Trends in using mesenchymal stromal/stem cells (MSCs) in treating corneal diseases

Since their inception in the 1960s–70s, mesenchymal stem/stromal cells (MSCs) have gained interest because of their differentiation potential, anti-inflammatory effects, and immune-modulating properties. Both cell-based and cell-free MSC treatments show healing capacity in injured tissues. Cell-based treatment comprises MSCs and all secreted products, whereas cell-free treatments include only the secreted products. MSCs are therapeutically administered to many damaged organs owing to their efficacy. Specifically, the eye is a unique organ system to study the effects of MSCs, as treatment is easily applied and measured owing to its external location. The eye holds an immune-privileged status, wherein inflammation and immune responses are innately down-regulated. As excessive inflammation in the cornea often leads to fibrosis and irreversible corneal hazing, many studies have investigated the anti-inflammatory and immune-modulating capacities of MSCs. Decades of research suggest that MSCs modulate the immune response by secreting cytokines, growth factors, and extracellular matrix proteins that inhibit the infiltration of inflammatory cells following injury and promote a healing phenotype via M2 macrophage polarization. MSCs have also shown trans-differentiation potential into cornea-specific cell types during the wound healing process, such as corneal epithelial, stromal, or endothelial cells. This review discusses recent investigations of MSC treatment in the cornea, focusing on therapeutic efficacy, mechanisms, and future directions.     

Preliminary study on blood coagulation and hemostatic effect and acute toxicity of Oxalis corniculata L. ethanol extract

In the present study, we aimed to observe the effects of ethanol extract of Oxalis corniculata L. on bleeding time (BT) and coagulation time (CT) and determine its acute toxicity in mice. Firstly, the ethanol extract of O. corniculata was administered to 40 mice, which were randomly divided into the normal control group, Yunnan Baiyao control group, low-dose group, and high-dose group. The low-dose (0.6 g/kg) and high-dose (1.2 g/kg) groups received ethanol extract of O. corniculata by gavage, and the BT and CT of the mice were measured by the slide method and tail clipping method, respectively. Secondly, the median lethal dose method was used with 50 mice to observe the physiological state, poisoning reaction, and death of the mice after intragastric administration. Finally, on the 14th day of the experiment, a necropsy was performed to observe any abnormality of the organs. In conclusion, in the coagulation and hemostasis tests, there were no statistical differences between the groups (P > 0.05), while there was a significant dose-response relationship, and the BT and CT were significantly shorter than those of the negative control group, which were comparable to the Yunnan Baiyao control group. Moreover, in the acute toxicity test, the median lethal dose of ethanol extract of O. corniculata was 6.0291 g/kg, with a 95% confidence interval of 5.3065–6.7829 mg/kg.     

Estimating Cost-Effectiveness Using Alternative Preference-Based Scores and Within-Trial Methods: Exploring the Dynamics of the Quality-Adjusted Life-Year Using the EQ-5D 5-Level Version and Recovering Quality of Life Utility Index

ObjectivesThis study aimed to explore quality-adjusted life-year (QALY) and subsequent cost-effectiveness estimates based on the more physical health–focused EQ-5D 5-level version (EQ-5D-5L) value set for England or cross-walked EQ-5D 3-level version UK value set scores or more mental health recovery-focused Recovering Quality of Life Utility Index (ReQoL-UI), when using alternative within-trial statistical methods. We describe possible reasons for the different QALY estimates based on the interaction between item scores, health state profiles, preference-based scores, and mathematical and statistical methods chosen.MethodsQALYs are calculated over 8 weeks from a case study 2:1 (intervention:control) randomized controlled trial in patients with anxiety or depression. Complete case and with missing cases imputed using multiple-imputation analyses are conducted, using unadjusted and regression baseline-adjusted QALYs. Cost-effectiveness is judged using incremental cost-effectiveness ratios and acceptability curves. We use previously established psychometric results to reflect on estimated QALYs.ResultsA total of 361 people (241:120) were randomized. EQ-5D-5L crosswalk produced higher incremental QALYs than the value set for England or ReQoL-UI, which produced similar unadjusted QALYs, but contrasting baseline-adjusted QALYs. Probability of cost-effectiveness <£30 000 per QALY ranged from 6% (complete case ReQoL-UI baseline-adjusted QALYs) to 64.3% (multiple-imputation EQ-5D-5L crosswalk unadjusted QALYs). The control arm improved more on average than the intervention arm on the ReQoL-UI, a result not mirrored on the EQ-5D-5L nor condition-specific (Patient-Health Questionnaire-9, depression; Generalized Anxiety Disorder-7, anxiety) measures.ConclusionsReQoL-UI produced contradictory cost-effectiveness results relative to the EQ-5D-5L. The EQ-5D-5L’s better responsiveness and “anxiety/depression” and “usual activities” items drove the incremental QALY results. The ReQoL-UI’s single physical health item and “personal recovery” construct may have influenced its lower 8-week incremental QALY estimates in this patient sample.     

