脂质纳米粒-mRNA递送系统及其在嵌合抗原受体T细胞治疗中的应用

尽管嵌合抗原受体（CAR）T细胞治疗在血液系统恶性肿瘤患者中取得了显著的临床疗效，但需要进一步优化。脂质纳米粒（LNP）-信使核糖核酸（mRNA）递送系统作为一种非病毒性基因载体运用于CAR-T细胞治疗研究中，一方面通过LNP将密封蛋白-6 mRNA靶向递送至抗原提呈细胞，从而实现抗原提呈细胞辅助性增强密封蛋白-6靶向的CAR-T细胞的功能，以进一步诱导对实体瘤的清除；另一方面，通过LNP将成纤维细胞激活蛋白（FAP）CARmRNA靶向递送至T细胞，实现体内FAP靶向的CAR-T细胞的制备，以通过阻断心脏纤维化过程达到治疗急性心肌损伤的目的。在CAR-T细胞研究和治疗中，LNP-mRNA递送系统具有不与细胞基因组整合、价格便宜、毒副作用小及可修饰等优点，亦存在蛋白瞬时表达导致调控细胞功能的持久性不足及制备等方面的技术局限性。本文综述了LNP-mRNA递送系统及其在CAR-T细胞治疗中的应用研究。     

Clinical analysis of peritoneal dialysis in the treatment of rapidly progressive glomerulonephritis

Objective To observe the clinical characteristics and prognosis of patients with rapidly progressive glomerulonephritis (RPGN) caused by lupus nephritis, antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitis, or primary glomerulonephritis who were treated with peritoneal dialysis (PD) and then withdrew PD because of renal recovery. Methods Data of the above patients were retrospectively analyzed. The patients were diagnosed as RPGN and received PD therapy in Kidney Disease Center, the First Affiliated Hospital, College of Medicine, Zhejiang University from February 2009 to August 2018. The patients were divided into early withdrawal group (PD time≤183 days, n=24) and late withdrawal group (PD time＞183 day, n=24). The differences of clinical characteristics between the two groups were compared. The cumulative incidence of adverse events in both groups was analyzed using Kaplan-Meier curves. Cox proportional hazards model was used to analyze the risk factors influencing the prognosis of patients. Results Forty-eight RPGN patients were included. The median time of maintaining PD was 178(76, 378) days. Compared with the late withdrawal group, the patients in early withdrawal group had lower levels of urine volume, serum albumin and parathyroid hormone, and lower rates of gross hematuria and hypertension at the beginning of PD, and received higher rates of methylprednisolone impulse, combined immunosuppressive agents, and hemodialysis or continuous renal replacement therapy (all P＜0.05). At the time of PD withdrawal, the levels of serum creatinine, serum calcium, serum albumin and parathyroid hormone in the early withdrawal group were significantly lower than those in the late withdrawal group (all P＜0.05). The Kaplan-Meier curves showed that there was no significant difference in the cumulative survival of patients in both groups (log-rank test χ2=3.485, P=0.062). Cox regression analysis revealed serum creatinine≥209 μmol/L at the time of PD withdrawal was an independent risk factor for poor prognosis (HR=5.253，95%CI 1.757-15.702, P=0.003). Conclusions PD can be used for RPGN patients caused by lupus nephritis, ANCA-associated vasculitis and primary nephritis. Serum creatinine≥209 μmol/L at the time of PD withdrawal is an independent risk factor for poor prognosis.     

Helicobacter pylori induces epithelial-mesenchymal transition in gastric carcinogenesis via the AKT/GSK3β signaling pathway

Helicobacter pylori (H. pylori) is a main risk factor for gastric cancer (GC). Epithelial-mesenchymal transition (EMT) is involved in the development and progression of H. pylori-associated GC. However, the exact molecular mechanism of this process remains unclear. The AKT/GSK3β signaling pathway has been demonstrated to promote EMT in several types of cancer. The present study investigated whether H. pylori infection induced EMT, and promoted the development and metastasis of cancer in the normal gastric mucosa, and whether this process was dependent on AKT activation. The expression levels of the EMT-associated proteins, including E-cadherin and N-cadherin, were determined in 165 gastric mucosal samples of different disease stages by immunohistochemical analysis. The expression levels of E-cadherin, N-cadherin, AKT, phosphorylated (p-)AKT (Ser473), GSK3β and p-GSK3β (Ser9) were further determined in H. pylori-infected Mongolian gerbil gastric tissues and cells co-cultured with H. pylori by immunohistochemical analysis and western blotting. The results indicated that the expression levels of the epithelial marker E-cadherin were decreased, whereas the expression levels of the mesenchymal marker N-cadherin were increased during gastric carcinogenesis. Their expression levels were associated with H. pylori infection. Furthermore, H. pylori infection resulted in downregulation of E-cadherin expression and upregulation of N-cadherin expression in Mongolian gerbils and GES-1 cells. In addition, an investigation of the associated mechanism of action revealed that p-AKT (Ser473) and p-GSK3β (Ser9) were activated in GES-1 cells following co-culture with H. pylori. Furthermore, following pretreatment of the cells with the AKT inhibitor VIII, the expression levels of E-cadherin, N-cadherin, p-AKT and p-GSK3β did not show significant differences between GES-1 cells that were co-cultured with or without H. pylori. The levels of p-AKT and p-GSK3β were increased in H. pylori-infected Mongolian gerbils. In conclusion, the present study demonstrated that H. pylori infection activated AKT and resulted in the phosphorylation and inactivation of GSK3β, which in turn promoted early stage EMT. These effects were AKT-dependent. This mechanism may serve as a prerequisite for GC development.     

