A new era of immuno-oncology in acute myeloid leukemia - antibody-based therapies and immune checkpoint inhibition

Acute myeloid leukemia (AML) remains a therapeutically challenging malignancy with high rate of relapse and poor outcomes. There has been increased understanding of the molecular characteristics of AML and the various roles of the immune system in its pathogenesis, the result of which has led to the study and development of multiple immune-based approaches for this disease. In this review, we aim to provide an overview of the recent advancements made in antibody-based approaches to the treatment of AML including monoclonal antibodies, antibody-drug conjugates, and immune checkpoint inhibition. In addition, we provide insight and discuss the promise of these agents, some of which may soon enter the therapeutic armamentarium we currently employ against this lethal disease.     

miR-21对视网膜母细胞瘤细胞增殖的影响及作用机制

目的　探讨MicroRNA-21（miR-21）对视网膜母细胞瘤(RB)细胞增殖的影响及作用机制。方法　采用Real-time PCR技术检测miR-21在正常视网膜组织和确诊RB组织中的表达情况；然后在转染的基础上运用MTT检查RB细胞存活率、流式细胞仪检测细胞凋亡率。Western blot法检测与细胞凋亡相关蛋白PDCD4、Bax、Bcl-2的表达。结果　与正常视网膜组织miR-21表达 （0.703±0.071）相比，RB组织miR-21为高表达（2.214±0.162），差异有统计学意义（P<0.01）。在Weri-Rb-1细胞中，与NC组(2.245±0.213)相比，miR-21抑制剂转染后明显降低了miR-21的表达水平，miR-21 inhibitor组为0.683±0.075,差异有统计学意义 （P<0.01）。两组细胞转染后24 h、48 h、72 h、96 h，MTT测定法检测细胞活力结果显示:两组24 h的A值比较，差异无统计学意义 （P>0.05），miR-21 inhibitor 组在 48 h、72 h、 96 h的A值均低于 NC 组，差异均有统计学意义 （均为P<0.01）。流式细胞术检测结果显示:NC组凋亡细胞在总细胞中百分比为（3.045±0.301）%和（4.832±0.493）%，miR-21 inhibitor组凋亡细胞在总细胞中百分比为（2.593±0.257）%和（40.167±4.014）%，miR-21 inhibitor组Weri-Rb-1细胞的凋亡率明显高于miR-21 NC组(P<0.01)。Western blot检测结果显示:NC组PDCD4表达（0.192±0.045）相比miR-21 inhibitor组（0.683±0.091）表达明显减少，NC组Bax的蛋白表达水平（0.143±0.036）相比miR-21 inhibitor组（1.192±0.054）也明显减少，差异均有统计学意义（P<0.01)，NC组Bcl-2蛋白表达（0.864±0.038）相比miR-21 inhibitor组（0.257±0.026）明显增多，差异有统计学意义（P<0.05)。结论　miR-21是RB的促癌基因，miR-21抑制剂可以通过降低miR-21表达抑制肿瘤细胞的增殖、促进细胞凋亡，这一过程与PDCD4、Bax、Bcl-2等凋亡相关蛋白有关。     

a-1, 2岩藻糖基转移酶II在肺鳞癌组织中的表达及对肺鳞癌细胞增殖的影响

目的：探讨a-1，2岩藻糖基转移酶II（FUT2）在肺鳞癌中的表达情况及其对肺鳞癌细胞增殖的影响。方法：应用UALCAN数据库分析FUT2在临床肺鳞癌患者组织中的差异表达及其与肺鳞癌患者临床TNM分期、年龄及性别因素的相关性；采用瞬时转染技术，将FUT2的shRNA质粒转染进H226细胞株中构建FUT2低表达的肺鳞癌细胞模型；利用CCK-8、平板克隆及Western blot实验检测FUT2对肺鳞癌细胞的生长、单克隆细胞团形成及PCNA蛋白表达的影响。结果：FUT2在肺鳞癌尤其是肺鳞癌TNM早中期呈现高表达趋势（P＜0.05），而与年龄和性别无关。低表达FUT2可以抑制肺鳞癌细胞的生长速率和单克隆细胞团的形成数，同时，可以抑制肺鳞癌细胞PCNA蛋白的表达水平，差异均有统计学意义（P＜0.05）。结论：FUT2在肺鳞癌中高表达且促进肺鳞癌细胞的增殖能力，有望成为肺鳞癌的早期诊断指标。     

