特发性血小板减少性紫癜患者调节性T细胞表达水平检测的临床意义

目的:分析特发性血小板减少性紫癜(Idiopathic thrombocytopenic purpura,ITP)患者调节性T细胞表达的变化,探讨其临床意义。方法:将我院2013年8月至2017年2月收治的117例ITP患者分别纳入初治组(n=54)、复发组(n=63);根据治疗结果再分为难治组、非难治组;选取同期30名健康体检者,纳入对照组。使用流式细胞术检测各组受试者外周血CD_4~+CD_(25)~+调节性T细胞、CD_4~+T细胞表达水平,并比较初治组治疗后不同临床疗效患者而外周血CD_4~+CD_(25)~+表达变化,分析调节性T细胞表达与ITP发生发展及预后结局的关联。结果:117例ITP患者中,初治54例,复发63例;将初治组、复发组治疗无效患者共计44例纳入难治组,将其余73例患者纳入非难治组。治疗前初治组、复发组CD_4~+CD_(25)~+、CD_4~+百分比及CD_4~+CD_(25)~+/CD_4~+比值均低于对照组,差异有统计学意义(P<0.05)。非难治组CD_4~+CD_(25)~+、CD_4~+百分比及CD_4~+CD_(25)~+/CD_4~+比值低于对照组,难治组CD_4~+CD_(25)~+、CD_4~+百分比及CD_4~+CD_(25)~+/CD_4~+比值低于非难治组,差异有统计学意义(P<0.05)。初治组显效、有效、进步者,其治疗后外周血CD4+CD25+百分比均较治疗前升高,治疗后外周血CD4+CD25+百分比由无效、进步、有效、显效依次升高,差异有统计学意义(P<0.05)。结论:ITP患者外周血CD_4~+CD_(25)~+、CD_4~+百分比及CD_4~+CD_(25)~+/CD_4~+比值的明显下降说明机体处于细胞免疫调控异常状态,调节性T细胞平衡紊乱的纠正有助于患者预后质量的改善。     

激光照射对H_2O_2处理的红细胞膜结构影响的AFM分析

本文首次利用原子力显微镜(atomic force microscope,AFM)的超高分辨率功能研究H_2O_2处理引起的红细胞膜超微结构的变化,及低强度He-Ne激光照射H_2O_2处理过的红细胞使膜结构恢复的情况。实验结果表明,无论是H_2O_2作用还是激光照射红细胞整体都能维持双凹圆盘状结构;但用AFM得到的超微结构图显示H_2O_2作用的红细胞膜表面高低起伏明显,面上平均粗糙度增加,出现较宽、较深的孔洞。随着激光照射剂量的增加,红细胞膜表面的高低起伏减弱,面上平均粗糙度下降,孔洞变浅、变窄,细胞表面趋于平滑。分析表明,H_2O_2与红细胞内游离铁离子发生类Fenton反应产生的羟自由基(·OH)损伤红细胞膜的结构,细胞的超微结构发生较明显的变化,细胞膜表面的平均粗糙度明显增大,出现较深、较宽的孔洞;激光照射消除部分自由基的作用,较小剂量的激光照射能使损伤部分减弱,膜表面的平均粗糙度降低,孔洞变浅、变窄;较大剂量的激光照射使膜表面平均粗糙度进一步降低,孔洞进一步变浅、变窄,损伤进一步恢复。     

高压传输线电磁场下细胞增殖状态的变化

高压传输线照射作用下的体外鸡胚纤维母细胞的增殖状态已经通过3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑)(MTT)比色法被研究,鸡胚纤维母细胞(CEF)被分成对照组和实验照射组,实验组暴露在4000V/m电场和0.09-0.1G磁场的220KV的高压传输线电磁场下照射了400天,得到的结果显示CEF的增殖被电磁场抑制,CEF的增长比率随着照射时间的增加而下降,相对于对照组,实验组CEF的细胞数量有所减少,这说明高压传 输线电磁场能够抑制体外 CEF 的增殖.     

