S100 protein expression during induced Schwann cell-like cell differentiation of rat bone marrow mesenchymal cells in vitro

BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.

Immune phenotypes of prostate cancer cells: Evidence of epithelial immune cell-like transition?

Prostate cancers(PCa)have been reported to actively suppress antitumor immune responses by creating an immune-suppressive microenvironment.There is mounting evidence that PCas may undergo an‘‘Epithelial Immune Cell-like Transition’’(EIT)by expressing molecules conventionally associated with immune cells(e.g.,a variety of cytokines/receptors,immune transcription factors,Ig motifs,and immune checkpoint molecules),which subsequently results in the suppression of anti-cancer immune activity within the tumor microenvironment.Recent progress within the field of immune therapy has underscored the importance of immune checkpoint molecules in cancer development,thus leading to the development of novel immunotherapeutic approaches.Here,we review the expression of select immune checkpoint molecules in PCa epithelial and associated immune cells,with particular emphasis on clinical data supporting the concept of an EIT-mediated phenotype in PCa.Furthermore,we summarize current advances in anti-immune checkpoint therapies,and provide perspectives on their potential applicability.

Stem cell-like memory T cells:Role in viral infections and autoimmunity

Stem cell-like memory T(TSCM)cells possess stem cell properties including multipotency and self-renewal and are being recognized as emerging players in various human diseases.Advanced technologies such as multiparametric flowcytometry and single cell sequencing have enabled their identification and molecular characterization.In case of chronic viral diseases such as human immunodeficiency virus-1,CD4+T_(SCM) cells,serve as major reservoirs of the latent virus.However,during immune activation and functional exhaustion of effector T cells,these cells also possess the potential to replenish the pool of functional effector cells to curtail the infection.More recently,these cells are speculated to play important role in protective immunity following acute viral infections such as coronavirus disease 2019 and might be amenable for therapeutics by ex vivo expansion.Similarly,studies are also investigating their pathological role in driving autoimmune responses.However,there are several gaps in the understanding of the role of T_(SCM) cells in viral and autoimmune diseases to make them potential therapeutic targets.In this minireview,we have attempted an updated compilation of the dyadic role of these complex T_(SCM) cells during such human diseases along with their biology and transcriptional programs.

Doublecortin-like kinase 1 exhibits cancer stem cell-like characteristics in a human colon cancer cell line

Objective： Colon cancer stem cells （CSCs） are implicated in colorectal cancer carcinogenesis, metastasis, and therapeutic resistance. The identification of these cells could help to develop novel therapeutic strategies. Doublecortin-like kinase 1 （DCLK1） has been viewed as a marker for gastrointestinal stem cells that fuel the self-renewal process, however others view them as a marker of Tuft cells or as an enteroendocrine subtype. The purpose of this study was to use a colon cancer cell line to identify and characterize the stem-like characteristics of the DCLKI＋ cell population. Methods： To enrich stem-like cells, HCT116 cells （derived from colon adenocarcinomas） were cultured using serum-free media to form spheres under both normal oxygen and hypoxia condition. DCLK1 transcript expression in the adherent parental cells and spheroids was quantified using quantitative real time reverse transcription- polymerase chain reaction [（q）RT-PCR]. DCLK1 protein expression was determined using flow cytometry. Self-renewal capability from adherent parental cells and spheroids was determined using extreme limiting dilution analysis （ELDA）. Results： Under both normal oxygen and hypoxia condition, the adherent parental cells were composed of cells that express low levels of DCLK1. However, spheroids exhibited an increased frequency of cells expressing DCLK1 on both mRNA and protein levels. Cells derived from spheroids also possess stronger self-renewal capability. Conclusions： The higher fraction of DCLK1 ＋ cells exhibited by spheroids and hypoxia reflects the stem- like characteristics of these cells. DCLK1 may represent an ideal marker to study and develop effective strategies to overcome chemo-resistance and relapse of colon cancer.

