“磨玻璃影”特征的早期肺腺癌中MPP成分的突变特征分析及ZNF469基因的探索

背景与目的目前,肺癌依然是我国发病率和死亡率最高的恶性肿瘤。而在早期肺腺癌(lung adenocarcinoma,LUAD)中,微乳头(micropapillary,MPP)成分尤其常见,且通常表现出高侵袭性,其与早期转移、淋巴浸润的风险以及患者的5年生存率显著相关。本研究旨在探究以磨玻璃影(ground-glass opacities,GGOs)为特征的早期LUAD中MPP成分和非MPP成分的异同,寻找MPP成分所特有的突变特征,并分析锌指蛋白家族的ZNF469基因与早期LUAD预后以及免疫浸润的关系。方法收集31例LUAD恶性肺结节,采用显微解剖法将其分为成对的MPP和非MPP成分。对早期恶性肺结节组分进行全外显子组测序(whole-exome sequencing,WES),利用maftools、非负矩阵分解(Nonnegative Matrix Factorization,NMF)法、Sigminer等方法进行突变特征分析,以揭示侵袭性LUAD中MPP组分相比于其他肿瘤组织所特有的基因组突变特征。利用癌症基因组图谱(The Cancer Genome Atlas,TCGA)的LUAD数据库中ZNF469的表达情况,探讨其与肺癌预后的关系;利用GeneMANIA数据库以及基因本体(Gene Ontology,GO)、京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析探索LUAD中与ZNF469相关基因的互作网络及信号通路;利用TIMER和TISIDB数据库分析ZNF469表达与LUAD中免疫细胞浸润水平的相关性。结果MPP成分具有较多的基因组变异,相比于非MPP成分的肿瘤组织,癌症体细胞突变目录(Catalogue of Somatic Mutations in Cancer,COSMIC)的13号突变特征(胞苷脱氨酶家族,APOBEC)是MPP成分所特有的,这提示其参与了MPP成分对LUAD早期侵袭过程的促进作用;并且APOBEC特征高的MPP样本具有更高的肿瘤突变负荷(tumor mutational burden,TMB),提示这些患者更能从免疫治疗中获益。LUAD中突变ZNF469的表达高于正常组织,与LUAD患者的不良预后有关。基因互作网络分析以及GO和KEGG富集分析发现,COL6A1、COL1A1、COL1A2、TGFB2、MMP2、COL8A2、C2CD4C等与ZNF469具有相互作用,且主要与编码胶原蛋白、参与细胞外基质构成有关。ZNF469表达与肿瘤的免疫浸润呈正相关。结论本研究揭示了中国人群侵袭性LUAD中MPP成分的特有突变特征,并发现突变ZNF469的高表达影响LUAD预后与免疫浸润,推测ZNF469可作为LUAD潜在的诊断及预后生物标志物。

肺腺癌中免疫微环境特征与EGFR突变状态的相关性研究

背景与目的在全球范围内肺癌发病率和死亡率一直居高不下,而肺腺癌是其中最主要的一个组织亚型,表皮生长因子受体(epidermal growth factor receptor,EGFR)突变是肺腺癌的一个重要驱动基因突变。近年来以程序性死亡受体1(programmed cell death 1,PD-1)及程序性死亡配体1(programmed cell death ligand 1,PD-L1)抑制剂为代表的免疫检查点抑制剂(immune checkpoint inhibitors,ICIs)治疗在部分肺癌患者中疗效显著。有研究表明EGFR突变的患者对免疫治疗的获益程度有限,本研究旨在讨论肺腺癌患者中EGFR突变状态与各类免疫细胞浸润数量、空间分布的关系。方法本研究纳入62例接受手术的肺腺癌患者。通过对手术切除的肿瘤组织进行不同区域多点采样,最终获取了223份肿瘤组织样本。通过基因检测获取了每份肿瘤组织的EGFR突变情况,包括突变丰度(variant allele frequency,VAF)以及突变亚型等信息。对肿瘤组织进行苏木精-伊红(hematoxylin-eosin,HE)染色、免疫组化染色、多重荧光免疫组化染色,并通过计算免疫组化评分获取各类免疫细胞在肿瘤组织中的浸润情况以及三级淋巴结构(tertiary lymphoid structure,TLS)的分布情况。结果与野生型患者相比,EGFR突变肺腺癌患者肿瘤组织中CD68^(+)巨噬细胞以及组织相容性复合物(major histocompatibility complex,MHC)II类抗原呈递细胞浸润更多,且MHC II类抗原呈递细胞空间分布的异质性较大,而CD56^(+)自然杀伤细胞以及CD8^(+)T细胞等浸润更低。EGFR VAF高的肿瘤组织与更低的CD3^(+)T细胞、CD20^(+)B细胞、CD56^(+)自然杀伤细胞、CD68^(+)巨噬细胞、CD8^(+)T细胞等细胞浸润相关,其中只有CD3^(+)T细胞分布的空间异质性较小。对于中国人群常见的两种EGFR突变亚型,EGFR外显子19缺失突变相较于EGFR外显子21 L858R突变肿瘤组织中有更低的免疫细胞浸润,但CD3^(+)T细胞、CD56^(+)自然杀伤细胞、CD68^(+)巨噬细胞以及CD8^(+)T细胞等免疫细胞分布的空间异质性更高。预后分析发现,CD3^(+)T细胞、CD20^(+)B细胞浸润程度高、TLS形成数量多以及CD8^(+)T细胞高分布异质性的EGFR突变患者拥有更长的无病生存期。结论EGFR突变肺腺癌具有独特的低免疫细胞浸润的“非炎性”肿瘤微环境,不同突变亚型以及VAF的肿瘤微环境之间也表现出异质性,这些差异不仅体现在免疫细胞的浸润数量上,也体现在免疫细胞的空间分布情况。因此对EGFR突变肺腺癌的免疫微环境进行进一步更深入的研究对未来提高EGFR突变肺腺癌患者的免疫治疗疗效具有重大意义。

