胃癌组织长链非编码RNA ZDHHC8P1、MEG3、TUG1表达与临床病理特征及预后的关系研究

目的:探讨胃癌组织长链非编码RNA(lncRNA)DHHC型锌指蛋白8假基因1(ZDHHC8P1)、母系表达基因3(MEG3)、牛磺酸上调基因1(TUG1)表达与临床病理特征和预后的关系。方法:选取2013年1月至2016年1月我院病理科收集的83例胃癌患者经手术切除或胃镜活检的癌组织及癌旁组织石蜡标本,检测胃癌和癌旁组织中ZDHHC8P1、MEG3、TUG1表达。分析ZDHHC8P1、MEG3、TUG1表达与胃癌临床病理特征的关系。随访所有患者,分析ZDHHC8P1、MEG3、TUG1表达与患者预后的关系。结果:胃癌组织中ZDHHC8P1、TUG1表达量均高于癌旁组织(P0.05),MEG3表达量低于癌旁组织(P0.05)。ZDHHC8P1表达与肿瘤直径、分化程度、浸润深度、TNM分期、淋巴结转移、远处转移有关(P0.05),MEG3、TUG1表达与分化程度、浸润深度、淋巴结转移有关(P0.05)。Kaplan-Meier生存曲线分析结果显示ZDHHC8P1、TUG1高表达患者无疾病进展生存(PFS)率、总生存(OS)率低于ZDHHC8P1、TUG1低表达患者(P0.05),MEG3低表达患者PFS、OS率低于MEG3高表达患者(P0.05)。Cox风险回归分析结果显示淋巴结转移、ZDHHC8P1高表达、TUG1高表达、MEG3低表达是胃癌患者不良预后的危险因素(HR=1.613、1.956、2.512、-0.824,P0.05)。结论:胃癌组织中ZDHHC8P1、TUG1呈高表达,MEG3呈低表达,ZDHHC8P1、TUG1、MEG3表达均与胃癌临床病理特征和预后有关,可作为胃癌患者预后评估的辅助指标。     

门冬胰岛素联合甘精胰岛素对新诊断2型糖尿病患者炎性因子、血糖及血脂水平的影响

目的:探讨门冬胰岛素联合甘精胰岛素对新诊断2型糖尿病患者血清炎性因子、血糖及血脂水平的影响。方法:选取我院2017年3月～2019年3月收治的100例新诊断2型糖尿病患者,将其随机分为研究组和对照组,每组50例患者。对照组患者给予门冬胰岛素治疗,研究组患者给予门冬胰岛素联合甘精胰岛素治疗,对比两组患者治疗前后血清炎性因子、血糖及血脂水平的变化。结果:治疗后,两组患者糖化血红蛋白(hemoglobin A1c,HbA1c)、空腹血糖(fasting plasma glucose,FPG)、餐后2h血糖(2h postprandial blod glucose,2hPBG)、血清肿瘤坏死因子(Tumor necrosis factor,TNF-α)、高敏C反应蛋白(high-sensitive C-reactive protein,hs-CRP)水平均较治疗前明显降低(P0.05),且研究组患者以上指标均显著低于对照组(P0.05);两组患者治疗后总胆固醇(Total Cholesterol,TC)、甘油三酯(Triglyceride,TG)、低密度脂蛋白胆固醇(Low-density lipoprotein cholesterol,LDL-C)水平均较治疗前明显降低,高密度脂蛋白胆固醇(High-density lipoprotein cholesterol,HDL-C)明显升高(P0.05),且研究组以上指标的改善程度均优于对照组(P0.05)。研究组患者的胰岛素用量明显少于对照组(P0.05),患者血糖首次达标时间明显短于对照组(P0.05)。结论:门冬胰岛素联合甘精胰岛素适用于新诊断2型糖尿病患者,可有效降低血糖、血脂、血清TNF-α和hs-CRP水平。     

