culture and electrophysiological detection of neurons

To enable the detection and modulation of modularized neural networks in vitro,this study proposes a microfluidic microelectrode array chip for the cultivation,compartmentalization,and control of neural cells.The chip was designed based on the specific structure of neurons and the requirements for detection and modulation.Finite-element analysis of the chip’s flow field was conducted using the COMSOL Multiphysics software,and the simulation results show that the liquid within the chip can flow smoothly,ensuring stable flow fields that facilitate the uniform growth of neurons within the microfluidic channels.By employing MEMS technology in combination with nanomaterial modification techniques,the microfluidic microelectrode array chip was fabricated successfully.Primary hippocampal neurons were cultured on the chip,forming a well-defined neural network.Spontaneous electrical activity of the detected neurons was recorded,exhibiting a 23.7%increase in amplitude compared to neuronal discharges detected on an open-field microelectrode array.This study provides a platform for the precise detection and modulation of patterned neuronal growth in vitro,potentially serving as a novel tool in neuroscience research.

A Fully-Integrated Memristor Chip for Edge Learning

It is still challenging to fully integrate computing in memory chip as edge learning devices.In recent work published on Science,a fully-integrated chip based on neuromorphic memristors was developed for edge learning as artificial neural networks with functionality of synapses,dendrites,and somas.A crossbar-array memristor chip facilitated edge learning including hardware realization,learning algorithm,and cycle-parallel sign-and threshold-based learning(STELLAR)scheme.The motion control and demonstration platforms were executed to improve the edge learning ability for adapting to new scenarios.

Simplified quantitative analysis method and its application in the insitu synthesized copper-based azide chips

Copper-based azide(Cu(N_(3))2 or CuN_(3),CA)chips synthesized by in-situ azide reaction and utilized in miniaturized explosive systems has become a hot research topic in recent years.However,the advantages of in-situ synthesis method,including small size and low dosage,bring about difficulties in quantitative analysis and differences in ignition capabilities of CA chips.The aim of present work is to develop a simplified quantitative analysis method for accurate and safe analysis of components in CA chips to evaluate and investigate the corresponding ignition ability.In this work,Cu(N_(3))2 and CuN_(3)components in CA chips were separated through dissolution and distillation by utilizing the difference in solubility and corresponding content was obtained by measuring N_(3)-concentration through spectrophotometry.The spectrophotometry method was optimized by studying influencing factors and the recovery rate of different separation methods was studied,ensuring the accuracy and reproducibility of test results.The optimized method is linear in range from 1.0-25.0 mg/L,with a correlation coefficient R^(2)=0.9998,which meets the requirements of CA chips with a milligram-level content test.Compared with the existing ICP method,component analysis results of CA chips obtained by spectrophotometry are closer to real component content in samples and have satisfactory accuracy.Moreover,as its application in miniaturized explosive systems,the ignition ability of CA chips with different component contents for direct ink writing CL-20 and the corresponding mechanism was studied.This study provided a basis and idea for the design and performance evaluation of CA chips in miniaturized explosive systems.

A novel integrated microfluidic chip for on-demand electrostatic droplet charging and sorting

On-demand droplet sorting is extensively applied for the efficient manipulation and genome-wide analysis of individual cells.However,state-of-the-art microfluidic chips for droplet sorting still suffer from low sorting speeds,sample loss,and labor-intensive preparation procedures.Here,we demonstrate the development of a novel microfluidic chip that integrates droplet generation,on-demand electrostatic droplet charging,and high-throughput sorting.The charging electrode is a copper wire buried above the nozzle of the microchannel,and the deflecting electrode is the phosphate buffered saline in the microchannel,which greatly simplifies the structure and fabrication process of the chip.Moreover,this chip is capable of high-frequency droplet generation and sorting,with a frequency of 11.757 kHz in the drop state.The chip completes the selective charging process via electrostatic induction during droplet generation.On-demand charged microdroplets can arbitrarilymove to specific exit channels in a three-dimensional(3D)-deflected electric field,which can be controlled according to user requirements,and the flux of droplet deflection is thereby significantly enhanced.Furthermore,a lossless modification strategy is presented to improve the accuracy of droplet deflection or harvest rate from 97.49% to 99.38% by monitoring the frequency of droplet generation in real time and feeding it back to the charging signal.This chip has great potential for quantitative processing and analysis of single cells for elucidating cell-to-cell variations.