神经母细胞瘤中Hsp27的表达及其与化疗药物关系的研究

目的:探讨分析热休克蛋白27(heat shock protein 27,Hsp27)与儿童神经母细胞瘤临床病理参数及化疗药物的关系。方法:应用免疫组织化学(immunohistochemistry,IHC)法检测90例神经母细胞瘤(neuroblastoma,NB)、24例节细胞神经母细胞瘤(gan?glioneuroblastoma,GNB)和20例节细胞神经瘤(ganglioneuroma,GN)组织中Hsp27蛋白的表达;实时荧光定量PCR(fluorescence quantitative PCR,RT-qPCR)检测59例NB/GNB、5例GN组织标本,及人神经母细胞瘤SH-SY5Y细胞和耐顺铂SH-SY5Y细胞中Hsp27 mRNA的表达;应用IHC或PCR扩增测序法检测其中18例NB/GNB肿瘤标本化疗药物相关性分子靶标。结果:Hsp27蛋白在NB/GNB中阳性表达率为64.9%,明显高于GN的0(χ^2=28.994,P<0.05);在NB/GNB中,Hsp27阳性表达与肿瘤的分期、危险度、转移和预后有关(P<0.05),与患儿年龄、性别、Shimada组织学分型、N-myc有无扩增无关(P>0.05);Hsp27 mRNA在NB/GNB中的相对表达量为3.763±1.854,高于GN的2.476±0.698(P>0.05);Hsp27 mRNA在耐顺铂细胞中的表达明显高于亲代细胞(F=16.246,P<0.01)。检测化疗药物相关性分子靶标7个,结果显示Hsp27的表达与铂类药物敏感性相关。结论:本研究初步显示Hsp27的表达与NB的分期、危险度、转移、预后相关,可能参与了NB/GNB的发生发展转移。Hsp27的表达与铂类敏感性相关,需进一步增大样本量来验证其相关性。

Impact of H_(2)S on Hg^(0)capture performance over nitrogen-doped carbon microsphere sorbent:experimental and theoretical insights

A nitrogen-doped carbon microsphere sorbent with a hierarchical porous structure was synthesized via aggregation-hydrothermal carbonization.The Hg^(0)adsorption performance of the nitrogen-doped carbon microsphere sorbent was tested and compared with that of the coconut shell activated carbon prepared in the laboratory.The effect of H_(2)S on Hg^(0)adsorption was also investigated.The nitrogen-doped carbon microsphere sorbent exhibited superior mercury removal performance compared with that of coconut shell activated carbon.In the absence of H_(2)S at a low temperature(≤100℃),the Hg^(0)removal efficiency of the nitrogen-doped carbon microsphere sorbent exceeded 90%.This value is significantly higher than that of coconut shell activated carbon,which is approximately 45%.H_(2)S significantly enhanced the Hg^(0)removal performance of the nitrogen-doped carbon microsphere sorbent at higher temperatures(100–180℃).The hierarchical porous structure facilitated the diffusion and adsorption of H_(2)S and Hg^(0),while the nitrogen-containing active sites significantly improved the adsorption and dissociation capabilities of H_(2)S,contributing to the generation of more active sulfur species on the surface of the nitrogen-doped carbon microsphere sorbent.The formation of active sulfur species and HgS on the sorbent surface was further confirmed using X-ray photoelectron spectroscopy and Hg^(0)temperature-programmed desorption tests.Density functional theory was employed to elucidate the adsorption and transformation of Hg^(0)on the sorbent surface.H_(2)S adsorbed and dissociated on the sorbent surface,generating active sulfur species that reacted with gaseous Hg^(0)to form HgS.