Phenotype-Directed DNA Nanomachine for in Situ Analysis of Stem Cell-like Subpopulation in Breast Cancer

The cancer stem cell hypothesis provides a basis for prediction of the recurrence and risk of metastasis in breast cancer.However,the unique expression pattern of stemness markers and the presence of nonstem-like cancer cells with varied phenotypes have brought great challenges to the characterization of breast cancer stem cells.To address these challenges,a phenotype-directed DNA nanomachine has been designed for high-accuracy labeling and in situ analysis of the stem cell-like subpopulation in breast cancer.The key for the design is to use cell surfaceanchored inputs to activate the nanomachine,which undergoes different branch migration pathways such that the signal strand can only be brought onto the cancer cells having the stem cell-like phenotype.Highly sensitive determination and single-step isolation of the stem cell-like subpopulation were achieved by incorporating functional groups into the signal strand such that the nanomachine was successfully applied in a tumor-bearing mouse model.Overall,the approach provides for a substantial improvement in capability for the analysis of the breast cancer stem cell-like subpopulation,and it is expected that the new approach will advance the use of DNA nanomachines in cancer-related studies.

Neural grafts containing exosomes derived from Schwann cell-like cells promote peripheral nerve regeneration in rats

Background:Schwann cell-like cells(SCLCs),differentiated from mesenchymal stem cells,have shown promising outcomes in the treatment of peripheral nerve injuries in preclinical studies.However,certain clinical obstacles limit their application.Hence,the primary aim of this study was to investigate the role of exosomes derived from SCLCs(SCLCs-exo)in peripheral nerve regeneration.Methods:SCLCs were differentiated from human amniotic mesenchymal stem cells(hAMSCs)in vitro and validated by immunofluorescence,real-time quantitative PCR and western blot analysis.Exosomes derived from hAMSCs(hAMSCs-exo)and SCLCs were isolated by ultracentrifugation and validated by nanoparticle tracking analysis,WB analysis and electron microscopy.A prefab-ricated nerve graft was used to deliver hAMSCs-exo or SCLCs-exo in an injured sciatic nerve rat model.The effects of hAMSCs-exo or SCLCs-exo on rat peripheral nerve injury(PNI)regeneration were determined based on the recovery of neurological function and histomorphometric variation.The effects of hAMSCs-exo or SCLCs-exo on Schwann cells were also determined via cell prolifer-ation and migration assessment.Results:SCLCs significantly expressed the Schwann cell markers glial fibrillary acidic protein and S100.Compared to hAMSCs-exo,SCLCs-exo significantly enhanced motor function recov-ery,attenuated gastrocnemius muscle atrophy and facilitated axonal regrowth,myelin forma-tion and angiogenesis in the rat model.Furthermore,hAMSCs-exo and SCLCs-exo were effi-ciently absorbed by Schwann cells.However,compared to hAMSCs-exo,SCLCs-exo signifi-cantly promoted the proliferation and migration of Schwann cells.SCLCs-exo also significantly upregulated the expression of a glial cell-derived neurotrophic factor,myelin positive regulators(SRY-box transcription factor 10,early growth response protein 2 and organic cation/carnitine transporter 6)and myelin proteins(myelin basic protein and myelin protein zero)in Schwann cells.Conclusions:These findings suggest that SCLCs-exo can more efficiently promote PNI regeneration than hAMSCs-exo and are a potentially novel therapeutic approach for treating PNI.

Deposition of bio-mimicking graphene sheets with lotus leaf-like and cell-like structures on the nickel substrate

Bio-mimicking graphene films,deposited on textured nickel substrates,were synthesized by the following method:replicating the surface textures of the lotus leaf by polymer duplication,fabricating textured nickel substrates by electroplating on the polymer coated with a Au film,preparing bio-mimicking graphene oxide films on the nickel substrates by vacuum filtration,and electrochemical reduction.By controlling the vacuum filtration,this replica method can not only replicate the lotus leaf structure by a graphene film,but also can achieve a novel cell-like graphene film.

