Norcantharidin anti-angiogenesis activity possibly through an endothelial cell pathway in human colorectal cancer

The present study was based on the unexpected discovery that norcantharidin exerted anti-angiogenesis activity when effects on growth of human colon cancer were studied. The aim was to further verify this finding and explore possible mechanisms using a tumor xenograft model in nude mice. We confirmed that norcantharidin (5 or 15 mg/kg) could inhibit angiogenesis of human colon cancer in vivo. In vitro, crossing river assay, cell adhesion assay and tube formation assay indicated that NCTD could reduce the migration, adhesion and vascular network tube formation ability of HUVECs. At the same time, the expression levels of VEGF and VEGFR-2 proteins which play important roles in angiogenesis were reduced as examined by western blotting analysis. Taken together, the results firstly showed NCTD could inhibit angiogenesis of human colon cancer in vivo, probably associated with effects on migration, adhesion and vascular network tube formation of HUVECs and expression levels of VEGF and VEGFR-2 proteins.     

Molecular and genetic biomarkers implemented from next-generation sequencing provide treatment insights in clinical practice for Waldenström macroglobulinemia

Waldenström macroglobulinemia (WM) is a distinct type of indolent lymphoplasmacytic lymphoma (LPL) with a high frequency of MYD88^L265P mutation. Treatment for WM/LPL is highly variable in clinic and ibrutinib (a Bruton tyrosine kinase inhibitor, BTKi) has become a new treatment option for WM. To investigate the clinical impact of genetic alterations in WM, we assembled a large cohort of 219 WMs and 12 LPLs dividing into two subcohorts: a training cohort, patients sequenced by a same targeted 29-gene next-generation sequencing (NGS) panel, and a validation cohort, patients sequenced by allele specific-PCR or other targeted NGS panels. In both training and validation subcohorts, MYD88^L265P and TP53 mutations showed favorable and adverse prognostic effects, respectively. CXCR4 nonsense/missense mutations (CXCR4^NS/MS), cytogenetic complex karyotypes, and a family history of lymphoma/leukemia in first-degree relatives were associated with significantly worse clinical outcomes only or more in the validation subcohort. We further investigated the efficacy of various treatments and interaction with genetic factors in the entire cohort. Upfront dexamethasone usage was associated with poorer clinical outcomes in patients who received non-proteasome-containing chemotherapy as first-line treatment independent of genetic factors. Maintenance rituximab was associated with better survival. Ibrutinib/BTKi showed potential benefit in relapsed/refractory patients and patients without CXCR4^NS/MS including those with TP53 mutations. In conclusion, genetic testing for MYD88^L265P, TP53, and CXCR4 mutations and cytogenetic analysis provide important information for prognosis prediction and therapy selection. The findings in these study are valuable for improving treatment decisions on therapies available for WM/LPL patients with integration of NGS in clinic.     

The pan-Bcl2 Inhibitor AT101 Activates the Intrinsic Apoptotic Pathway and Causes DNA Damage in Acute Myeloid Leukemia Stem-Like Cells

Background

Leukemia stem cells (LSCs) are considered to be the cause of treatment failure and relapse in acute myeloid leukemia (AML). Overexpression of the Bcl-2 family of anti-apoptotic proteins such as Bcl-2, Bcl-xl, and Mcl-1 accounts for survival and self-renewal of LSCs. AT101 binds to the BH3 motif of all Bcl-2 family anti-apoptotic proteins and demonstrates anti-tumor activity in multiple types of tumor. Thus, we hypothesized that this agent might have the potential to deplete LSCs.

Objective

The present study aims to investigate if and by what mechanism AT101 is able to target AML stem-like cells.

Methods

As LSCs and hematopoietic stem cells (HSCs) are enriched in CD34⁺CD38^? populations, CD34⁺CD38^? cells from KG1α and Kasumi-1 cell lines as well as CD34⁺ blasts from AML patients were used as LSC models, while CD34⁺ cells from healthy donors were used as normal hematopoietic cells. Cell proliferation and apoptosis were assessed by a cell counting kit-8 (CCK8) assay and an Annexin V/PI assay using flow cytometry, respectively. Colony-forming units experiments were performed to monitor the stemness features of AML cells. Western blot and quantitative real-time polymerase chain reaction (qPCR) analysis were performed to examine the levels of proteins and mRNAs related to either the intrinsic apoptotic pathway or DNA damage response.

Results

AT101 inhibited proliferation and induced apoptosis in CD34⁺CD38^? KG1α and Kasumi-1 cells in a dose- and time-dependent manner. Exposure to AT101 for 24 h resulted in apoptosis in primary CD34⁺ AML blasts (EC₅₀ [concentration needed for a 50% maximal effect] = 2.45–76.00 μmol/L), while it only had a modest effect on normal CD34⁺ hematopoietic cells. Mechanistically, AT101 activated the intrinsic apoptotic pathway by inhibition of Bcl-2 anti-apoptotic proteins, reflected by a decrease in mitochondrial membrane potential. Moreover, AT101 caused DNA damage (e.g., increased γH2AX phosphorylation), which might also contribute to its anti-leukemic effects. Interestingly, the ex vivo efficacy of AT101 in primary AML samples significantly correlated to hyperleukocytosis and FLT3-ITD mutations. AT101 was also effective against CD34⁺ blasts isolated from elderly patients and patients who did not achieve complete remission after induction therapy.

Conclusions

AT101 effectively eliminates LSCs in vitro through the induction of DNA damage and activation of the intrinsic apoptotic pathway. AT101 is effective towards leukemic cells from patients with adverse prognostic factors, suggesting that AT101 could have the potential as an alternative salvage therapy for the treatment of relapsed and refractory AML.

