人类胎盘基因CpG岛甲基化水平与自然流产的相关性

目的探讨人类胎盘基因CpG岛甲基化水平与孕妇自然流产的相关性。方法选取接受常规产前检查的孕妇55例为研究对象,其中25例自然流产(流产组),30例正常分娩(对照组)。收集入组孕妇的临床资料及人类胎盘组织样本。比较2组临床一般资料。分析相关胎盘基因的甲基化水平对孕妇自然流产结局的预测价值。采用Logistic回归分析法分析孕妇自然流产的影响因素。结果2组在孕妇年龄、流产儿/新生儿出生体质量、孕妇妊娠天数和流产儿/新生儿性别方面比较,差异有统计学意义(P<0.05)。MECP2-1、MECP2-4、HSD11B2、MECP2-3和MECP2-2基因甲基化与自然流产相关。流产组的MECP2-1、MECP2-4、HSD11B2和MECP2-3基因甲基化率高于对照组,差异有统计学意义(P<0.05)。MECP2-1、MECP2-4、HSD11B2、MECP2-3和MECP2-2的曲线下面积(AUC)分别为0.773、0.737、0.700、0.663和0.627。Logistic回归分析结果显示,MECP2-1是孕妇发生流产的独立影响因素(P<0.05)。结论MECP2-1、MECP2-4、HSD11B2、MECP2-3和MECP2-2基因甲基化水平和孕妇的自然流产结局有关。MECP2-1基因是自然流产发生的独立影响因素。

基于DIC及CPG技术的热冷循环后花岗岩Ⅰ型断裂特性

深部储层岩石的热力学特性,尤其是循环热冷作用下损伤破坏特性,对于增强型地热系统井壁稳定性分析及地热开采效率评估具有重要意义。针对中心直裂纹半圆盘(NSCB)花岗岩试样,首先进行不同热冷循环处理(加热温度400℃,最高循环次数13次),然后开展3点弯Ⅰ型断裂韧度特性试验。基于裂纹扩展计(CPG)和数字图像相关(DIC)测试技术,研究了热冷循环作用下花岗岩Ⅰ型断裂韧度、断裂过程区(FPZ)、裂纹扩展速率及断裂轮廓特征的影响规律。试验结果表明:当热冷循环次数达到10次以上,花岗岩试样脆性明显减弱,而峰前软化特性和峰后延性增强;花岗岩的断裂过程区由裂缝尖端开始逐步孕育,断裂过程区长度随荷载增大呈先增大后减小的趋势,Ⅰ型断裂韧度、最大断裂过程区长度及裂纹平均扩展速度随热冷循环次数增大而指数减小,Ⅰ型断裂面随热冷循环次数增大越来越不平整。最后,基于X-ray衍射(XRD)和扫描电镜(SEM)等微观测试技术,研究了热冷循环作用对花岗岩矿物成分及微观结构的影响规律,结果表明花岗岩4种矿物成分的峰值衍射强度及矿物质量分数均随热冷循环次数增大而降低,而微裂纹大小及数量随热冷循环次数增大而增大。热冷循环作用下花岗岩的损伤劣化机理包含了多次高温热损伤、水冷冲击及水弱化等3方面的联合作用结果。

尾鳍驱动仿生机器鱼的CPG控制

随着人们对海洋资源关注度的增加,对仿生机器鱼的游动性能提出更高要求,为了使机器鱼游动更贴近真实鱼的游动效果,根据Light Hill的大摆幅细长体理论的鱼体波曲线对水下仿生机器鱼的游动模式进行了研究,并采用HOPF神经元振荡器模型建立非线性振荡器模型,运用最近相邻耦合关系构建了CPG控制网络拓扑图,建立了CPG控制模型,通过调节CPG控制参数实现了机器鱼多运动模态的控制与切换。针对鲹科仿生机器鱼的游动方式进行SolidWorks三维建模,在ADAMS中进行运动仿真,并搭建实体样机。实验结果表明,采用基于HOPF的链式耦合中枢模式发生器对舵机的驱动进行控制,机器鱼尾鳍的摆动具有很好的运动柔顺性,易于实现运动模态平滑切换,游动更加灵活。这为机器鱼在海洋执行更复杂任务的研究打下了坚实基础。