Single event effects evaluation on convolution neural network in Xilinx 28 nm system on chip

Convolutional neural networks(CNNs) exhibit excellent performance in the areas of image recognition and object detection, which can enhance the intelligence level of spacecraft. However, in aerospace, energetic particles, such as heavy ions, protons, and alpha particles, can induce single event effects(SEEs) that lead CNNs to malfunction and can significantly impact the reliability of a CNN system. In this paper, the MNIST CNN system was constructed based on a 28 nm systemon-chip(SoC), and then an alpha particle irradiation experiment and fault injection were applied to evaluate the SEE of the CNN system. Various types of soft errors in the CNN system have been detected, and the SEE cross sections have been calculated. Furthermore, the mechanisms behind some soft errors have been explained. This research will provide technical support for the design of radiation-resistant artificial intelligence chips.

Rapid fabrication of modular 3D paper-basedmicrofluidic chips using projection-based 3D printing

Paper-based microchips have different advantages,such as better biocompatibility,simple production,and easy handling,making them promising candidates for clinical diagnosis and other fields.This study describes amethod developed to fabricate modular three-dimensional(3D)paper-based microfluidic chips based on projection-based 3D printing(PBP)technology.A series of two-dimensional(2D)paper-based microfluidic modules was designed and fabricated.After evaluating the effect of exposure time on the accuracy of the flow channel,the resolution of this channel was experimentally analyzed.Furthermore,several 3D paper-based microfluidic chips were assembled based on the 2D ones using different methods,with good channel connectivity.Scaffold-based 2D and hydrogel-based 3D cell culture systems based on 3D paper-based microfluidic chips were verified to be feasible.Furthermore,by combining extrusion 3D bioprinting technology and the proposed 3D paper-based microfluidic chips,multiorgan microfluidic chips were established by directly printing 3D hydrogel structures on 3D paperbased microfluidic chips,confirming that the prepared modular 3D paper-based microfluidic chip is potentially applicable in various biomedical applications.

Efficient stochastic parallel gradient descent training for on-chip optical processor

In recent years,space-division multiplexing(SDM)technology,which involves transmitting data information on multiple parallel channels for efficient capacity scaling,has been widely used in fiber and free-space optical communication sys-tems.To enable flexible data management and cope with the mixing between different channels,the integrated reconfig-urable optical processor is used for optical switching and mitigating the channel crosstalk.However,efficient online train-ing becomes intricate and challenging,particularly when dealing with a significant number of channels.Here we use the stochastic parallel gradient descent(SPGD)algorithm to configure the integrated optical processor,which has less com-putation than the traditional gradient descent(GD)algorithm.We design and fabricate a 6×6 on-chip optical processor on silicon platform to implement optical switching and descrambling assisted by the online training with the SPDG algorithm.Moreover,we apply the on-chip processor configured by the SPGD algorithm to optical communications for optical switching and efficiently mitigating the channel crosstalk in SDM systems.In comparison with the traditional GD al-gorithm,it is found that the SPGD algorithm features better performance especially when the scale of matrix is large,which means it has the potential to optimize large-scale optical matrix computation acceleration chips.

Effects of Using Softwood Pellet Biochar Prepared at Different Temperatures with Grass Chippings on Retention of Heavy Metals in Contaminated Soils

Heavy metals have been viewed as hazardous environmental pollutants, and anthropogenic activities due to their high toxicity and persistent nature in the environment. Anthropogenic activities such as artisanal mining, industrial activities, improper usage of fertilizers and pesticides, and indiscriminate open waste disposal bring about an increase in the presence of heavy metals in the environment. In the Keffi Metropolis, different elements lead to land contamination which debilitates soil quality, plant survival, human well-being, and the environment as a result of extensive dispersion or quantity of heavy metals in the soil and water. In recent years, biochar has emerged as a promising soil amendment for mitigating heavy metal pollution due to its unique physicochemical properties. This paper provides the effects of softwood pellet biochar on the retention of heavy metals in contaminated soils. A microcosm experiment was carried out to investigate the effects of biochar on the retention of heavy metals in contaminated soils. This research aimed to give an overview of the effects of softwood biochar at different temperatures (550˚C and 700˚C) on the retention of heavy metals and metalloids released from the soil during water inundation. The results show that the addition of organic matter (grass chippings) minimizes heavy metal mobilization. Also, biochar at high temperatures is more effective than those at low temperatures. The expected outcome of the research analysis includes providing insights into the role of biochar in retaining heavy metal contamination and further understanding the use of biochar as a sorbent for the management of contaminated soil.