弱激光对神经元瞬时外向钾通道电流特性的影响

应用全细胞膜片钳技术,研究了急性分离的大鼠海马CA3区锥体神经细胞在波长670 nm、功率5 mW的半导体激光器照射时,其瞬时外向钾通道电流特性。实验发现:弱激光对瞬时外向钾电流I_A有抑制作用,5 min激光抑制作用达到稳定,去极化至+50 mV时抑制百分比为(40.13±5.19)%(n=10);弱激光对I_A的抑制作用呈现电压依赖性和可逆性,对照组、照射组和恢复组的最大激活电流密度分别为(398.55±36.49)pA/pF、(238.62±30.78)pA/pF(n=10,P<0.01)和(354.08±35.16)pA/pF(n=10,P>0.05);激光作用可显著地影响瞬时外向钾通道电流的稳态激活和失活过程,对照组和激光照射组通道的半数激活电压分别为(-27.05±4.53) mV和(-2.10±3.14) mV(n=10, P<0.01),斜率因子分别为(-26.71±6.15) mV和(-20.70±4.38) mV(n=10,P<0.05),半数失活电压分别为(-70.49±7.21) mV和(-81.27±6.26) mV(n=10, P<0.01),斜率因子分别为(9.47±3.54) mV和(9.58±3.02) mV(n=10,P>0.05)。结果表明:弱激光作用海马神经元可以改变其瞬时外向钾通道特性,从而影响动作电位的形成和发放,调节神经元的生理功能,有利于受损神经元的恢复和再生。     

GPR119受体激动剂GSK1292263对糖尿病的作用及机制探讨

目的 :研究GPR11 9受体激动剂GSK1292263对糖尿病大鼠的影响及其作用机制。方法 :SD大鼠随机取20只作为正常对照组(N组),糖尿病建模成功后,随机分为糖尿病组(S组)、糖尿病治疗组(S+G组)。糖尿病治疗组给予GSK1292263连续灌胃12周,每2周称重1次,根据体重调整给药量。正常对照组和糖尿病模型组大鼠给予生理盐水灌胃。12周后观察3组大鼠体重、空腹血糖、空腹胰岛素的变化。免疫组化法检测胰腺组织中胰岛细胞及胰岛素分泌情况。Western-Blot检测胰腺组织中增殖细胞核抗原(PCNA)蛋白表达水平。结果:12周后,S组与S+G组体重差异无统计学意义,2组体重均明显低于N组体重。S+G组FPG和FINS低于S组,组间比较差异有统计学意义。S+G组FPG高于N组,组间比较差异有统计学意义,FINS高于N组,组间差异无统计学意义。S+G组大鼠胰腺组织受损情况轻于S组,胰岛素分泌量较S组增加,但低于同时段N组。S+G组的PCNA蛋白表达较同时间点S组高。结论:GPR119受体激动剂GSK129 2263能够有效降低大鼠体内血糖水平,通过提高胰腺组织PCNA表达水平,减少胰腺组织损害。     

不同内侧柱支撑方式固定老年肱骨距缺失的肱骨近端骨折的有限元分析

目的 通过有限元分析比较三种不同内侧柱支撑方式固定肱骨距缺失的肱骨近端骨折的力学稳定性,为临床此类骨折的内固定选择提供参考。方法 选取一名63岁老年女性肱骨近端骨折患者的右侧肱骨及左侧胫腓骨的CT平扫的影像学资料存储为DICOM格式。利用有限元分析软件重建肱骨、腓骨及骨水泥模型。利用逆向扫描及重建建立肱骨近端钛合金锁定板模型,按照肱骨距缺失的骨折形式截骨,并按照不同工况组装模型。将模型分为A、B、C三组即单纯钛合金锁定板组、钛合金锁定板联合自体腓骨植骨组、钛合金锁定板联合骨水泥组。并按螺钉数量不同分组(AE组、ACE组、全螺钉组)。利用Workbench 2021 R1做力学分析,设置各模型弹性模量、泊松比、接触方式后,计算肱骨模型的最大位移及钛合金锁定板的最大等效应力。结果 A组中A1肱骨最大位移为4.6936 mm, A2肱骨最大位移为4.6065 mm, A3肱骨最大位移为4.5814 mm; A组中A1钛合金锁定板最大等效应力为238.69 MPa, A2钛合金锁定板最大等效应力为246.85 MPa, A3钛合金锁定板最大等效应力为247.33 MPa。B组中B1肱骨最大位移...     