Self-assembly with varying hydrophobic centers:Synthesis of red blood cell-like basic magnesium carbonate microspheres

Basic magnesium carbonate microspheres with a red blood cell （RBC）-like appearance and diameters of ~3μm were synthesized by amphiphilic molecule-participated self-assembly under hydrothermal conditions, In the self-assembly, sodium dodecyl benzene sulfonate served as a template for the formation of Mg（OH）2 spherical micelles and also as a reactant precursor that releases CO2 to react with Mg（OH）2. The growth of the microspheres is driven by the continuous generation of new hydrophobic centers because of the consumption of hydrophilic poles （--SO3-）. The surfactant-directed self-assembly can be applied to the synthesis of other carbonate or metallic oxide self-assemblies, indicating that it is a universal self-assembly method for amphiphilic molecules.

Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells

During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions.However,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported.In this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions.By using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced differentiation.We obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 down-regulated.We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other proteins.These proteins also include neural and glial proteins,such as BDNF,CNTF and GFAP.The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.

基于NOD样受体3炎性小体通路对利拉鲁肽在氧化低密度脂蛋白诱导内皮细胞损伤的作用机制研究

背景动脉粥样硬化是世界范围内引起心脑血管疾病最主要的原因,炎症是目前研究热点,其中NOD样受体3(NLRP3)是研究最为深入的炎症小体。胰高糖素样肽1(GLP-1)受体激动剂有抗动脉粥样硬化作用,具体机制尚不明确。目的研究利拉鲁肽通过拮抗氧化低密度脂蛋白(ox-LDL)诱导的内皮细胞损伤的作用机制。方法2022-03-25—05-19培养人脐静脉内皮细胞(HUVEC),取HUVEC加空白血清作为对照组,100μg/mL的ox-LDL干预HUVEC 48 h作为模型组,100μg/mL的ox-LDL干预HUVEC 24 h后分别加入100、200、400 nmol/L利拉鲁肽处理24 h作为利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组。CCK-8法计算细胞增殖率。通过扫描电镜观察焦亡细胞形态。检测乳酸脱氢酶(LDH)活力。酶联免疫吸附试验(ELISA)检测白介素(IL)-1β、IL-18表达水平。蛋白质免疫印迹试验(Western blot)检测NLRP3、接头蛋白凋亡相关斑点样蛋白(ASC)、天冬氨酸蛋白水解酶1(Caspase-1)、焦亡执行蛋白(GSDMD)、N端结构域的焦亡执行蛋白(N-GSDMD)表达水平。结果模型组、利拉鲁肽低浓度组和利拉鲁肽中浓度组细胞增殖率低于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组细胞增殖率高于模型组(P<0.05)。细胞扫描电镜结果示模型组细胞焦亡明显,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组细胞焦亡情况明显改善。模型组、利拉鲁肽低浓度组LDH活力高于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组低于模型组(P<0.05)。模型组、利拉鲁肽低浓度组IL-1β表达水平高于对照组,利拉鲁肽中浓度组、利拉鲁肽高浓度组IL-1β表达水平低于模型组(P<0.05);模型组IL-18表达水平高于对照组,利拉鲁肽低浓度组、利拉鲁肽中浓度组、利拉鲁肽高浓度组IL-18表达水平低于模型组(P<0.05)。模型组NLRP3、ASC、Caspase-1、GSDMD、N-GSDMD表达水平高于对照组,利拉鲁肽低浓度组ASC、Caspase-1表达水平高于对照组,利拉鲁肽中浓度组NLRP3、ASC表达水平低于模型组,利拉鲁肽高浓度组NLRP3、ASC、Caspase-1表达水平低于模型组(P<0.05)。结论利拉鲁肽显著抑制ox-LDL诱导的内皮细胞NLRP3炎性小体活化,并且能够抑制内皮细胞的焦亡,具有抗动脉粥样硬化作用。