Flip Chip技术在集成电路封装中的应用

阐述从集成电路封装发展现状、Flip Chip技术内涵、Flip Chip技术在集成电路封装中的应用剖析、市场发展展望等多个角度,探讨在集成电路封装中,应用Flip Chip技术的必要性和重要性。

An electrochemiluminescent magneto-immunosensor for ultrasensitive detection of hs-cTnI on a microfluidic chip

Sensitive detection and precise quantitation of trace-level crucial biomarkers in a complex sample matrix has become an important area of research.For example,the detection of high-sensitivity cardiac troponin I(hs-cTnI)is strongly recommended in clinical guidelines for early diagnosis of acute myocardial infarction.Based on the use of an electrode modified by single-walled carbon nanotubes(SWCNTs)and a Ru(bpy)32+-doped silica nanoparticle(Ru@SiO2)/tripropylamine(TPA)system,a novel type of electrochemiluminescent(ECL)magneto-immunosensor is developed for ultrasensitive detection of hs-cTnI.In this approach,a large amount of[Ru(bpy)3]2+is loaded in SiO2(silica nanoparticles)as luminophores with high luminescent efficiency and SWCNTs as electrode surface modification material with excellent elec-trooxidation ability for TPA.Subsequently,a hierarchical micropillar array of microstructures is fabricated with a magnet placed at each end to efficiently confine a single layer of immunomagnetic microbeads on the surface of the electrode and enable 7.5-fold signal enhancement.In particular,the use of transparent SWCNTs to modify a transparent ITO electrode provides a two-order-of-magnitude ECL signal amplifi-cation.A good linear calibration curve is developed for hs-cTnI concentrations over a wide range from 10 fg/ml to 10 ng/ml,with the limit of detection calculated as 8.720 fg/ml(S/N 3).This ultrasensitive immunosensor exhibits superior detection performance with remarkable sta-=bility,reproducibility,and selectivity.Satisfactory recoveries are obtained in the detection of hs-cTnI in human serum,providing a potential analysis protocol for clinical applications.

Chiplet技术推动半导体产业可持续发展的研究

随着半导体产业面临莫尔定律的瓶颈,寻找新的技术推动力变得至关重要。文章将焦点放在Chiplet技术,这是一种可突破物理和经济限制、进一步提升性能的新兴技术。Chiplet技术利用模组化策略,不仅能提供更高的效能,且符合可持续发展的要求。文章深入探讨了Chiplet技术如何在莫尔定律遇到挑战的情况下推动半导体产业的可持续发展,并期望提供新的思维和发展方向。

副溶血弧菌QsvR ChIP-qPCR方法的建立

目的:建立副溶血弧菌QsvR的染色质免疫共沉淀实时定量PCR(ChIP-qPCR)实验方法。方法:通过热激转化和转化结合将标记2×Flag标签的qsvR片段转入qsvR突变株(ΔqsvR)中,阿拉伯糖诱导表达QsvR-2×Flag蛋白,通过甲醛与基因组DNA进行交联形成2×Flag-QsvR-DNA复合物,将基因组DNA超声裂解为100~1000 bp大小不等的片段,通过特异性抗原抗体反应将蛋白质-DNA复合物沉淀下来,高盐和加热解交联,回收DNA进行qPCR定量DNA,分析副溶血弧菌体内QsvR与靶基因的结合情况。结果:成功构建出实验菌株ΔqsvR/pBAD33-qsvR-2×Flag,以不加阿拉伯糖诱导为对照,经阿拉伯糖诱导菌株的aphA、opaR、exsB和vtrA的免疫共沉淀量明显较高,表明在副溶血弧菌体内QsvR与aphA、opaR、exsB和vtrA具有结合作用。结论:成功建立了副溶血弧菌QsvR的ChIP-qPCR实验方法,可用于原核生物体内蛋白质-DNA相互作用的研究。

CHIP蛋白在疾病中的作用研究进展

CHIP(carboxyl terminus of Hsc70-interacting protein)又称STUB1(STIP1 homology and U-box containing protein 1),是一种分子伴侣结合蛋白,具有E3泛素连接酶活性.研究表明,CHIP蛋白在细胞体内通过蛋白质量控制系统调控细胞代谢、分化、增殖、迁移和肿瘤发生发展等多种生理病理过程.本文对CHIP在癌症及多种常见疾病中的作用进行综述,以期为临床治疗和药物开发提供潜在突破点.