放疗干预对宫颈癌荷瘤小鼠的抑瘤作用及对Th1/Th2免疫细胞比例的影响

目的探讨放疗干预对宫颈癌荷瘤小鼠的抑瘤作用及其对辅助性T细胞1(Th1)/Th2细胞比例的影响。方法建立宫颈癌荷瘤小鼠模型,随机分为荷瘤组和放疗组,每组各10只,另设对照组10只。放疗组进行放疗干预14天,荷瘤组和对照组不治疗。放疗后4、6、8、10、12、14天测量肿瘤体积;末次治疗后,ELISA法测定血清干扰素-γ(IFN-γ)、白介素-2(IL-2)、IL-4、IL-10含量;计算抑瘤率、胸腺指数和脾脏指数;HE染色观察肿瘤组织学变化;TUNEL染色观察肿瘤组织细胞凋亡情况;流式细胞术检测脾脏Th1/Th2细胞比例;RT-qPCR和Western blot检测脾脏T盒子转录因子(T-bet)、GATA结合蛋白3(GABA-3)mRNA和蛋白表达。结果与荷瘤组比较,放疗组小鼠4、6、8、10、12、14天肿瘤体积及瘤质量减小,血清IL-2、IFN-γ升高,IL-4、IL-10降低,胸腺指数、脾脏指数升高,Th1细胞比例、Th1/Th2增加,Th2细胞比例减少,T-bet mRNA和蛋白及T-bet/GATA-3表达升高,GATA-3 mRNA和蛋白表达降低(P<0.05)。HE染色显示,荷瘤组肿瘤细胞数量较多,核大深染,无明显坏死;放疗组肿瘤细胞数量减少,出现大量坏死组织。TUNEL染色显示,荷瘤组TUNEL阳性细胞较少,放疗组TUNEL阳性细胞明显增多。结论放疗对宫颈癌荷瘤小鼠具有明显抑瘤作用,可能是通过调节T-bet/GATA-3表达,促进Th1/Th2分化平衡,增强机体免疫功能发挥作用。     

CD19+CD24hiCD38hi B细胞比例预测肝移植术后急性细胞性排斥反应发生的临床研究

目的分析肝移植术后受者外周血CD19⁺CD24^hiCD38^hi B细胞占单个核细胞比例变化情况及其与急性细胞性排斥反应(ACR)之间的关系。 方法回顾性分析2013年12月至2015年12月在浙江大学医学院附属第一医院接受心脏死亡器官捐献肝移植的80例成人受者临床资料，根据术后是否发生ACR，将受者分为ACR组(25例)和非ACR组(55例)。术前、术后各个时间点抽取参加研究者静脉血并分离外周血单个核细胞，加入异硫氰酸荧光素-单克隆鼠抗人CD19抗体、藻红蛋白-单克隆鼠抗人CD24抗体和别藻蓝蛋白-单克隆鼠抗人CD38抗体，流式细胞仪检测各组CD19⁺CD24^hiCD38^hi B细胞百分比。采用t检验和单因素方差分析比较正态分布计量资料，采用χ²检验比较计数资料，采用Kaplan-Meier法绘制受试者工作特征曲线(ROC曲线)。P<0.05为差异有统计学意义。 结果ACR组、非ACR组受者术前外周血平均CD19⁺CD24^hiCD38^hi B细胞比例分别为(3.13±0.91)%、(3.49±0.83)%，差异无统计学意义(t＝1.636，P>0.05)。ACR组术后发生ACR前外周血平均CD19⁺CD24^hiCD38^hi B细胞比例为(1.87±0.70)%。非ACR组受者术后3个月、6个月和1年外周血平均CD19⁺CD24^hiCD38^hi B细胞比例分别为(1.64±0.52)%、(1.63±0.56)%和(2.04±1.24)%，术后3、6个月平均值均低于术前和术后1年，差异均有统计学意义(P均<0.05)。ACR组受者发生ACR时外周血平均CD19⁺CD24^hiCD38^hi B细胞比例为(0.8±0.5)%，低于发生ACR前的平均水平(t＝5.752，P<0.05)，且低于非ACR组术后3个月、6个月和1年的平均水平(P<0.05)。ACR组受者接受抗排斥反应治疗后，CD19⁺CD24^hiCD38^hi B细胞比例也逐渐增加，ACR发生后7 d为(0.84±0.08)%，与ACR发生时相比差异无统计学意义(P>0.05)；而发生30 d后达(1.65±0.18)%，与ACR发生时相比差异有统计学意义(P<0.05)。当截断值为1.015%时，CD19⁺CD24^hiCD38^hi B细胞比例预测ACR发生的敏感度和特异度分别为0.786和0.702，ROC曲线下面积为0.775(95%CI: 0.671~0.879，P<0.05)。 结论CD19⁺CD24^hiCD38^hi B细胞比例下降与肝移植术后ACR反应发生有关，并可作为预测ACR发生的细胞标志物。     