血清胶质纤维酸性蛋白、甲基化CpG结合蛋白2对脑胶质瘤的诊断价值及其与病理特征的相关性分析

目的探讨血清胶质纤维酸性蛋白(GFAP)、甲基化CpG结合蛋白2(MeCP2)对脑胶质瘤的诊断价值及其与病理特征的相关性。方法回顾性选取2021年1月至2024年1月南京医科大学附属脑科医院(南京脑科医院)收治的80例脑胶质瘤患者,将其作为胶质瘤组,选取本院同期收治的80例非胶质瘤颅内良性肿瘤患者作为对照组。应用双抗体夹心法检测两组患者血清GFAP表达水平,应用荧光定量聚合酶链反应法检测肿瘤组织MeCP2表达水平。比较脑胶质瘤患者与对照组的GFAP、MeCP2表达水平,并建立ROC曲线分析GFAP、MeCP2对脑胶质瘤的诊断价值。分析不同病理特征脑胶质瘤患者GFAP、MeCP2表达水平,并采用Spearman相关性分析GFAP、MeCP2与脑胶质瘤病理特征的相关性。结果胶质瘤组患者GFAP、MeCP2表达水平分别为(81.74±7.47)ng/L、2.31±0.31,均高于对照组[(5.35±1.32)ng/L、0.72±0.16],差异均有统计学意义(P<0.05)。绘制ROC曲线结果显示,GFAP、MeCP2、两者联合诊断脑胶质瘤的曲线下面积(AUC)分别为0.721、0.698、0.897。GFAP、MeCP2两者联合对脑胶质瘤的诊断AUC高于两者单一诊断。病理分级Ⅳ级、远处转移患者GFAP水平均高于其他分级、未远处转移,差异均有统计学意义(P<0.05)。病理分级Ⅳ级、肿瘤大小≥3 cm、远处转移患者MeCP2水平均高于其他分级、肿瘤大小<3 cm、未远处转移患者,差异均有统计学意义(P<0.05)。Spearman相关分析结果表明,GFAP与脑胶质瘤不同病理分级、远处转移均呈正相关(r=0.579、0.378,P<0.05);MeCP2与脑胶质瘤不同病理分级、远处转移、脑胶质瘤肿瘤大小均呈正相关(r=0.657、0.457、0.384,P<0.05)。结论胶质纤维酸性蛋白、甲基化CpG结合蛋白2对于脑胶质瘤均具有重要诊断参考价值,两者联合诊断敏感度、特异度更高,且两者与脑胶质瘤的病理分级、肿瘤大小、远处转移情况密切相关。

Research on the development methodology for clinical practice guidelines for organic integration of traditional Chinese medicine and Western medicine

Integrated traditional Chinese medicine(TCM)and Western medicine(WM)is a new medical science grounded in the knowledge bases of both TCM and WM,which then forms a unique modern medical system in China.Integrated TCM and WM has a long history in China,and has made important achievements in the process of clinical diagnosis and treatment.However,the methodological defects in currently published clinical practice guidelines(CPGs)limit its development.The organic integration of TCM and WM is a deeper integration of TCM and WM.To realize the progression of"integration"to"organic integration",a targeted and standardized guideline development methodology is needed.Therefore,the purpose of this study is to establish a standardized development procedure for clinical practice guidelines for the organic integration of TCM and WM to promote the systematic integration of TCM and WM research results into clinical practice guidelines in order to achieve optimal results as the whole is greater than the sum of the parts.

基于Hopf振荡器的Spiking-CPG六足机器人步态运动控制

中枢模式发生器(central pattern generator,CPG)在六足机器人的运动步态控制中起着至关重要的作用。为能够更高效、更低能耗地控制六足机器人的步态运动,提出了一种Spiking-CPG(SCPG)神经网络作为六足机器人的仿生控制系统,其结合Hopf振荡器与以时疏的spiking信号传递信息的仿生神经元LIF(leaky integrate-and-fire),采用环型拓扑结构,使用六组各2000个LIF神经元组成的集合相互连接而成。该SCPG控制系统能够生成六足机器人常见的波动步态、四足步态、三足步态,通过调节相位差参数实现快速、顺滑、稳定的切换运动步态,实时调整所需的频率、振幅,在面对外界干扰时能够在很短的时间内恢复原状,具有很好的鲁棒性。在Webots平台上搭建了一个三维的六足机器人模型,将SCPG的信号输出并经过关节映射函数变换后,来控制六足机器人的运动,验证了所设计六足机器人的运动稳定性和SCPG控制方案的可行性与有效性。最后,在Intel的Loihi芯片上移植了SCPG神经网络控制器,结果表明,其具备高效的执行速度和更低的能耗,在六足机器人的运动控制中具备良好的应用前景。