槲皮素对紫外线损伤NIH3T3细胞的保护作用研究

目的：研究紫外对NIH3T3细胞的影响,并探讨槲皮素对NIH3T3细胞的紫外损伤保护作用。方法：建立紫外对NIH3T3细胞的损伤模型。通过不同浓度的槲皮素（1μmol／L、10μmol／L、20μmol／L）的预处理,用MTT方法检测细胞的存活率以及激光共聚焦显微镜检测细胞内活性氧含量和线粒体膜电位变化。结果紫外损伤后的细胞内活性氧自由基含量明显增加（p〈0．05）,细胞膜电位下降（p〈0．05）,细胞凋亡后细胞存活率下降（p〈0．05）;槲皮素预处理组的细胞内活性氧自由基含量、线粒体膜电位、细胞存活率都呈现出较强的剂量依赖性。结论：紫外损伤可诱导细胞内产生大量的活性氧自由基,使细胞线粒体膜电位下降,降低细胞的存活率;槲皮素预处理后,通过清除细胞内的活性氧自由基,维持细胞的线粒体膜电位,抑制细胞凋亡。     

脂质质谱成像揭示三氯生促进三维肝癌肿瘤细胞球增长的相关机理

三氯生(triclosan, TCS)作为一种广谱抗菌剂被广泛应用于多种日常消费品中。当前研究表明，TCS暴露可以促进肝癌肿瘤增长，但涉及的相关脂质代谢机理尚不完全清楚。基于此，本研究建立了TCS暴露的肝癌肿瘤细胞球模型，利用质谱成像技术分析对照组和暴露组间脂质小分子在肿瘤细胞球内丰度和分布的差异。结果表明，6μmol/L TCS暴露可以显著促进肝癌肿瘤细胞球的增长，并引起脂质代谢的变化。27种脂质代谢小分子(含19种甘油磷脂、3种甘油酯和5种鞘脂)的丰度在对照组和暴露组的肿瘤细胞球内发生了显著性的变化。在暴露组的19种甘油磷脂中，16种在肿瘤细胞球外围增殖区发生了显著性上调；在3种甘油酯中，1种在外围增殖区发生了显著性上调，2种在内部凋亡区发生了显著性下调；5种鞘脂在肿瘤细胞球内部凋亡区均发生了显著性下调。可见，TCS可能是通过促进肿瘤细胞球外围细胞的增殖以及抑制内层细胞的凋亡来促进三维肿瘤细胞球的增长。该结果可为进一步探讨环境污染物对肿瘤发展影响的分子机制提供参考。     

血清miR-297a、FGF23水平与高血压患者冠状动脉钙化的相关性研究

目的 :观察高血压患者miR-297a、成纤维细胞生长因子23(Fibroblast growth factor23,FGF23)表达水平与冠状动脉钙化的相关性。方法 :选取95例住院高血压患者作为研究对象,另选取同期在进行体检的40例无冠状动脉钙化健康者作为对照组。所有研究对象均完成血管内超声。按照血管内超声结果,分为无钙化组、轻度钙化组、重度钙化组。采用qRT-PCR检测研究对象血清miR-297a水平,采用酶联免疫吸附试验(ELISA)检测研究对象血清FGF23水平及成骨细胞碱性磷酸酶(alkaline phosphatase,ALP)、骨形态发生蛋白2(Bone morphogenesis pale,BMP-2)、基质Gla蛋白(matrix gla protein,MGP);分析血清miR-297a、FGF23水平与ALP、BMP-2、MGP水平的关系;绘制受试者工作特征曲线(ROC),评估miR-297a、FGF23对高血压患者冠状动脉钙化的诊断价值;采用二元logistic回归分析影响高血压患者冠状动脉钙化的危险因素。结果:轻度钙化组和重度钙化组miR-297a表达低于非钙化组和健康对照组,差异有统计学意义(P<0.05),轻度钙化组和重度钙化组FGF23水平高于非钙化组和健康对照组(P <0.05);与轻度钙化组相比,重度钙化组miR-297a表达降低,组间差异有统计学意义(P <0.05),轻度钙化组与重度钙化组FGF23水平相比,差异无统计学意义(P>0.05)。轻度钙化组和重度钙化组的ALP、MGP高于非钙化组和健康对照组,差异有统计学意义(P <0.05)。轻度钙化组、重度钙化组、非钙化组BMP-2高于健康对照组,组间差异有统计学意义(P <0.05)。miR-297a与ALP、BMP-2呈负相关,与MGP呈正相关;FGF23与ALP、BMP-2呈正相关,与MGP呈负相关;ROC曲线显示,miR-297a预测高血压患者冠状动脉钙化曲线下面积为0.859(敏感度为88.7%,特异性为86.8%);FGF23水平预测高血压患者冠状动脉钙化的曲线下面积为0.906(敏感度为92.5%,特异性为89.5%);miR-297a高表达是影响血管钙化的保护因素,FGF23高表达是影响冠状动脉钙化的危险因素。结论:血清miR-297a表达降低、FGF23水平升高与高血压患者发生冠状动脉钙化有关,检测血清miR-297a、FGF23水平可评估高血压患者发生血管钙化的风险。     

