外泌体对脂多糖诱导的牙髓干细胞成骨分化的作用及代谢机制

目的:探究外泌体对脂多糖(LPS)诱导的牙髓干细胞(DPSCs)成骨分化的影响.方法:收集健康成人和儿童的脱落乳牙牙髓干细胞(SHED),分离、培养DPSCs和SHED并用流式细胞仪鉴定.收集SHED上清的外泌体,使用透射电镜、Western blot和粒径分析进行鉴定;检测对照组(10 mg/L PBS)、LPS组(10 mg/L LPS)和LPS+外泌体组(10 mg/L LPS+10 mg/L外泌体)的DPSCs细胞外酸化率(ECAR)和耗氧率(OCR)水平;加入线粒体呼吸酶抑制剂抗霉素A后,实时定量RT?PCR和茜素红染色检测各组DPSCs的成骨分化能力.结果:成功获得DPSCs和SHED来源外泌体;LPS可以提高DPSCs的ECAR水平,使OCR/ECAR水平下降,而加入外泌体后,DPSCs的OCR水平升高,OCR/ECAR水平得到恢复;外泌体可以恢复LPS诱导的成骨分化下降,而加入抗霉素A可以抑制外泌体的促成骨分化能力.结论:SHED来源的外泌体通过提高OCR/ECAR水平,来促进炎症微环境下DPSCs的成骨分化能力.     

Radiological manifestations of renal osteodystrophy in the orofacial region: a case report and literature review

Renal osteodystrophy (ROD) is a metabolic skeletal pathology that frequently affects the orofacial region and presents with characteristic radiological manifestations. This paper reports a case of ROD causing diffused orofacial bone alterations in a female patient with end-stage chronic kidney disease. The radiological features on panoramic, posteroanterior and lateral skull and computed tomography radiographs are described for this case. Previous reports in the literature are reviewed and discussed with a focus on radiological characteristics and differential diagnoses.     

Hearing impairment and hearing aid usage in Singapore

Objective: To examine the epidemiological and audiological profile, pattern of hearing aid (HA) fitting and usage among HA users in Singapore. Design: Retrospective cross-sectional study. Study sample: Cohort of 1068 subjects issued with HAs at a tertiary hospital between 2001 and 2013. Results: Mean age was 70?years with 50.8% male. Mean hearing loss (HL) was 63.1?dB and 69.5% had at least moderate-severe HL. Sensorineural HL was present in 66.4% and 69.8% had symmetrical HL. “Behind-the-Ear” HAs were cheaper than “In-Ear” HAs. Standard “Behind-The-Ear” HAs were also cheaper than “Receiver-in-Canal” HAs. Among In-Ear HAs, “In-the-Canal” and “Completely-in-Canal” were more popular than “In-the-Ear” HAs despite costing more. HA was used ≥4?days/week by 85.6% but >7?h per day by only 35.7%. Only 18% received bilateral first HA fitting. In multivariate analysis, younger age and symmetrical HL were predictors of bilateral HA uptake while better Pure-Tone-Audiometry of aided ear and >7?h of daily HA usage were predictors of successive HA fitting. Conclusions: HA users in Singapore were elderly and presented with advanced HL. Bilateral HA adoption and average daily use were low compared to other developed countries. Future research on understanding the suboptimal HA usage should explore patients' motivation, hearing disability, and HA effectiveness.     

磷脂酰肌醇3激酶/蛋白激酶B通过上调内皮型一氧化氮合酶参与远端缺血后处理的脑保护作用

目的 探讨内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及磷脂酰肌醇3激酶/蛋白激酶B(phosphoinositide-3 kinase/protein kinase B,PI3K/Akt)通路在远端缺血后处理(remote ischemic postconditioning,RIPoC)减少大鼠全脑缺血/再灌注(ischemia/repeffusion,I/R)损伤中的作用.方法 成年雄性SD大鼠100只,体重为200 g～250 g,按随机数字表法随机分为5组(每组20只):假手术组(S组)、缺血/冉灌注组(I/R组)、缺血/冉灌注+远端缺血后处理组(I/R+RIPoC组)、左旋硝基精氨酸甲酯( L-NAME)+缺血/再灌注+远端缺血后处理组(L-NAME+I/R+RIPoC组),以及LY294002+缺血/再灌注+远端缺血后处理组(LY+I/R+RIPoC组).采用四动脉阻断法建立大鼠全脑I/R模型.S组不制备全脑I/R模型；I/R+RIPoC组、L-NAME+I/R+RIPoC组及LY+I/R+RIPoC组于再灌注开始行双侧股动脉缺血15 min,再灌注15 min,共3个循环.L-NAME+I/R+RIPoC组于脑缺血前10 min腹腔注射非选择性一氧化氮合酶(nitric oxide synthase,NOS)抑制剂L-NAME,LY+I/R+RIPoC组于脑缺血前10 min侧脑室注射PI3K特异性抑制剂LY294002.脑再灌注48 h时行海马CA1区DNA原位末端缺口标记技术(terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling,TUNEL)阳性细胞计数,测定海马CA1区抗磷酸化的eNOS抗体(p-eNOS)、eNOS、p-Akt及Akt的蛋白水平,再灌注4d时行Morris水迷宫实验,再灌注7d时计算海马CA1区神经元密度.结果 与S组比较,I/R组、I/R+RIPoC组、L-NAME+I/R+RIPoC组及LY+I/R+RIPoC组再灌注时海马CA1区凋亡细胞[(0.8±0.8)、(84.7±6.8)、(52.8±7.8)、(74.3±9.0)、(79.5±7.3)个/mm]增加(P＜0.01),行为学损伤增加(P＜0.01),神经元密度[(193±7)、(10±7)、(91±11)、(38±7)、(26±7)个/mm]降低(P＜0.01).与I/R组比较,I/R+RIPoC组再灌注时凋亡细胞减少(P＜0.01),行为学损伤减少(P＜0.01),神经元密度增加(P＜0.01).与I/R+RIPoC组比较,L-NAME+I/R+RIPoC组及LY+I/R+RIPoC组再灌注时凋亡细胞增加(P＜0.01),行为学损伤增加(P＜0.01),神经元密度降低(P＜0.01).L-NAME能够抑制RIPoC后p-eNOS( 0.48±0.03、0.23 ±0.04)和eNOS (0.91±0.07、0.64±0.06)的升高(P＜0.01),LY294002不仅能抑制RIPoC后p-Akt (0.74±0.06、0.44±0.04)的升高(P＜0.01),而且能抑制RIPoC后p-eNOS( 0.48±0.03、0.23±0.04)和eNOS( 0.91±0.07、0.63±0.06)的升高(P＜0.01).结论 RIPoC能够减轻大鼠全脑I/R损伤,其作用机制与PI3K/Akt途径介导的eNOS激活和上调有关.     