Levels of evidence and grades of recommendation supporting European society for medical oncology clinical practice guidelines

Background:The European Society for Medical Oncology(ESMO)guidelines are among the most comprehensive and widely used clinical practice guidelines(CPGs)globally.However,the level of scientific evidence supporting ESMO CPG recommendations has not been systematically investigated.This study assessed ESMO CPG levels of evidence(LOE)and grades of recommendations(GOR),as well as their trends over time across various cancer settings.Methods:We manually extracted every recommendation with the Infectious Diseases Society of America(IDSA)classification from each CPG.We examined the distribution of LOE and GOR in all available ESMO CPG guidelines across different topics and cancer types.Results:Among the 1,823 recommendations in the current CPG,30%were classified as LOEⅠ,and 43%were classified as GOR A.Overall,there was a slight decrease in LOEⅠ(−2%)and an increase in the proportion of GOR A(+1%)in the current CPG compared to previous versions.The proportion of GOR A recommendations based on higher levels of evidence such as randomized trials(LOEⅠ–Ⅱ)shows a decrease(71%vs.63%,p=0.009)while recommendations based on lower levels of evidence(LOEⅢ–Ⅴ)show an increase(29%vs.37%,p=0.01)between previous and current version.In the current versions,the highest proportion of LOEⅠ(42%)was found in recommendations related to pharmacotherapy,while the highest proportion of GOR A recommendations was found in the areas of pathology(50%)and diagnostic(50%)recommendations.Significant variability in LOEⅠand GOR A recommendations and their changes over time was observed across different cancer types.Conclusion:One-third of the current ESMO CPG recommendations are supported by the highest level of evidence.More well-designed randomized clinical trials are needed to increase the proportion of LOEⅠand GOR A recommendations,ultimately leading to improved outcomes for cancer patients.

Systematic Security Guideline Framework through Intelligently Automated Vulnerability Analysis

This research aims to propose a practical framework designed for the automatic analysis of a product’s comprehensive functionality and security vulnerabilities,generating applicable guidelines based on real-world software.The existing analysis of software security vulnerabilities often focuses on specific features or modules.This partial and arbitrary analysis of the security vulnerabilities makes it challenging to comprehend the overall security vulnerabilities of the software.The key novelty lies in overcoming the constraints of partial approaches.The proposed framework utilizes data from various sources to create a comprehensive functionality profile,facilitating the derivation of real-world security guidelines.Security guidelines are dynamically generated by associating functional security vulnerabilities with the latest Common Vulnerabilities and Exposure(CVE)and Common Vulnerability Scoring System(CVSS)scores,resulting in automated guidelines tailored to each product.These guidelines are not only practical but also applicable in real-world software,allowing for prioritized security responses.The proposed framework is applied to virtual private network(VPN)software,wherein a validated Level 2 data flow diagram is generated using the Spoofing,Tampering,Repudiation,Information Disclosure,Denial of Service,and Elevation of privilege(STRIDE)technique with references to various papers and examples from related software.The analysis resulted in the identification of a total of 121 vulnerabilities.The successful implementation and validation demonstrate the framework’s efficacy in generating customized guidelines for entire systems,subsystems,and selected modules.

A review of public health guidelines for postpartum physical activity and sedentary behavior from around the world

Background:The period following pregnancy is a critical time window when future habits with respect to physical activity(PA) and sedentary behavior(SB) are established;therefore,it warrants guidance.The purpose of this scoping review was to summarize public health-oriented country-specific postpartum PA and SB guidelines worldwide.Methods:To identity guidelines published since 2010,we performed a(a) systematic search of 4 databases(CINAHL,Global Health,PubMed,and SPORTDiscus),(b) structured repeatable web-based search separately for 194 countries,and(c) separate web-based search.Only the most recent guideline was included for each country.Results:We identified 22 countries with public health-oriented postpartum guidelines for PA and 11 countries with SB guidelines.The continents with guidelines included Europe(n=12),Asia(n=5),Oceania(n=2),Africa(n=1),North America(n=1),and South America(n=1).The most common benefits recorded for PA included weight control/management(n=10),reducing the risk of postpartum depression or depressive symptoms(n=9),and improving mood/well-being(n=8).Postpartum guidelines specified exercises to engage in,including pelvic floor exercises(n=17);muscle strengthening,weight training,or resistance exercises(n=13);aerobics/general aerobic activity(n=13);walking(n=11);cycling(n=9);and swimming(n=9).Eleven guidelines remarked on the interaction between PA and breastfeeding;several guidelines stated that PA did not impact breast milk quantity(n=7),breast milk quality(n=6),or infant growth(n=3).For SB,suggestions included limiting long-term sitting and interrupting sitting with PA.Conclusion:Country-specific postpartum guidelines for PA and SB can help promote healthy behaviors using a culturally appropriate context while providing specific guidance to public health practitioners.