矿物药青礞石对PTZ点燃癫痫大鼠脑组织中氨基酸神经递质含量的影响

建立了超高效液相色谱-三重四极杆-线性离子阱质谱（Qtrap-UPLC-MS/MS）法同时测定大鼠脑组织中4种氨基酸类神经递质,研究了矿物药青礞石原药材粉末、水煎液、药渣对戊四氮（Pentylenetetrazol,PTZ）点燃癫痫模型大鼠脑组织中氨基酸类神经递质含量的影响。在ESI⁺电离模式下,采用多反应监测扫描方式测定脑组织（皮层,海马）中谷氨酸（Glu）、甘氨酸（Gly）、天冬氨酸（Asp）及γ-氨基丁酸（GABA）的含量。结果表明：4种氨基酸的线性关系良好,精密度、准确度均符合生物样品的分析要求。大鼠给予青礞石不同样品后,脑组织皮层和海马中兴奋性神经递质Glu、Asp水平显著降低。与模型组相比,皮层中Gly占氨基酸总含量的百分比均增加,Asp/Gly值、Glu/Gly值均降低;海马中,各组Glu、Asp、Gly、GABA的浓度均降低。这说明,矿物药青礞石能有效降低PTZ点燃大鼠脑内兴奋性氨基酸递质含量,调节皮层和海马区Gly含量。     

Leptin promotes proliferation and invasion of osteosarcoma cells by upregulating the expression of SIRT1

Osteosarcoma (OS) is a primary high-grade malignant bone neoplasm, and the prognosis of OS remains poor due to early metastasis. Leptin plays an essential role in tumorigenesis, but the role of leptin in the development of OS is still not fully understood. In this study, we used a human osteosarcoma MG-63 cell line as an experimental model. MG-63 cells were treated with leptin, and cell proliferation, apoptosis, adhesion, invasion, and gene expression, were evaluated. The results showed that leptin promoted proliferation, decreased adhesion, suppressed apoptosis, and promoted invasion, of MG-63 cells. Moreover, the expression of SIRT1 was upregulated in MG-63 cells exposed to leptin. Furthermore, MMP-2, 8, and 9 were significantly upregulated by SIRT1, while SIRT1 knockdown inhibited the proliferation and invasion of MG-63 cells. In conclusion, our results suggest that leptin promotes OS cell proliferation and invasion by inducing the expression of SIRT1.     

CDK5 inhibition promotes osteoblastic differentiation of MSCs and blocks the migration of osteosarcoma MG-63 cells

CDK5 belongs to the cyclin-dependent kinase family. CDK5 is multifunctional and plays an important role in neural differentiation. However, the role of CDK5 in osteoblastic differentiation remains unclear. The present study investigated functions and molecular mechanism of CDK5 in osteoblastic differentiation. It was found that, CDK5 inhibition promoted the expression of Runx2, ALP, OCN and OPN of MSCs and the mineralization of MC-3T3E1 cells and MSCs. CDK5 inhibition enhanced the development of F-actin, nuclear localization of β-catenin and YAP, as well as the expression of RMRP RNA. When F-actin was suppressed by Blebbistatin, the nuclear localization of YAP and β-catenin, and expression of RMRP RNA as well as Runx2 and ALP were decreased. These indicate Seliciclib promotes osteoblastic differentiation mainly by F-actin. Moreover, Seliciclib also suppressed the migration of MG-63, suggesting a potential application for Seliciclib in bone defect repair and inhibition of the migration of osteosarcoma cells after osteosarcoma surgical resection.     