NK4基因转染对人宫颈癌细胞CaSki生物学特性的影响

 目的 通过在人宫颈癌细胞CaSki中转染NK4 cDNA片段，研究NK4对人宫颈癌的潜在治疗作用。方法 将NK4和荧光霉素表达质粒分别稳定转染人宫颈癌细胞CaSki。采用Western blot检测细胞培养基中NK4蛋白以及经不同培养基（CaSki, CaSki/LUC 及CaSki/NK4培养上清）培养后CaSki细胞中c-Met和磷酸化c-Met的表达。采用MTT法绘制细胞生长曲线。采用细胞划痕试验检测NK4对细胞转移的作用。结果 在CaSki/NK4培养基中有NK4蛋白表达，对照细胞（CaSki, CaSki/LUC）中无表达。磷酸化c-Met的表达在CaSki/NK4培养基培养的CaSki细胞中被抑制，而在对照细胞（CaSki, CaSki/LUC）培养基培养的CaSki细胞中正常表达。各组c-Met表达无差异。3种细胞体外生长曲线差异无统计学意义（P >0.05）。细胞划痕实验结果表明，CaSki/NK4划痕区域的细胞个数明显少于CaSki和CaSki/LUC。结论 NK4能抑制人宫颈癌细胞c-Met受体磷酸化，并能抑制宫颈癌细胞体外活动能力。     

Effect of inflammatory cytokine expression in adenovirus vector-mediated human receptor activity modifying protein-1 after carotid artery saccule injury on proliferative intima

[目的]探讨腺病毒载体介导人受体活性修饰蛋白-1(receptor activity modifying protein-1,hRAMPI)基因对兔颈动脉粥样硬化并球囊成形术后炎性细胞因子表达的影响.[方法]建立兔动脉粥样硬化狭窄模型并行球囊损伤血管(简称血管成形术),随机抽样分为RAMP1组(n=18)和对照组(n=18),经球囊局部注射携带hRAMP1基因腺病毒载体(pAd2-GFP-RAMP1)或PBS,于注射后7、14d和28d,应用ELISA法检测肿瘤坏死因子-α(TNF-α)和C-反应蛋白(CRP)表达水平；Western blot检测局部hRAMP1目的基因表达；免疫组织化学染色测定血管局部TNF-α表达,HE染色检测血管形态学.[结果]血管成形术后不同时间点TNF-α表达增加[7d:(74.13±4.99),14d:(93.40±6.69),28d:(67.46±6.57)],外源hRAMP1注射后TNF-α表达下降[7d:(64.95±6.77),14d:(75.29±4.73),28d:(45.08±5.00),P＜O.05],病毒注射后7d和14d RAMP1组CRP水平[7d:(29.27±1.57),14d:(9.68±1.60)]与对照组比较[7d:(43.96±7.88),14d:(13.51±1.68)]显著下降(P＜0.05),28d2组间无差异性；腺病毒注射后28d损伤血管局部仍检测到hRAMP1蛋白表达,同时RAMP1组局部TNF-α表达与对照组比较明显下降,HE染色显示:RAMP1组新生内膜面积[7d:(0.07±0.18),14d:(0.15±0.05),28d:(0.35±0.05)]与对照组[7d:(0.14±0.02),14d:(0.39±0.09),28d:(0.56±0.05]比较明显降低(P＜0.05).[结论]外源hRAMPI基因调节兔动脉粥样硬化并血管成形术后CRP和TNF-α的表达,抑制血管成形术后再狭窄.     

血清CAl9-9对诊断和治疗胆胰系统疾病的临床意义

在胆胰系统疾病中，血清CAl9—9是常规的检测指标之一，其数值的高低，对胆胰系统疾病的诊断、治疗及预后有重要的指导意义。随着医学技术的不断发展与进步，学者们对于血清CAl9—9的认识日趋成熟。胆胰系统良恶性疾病可以通过血清CAl9—9的数值变化得到鉴别，在胆胰系统恶性肿瘤疾病中，血清CAl9—9有较高的敏感度。血清CAl9-9水平可作为肿瘤可切除性，以及患者预后的生存时间和生命质量的独立评估因素。本文阐述了血清CAl9—9在胆胰系统疾病中的诊断、治疗及预后评估中的重要作用。