血管生成素样蛋白4通过调节成纤维细胞和内皮细胞功能影响糖尿病足进程

背景:研究表明,血管因素对糖尿病足的发生具有重要影响。目的:探讨血管生成素样蛋白4在糖尿病足形成中的重要作用。方法:①对糖尿病足患者的基因表达谱数据进行生物信息学分析,找到关键基因。收集糖尿病足患者以及无糖尿病健康人的皮肤标本进行苏木精-伊红染色、免疫组化染色以及qRT-PCR实验,检测血管生成素样蛋白4表达情况。②培养人永生化皮肤成纤维细胞系和原代人脐静脉内皮细胞,将2种细胞分别分为对照组和外源性补充血管生成素样蛋白4组,通过划痕实验以及CCK-8实验分别检测成纤维细胞的迁移能力和增殖能力,通过Ki67实验检测内皮细胞的增殖能力。结果与结论:①生信分析发现,血管生成素样蛋白4基因的下调可能是导致糖尿病足形成的关键基因。②苏木精-伊红染色结果显示,与正常皮肤相比,血管生成素样蛋白在糖尿病足皮肤内弱表达,且其mRNA水平相对表达量降低(P<0.01)。③划痕实验结果显示,与对照组相比,血管生成素样蛋白4组成纤维细胞迁移能力明显增强;CCK-8细胞增殖实验显示,血管生成素样蛋白4组成纤维细胞的吸光度值在24,48 h均高于对照组(P<0.01,P<0.001);提示血管生成素样蛋白4可增强高糖处理的成纤维细胞迁移及增殖能力。④Ki67实验结果显示,与对照组相比,血管生成素样蛋白4组内皮细胞Ki67阳性细胞数目明显多于对照组;CCK-8细胞增殖实验显示,血管生成素样蛋白4组内皮细胞的吸光度值在24,48 h均高于对照组(P<0.05,P<0.001)。(5)以上结果均提示血管生成素样蛋白4可增强高糖处理内皮细胞的增殖能力。

腺病毒介导SDF-1/NELL-1双基因转染ADSCs复合Nano-n HA支架对犬下颌骨缺损修复的实验研究

目的:构建腺病毒介导的基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)和尼尔样-1型分子(Nell-like molecule-l,Nell-1)双基因转染犬ADSCs复合Nano-n HA支架,观察其对犬下颌骨缺损的修复作用。方法:构建携SDF-1及NELL-1目的基因片段的腺病毒表达载体,分组转染犬ADSCs后行体外成骨分化诱导,ELISA法检测目的基因转染ADSCs后结合支架体内外生长各期目的蛋白表达。20只比格犬随机分为5组,A组为空白组(无支架置入),B组为单纯支架组,C组为SDF-1/Nano-n HA组,D组为Nell-1/Nano-n HA组,E组为SDF-1/Nell-1/Nano-n HA组。CM-Dil细胞标记后构建ADSCs-Nano-n HA支架骨组织工程复合体,制备犬双侧下颌骨缺损模型,将不同细胞支架复合体分组植入下颌骨缺损区。术后第4、8、12周取材行大体观察、CT、扫描电镜、细胞示踪实验及组织学检测,比较各组缺损区新骨形成情况,行统计学分析。结果:ADSCs传代培养及成骨诱导分化状态良好,荧光显微镜下观察SDF-1、Nell-1及SDF-1/Nell-1重组腺病毒均能稳定转染ADSCs,各组目的蛋白表达体内外实验表达有显著性差异。通过大体观察及X线、CT扫描、ECM检测发现转染组骨缺损区新骨形成情况优于未转染组,且共转染组成骨速度及质量优于其他各组。组织学染色可见转染组新骨形成及血管生成情况均优于未转染组,且共转染组新生骨小梁面积及骨成熟度均优于其他各组。结论:SDF-1、Nell-1均可转染ADSCs并可稳定表达,目的基因转染ADSCs复合Nano-n HA支架后可显著促进下颌骨缺损的成骨修复,为组织工程修复成骨提供了新路径。