Impact of glucagon-like peptide receptor agonists on endoscopy and its preoperative management: Guidelines, challenges, and future directions

Glucagon-like peptide receptor agonists(GLP-1RA)are used to treat type 2 diabetes mellitus and,more recently,have garnered attention for their effect-iveness in promoting weight loss.They have been associated with several gastrointestinal adverse effects,including nausea and vomiting.These side effects are presumed to be due to increased residual gastric contents.Given the potential risk of aspiration and based on limited data,the American Society of Anesthesi-ologists updated the guidelines concerning the preoperative management of patients on GLP-1RA in 2023.They included the duration of mandated cessation of GLP-1RA before sedation and usage of“full stomach”precautions if these medications were not appropriately held before the procedure.This has led to additional challenges,such as extended waiting time,higher costs,and increased risk for patients.In this editorial,we review the current societal guidelines,clinical practice,and future directions regarding the usage of GLP-1RA in patients undergoing an endoscopic procedure.

Fever assessment in children under five: Are we following the guidelines?

BACKGROUND Fever is a common cause of medical consultation and hospital admission,particularly among children.Recently,the United Kingdom’s National Institute for Health and Care Excellence(NICE)updated its guidelines for assessing fever in children under five years of age.The efficient assessment and management of children with fever are crucial for improving patient outcomes.AIM To evaluate fever assessment in hospitalized children and to assess its adherence with the NICE Fever in under 5s guideline.METHODS We conducted a retrospective cohort review of the electronic medical records of children under five years of age at the Department of Pediatrics,Salmaniya Medical Complex,Bahrain,between June and July 2023.Demographic data,vital signs during the first 48 h of admission,route of temperature measurement,and indications for admission were gathered.Fever was defined according to the NICE guideline.The children were divided into five groups according to their age(0-3 months,>3-6 months,>6-12 months,>12-36 months,and>36-60 months).Patients with and without fever were compared in terms of demography,indication for admission,route of temperature measurement,and other vital signs.Compliance with the NICE Fever in the under 5s guideline was assessed.Full compliance was defined as>95%,partial compliance as 70%-95%,and minimal compliance as≤69%.Pearson’sχ^(2),Student’s t test,the Mann-Whitney U test,and Spearman’s correlation coefficient(rs)were used for comparison.RESULTS Of the 136 patients reviewed,80(58.8%)were boys.The median age at admission was 14.2[interquartile range(IQR):1.7-44.4]months,with the most common age group being 36-60 months.Thirty-six(26.4%)patients had fever,and 100(73.6%)were afebrile.The commonest age group for febrile patients(>12-36 months)was older than the commonest age group for afebrile patients(0-3 months)(P=0.027).The median weight was 8.3(IQR:4.0-13.3)kg.Patients with fever had higher weight than those without fever[10.2(IQR:7.3-13.0)vs 7.1(IQR:3.8-13.3)kg,respectively](P=0.034).Gastrointestinal disease was the leading indication for hospital admission(n=47,34.6%).Patients with central nervous system diseases and fever of unknown etiology were more likely to be febrile(P=0.030 and P=0.011,respectively).The mean heart rate was higher in the febrile group than the afebrile group(140±24 vs 126±20 beats per minute,respectively)[P=0.001(confidence interval:5.8-21.9)]with a positive correlation between body temperature and heart rate,r=0.242,n=136,P=0.004.A higher proportion of febrile patients received paracetamol(n=35,81.3%)compared to the afebrile patients(n=8,18.6%)(P<0.001).The axillary route was the most commonly used for temperature measurements(n=40/42,95.2%),followed by the rectal route(n=2/42,4.8%).The department demonstrated full compliance with the NICE guideline for five criteria:the type of thermometer used,route and frequency of temperature measurement,frequency of heart rate measurement,and use of antipyretics as needed.Partial compliance was noted for two criteria,the threshold of fever at 38°C or more,and the respiratory rate assessment in febrile patients.Minimal compliance or no record was observed for the remaining three criteria;routine assessment of capillary refill,temperature reassessment 1-2 h after each antipyretic intake,and refraining from the use of tepid sponging.CONCLUSION This study showed that fever assessment in hospitalized children under five years of age was appropriate,but certain areas of adherence to the NICE guideline still need to be improved.