Atomic force microscopy can be used to mechanically stimulate osteoblasts and evaluate cellular strain distributions

In this study, atomic force microscopy (AFM) was used to mechanically stimulate primary osteoblasts. In response to mechanical force applied by the AFM, the indented cell increased its intracellular calcium concentration. The material properties of the cell could be estimated and the membrane strains calculated. We proceeded to validate this technique experimentally and a 20% error was found between the predicted and the measured diameter of indentation. We also determined the strain distributions within the cell that result from AFM indentation using a simple finite element model. This enabled us to formulate hypotheses as to the mechanism through which cells may sense the applied mechanical strains. Finally, we report the effect of the Poisson ratio and the cell thickness on the strain distributions. Varying the Poisson ratio did not change the order of magnitude of the strains; whereas the cellular thickness dramatically changed the order of magnitude of the cellular strains. We conclude that AFM can be used for controlled mechanical stimulation of osteoblasts and that cellular strain distributions can be computed with a good accuracy when the cell is indented in its highest part.     

Proliferation study and microscopy evaluation on the effects of tannic acid in human fetal osteoblast cell line (hFOB 1.19)

Tannic acid (TA) is a phenolic compound that might act directly on osteoblast metabolism. The study was performed to investigate the effects of TA on the proliferation, mineralization, and morphology of human fetal osteoblast cells (hFOB 1.19). The cells were divided into TA‐treated, untreated, and pamidronate‐treated (control drug) groups. Half maximal effective concentration (EC₅₀) values for TA and pamidronate were measured using MTT assay. The EC₅₀ of hFOB 1.19 cells treated with TA was 2.94 M. This concentration was more effective compared to the pamidronate (15.27 M). Cell proliferation assay was performed to compare cell viability from Day 1 until Day 14. The morphology of hFOB 1.19 was observed via inverted microscope and scanning electron microscope. Calcium (Ca) and phosphate (P) were assessed using energy‐dispersive X‐ray (EDX) analysis. Furthermore, the mineralization of hFOB 1.19 was determined by von Kossa staining (P depositions) and Alizarin Red S staining (Ca depositions). The number of cells treated with TA was significantly higher than the two control groups at Day 10 and Day 14. The morphology of cells treated with TA was uniformly fusiform‐shaped with filopodia extensions. Besides, globular‐like structures of deposited minerals were observed in the TA‐treated group. In line with other findings, EDX spectrum analysis confirmed the presence of Ca and P. The cells treated with TA had significantly higher percentage of both minerals at Day 3 and Day 10 compared to the two control groups. In conclusion, TA enhances cell proliferation and causes cell morphology changes, as well as improved mineralization.     

Thermally defined cryomicroscopy and some applications on human leucocytes

A freezing-stage has been developed for use on a standard light-microscope, which can provide reproducible, precisely linear cooling and warming rates in the range from 0·1 to 10,000 K/min. Biological cells in aqueous solutions can be observed during the freeze-thaw cycle; the volume loss due to osmotic efflux of water and the intracellular crystallization of water are detected by video-monitoring. The temperature field generated in the observed samples is comparable to extended cylindrical probes and allows the transfer of cryomicroscopic data to technically used vial geometries. Lymphocytes and granulocytes were observed during freezing using the system described. They were separated and washed, and then frozen on the cold stage of the cryomicroscope at cooling rates ranging from 2 to 500 K/min. Shrinkage of the cells was observed up to 100 K/min and intracellular ice formation could be detected starting at 10 K/min. The results show that human leucocytes show excessive shrinkage up to 36% of their initial volume; the probability of intracellular ice formation exhibits a sharp increase from 10 to 100 K/min where nearly all cells contain ice.     