N6-甲基腺苷甲基化相关基因IGF2BP3在肾透明细胞癌的作用研究

目的筛选肾透明细胞癌(ccRCC)中的关键N6-甲基腺苷(m^(6)A)甲基化相关基因,并研究其与ccRCC预后、ccRCC细胞的迁移和侵袭的关系。方法从癌症基因组图谱(TCGA)和基因型组织表达(GTEx)数据库中下载ccRCC和癌旁组织的RNA测序数据和临床数据,采用R4.1.1分析表达谱和预后,并筛选关键基因。收集10例ccRCC手术临床标本,采用定量PCR(qPCR)和免疫组织化学法分别检测基因mRNA和蛋白表达。在人ccRCC细胞系RCC23中,通过SiRNA敲减关键基因,并用CCK-8检测细胞的存活率,采用划痕试验和Transwell试验分别检测细胞的迁移和侵袭。结果19个m^(6)A甲基化相关基因中仅有胰岛素样生长因子ⅡmRNA结合蛋白3(IGF2BP3)在ccRCC组织中高表达,且IGF2BP3高表达与ccRCC患者预后不良呈正相关。通过qPCR和免疫组织化学法在临床标本中验证了IGF2BP3的高表达。通过小干扰RNA(siRNA)将IGF2BP3敲减后发现,RCC23细胞的存活率明显下降,且细胞的迁移和侵袭能力下降。结论IGF2BP3可能是预测ccRCC患者预后的生物标志物和潜在的药物治疗靶点。

人脐带间充质干细胞抑制脓毒症小鼠肺组织细胞的焦亡

背景:急性肺损伤是脓毒症患者的常见严重并发症,其发病机制复杂,目前尚无有效的药物治疗方案。人脐带间充质干细胞通过抑制细胞过度焦亡从而改善脓毒症急性肺损伤的作用机制愈发受到重视。目的:探索人脐带间充质干细胞对脓毒症所致急性肺损伤小鼠肺组织细胞焦亡的影响及机制。方法:48只Balb/c雄性小鼠按随机数字法分为假手术组、模型组、治疗组(n=16)。模型组、治疗组行盲肠结扎穿孔建立脓毒症所致急性肺损伤模型,假手术组仅开腹后缝合。治疗组术后6 h尾静脉注射200μL人脐带间充质干细胞悬液(5×105个细胞),模型组及假手术组术后6 h尾静脉注射200μL生理盐水。术后28 h,各组随机取5只小鼠予以尾静脉注射伊文思蓝检测肺血管通透性;收集各组剩余小鼠肺泡灌洗液和肺组织,ELISA检测肺泡灌洗液中白细胞介素1β、白细胞介素18水平,计数肺泡灌洗液中细胞总数及巨噬细胞数;苏木精-伊红染色观察肺组织病理学形态,TUNEL染色观察肺组织细胞焦亡情况,RT-PCR、Western blot检测肺组织中Toll样受体4、GSDMD及Caspase-11的mRNA和蛋白表达。结果与结论:与模型组相比,治疗组术后28 h肺血管通透性显著下降(P<0.05),肺泡灌洗液中白细胞介素1β、白细胞介素18水平降低(P<0.05),细胞总数及巨噬细胞数降低(P<0.05),肺损伤程度减轻,肺组织细胞焦亡指数降低(P<0.05),肺组织Toll样受体4、GSDMD、GSDMD-N及Caspase-11 mRNA及蛋白表达水平均显著下降(P<0.05)。结果表明,人脐带间充质干细胞可能通过抑制Toll样受体4/Caspase-11/GSDMD信号通路活化,改善肺组织细胞焦亡,从而在一定程度上改善脓毒症小鼠肺损伤的严重程度。