CPG Human Motion Phase Recognition Algorithm for a Hip Exoskeleton with VSA Actuator

Due to the dynamic stiffness characteristics of human joints, it is easy to cause impact and disturbance on normal movements during exoskeleton assistance. This not only brings strict requirements for exoskeleton control design, but also makes it difficult to improve assistive level. The Variable Stiffness Actuator (VSA), as a physical variable stiffness mechanism, has the characteristics of dynamic stiffness adjustment and high stiffness control bandwidth, which is in line with the stiffness matching experiment. However, there are still few works exploring the assistive human stiffness matching experiment based on VSA. Therefore, this paper designs a hip exoskeleton based on VSA actuator and studies CPG human motion phase recognition algorithm. Firstly, this paper puts forward the requirements of variable stiffness experimental design and the output torque and variable stiffness dynamic response standards based on human lower limb motion parameters. Plate springs are used as elastic elements to establish the mechanical principle of variable stiffness, and a small variable stiffness actuator is designed based on the plate spring. Then the corresponding theoretical dynamic model is established and analyzed. Starting from the CPG phase recognition algorithm, this paper uses perturbation theory to expand the first-order CPG unit, obtains the phase convergence equation and verifies the phase convergence when using hip joint angle as the input signal with the same frequency, and then expands the second-order CPG unit under the premise of circular limit cycle and analyzes the frequency convergence criterion. Afterwards, this paper extracts the plate spring modal from Abaqus and generates the neutral file of the flexible body model to import into Adams, and conducts torque-stiffness one-way loading and reciprocating loading experiments on the variable stiffness mechanism. After that, Simulink is used to verify the validity of the criterion. Finally, based on the above criterions, the signal mean value is removed using feedback structure to complete the phase recognition algorithm for the human hip joint angle signal, and the convergence is verified using actual human walking data on flat ground.

Chi-CpG NP的制备及其对rNMB0315蛋白的免疫效果

目的制备黏膜免疫佐剂Chi-CpG NP,并检测其对重组B群脑膜炎奈瑟菌0315蛋白(rNMB0315)的免疫效果。方法用壳聚糖纳米颗粒(Chi NP)包裹免疫刺激分子(CpG-ODN)合成Chi-CpG NP,随后检测其理化性质和安全性。将不同黏膜免疫佐剂与rNMB0315充分混合、吸附,滴鼻免疫雌性BALB/c小鼠,检测PBS组、rNMB0315组、rNMB0315+CpG组、rNMB0315+Chi NP组、rNMB0315+Chi-CpG NP组小鼠免疫42天后的体液免疫(IgG、IgG1、IgG2a、sIgA)、细胞免疫水平[白细胞介素(IL)-4、γ干扰素(IFN-γ)、IL-17A]、免疫保护效果以及血清体外杀菌活性。结果成功制备的Chi-CpG NP呈球形,大小均一,有较好的吸附性能和良好的安全性。小鼠免疫42天后,体液免疫和细胞免疫水平rNMB0315+Chi-NP组、rNMB0315+Chi-CpG NP组高于rNMB0315组(P<0.05);rNMB0315+Chi-CpG NP组高于rNMB0315+CpG组和rNMB0315+Chi-NP组(P<0.05)。致死剂量B群脑膜炎奈瑟菌攻击各组小鼠后,rNMB0315+Chi-CpG NP组小鼠存活率最高;血清体外杀菌抗体滴度为1∶4。结论成功制备了Chi-CpG NP。Chi-CpG NP能增强rNMB0315蛋白的免疫效果,提高小鼠的存活率。