Cyclin D1启动子在养精胶囊促进小鼠精原干细胞增殖中的作用※

目的 研究养精胶囊促进小鼠精原干细胞（SSCs）增殖的分子机制。方法 将不同浓度的养精胶囊提取物加入SSCs中培养48 h，CCK-8检测细胞的增殖活性，流式细胞仪检测细胞周期，荧光素酶报告基因检测Cyclin D1启动子的活性，qRT-PCR以及免疫荧光检测Cyclin D1的表达。之后进行阻断实验，在加入养精胶囊前预先加入siRNA-Cyclin D1，同样的方法检测细胞的增殖活性、细胞周期、Cyclin D1启动子的活性以及Cyclin D1的表达情况。结果 低、中、高浓度的养精胶囊可以促进SSCs的增殖，提高S期细胞的比例，增强Cyclin D1启动子的活性，促进Cyclin D1的表达。阻断Cyclin D1后，SSCs的增殖活性降低，S期细胞比例减少，Cyclin D1启动子的活性降低，Cyclin D1的表达减少。结论 养精胶囊通过增强Cyclin D1启动子的活性提高Cyclin D1的转录和翻译水平，进而促进SSCs增殖。     

枳术丸水煎液及其拆方含药血清对大鼠结肠Cajal间质细胞增殖和凋亡的影响

目的:探讨枳术丸水煎液及其拆方对大鼠结肠Cajal间质细胞(ICC)增殖和凋亡的影响及其分子机制,并从细胞分子层面分析中医攻补兼施法治疗慢传输型便秘可能的机制。方法:将40只大鼠随机分成白术组、枳实组、枳术丸组、空白血清组,每组10只。白术组灌以17.7 g·kg-1·d-1白术水煎液,枳实组灌以8.9 g·kg-1·d-1枳实水煎液,枳术丸组灌以26.4 g·kg-1·d-1枳术丸水煎液,空白血清组灌以无菌蒸馏水3 mL,每日1次,连续7 d,取血制备含药血清和空白血清。将4组含药血清分别配成5%,10%,15%,20%4个体积分数,干预24 h,观察ICC的数量、形态,用细胞增殖毒性检测试剂盒(CCK-8)检测各组细胞增殖情况,测定各组最佳干预体积分数。将大鼠结肠ICC细胞分为正常组、空白血清组、白术组、枳实组、枳术丸组,5-乙炔基-2'脱氧尿嘧啶核苷(EdU)检测各组ICC的增殖情况,流式细胞仪检测各组ICC的凋亡情况;蛋白免疫印迹法(Western blot)检测各组ICC中X连锁凋亡抑制蛋白(XIAP),增殖细胞核抗原(PCNA)蛋白表达情况。结果:与正常组比较,空白血清组、白术组、枳实组最佳干预体积分数均为10%,枳术丸组最佳干预体积分数为5%。与正常组比较,所有实验中空白血清组对ICC增殖和凋亡的影响没有明显差异;与正常组及空白血清组比较,白术组、枳实组、枳术丸组ICC增殖率有显著差异(P0.05,P0.01),与白术组和枳实组比较,枳术丸组对ICC增殖率明显增加(P0.05,P0.01);与枳实组比较,白术组对ICC增殖率变化不明显。与正常组及空白血清组比较,白术组、枳实组、枳术丸组ICC的凋亡率无明显差异;与正常组及空白血清组比较,白术组、枳实组、枳术丸组内的XIAP,PCNA蛋白水平均明显上调(P0.05,P0.01),三组间比较差异不明显。结论:枳术丸、枳实、白术在一定浓度下均能通过增加XIAP,PCNA蛋白表达促进ICC的增殖,且对ICC的凋亡无明显影响;以攻补兼施法立法的方剂枳术丸能促进ICC的增殖,并且其增殖效果优于攻伐单药枳实和补益单药白术。     

羌活中的香豆素类成分及其抑制脂多糖诱导的RAW 264.7细胞NO生成活性的研究

目的研究羌活Notopterygium incisum的香豆素类成分及其抗炎活性。方法采用硅胶、HPLC等柱色谱方法进行分离纯化,通过质谱、核磁共振波谱数据鉴定化合物的结构;采用脂多糖(LPS)诱导的小鼠巨噬细胞RAW 264.7炎症反应模型,考察羌活中香豆素类成分对炎症反应模型一氧化氮(NO)生成的影响。结果从羌活甲醇提取物分离得到24个香豆素类化合物,分别鉴定为异欧前胡素(1)、川白芷素(2)、补骨脂素(3)、香柑内酯(4)、茵陈素(5)、欧芹酚(6)、5-去氢羌活醇(7)、环氧脱水羌活醇(8)、7″-O-甲基异羌活醇(9)、佛手柑素(10)、7-异戊烯氧基-6-甲氧基-香豆素(11)、栓翅芹烯醇(12)、羌活醇(13)、去甲呋喃羽叶芸香素(14)、异羌活醇(15)、蛇床夫内酯(16)、6-异戊烯氧基伞形花内酯(17)、紫花前胡苷元(18)、异虎耳草素(19)、紫花前胡苷(20)、前胡苷V(21)、前胡苷I(22)、印枳苷元-11-O-β-D-吡喃葡萄糖基-(1→6)-β-D-吡喃葡萄糖苷(23)、羌活苷(24)。化合物7～10、13和15抑制LPS诱导的RAW 264.7细胞NO生成活性最强,最大半数抑制浓度(IC_(50))值为8.50～35.12μmol/L。结论化合物7为新的天然产物,化合物17为首次从羌活中分离得到;C-5位上具有多烯烃结构的香豆素抑制LPS诱导的RAW 264.7细胞NO生成活性较强。     