组织工程化口腔黏膜等效物的血管化特征

背景:前期研究中三维细胞重建组织工程化口腔黏膜等效物结构类似于正常口腔黏膜,即存在类上皮样结构、类固有层样结构、类血管腔样结构,并已初步实现了等效物的血管化建立,但其血管化特征尚不十分明确。目的:采用血管内皮细胞特异性标志物表达谱关联激光捕获显微切割系统靶向获取血管化口腔黏膜等效物的血管样结构,评价其成血管能力,揭示其血管化特征。方法:分别从人牙龈上皮组织和固有层组织原代培养人牙龈上皮细胞、人牙龈成纤维细胞、人牙龈间充质干细胞,人牙龈间充质干细胞经单克隆扩增培养后诱导分化形成血管内皮样细胞。将人牙龈上皮细胞、人牙龈成纤维细胞、血管内皮样细胞分层负载于脱细胞血管基质-0.25%类人Ⅰ型胶原支架上,构建血管化口腔黏膜等效物。将血管化口腔黏膜等效物(实验组)与脱细胞血管基质-0.25%类人Ⅰ型胶原支架(对照组)分别植入裸鼠背部皮下,14 d后两组切口表面涂布生物胶,实验组生物胶表面接种人牙龈上皮细胞,对照组不接种细胞,继续饲养14 d后取材,利用形态学观察口腔黏膜等效物分层结构;采用较为全面的血管内皮细胞特异性标志物表达谱对口腔黏膜等效物中的新生血管样结构进行免疫组化、免疫荧光标记,进行血管化特征分析;采用激光捕获显微切割系统靶向捕获免疫组化特异性标记的口腔黏膜等效物中新生血管样结构,靶向分析其血管化特征。结果与结论:(1)形态学观察显示口腔黏膜等效物细胞层次清晰,结构类似于正常口腔黏膜,即存在类上皮样结构、类固有层样结构、类血管腔样结构,类血管腔样结构内存在散在红细胞;(2)口腔黏膜等效物组中EdU Apollo示踪种子细胞结果显示:EdU Apollo 488标记的人牙龈上皮细胞呈绿色荧光表达;DAPI标记的人牙龈成纤维细胞呈蓝色荧光表达,体内形成类固有层样结构;Ed U Apollo 567标记的血管内皮样细胞呈红色荧光表达,体内形成类血管样结构;(3)血管内皮细胞特异性标志物表达谱免疫荧光标记血管结构显示,与正常口腔黏膜相比,口腔黏膜等效物中CD31、CD51、CD54、CD105、Tie-2、VWF、血管内皮生长因子受体1、血管内皮生长因子受体2表达升高(P<0.0001),CD34表达无明显变化(P>0.05);(4)与特异性标记的口腔黏膜血管结构相比,激光捕获显微切割系统靶向捕获的口腔黏膜等效物血管样结构中CD51、CD54、CD105、Tie-2、VWF、血管内皮生长因子受体1、血管内皮生长因子受体2表达升高(P<0.0001),CD31、CD34表达无明显变化(P>0.05);(5)结果表明,通过三维细胞分层重建的口腔黏膜等效物能够实现良好的血管化,其血管化特征符合新生血管生成的免疫学功能及特点;血管化助力三维细胞分层重建的口腔黏膜等效物再生。