原发性干燥综合征合并自身免疫性肝病的临床特点及预后分析

目的:分析原发性干燥综合征(primary Sjögren’s syndrome,pSS)合并自身免疫性肝病(autoimmune liver disease,ALD)的临床特点和预后。方法:回顾性分析浙江大学医学院附属第一医院2014年2月至2017年12月住院的pSS患者,比较pSS伴或不伴ALD两组间的临床表现和预后的差异。结果:共纳入203例pSS患者,其中合并ALD者68例(自身免疫性肝炎31例,原发性胆汁性胆管炎37例),不合并ALD者135例。两组间年龄、性别比例、口干、眼干、疼痛、乏力、淋巴结肿大、涎腺肿大、皮疹、肺部病变、肾脏受累等临床表现,其他免疫性疾病(如自身免疫性甲状腺疾病、类风湿关节炎和血管炎)的发生率,抗核抗体(antinuclear antibody,ANA)滴度,抗干燥综合征A抗体(Sjögren’s syndrome A antibody,SSA)、SSA52、抗干燥综合征B抗体(Sjögren’s syndrome B antibody,SSB)阳性率,红细胞沉降率和C反应蛋白水平差异均无统计学意义。pSS合并ALD的患者病程较短、抗线粒体-M2抗体 (anti-mitochondrial M2 antibody, AMA-M2)和抗着丝点抗体阳性率高,IgG、IgM水平高,C3水平低,血细胞减少,肝脏相关血清学指标(如谷丙转氨酶、谷草转氨酶、谷氨酰转肽酶、碱性磷酸酶、总胆红素、直接胆红素、间接胆红素)水平升高,肝硬化的比例增加,死亡事件明显增多(死亡率13.24% vs. 2.96%, P=0.013),预后更差。二元Logistic回归分析发现,pSS合并ALD的患者出现死亡事件的不良因素为肝硬化、EULAR干燥综合征疾病活动性指数评分(the EULAR Sjögren’s syndrome disease activity index,ESSDAI)和总胆红素水平。Kaplan-Meier生存曲线提示合并ALD的患者生存率低于对照组。结论:pSS合并ALD的患者病情更重、死亡事件发生率高,需要临床重视并加强对症治疗。     

儿童急性淋巴细胞白血病患者接种流感疫苗的有效性和安全性系统评价

目的通过检索已出版的文献,回顾有关儿童急性淋巴细胞白血病(ALL)患者接种流感疫苗有效性和安全性,探索最优的接种策略,为临床提供参考。方法按照PICOS原则制定研究主题的检索策略,检索Pub Med、Em Base等英文文献数据库,由两位研究者独立使用PRISMA流程图筛选符合标准的文献,提取研究数据并进行评价。结果共检索到文献186篇,经筛选获有效文献29篇。评价研究文献显示,多数ALL患儿接种灭活流感疫苗可产生中等强度的免疫应答,各亚型/系的血清抗体保护率可分别达到33%~85%(A/HIN1)、38%~92%(A/H3N2)、14%~92%(B)及37%~88%(pH1N1);血清阳转率可分别达到0%~65%(A/HIN1)、20%~85%(A/H3N2)、0%~92%(B)及11%~81%(pH1N1),低于健康儿童或实体肿瘤患儿。疫苗预防实验室确诊的流感或流感样病例的临床使用有效性的研究有限,研究结果有争议。接种时有较高的基线淋巴细胞计数、Ig G水平、结束化疗或低化疗强度阶段、高年龄段患儿以及给予加强剂次可能会有较强的免疫应答;减活流感疫苗、高剂量流感疫苗、含佐剂流感疫苗均不能改善ALL患儿的免疫应答。疫苗在各治疗阶段接种均有较好的耐受性,不良反应轻微。结论 ALL患儿接种流感疫苗安全性好,可产生中等强度的血清学保护作用,建议维持治疗阶段接种标准剂量灭活流感疫苗;疫苗的临床有效性还有待设计良好的临床试验验证。     

新型冠状病毒肺炎疫情下抗凝药学服务的策略及应用研究

目的：新型冠状病毒肺炎疫情期间采用线上程序远程的抗凝药学模式服务服用华法林的患者，以保证华法林抗凝治疗的有效性和安全性。方法：采用回顾性调查及问卷调查的方式，分析疫情期间2020年1.20-2.19实施远程抗凝药学服务以来，患者治疗窗内时间（time in therapeutic range，TTR）、不良事件、患者咨询、患者依从性、患者INR监测率、患者满意度等数据，并与去年同期线下门诊进行对比。结果：在实施远程抗凝管理的药学服务后，远程线上服务人次占比（90.63%）同比去年（13.57%）上涨明显（P<0.001），患者TTR （79.57%）同比去年（58.90%）明显提高（P=0.004），不良事件发生率降低，华法林按时服药率（96.77%）明显升高（P=0.003），国际标准化比值（International Normalized Ratio，INR）监测率（93.55%）显著提高（P=0.002），患者满意度极高，明显减少患者在院停留时间及经济成本。结论：在新冠肺炎疫情期间，该抗凝药学模式的实施解决了抗凝患者密集的药学服务需求和其减少患者暴露避免潜在交叉感染风险之间的矛盾，构建起临床药师抗击疫情的新型抗凝药学服务模式。