Sensory evoked potentials in patients with Rett syndrome through the lens of animal studies: Systematic review

ObjectiveSystematically review the abnormalities in event related potential (ERP) recorded in Rett Syndrome (RTT) patients and animals in search of translational biomarkers of deficits related to the particular neurophysiological processes of known genetic origin (MECP2 mutations).MethodsPubmed, ISI Web of Knowledge and BIORXIV were searched for the relevant articles according to PRISMA standards.ResultsERP components are generally delayed across all sensory modalities both in RTT patients and its animal model, while findings on ERPs amplitude strongly depend on stimulus properties and presentation rate. Studies on RTT animal models uncovered the abnormalities in the excitatory and inhibitory transmission as critical mechanisms underlying the ERPs changes, but showed that even similar ERP alterations in auditory and visual domains have a diverse neural basis. A range of novel approaches has been developed in animal studies bringing along the meaningful neurophysiological interpretation of ERP measures in RTT patients.ConclusionsWhile there is a clear evidence for sensory ERPs abnormalities in RTT, to further advance the field there is a need in a large-scale ERP studies with the functionally-relevant experimental paradigms.SignificanceThe review provides insights into domain-specific neural basis of the ERP abnormalities and promotes clinical application of the ERP measures as the non-invasive functional biomarkers of RTT pathophysiology.     

Inactivated influenza vaccine formulated with single-stranded RNA-based adjuvant confers mucosal immunity and cross-protection against influenza virus infection

Influenza vaccination is considered the most valuable means to prevent and control seasonal influenza infections, which causes various clinical symptoms, ranging from mild cough and fever to even death. Among various influenza vaccine types, the inactivated subunit type is known to provide improved safety with reduced reactogenicity. However, there are some drawbacks associated with inactivated subunit type vaccines, with the main ones being its low immunogenicity and the induction of Th2-biased immune responses. In this study, we investigated the role of a single-stranded RNA (ssRNA) derived from the intergenic region in the internal ribosome entry site of the Cricket paralysis virus as an adjuvant rather than the universal vaccine for a seasonal inactivated subunit influenza vaccine. The ssRNA adjuvant stimulated not only well-balanced cellular (indicated by IgG2a, IFN-γ, IL-2, and TNF-α) and humoral (indicated by IgG1 and IL-4) immune responses but also a mucosal immune response (indicated by IgA), a key protector against respiratory virus infections. It also increases the HI titer, the surrogate marker of influenza vaccine efficacy. Furthermore, ssRNA adjuvant confers cross-protective immune responses against heterologous influenza virus infection while promoting enhanced viral clearance. Moreover, ssRNA adjuvant increases the number of memory CD4⁺ and CD8⁺ T cells, which can be expected to induce long-term immune responses. Therefore, this ssRNA-adjuvanted seasonal inactivated subunit influenza vaccine might be the best influenza vaccine generating robust humoral and cellular immune responses and conferring cross-protective and long-term immunity.     

白黎芦醇对GH3细胞增殖和PRL合成的影响

目的 探讨白黎芦醇对垂体腺瘤GH3细胞增殖和泌乳素合成的影响，及其对雌激素的拮抗作用。方法 在无血清无酚红的培养条件下，白黎芦醇单独或与雌二醇联合作用于GH3细胞，用MTT法测定细胞增殖，用免疫荧光法、RT-PCR和Western印迹法测定泌乳素的表达情况。结果 白黎芦醇对GH3细胞增殖具有刺激和抑制双相作用，呈时间一剂量依赖性。并且白黎芦醇使GH3细胞中PRL阳性细胞比例下降。同时，白黎芦醇抑制泌乳素的合成。白黎芦醇对雌二醇诱发的细胞增殖和泌乳素合成均有拮抗作用．但对泌乳素合成的拮抗作用较强，而雌二醇刺激细胞增殖作用较其诱发泌乳素合成作用强。结论 白黎芦醇对GH3细胞增殖和泌乳素合成均有抑制作用，从两方面发挥着抗肿瘤作用。