加味八珍汤联合PFNA术治疗老年股骨转子间骨折患者的临床效果及对血液流变学指标的影响

目的:研究加味八珍汤联合股骨近端防旋髓内钉(Proximal Femoral Nail Antirotation,PFNA)对老年股骨转子间骨折患者的临床效果及对术后隐性失血和血液流变学的影响。方法:选择2014年3月至2017年3月我院收治的68例接受PFNA治疗的转子间骨折者,将其随机平均分为观察组(34例,术后口服加味八珍汤颗粒7天)、对照组(34例)。对比两组总失血量、隐性失血量、隐性失血比例、血液流变学指标(血浆粘度及全血黏度低、中、高切)、不良反应等。结果:⑴两组各剔除2例,94.12%(64/68)的受试者完成研究。⑵两组总失血量相当,差异无统计学意义(P0.05);与对照组相比,观察组隐性失血量及隐性失血发生率均显著降低(P0.05)。⑶治疗前,两组血浆粘度及低、中、高切全血黏度比较差异无统计学意义(P0.05);治疗后,两组血浆粘度及低、中、高切全血黏度均显著低于本组治疗前(P0.05),且观察组以上指标均明显低于对照组(P0.05)。⑷观察组发生肌间静脉血栓1例,无药物过敏及毒副反应;对照组发生1例深静脉血栓、4例肌间静脉血栓。观察组不良反应率为3.125%,显著低于对照组(15.625%,P0.05)。结论:加味八珍汤既可显著减少老年股骨转子间骨折患者PFNA术后隐性失血,又能改善血流动力学、减少并发症。     

滇川桂3省暗褐网柄牛肝菌菌腔虫瘿的寄主植物与介壳虫种类调查

暗褐网柄牛肝菌Phlebopus portentosus与介壳虫形成的菌腔虫瘿是该菌营养机制研究的关键环节。本研究先后在云南、四川和广西3省区暗褐网柄牛肝菌产区的16个地点,对菌腔虫瘿的生态和生物学进行了大量的野外调查。发现根部着生菌腔虫瘿的寄主植物有31种,涉及16个科的28个属。与暗褐网柄牛肝菌形成菌腔虫瘿的介壳虫种类有12种,其中10种隶属粉蚧科Pseudococcidae、绵蚧科Monophlebidae、蚧科Coccidae各1种。在不同的寄主植物上菌腔虫瘿的寄生位置和形状会有所不同,与暗褐网柄牛肝菌菌丝形成菌腔虫瘿的寄主植物和介壳虫之间不存在专一性。上述研究结果为暗褐网柄牛肝菌的仿生栽培奠定了基础。     

Molecular phylogeny and species delimitation of Stachyuraceae: Advocating a herbarium specimen‐based phylogenomic approach in resolving species boundaries

Species concept and delimitation are fundamental to taxonomic and evolutionary studies. Both inadequate informative sites in the molecular data and limited taxon sampling have often led to poor phylogenetic resolution and incorrect species delineation. Recently, the whole chloroplast genome sequences from extensive herbarium specimen samples have been shown to be effective to amend the problem. Stachyuraceae are a small family consisting of only one genus Stachyurus of six to 16 species. However, species delimitation in Stachyurus has been highly controversial because of few and generally unstable morphological characters used for classification. In this study, we sampled 69 individuals of seven species (each with at least three individuals) covering the entire taxonomic diversity, geographic range, and morphological variation of Stachyurus from herbarium specimens for genome‐wide plastid gene sequencing to address species delineation in the genus. We obtained high‐quality DNAs from specimens using a recently developed DNA reconstruction technique. We first assembled four whole chloroplast genome sequences. Based on the chloroplast genome and one nuclear ribosomal DNA sequence of Stachyurus, we designed primers for multiplex polymerase chain reaction and high throughput sequencing of 44 plastid loci for species of Stachyurus. Data of these chloroplast DNA and nuclear ribosomal DNA internal transcribed spacer sequences were used for phylogenetic analyses. The phylogenetic results showed that the Japanese species Stachyurus praecox Siebold & Zucc. was sister to the rest in mainland China, which indicated a typical Sino‐Japanese distribution pattern. Based on diagnostic morphological characters, distinct distributional range, and monophyly of each clade, we redefined seven species for Stachyurus following an integrative species concept, and revised the taxonomy of the family based on previous reports and specimens, in particular the type specimens. Furthermore, our divergence time estimation results suggested that Stachyuraceae split from its sister group Crossosomataceae from the New World at ca. 54.29 Mya, but extant species of Stachyuraceae started their diversification only recently at ca. 6.85 Mya. Diversification time of Stachyurus in mainland China was estimated to be ca. 4.45 Mya. This research has provided an example of using the herbarium specimen‐based phylogenomic approach in resolving species boundaries in a taxonomically difficult genus.     