人脐带间充质干细胞共培养联合人参皂苷Rg1对心力衰竭细胞模型的影响

背景:目前,如何促进细胞扩增、减少细胞损失、提高细胞归巢率、减少细胞凋亡是人脐带间充质干细胞临床前研究的主要问题。人参皂苷Rg1可在体外或体内促进间充质干细胞在不同微环境中的增殖、分化,其可能是提高人脐带间充质干细胞移植效率的候选药物。目的:探讨人脐带间充质干细胞共培养联合人参皂苷Rg1对戊巴比妥钠诱导的心衰细胞模型的影响。方法:将H9C2细胞分为5组:对照组、戊巴比妥钠组、人脐带间充质干细胞组、人参皂苷Rg1组、人脐带间充质干细胞+人参皂苷Rg1组。对照组H9C2细胞用正常DMEM培养基培养24 h,其他组H9C2细胞用含0.8%戊巴比妥钠的DMEM培养基培养7 min构建心衰细胞模型。建模后,分别用人脐带间充质干细胞、人参皂苷Rg1单独或共同处理。采用CCK-8法和EdU染色检测细胞增殖情况,TUNEL法检测细胞凋亡情况,按试剂盒说明检测Na^(+)-K^(+)-ATP酶和Ca^(2+)-Mg^(2+)-ATP酶活性,ELISA测定上清液中肿瘤坏死因子α、白细胞介素1β和白细胞介素6水平,RT-qPCR测定细胞中肿瘤坏死因子α、白细胞介素1β、白细胞介素6、Bax和Bcl-2 mRNA表达,Western blot测定细胞中Bax、Bcl-2、Toll样受体4、p65和p-p65蛋白表达。结果与结论:①与戊巴比妥钠组比较,人脐带间充质干细胞组、人参皂苷Rg1组和人脐带间充质干细胞+人参皂苷Rg1组H9C2细胞活力和EdU阳性率升高,TUNEL阳性率以及Bax mRNA和蛋白表达降低,Bcl-2 mRNA和蛋白表达升高,Na^(+)-K^(+)-ATP酶活性降低,Ca^(2+)-Mg^(2+)-ATP酶活性升高,H9C2细胞上清液中肿瘤坏死因子α、白细胞介素1β和白细胞介素6水平降低,H9C2细胞中肿瘤坏死因子α、白细胞介素1β和白细胞介素6 mRNA表达降低,Toll样受体4和p-p65蛋白表达降低,差异均有显著性意义(P<0.05);②与人脐带间充质干细胞组和人参皂苷Rg1组比较,人脐带间充质干细胞+人参皂苷Rg1组上述指标进一步改善(P<0.05)。结果表明:人脐带间充质干细胞联合人参皂苷Rg1提高了戊巴比妥钠诱导的心衰细胞活力,并抑制了Toll样受体4/核因子κB通路介导的炎症。

基于蜂窝仿生流场的质子交换膜燃料电池的性能研究及优化

为了增强质子交换膜燃料电池(PEMFCs)内部燃料分布的均匀性与水管理能力,本文研究了一种新型的基于仿生结构的蜂窝仿生流场。首先,根据蜂窝结构特征,搭建出三维多相流非等温几何模型并确立PEMFC的数学模型;其次,采用计算流体动力学Fluent软件对模型进行无关性与有效性验证,并对蜂窝仿生流场和常规平行流场的电池性能和燃料分布进行了模拟分析;最后,对比极化性能、气体流速分布、燃料与液态水组分浓度和电流密度分布,验证了蜂窝仿生流场设计的有效性。研究结果表明:采用蜂窝仿生流场的电池峰值功率密度比常规平行流场高15.6%,其内部水管理能力和燃料分布均匀性均强于平行流场;左端进气的蜂窝仿生流场比采用上端进气的蜂窝仿生流场在气体通路内具有更均匀的气体压力,可有效避免流道内产生局部涡流,其峰值功率密度可以达到0.3933 W/cm2。

番泻苷B通过Wnt/β-catenin通路抑制视网膜母细胞瘤HXO-Rb44细胞增殖、凋亡和侵袭

目的探究番泻苷B对视网膜母细胞瘤HXO-Rb44细胞增殖、凋亡、侵袭及Wnt/β-catenin信号通路的影响。方法通过采用不同剂量(0、5、10、20μmol/L)番泻苷B处理HXO-Rb44细胞,将细胞随机分为4组:Control组,番泻苷B 5、10、20μmol/L组。MTT法检测细胞活力;克隆形成实验检测克隆形成率;流式细胞仪检测细胞凋亡率;Transwell检测侵袭细胞数;细胞成球实验检测细胞成球直径和细胞成球数目;蛋白质印迹检测cleaved caspase-3、caspase-3、MMP-2、MMP-9、SOX2、OCT4、CD44、Wnt1、β-catenin蛋白表达。结果与Control组比较,番泻苷B 10、20μmol/L组细胞活力、克隆形成率显著降低(P<0.05),细胞凋亡率和cleaved caspase-3/caspase-3表达显著升高(P<0.05),侵袭细胞数和MMP-2、MMP-9蛋白表达显著降低(P<0.05),细胞成球直径、细胞成球数目和SOX2、OCT4、CD44蛋白表达显著降低(P<0.05),Wnt1、β-catenin蛋白表达显著降低(P<0.05)。结论番泻苷B可抑制视网膜母细胞瘤HXO-Rb44细胞增殖、侵袭和干细胞样特性,诱导细胞凋亡,抑制Wnt/β-catenin信号通路的活化。