     

Generation of panels of anti-idiotypic single-chain variable fragments mimicking Cry2Aa toxin using the chain shuffling technique

Here, we firstly reported to generate panels of anti-idiotypic single-chain variable fragments (scFvs) of Bacillus thuringiensis Cry2Aa toxin by chain shuffling method. Light- and heavy-chain-shuffled libraries based on two parental clones were constructed. The signal of the light-chain-shuffled library was 4.2-fold higher than the heavy one. Solely after one round panning, eight mutants were rapidly isolated from the chain-shuffled libraries. The apparent affinity of the best clones was 2.65?×?10⁶ M^?1, a 2.59-fold improvement over the parental clone. The binding abilities between anti-idiotypic scFvs with brush border membrane vesicle of Plutella xylostella larvae were also determined. This work demonstrated that chain shuffling could rapidly generate panels of antibodies with similar binding abilities but of the distinct idiotypic composition. The serials of mutants can be used to map the binding epitope of the receptors in target insects instead of Cry2Aa toxin.     

Eleven novel polymorphic microsatellite markers for genetic analysis of the venus clam Cyclina sinensis Gmelin, 1791

The venus clam Cyclina sinensis is one of the most commercially important species in marine clam aquaculture on the coast of China. In this study, 11 novel polymorphic microsatellite loci were isolated and characterized from a wild stock of the venus clam found on the Dalian coast of China. The number of alleles varied between three and six, and the observed and expected heterozygosity at the population level ranged from 0.8276 to 1.0000–0.6672 to 0.7876, respectively. No single locus deviated significantly from Hardy–Weinberg equilibrium (P > 0.01).These microsatellite markers will be useful in studies of parentage, population genetics and genome mapping in this species.     

Neural representations of the amount and the delay time of reward in intertemporal decision making

Numerous studies have examined the neural substrates of intertemporal decision‐making, but few have systematically investigated separate neural representations of the two attributes of future rewards (i.e., the amount of the reward and the delay time). More importantly, no study has used the novel analytical method of representational connectivity analysis (RCA) to map the two dimensions'' functional brain networks at the level of multivariate neural representations. This study independently manipulated the amount and delay time of rewards during an intertemporal decision task. Both univariate and multivariate pattern analyses showed that brain activity in the dorsomedial prefrontal cortex (DMPFC) and lateral frontal pole cortex (LFPC) was modulated by the amount of rewards, whereas brain activity in the DMPFC and dorsolateral prefrontal cortex (DLPFC) was modulated by the length of delay. Moreover, representational similarity analysis (RSA) revealed that even for the regions of the DMPFC that overlapped between the two dimensions, they manifested distinct neural activity patterns. In terms of individual differences, those with large delay discounting rates (k) showed greater DMPFC and LFPC activity as the amount of rewards increased but showed lower DMPFC and DLPFC activity as the delay time increased. Lastly, RCA suggested that the topological metrics (i.e., global and local efficiency) of the functional connectome subserving the delay time dimension inversely predicted individual discounting rate. These findings provide novel insights into neural representations of the two attributes in intertemporal decisions, and offer a new approach to construct task‐based functional brain networks whose topological properties are related to impulsivity.     

信迪利单抗和贝伐珠单抗联合治疗肺腺癌致免疫性输尿管炎/膀胱炎1例报告及文献复习

免疫检查点抑制剂（ICI）延长了晚期癌症患者的生存期,然而随着ICI 在抗癌治疗中的广泛应用,却涌现出新的、罕见的免疫相关不良反应（irAE）。本文报道1例由程序性死亡受体-1（PD-1）抑制剂信迪利单抗所致的罕见irAE—免疫性输尿管炎/膀胱炎。患者为55岁肺腺癌（T4N3M1,Ⅳ期）,女性,在给予“贝伐珠单抗”联合“信迪利单抗”治疗4个疗程后,出现尿频、尿急、尿痛及肉眼血尿等症状治疗。尿常规显示红细胞和白细胞明显增多,尿液培养和细胞学检查均为阴性,泌尿系统彩超及全程CT显示双侧输尿管全程扩张、管壁增厚、周围多发渗出、膀胱壁增厚、周围多发渗出。经多学科会诊（MDT）及文献检索后,患者被诊断为免疫性输尿管炎/膀胱炎。本文还回顾了文献报道的病例,以阐明其临床特征,为免疫性输尿管炎/膀胱炎的临床早诊断、适当处理及治疗提供有益的帮助。     

替罗非班治疗MRI筛选的轻中度非心源性缺血性卒中的安全性和有效性研究

目的 探讨替罗非班治疗MRI筛选的轻中度急性非心源性缺血性卒中的安全性及临床疗效.方法 前瞻性连续纳入2020年1月-2021年1月在青岛大学附属烟台毓璜顶医院住院治疗的,发病6～72 h的,完善MRI检查,非大动脉闭塞,未进行再灌注治疗的轻中度非心源性缺血性卒中患者60例作为研究对象.头颅MRI和MRA排除大面积脑梗...     

经阴道超声引导下减胎术在输卵管间质部妊娠合并宫内孕中的应用——减胎成功2例报告

目的探讨孕早期采用经阴道超声引导下减胎术减灭宫外妊娠胚芽的临床治疗效果。方法通过阴道超声减胎抽吸宫外妊娠组织。结果对2例诊断为未破裂输卵管间质部的早期异位活胎妊娠,通过经阴道超声引导下减胎穿刺治疗,被保留的宫内胎儿持续妊娠至足月分娩。结论孕早期采用经阴道超声引导下减胎术选择性抽吸输卵管间质部胚芽,可获得理想的临床治疗效果。但是本方法仅去除胚芽组织,对残余的绒毛组织,需密切追踪随访。