Knockdown of DNA‐binding protein A enhances the chemotherapy sensitivity of colorectal cancer via suppressing the Wnt/β‐catenin/Chk1 pathway

DNA‐binding protein A (dbpA) is reported to be upregulated in many cancers and associated with tumor progress. The present study aimed to investigate the role of dbpA in 5‐fluorouracil (5‐FU)‐resistant and oxaliplatin (L‐OHP)‐resistant colorectal cancer (CRC) cells. We found that 5‐FU and L‐OPH treatment promoted the expression of dbpA. Enhanced dbpA promoted the drug resistance of SW620 cells to 5‐FU and L‐OHP. DbpA knockdown inhibited cell proliferation, induced cell apoptosis, and cell cycle arrested in SW620/5‐FU and SW620/L‐OHP cells. Besides, dbpA short hairpin RNA (shRNA) enhanced the cytotoxicity of 5‐FU and L‐OHP to SW620/5‐FU and SW620/L‐OHP cells. Meanwhile, dbpA shRNA inhibited the activation of the Wnt/β‐catenin pathway that induced by 5‐FU stimulation in SW620/5‐FU cells. Activation of the Wnt/β‐catenin pathway or overexpression of checkpoint kinase 1 (Chk1) abrogated the promoting effect of dbpA downregulation on 5‐FU sensitivity of CRC cells. Importantly, downregulation of dbpA suppressed tumor growth and promoted CRC cells sensitivity to 5‐FU in vivo. Our study indicated that the knockdown of dbpA enhanced the sensitivity of CRC cells to 5‐FU via Wnt/β‐catenin/Chk1 pathway, and DbpA may be a potential therapeutic target to sensitize drug resistance CRC to 5‐FU and L‐OHP.     

Atg3 promotes Atg8 lipidation via altering lipid diffusion and rearrangement

Atg3‐catalyzed transferring of Atg8 to phosphatidylethanolamine (PE) in the phagophore membrane is essential for autophagy. Previous studies have demonstrated that this process requires Atg3 to interact with the phagophore membrane via its N‐terminal amphipathic helix. In this study, by using combined biochemical and biophysical approaches, our data showed that in addition to binding to the membranes, Atg3 attenuates lipid diffusion and enriches lipid molecules with smaller headgroup. Our data suggest that Atg3 promotes Atg8 lipidation via altering lipid diffusion and rearrangement.     

Uncovering the evolutionary origin of blue anthocyanins in cereal grains

Functional divergence after gene duplication plays a central role in plant evolution. Among cereals, only Hordeum vulgare (barley), Triticum aestivum (wheat) and Secale cereale (rye) accumulate delphinidin‐derived (blue) anthocyanins in the aleurone layer of grains, whereas Oryza sativa (rice), Zea mays (maize) and Sorghum bicolor (sorghum) do not. The underlying genetic basis for this natural occurrence remains elusive. Here, we mapped the barley Blx1 locus involved in blue aleurone to an approximately 1.13 Mb genetic interval on chromosome 4HL, thus identifying a trigenic cluster named MbHF35 (containing HvMYB4H, HvMYC4H and HvF35H). Sequence and expression data supported the role of these genes in conferring blue‐coloured (blue aleurone) grains. Synteny analyses across monocot species showed that MbHF35 has only evolved within distinct Triticeae lineages, as a result of dispersed gene duplication. Phylogeny analyses revealed a shared evolution pattern for MbHF35 in Triticeae, suggesting that these genes have co‐evolved together. We also identified a Pooideae‐specific flavonoid 3′,5′‐hydroxylase (F3′5′H) lineage, termed here Mo_F35H2, which has a higher amino acid similarity with eudicot F3′5′Hs, demonstrating a scenario of convergent evolution. Indeed, selection tests identified 13 amino acid residues in Mo_F35H2 that underwent positive selection, possibly driven by protein thermostablility selection. Furthermore, through the interrogation of barley germplasm there is evidence that HvMYB4H and HvMYC4H have undergone human selection. Collectively, our study favours blue aleurone as a recently evolved trait resulting from environmental adaptation. Our findings provide an evolutionary explanation for the absence of blue anthocyanins in other cereals and highlight the importance of gene functional divergence for plant diversity and environmental adaptation.