微互连接头的蠕变数值模拟及失效寿命预测

电子封装中的微互连接头失效是电子设备的主要失效原因之一.文中根据实际封装期间建立了一个1/8的板级封装组件有限元模型.对微互连接头的蠕变变形进行了数值模拟,并依据疲劳寿命模型进行了失效预测.模拟中的互连接头分别采用了两种钎料（Sn-3.8Ag-0.7Cu和63Sn-37Pb）进行定义,对比了两种钎料的变形及寿命.     

Calibration for quantitative X-ray microanalysis of skeletal muscle cells in culture.

Skeletal muscle in culture is used to demonstrate that the intracellular concentration of elements in tissue culture cells, especially of diffusible ions such as sodium, chlorine, potassium, and calcium can be measured by X-ray microanalysis in the scanning electron microscope. Quantitative analysis is possible by comparison with X-ray spectra of cells incubated in electrolyte solutions of known concentration. The minimum detectable concentration is approximately 2 mM.     

Platelet rich plasma (PRP) induces autophagy in osteoblast precursor 3T3-L1

Autophagy is an essential cellular homeostatic mechanism by which intracellular components are delivered into the lysosomes for degradation and recycling. Autophagy has been related with a diversity of pathological or physiological dentary processes such as bone remodeling, skeletal aging, osteoclastogenesis, osteoblastogenesis and different types of oral cancer. Platelet-rich plasma (PRP), isolated from autologous blood, is a plasma preparation containing a higher concentration of platelets which contains numerous different growth factors and cytokines that activate several cellular signaling cascades. The purpose of this study is to investigate the effect of PRP on autophagy stimulation in both osteoblast precursor 3T3-L1 and non-related osteoblastic cells. Our results showed that PRP can increase the number of autophagic structures in 3T3-L1 and HeLa (cervical cancer cells) cells. Moreover, we have determined by Western blot a rise in the lipidated form of the autophagic protein LC3 (i.e. LC3-II) upon PRP treatment. Taken together, our results suggest that PRP is able to induce a strongly autophagy response in osteoblast precursor and, to a lesser extent, in non-related osteoblastic cells, suggesting that PRP could be a potential therapeutic tool for some autophagy-related diseases associated with bone homeostasis.     

阿托伐他汀钙联合普罗布考治疗冠心病合并高脂血症疗效观察

目的观察分析联合阿托伐他汀钙与普罗布考治疗冠心病合并高脂血症的临床疗效。方法将符合标准的冠心病合并高脂血症患者76例随机分为对照组38例和实验组38例,对照组口服阿托伐他汀钙20 mg/d,1次/d;观察组在此基础上加服普罗布考500mg/d,2次/d。2组均连续服药8周。治疗前后对2组患者的TC、TG、LDC-L水平进行检测。结果治疗后2组间血脂指标TC、TG、LDC-L间差异具有统计学意义（P〈0.05）。观察组与对照组完成治疗后总有效率相比,差异具有统计学意义（P〈0.05）,2组均未见明显不良反应。结论联合阿托伐他汀钙与普罗布考联合治疗冠心病合并高脂血症临床效果优于单用阿托伐他汀钙,值得推广采用。     

Numerical simulation on the local stress and local deformation in multi-point stretch forming process

Multi-point stretch forming (MPSF) is a new flexible forming technique to form aircraft outer skin parts. The multi-point stretching die (MPSD) replaces the traditional fixed shape stretching die, and the sheet metal is formed over a MPSD composed by the punch element. The MPSD is a discontinuous surface of discrete stretching die, and the stress concentration and local strain occur on formed parts. These lead to generate dimples on the surface of formed part. In this paper, a series of numerical simulations on MPSF processes for stretching parabolic cylindrical, spherical, and saddle-shaped parts were carried out. The local stress and local strain in thickness distribution of MPSF part were analyzed by dispersed the blank into solid elements. The forming results of MPSF were compared with those that use traditional stretch forming, and the influences of thickness of elastic cushion and the size of punch element on the stress concentration and local strain were surveyed. The simulation results show the distribution of local stress and local deformation in different layers, and the elastic cushion and the small size of punch element can reduce the stress concentration and local deformation. The results may understand the stress distribution on the sheet and prevent the defect of dimple.