血清sTLT-1、SP-D、NF-κB水平在脓毒症急性肺损伤预后评估中的价值

目的 探究血清可溶性髓样细胞触发受体样转录因子-1(sTLT-1)、表面活性蛋白-D(SP-D)、核转录因子-κB(NF-κB)水平在脓毒症急性肺损伤(ALI)预后评估中的价值。方法 选择2021年4月—2023年4月来我院就诊脓毒症患者36例作为脓毒症组,选择同期脓毒症并ALI患者80例作为ALI组。比较两组的一般资料、血清sTLT-1、SP-D、NF-κB水平、APACHEⅡ评分、LIPS评分。ALI组患者根据1个月内生存情况,分为死亡组(n=32)和存活组(n=48)。比较死亡组和存活组的一般资料、血清sTLT-1、SP-D、NF-κB水平、APACHEⅡ评分、LIPS评分,分析ALI组预后不良的影响因素及血清学sTLT-1、SP-D、NF-κB指标对ALI预后评估价值。结果 ALI组患者的sTLT-1、SP-D、NF-κB、APACHEⅡ评分、LIPS评分均高于脓毒症组(P<0.05);单因素分析结果显示,ALI患者中死亡组及存活组的性别、年龄、感染部位、吸烟例数及BMI水平比较,差异无统计学意义(P>0.05);ALI患者中死亡组血清sTLT-1、SP-D、NF-κB、APACHEⅡ评分、LIPS评分均高于存活组(P<0.05);多因素分析显示,血清sTLT-1、SP-D、NF-κB、APACHEⅡ评分、LIPS评分是ALI患者预后不良的独立危险因素;采用ROC曲线分析,sTLT-1、SP-D、NF-κB联合诊断ALI预后不良的AUC为0.914,均高于单一sTLT-1、SP-D、NF-κB的0.832、0.796、0.816。结论 血清sTLT-1、SP-D、NF-κB水平与脓毒症ALI的发生及发展相关,同时是其预后不良的独立危险因素,临床联合检测sTLT-1、SP-D、NF-κB三项指标有助于指导ALI预后评估。

吉林地区朝、满、汉3个民族KIR基因的分布频率研究

目的通过分析吉林地区汉族、满族及朝鲜族的杀伤细胞免疫球蛋白样受体(KIR)基因的频率与多态性,为探究KIR基因与疾病关联提供基础数据支持。方法采用聚合酶链反应-序列特异性引物(PCR-SSP)技术对吉林地区129名满族、198名朝鲜族及201名汉族人群的KIR基因进行分型。结果KIR3DL2、KIR3DL3、KIR3DP1及KIR2DL4在所有检测对象中均为100%检出。KIR2DL1、KIR2DL3、KIR2DS4、KIR3DL1及KIR2DP1等基因在这3个民族中的检出频率较高,介于93%~98%之间。相比之下,KIR2DL2、KIR2DL5、KIR3DS1、KIR2DS1、KIR2DS2、KIR2DS3及KIR2DS5的检出率较低,分布在13%~45%。特别是满族KIR2DL5(17.83%)与KIR2DS1(17.83%)的基因检出频率显著低于吉林地区朝鲜族(42.93%、47.47%)和汉族(33.83%、33.33%);吉林地区朝鲜族KIR2DL5(42.93%)与KIR2DS1(47.47%)的检出频率显著高于汉族(33.83%、33.33%)和满族(17.83%、17.83%);吉林地区汉族单体型KIRAA的频率在吉林3个受检民族中是最高的为61.19%,显著高于朝鲜族(42.93%),以上各组差异比较P<0.05,经Bonferroni校正后,Pc<0.05。结论吉林地区的朝鲜族、满族与汉族在KIR基因分布方面既体现了中国人群KIR基因多态性,又各自展示了独特的民族遗传特性和地域性。