基于CPG网络的机器人运动关节控制

机器人技术在现代产业中的应用越来越广泛,如何适应复杂环境是当前机器人技术的一项热点研究,足式机器人由于其仿生特性能够较好地适应环境。本文研究了一种基于CPG(Certral Pattern Generators,中枢模型发生器)网络的机器人关节运动控制,以Hopf振荡器作为CPG网络的振荡单元,并通过仿真验证了其在平面和上坡的控制有效性。仿真结果表明使用该控制方式能够较好地控制机器人各个关节,机器人在行走过程中较为平稳。

基于CPG的六足机器人运动控制方法研究综述

以六足机器人为研究对象,分析了中枢模式发生器(CPG)的国内外发展现状,揭示其主要是以振荡器作为控制器进行信号输出。针对如何搭建CPG模型,以Hopf振荡器为例进行模型搭建和改进,搭建六足机器人实验平台验证CPG模型控制六足机器人步态运动是可行的。最后对基于CPG的六足机器人发展进行了展望。

启动子上的增强子和CpG岛在转基因沉默和位置效应中的作用

目的探讨哺乳动物细胞表达载体启动子上增强子和CpG岛调控元件在转基因沉默和位置效应中的作用。方法应用带增强子的八聚体结合转录因子4(OCT4)基因启动子、带CpG岛的性别决定区Y框蛋白2(SOX2)基因启动子、带增强子和CpG岛的巨细胞病毒(CMV)基因启动子以及不含近端增强子和CpG岛的NANOG基因启动子构建哺乳动物细胞表达载体,并分别转染中国仓鼠卵巢细胞株CHO-K1细胞和人胚胎肾细胞(HEK293)。应用Image J软件中Mean算法计算OCT4、SOX2、CMV和NANOG基因启动子在CHO-K1细胞和HEK293细胞介导的增强绿色荧光蛋白(EGFP)表达的荧光强度,应用Image J软件中Mininum、Maxentropy、Mean 3种算法分别计算OCT4、SOX2、CMV和NANOG基因启动子4种载体稳定转染CHO-K1细胞后形成的多个单细胞克隆混合生长样品在同一个样本中最大荧光强度的细胞、大于平均荧光强度的所有细胞和最小荧光强度的所有细胞的EGFP平均荧光强度;应用有限稀释法从G418筛选后获得的稳定转染的多个单克隆混合的CHO-K1细胞中,分别筛选含EGFP的pE-C1、pE-Oct4、pE-Sox2、pE-Nanog质粒稳定转染CHO-K1细胞的单克隆细胞,每个质粒载体随机筛选3个单克隆细胞,应用Image J软件分析EGFP平均荧光强度;应用酶联免疫吸附法检测转染第20、30天CHO-K1细胞中EGFP蛋白表达水平,应用实时荧光定量聚合酶链式反应法检测转染第20、30天CHO-K1细胞中EGFP mRNA表达水平。结果SOX2基因启动子在CHO-K1细胞和HEK293细胞中介导表达的EGFP平均荧光强度比较差异无统计学意义(t=0.770,P>0.05)。OCT4、CMV、NANOG基因启动子在CHO-K1细胞中介导表达的EGFP平均荧光强度显著高于HEK293细胞(t=7.000、11.100、4.900,P<0.05)。SOX2基因启动子于转染第20、30天在CHO-K1细胞中介导表达的EGFP蛋白水平比较差异无统计学意义(t=0.330,P>0.05)。CMV基因启动子于转染第20天在CHO-K1细胞中介导表达的EGFP蛋白水平显著低于转染第30天(t=3.770,P<0.05);OCT4基因启动子于转染第20、30天在CHO-K1细胞中介导表达的EGFP蛋白水平比较差异无统计学意义(t=2.500,P>0.05);NANOG基因启动子于转染第20、30天在CHO-K1细胞中介导表达的EGFP蛋白表达水平比较差异无统计学意义(t=0.014,P>0.05)。转染第20天与转染第30天SOX2、CMV、NANOG基因启动子在CHO-K1细胞中介导表达的EGFP平均荧光强度值比较差异均无统计学意义(t=0.130、0.830、0.210,P>0.05);转染第30天,OCT4基因启动子在CHO-K1细胞中介导表达的EGFP平均荧光强度值显著低于转染第20天(t=5.750,P<0.05)。SOX2、CMV、OCT4、NANOG基因启动子在CHO-K1细胞中介导表达的EGFP蛋白水平比较差异无统计学意义(F=4.070,P>0.05)。CMV、OCT4、NANOG基因启动子在CHO-K1细胞中介导的EGFP mRNA相对表达量显著高于SOX2基因启动子(t=5.440、5.000、5.740,P<0.05);CMV、OCT4基因启动子在CHO-K1细胞中介导的EGFP mRNA相对表达量显著高于NANOG基因启动子(t=3.220、4.270,P<0.05);CMV基因启动子在CHO-K1细胞中介导的EGFP mRNA相对表达量高于OCT4基因启动子,但差异无统计学意义(t=1.270,P>0.05)。SOX2基因启动子介导表达的EGFP在转录水平和转录后水平差异最大(t=16.900,P<0.05);NANOG基因启动子介导表达的EGFP在转录水平和转录后水平差异次之(t=14.930,P<0.05);OCT4和CMV基因启动子介导表达的EGFP在转录水平和转录后水平差异最小(t=2.060、0.430,P>0.05)。应用Mininum、Maxentropy、Mean 3种算法计算OCT4基因启动子在多克隆下介导表达的EGFP的荧光强度比较差异无统计学意义(F=3.720,P>0.05),SOX2、CMV和NANOG基因启动子在多克隆下介导EGFP表达的荧光强度比较差异均有统计学意义(F=516.400、428.500、28.120,P<0.05)。不同单克隆细胞中的差异分析显示,4种载体在不同单克隆细胞中介导表达的EGFP荧光强度值的标准误差相比,从高到低依次为NANOG基因启动子在不同单克隆细胞中介导表达的EGFP荧光强度值标准误差、SOX2基因启动子在不同单克隆细胞中介导表达的EGFP荧光强度值标准误差、OCT4基因启动子在不同单克隆细胞中介导表达的EGFP荧光强度值标准误差、CMV基因启动子在不同单克隆细胞中介导表达的EGFP荧光强度值标准误差,且OCT4基因启动子与SOX2、CMV基因启动子在单克隆细胞中介导表达的EGFP平均荧光强度比较差异有统计学意义(t=3.070、4.360,P<0.05)。结论带CpG岛的启动子能在转录后克服转基因沉默效应,带增强子的启动子具有克服位置效应的作用;二者的恰当组合可用于设计在哺乳动物细胞中高效稳定表达的启动子。