Dual ultrasound-guided totally implantable venous access ports via the right internal jugular vein in pediatric patients with cancer:a preliminary experience in a single institution

Objective To assess the efficacy and safety of dual ultrasound-guided(DUG)totally implantable venous access port(TIVAP)implantation(namely,using ultrasoundguided percutaneous puncture with transesophageal echocardiography-guided catheterization)via the right internal jugular vein(IJV)in pediatric patients with cancer.Methods Fifty-five children with cancer requiring chemotherapy underwent DUG-TIVAP implantation via the right IJV.Clinical data were recorded,including the procedure success rate,first attempt success rate,and perioperative and postoperative complications.Results All 55 cases were successfully operated on.The first puncture success rate was 100%.The operation time was 22–41min,with a mean time of 30.8±5.5min.The mean TIVAP implantation time was 253±145 days(range 42–520 days).There were no perioperative complications.The postoperative complication rate was 5.4%(3/55),including skin infections around the port in one case,catheter-related infection in one case,and fibrin sheath formation in one case.The ports were all preserved after anti-infection or thrombolytic therapy.No unplanned port withdrawal was recorded in this study.Conclusions DUG-TIVAP implantation is a technique with a high success rate and a low complication rate;therefore,it provides an alternative for children with cancer.Further randomized controlled studies are needed to confirm the efficacy and safety of DUG-TIVAP via the right IJV in children.

Comprehensive geophysical profile and lithosphere structures and geodynamics in western Qinghai-Xizang (Tibetan) Plateau

A comprehensive study in this paper for the lithosphere velocity structure, conductivity structure, density and magnetism structure at the Qinghai-Xizang(Tibetan) Plateau has been completed based on the data of gravity, geomagnetism, and magneto-telluric sounding (MTS), explosion seismology acquired on the comprehensive geophysical profile along Gyirong-Sange-

Gallium bismuth halide GaBi-X2 （X = I, Br, CI） monolayers with distorted hexagonal framework： Novel room- temperature quantum spin Hall insulators

Quantum spin Hall （QSH） insulators with a large topologically nontrivial bulk gap are crucial for future applications of the QSH effect. Among these, group III-V monolayers and their halides, which have a chair structure （regular hexagonal framework）, have been widely studied. Using first-principles calculations, we formulate a new structure model for the functionalized group III-V monolayers, which consist of rectangular GaBi-X2 （X = I, Br, C1） monolayers with a distorted hexagonal framework （DHF）. These structures have a far lower energy than the GaBi-X2 monolayers with a chair structure. Remarkably, the DHF GaBi-X2 monolayers are all QSH insulators, which exhibit sizeable nontrivial band gaps ranging from 0.17 to 0.39 eV. The band gaps can be widely tuned by applying different spin-orbit coupling strengths, resulting in a distorted Dirac cone.

Sorption of triclosan on electrospun fibrous membranes: Effects of pH and dissolved organic matter

Three hydrophobic and polyporous electrospun fibrous membranes(EFMs)were prepared by electrospinning methoxy polyethylene glycol-poly(lactide-co-glycolide)(MPEG-PLGA),poly(D,L-lactide-co-glycolide)(PLGA)and poly(D,L-lactide)(PDLLA).The effects of pH and dissolved organic matter(DOM)on triclosan(TCS)sorption by EFMs in aqueous solution were investigated.The results indicated that hydrogen bonding,hydrophobic and π-π bonding interactions led to fast adsorption,which governed the adsorption rates of TCS onto EFMs.The maximum sorption capacities of MPEG-PLGA,PLGA and PDLLA reached 130,93 and 99 mg g^-1,respectively,which were in positive correlation with their pore volumes and influenced by pore filling processes.The solution pH could significantly influence the TCS sorption by EFMs.In acid condition,protonated TCS facilitated their sorption onto EFMs.No obvious sorption was observed in alkaline condition due to repulsive forces between negatively charged EFMs and deprotonated TCS(pKa=7.9).The presence of DOM inhibited TCS sorption onto EFMs due to competitive adsorption.The results could be due to the occupation of the adsorption sites and the blockage of the pore entrance by DOM.