泛素E3连接酶CHIP介导RCC2的泛素化及降解

目的 本课题组前期构建了一种正交泛素传递方法,并发现染色体浓缩调节蛋白2(RCC2)是泛素连接酶CHIP的潜在底物。本实验旨在验证CHIP是否能介导RCC2的泛素化及降解,以期通过CHIP来泛素化调控RCC2的表达和功能。方法 首先构建CHIP及其突变体质粒,验证CHIP与RCC2是否有相互作用;然后构建pLVX-RCC2-FLAG重组质粒并转染进CHIP敲减细胞株(shCHIP)、HEK293细胞株、CHIP过表达细胞株(OE CHIP),通过免疫共沉淀下拉富集RCC2-FLAG并检测其泛素化水平;接下来在HEK293细胞中转染不同量的CHIP,以及设置CHX chase蛋白稳定性实验,检测RCC2的蛋白水平变化;最后利用泛素突变体K48R-Ub及K11R-Ub进行CHIP介导RCC2上形成泛素链的实验检测。结果 在HEK293细胞中,CHIP能够介导RCC2的泛素化,并在RCC2上形成K48和K11多聚泛素链,促进其通过蛋白酶体发生降解。结论 泛素连接酶CHIP能够介导RCC2的泛素化及降解,为实现通过CHIP来泛素化调控RCC2提供了理论基础。

Flip Chip结构的IC信号完整性仿真分析

针对Flip Chip封装型芯片设计过程中存在的传输线阻抗不连续与串扰过大等问题,从层叠设置与板材介质厚度两个角度提出了一种基于阻抗、串扰的仿真分析设计方法,主要涉及两个方面:对于由信号参考平面被分割而造成的阻抗突变问题,通过添加参考平面而使信号具有完整的回流路径,使得传输线的阻抗在平面分割处由167.5Ω降至52.5Ω;对于因布线密度过大而造成的走线之间的串扰系数偏高的问题,通过减小板材的介质厚度使传输线间串扰系数的最大值从17.26%降至14.01%。仿真结果表明,此设计方法有效降低了芯片设计中潜在的信号完整性风险,提高了芯片的可靠性和稳定性。

Ultrasensitive quantification of trace amines based on N-phosphorylation labeling chip 2D LC-QQQ/MS

Trace amines(TAs)are metabolically related to catecholamine and associated with cancer and neurological disorders.Comprehensive measurement of TAs is essential for understanding pathological processes and providing proper drug intervention.However,the trace amounts and chemical instability of TAs challenge quantification.Here,diisopropyl phosphite coupled with chip two-dimensional(2D)liquid chromatography tandem triple-quadrupole mass spectrometry(LC-QQQ/MS)was developed to simultaneously determine TAs and associated metabolites.The results showed that the sensitivities of TAs increased up to 5520 times compared with those using nonderivatized LC-QQQ/MS.This sensitive method was utilized to investigate their alterations in hepatoma cells after treatment with sorafenib.The significantly altered TAs and associated metabolites suggested that phenylalanine and tyrosine metabolic pathways were related to sorafenib treatment in Hep3B cells.This sensitive method has great potential to elucidate the mechanism and diagnose diseases considering that an increasing number of physiological functions of TAs have been discovered in recent decades.

CHIP促进衰老内皮细胞凋亡的实验研究

目的:观察CHIP促进复制性衰老人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)凋亡的作用。方法:通过连续培养传代方法建立原代HUVECs复制性衰老模型,根据细胞形态的变化,以及衰老相关β-半乳糖苷酶染色阳性率及细胞上清液中衰老相关标志物评估HUVECs衰老情况;采用RT-PCR和Western blot检测HUVECs衰老前后CHIP mRNA及蛋白表达变化,Western blot检测过表达CHIP后凋亡相关蛋白裂解的胱天蛋白酶3(Cleaved Caspase-3,CCASP3)、CCASP9表达变化,流式细胞术检测细胞凋亡情况。结果:衰老HUVECs的细胞形态发生改变、β-半乳糖苷酶染色阳性细胞增加,细胞上清液中白细胞介素-6(Interleukin-6,IL-6)及IL-8含量增加;CHIP mRNA及蛋白表达降低;过表达CHIP后,与对照组相比,CHIP过表达组CCASP3、CCASP9表达增加、凋亡细胞比例增加。结论:复制性衰老HUVECs中CHIP表达降低,过表达CHIP可促进复制性衰老HUVECs凋亡。

Pattern recognition in multi-synaptic photonic spiking neural networks based on a DFB-SA chip

Spiking neural networks(SNNs)utilize brain-like spatiotemporal spike encoding for simulating brain functions.Photonic SNN offers an ultrahigh speed and power efficiency platform for implementing high-performance neuromorphic computing.Here,we proposed a multi-synaptic photonic SNN,combining the modified remote supervised learning with delayweight co-training to achieve pattern classification.The impact of multi-synaptic connections and the robustness of the network were investigated through numerical simulations.In addition,the collaborative computing of algorithm and hardware was demonstrated based on a fabricated integrated distributed feedback laser with a saturable absorber(DFB-SA),where 10 different noisy digital patterns were successfully classified.A functional photonic SNN that far exceeds the scale limit of hardware integration was achieved based on time-division multiplexing,demonstrating the capability of hardware-algorithm co-computation.