自制ISCOMATRIX与CpG联合的HCA587蛋白疫苗的制备及其免疫效应评估

目的确定自制免疫刺激复合物(ISCOMATRIX)作为HCA587蛋白疫苗佐剂的有效剂量,并探讨其与CpG联合作为HCA587蛋白疫苗佐剂的免疫效力和肿瘤治疗效果。方法应用透析法将Quil A、胆固醇和磷脂制备成ISCOMATRIX,经磷钨酸负染并采用透射电子显微镜观察自制ISCOMATRIX的结构。建立小鼠免疫模型,将联合CpG的HCA587蛋白疫苗与不同剂量的自制ISCOMATRIX佐剂混合,经尾根部皮下免疫小鼠;采用酶联免疫斑点试验(ELISpot)检测产生IFN-γ的脾细胞频数,采用酶联免疫吸附法(ELISA)检测血清HCA587特异性抗体水平。在小鼠右侧胁腹部皮下接种B16-HCA587肿瘤细胞建立荷瘤小鼠模型,按照处理方式不同将小鼠分为HCA587蛋白疫苗治疗组(接种100μL包含CpG与最佳剂量的自制ISCOMATRIX的HCA587蛋白疫苗)与PBS对照组(注射100μL的无菌PBS),使用游标卡尺测量肿瘤大小。结果自制ISCOMATRIX佐剂呈典型的蜂窝状结构,直径20~30 nm,大小均一。12、36μg(QuilA浓度分别为0.012%、0.036%)的自制ISCOMATRIX组小鼠脾细胞分泌IFN-γ的频数与商品化ISCOMATRIX阳性对照组比较,差异无统计学意义(均P>0.05),且显著高于CpG+36μg ISCOM对照组与PBS组(均P<0.0001);5μg与18μg自制ISCOMATRIX组小鼠分泌IFN-γ的脾细胞频数,高于CpG+36μg ISCOM对照组与PBS组(均P<0.0001),但低于商品化ISCOM组(P=0.004)。12、18与36μg自制ISCOMATRIX组的血清HCA587抗体水平与商品化ISCOM阳性对照组比较,差异无统计学意义(均P>0.05);与PBS组相比,12μg自制ISCOMATRIX组1×10^(4)倍稀释血清的抗体滴度显著升高(P=0.0062)。与PBS对照组相比,HCA587蛋白疫苗治疗组肿瘤生长速度减缓,且第35天测量时肿瘤大小显著较小(P=0.0181)。结论以12μg自制ISCOMATRIX作为佐剂联合CpG制备的HCA587蛋白疫苗能诱导有效的免疫应答,并抑制荷瘤小鼠的肿瘤的生长,具有一定的肿瘤治疗作用。