朝鲜淫羊藿酸性多糖的理化特性及对HepG2细胞脂质堆积的改善作用＊

目的 对比研究朝鲜淫羊藿酸性多糖酯化还原前后的理化特性，并探讨其改善油酸诱导的肝癌HepG2细胞脂质堆积活性的差异。方法 采用高效凝胶渗透色谱法测定朝鲜淫羊藿酸性多糖（EFPA）的均一性和分子量，高效液相色谱法测定EFPA和酯化还原后朝鲜淫羊藿酸性多糖（EFPA-R）的单糖组成；采用油酸（OA）处理HepG2细胞诱导建立脂质蓄积模型，不同浓度EFPA与EFPA-R（10、30、100、300 μg·mL^-1）分别和OA共同作用于细胞24 h，采用CCK-8试剂盒测定细胞存活率，油红O染色观察细胞内脂滴蓄积情况，并采用试剂盒测定细胞内总胆固醇（TC）、甘油三酯（TG）含量。结果 EFPA为成分均一的多糖组分，分子量为125.8 kDa，由甘露糖、葡萄糖、半乳糖、葡萄糖醛酸和阿拉伯糖组成，摩尔比为1.7∶7.4∶1.4∶1.8∶1.0，葡萄糖占比最大，EFPA-R由甘露糖、葡萄糖、半乳糖和阿拉伯糖组成，摩尔比为0.8∶10.6∶2.1∶1.0；在10-300 μg·mL^-1范围内，EFPA和EFPA-R对HepG2细胞的抑制作用较弱，作为给药浓度；与空白组相比，模型组细胞中TC、TG含量显著升高（P < 0.01），细胞内红色脂滴显著增多，与模型组相比，EFPA可显著降低细胞中TC、TG含量（P < 0.01），明显减少细胞内红色脂滴（P < 0.05或P < 0.01），EFPA-R干预后细胞则无明显变化。结论 EFPA可明显改善HepG2细胞脂质堆积情况，且呈现剂量依赖性，而半乳糖醛酸（GalA）的存在可能是其抑制HepG2细胞脂质蓄积的关键因素。     

外泌体在肿瘤细胞与TAMs之间相互作用中“桥梁”和调控作用的研究进展北大核心

外泌体是一类直径为30~100 nm的圆盘囊泡,其内包含许多组分,诸如复杂RNA和蛋白质等,主要参与细胞间的信号转导。肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)是肿瘤微环境中普遍存在的巨噬细胞,通过对肿瘤生长、免疫逃逸、侵袭和转移、耐药性等多方面的作用影响肿瘤进程。外泌体在肿瘤相关巨噬细胞的招募、极化及抗肿瘤免疫调控等方面发挥着重要的调节功能。同时,TAMs以外泌体为媒介作用于肿瘤细胞,从而构成了外泌体、TAMs与肿瘤细胞之间相互作用的调控通路。综上所述,本文旨在阐明肿瘤细胞与TAMs之间,以外泌体为“桥梁”相互影响的潜在机制,以及靶向肿瘤细胞和TAMs来源的外泌体在恶性肿瘤治疗中的展望。