RGD修饰的低分子量鱼精蛋白/CpG ODN复合物的制备及抗肿瘤活性

目的利用RGD修饰的低分子量鱼精蛋白(low molecular weight protamine,LMWP)包裹CpG寡聚脱氧核苷酸(CpG oligodeoxynucleotide,CpG ODN)制备复合物,评价其对CpG ODN抗肿瘤活性的影响。方法基于静电引力将RGD-LMWP与CpG ODN以不同质量比共孵育制备复合物,并利用凝胶阻滞实验分析RGD-LMWP对CpG的缩合能力;使用zeta电位/粒度仪对RGD-LMWP/CpG复合物的粒径和zeta电位进行表征;采用体外细胞实验评价RGD-LMWP/CpG复合物对肿瘤细胞的摄取效率和细胞毒性。结果采用共孵育法成功制备了RGD-LMWP/CpG复合物。当RGD-LMWP与CpG的质量比为3∶1时,RGD-LMWP能完全压缩CpG。此时,复合物的平均粒径和zeta电位分别为119.7±1.2nm和47.9±0.5mV。体外细胞实验表明,与游离CpG比较,RGD-LMWP/CpG(3∶1)复合物能显著提高人子宫颈癌细胞株(HeLa)细胞对CpG的摄取效率,抑制HeLa细胞的增殖。结论RGD-LMWP与CpG结合形成的复合物可以有效地将CpG递送至肿瘤细胞内并诱导肿瘤细胞凋亡,是一种具有研究前景的抗肿瘤给药方案。

Management of regional citrate anticoagulation for continuous renal replacement therapy:guideline recommendations from Chinese emergency medical doctor consensus

Continuous renal replacement therapy(CRRT)is widely used for treating critically-ill patients in the emergency department in China.Anticoagulant therapy is needed to prevent clotting in the extracorporeal circulation during CRRT.Regional citrate anticoagulation(RCA)has been shown to potentially be safer and more effective,and is now recommended as the preferred anticoagulant method for CRRT.However,there is still a lack of unified standards for RCA management in the world,and there are many problems in using this method in clinical practice.The Emergency Medical Doctor Branch of the Chinese Medical Doctor Association(CMDA)organized a panel of domestic emergency medicine experts and international experts of CRRT to discuss RCA-related issues,including the advantages and disadvantages of RCA in CRRT anticoagulation,the principle of RCA,parameter settings for RCA,monitoring of RCA(mainly metabolic acid-base disorders),and special issues during RCA.Based on the latest available research evidence as well as the paneled experts'clinical experience,considering the generalizability,suitability,and potential resource utilization,while also balancing clinical advantages and disadvantages,a total of 16 guideline recommendations were formed from the experts'consensus.

International experts consensus guidelines on robotic liver resection in 2023

The robotic liver resection(RLR)has been increasingly applied in recent years and its benefits shown in some aspects owing to the technical advancement of robotic surgical system,however,controversies still exist.Based on the foundation of the previous consensus statement,this new consensus document aimed to update clinical recommendations and provide guidance to improve the outcomes of RLR clinical practice.The guideline steering group and guideline expert group were formed by 29 international experts of liver surgery and evidence-based medicine(EBM).Relevant literature was reviewed and analyzed by the evidence evaluation group.According to the WHO Handbook for Guideline Development,the Guidance Principles of Development and Amendment of the Guidelines for Clinical Diagnosis and Treatment in China 2022,a total of 14 recommendations were generated.Among them were 8 recommendations formulated by the GRADE method,and the remaining 6 recommendations were formulated based on literature review and experts’opinion due to insufficient EBM results.This international experts consensus guideline offered guidance for the safe and effective clinical practice and the research direction of RLR in future.