miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。     

超声引导下腔内射频消融联合点式剥脱治疗下肢静脉曲张的临床效果观察

目的:探讨超声引导下腔内射频消融(radiofrequency endovenous obliteration,RFO)联合点式剥脱治疗下肢静脉曲张的临床疗效及安全性。方法:选取2017年10月至2018年10月哈尔滨医科大学附属第一医院收治的下肢静脉曲张患者共320例(320条肢体),将其随机分为观察组200例和对照组120例。观察组200例(200条肢体)采用超声引导下腔内射频消融联合点式剥脱进行治疗,对照组120例(120条肢体)采用传统大隐静脉高位结扎剥脱、曲张静脉点式剥脱治疗。比较两组患者平均住院时间、术后并发症的发生情况及术后1年下肢静脉的彩超随访情况。结果:观察组患者平均住院时间较对照组显著缩短,术后皮肤瘀斑、皮下血肿、隐神经损伤、切口感染的发生率显著低于对照组(P0.05),两组患者术后1年复查,其复发率及大隐静脉主干闭塞情况比较差异没有统计学意义(P0.05)。结论:与传统大隐静脉高位结扎剥脱、曲张静脉点式剥脱治疗相比,超声引导下腔内射频消融联合点式剥脱治疗下肢静脉曲张的疗效及安全性均更高,有利于患者术后康复。     

RPN2与肝癌患者预后及对肝癌细胞生长的作用

目的:探讨核糖蛋白2(ribophorin II,RPN2)在肝细胞肝癌(HCC)组织中的表达和对HCC患者生存的影响,同时分析RPN2对肝癌HepG2细胞生长和克隆形成的作用。方法:应用免疫组化方法和HCC公共芯片数据,从蛋白和m RNA水平检测HCC组织中RPN2的表达,同时分析RPN2与HCC患者临床参数的关系及预后相关性;进一步利用MTS法和克隆形成实验在肝癌HepG2细胞中检测RPN2对细胞生长的作用。结果:98例肝癌组织中,RPN2阳性表达率88.78%,对应癌旁肝组织中,RPN2阳性表达率74.49%;癌组织中RPN2染色评分为5.80±3.15,癌旁肝组织RPN2染色评分为2.13±1.59,肝癌组织中RPN2表达显著上调(P0.001)。3个肝癌公共芯片数据(共522例肝癌)中RPN2的m RNA表达水平同样显著升高(均P0.001)。98例肝癌患者RPN2表达水平与肿瘤直径(P=0.004)、门脉侵袭(P=0.012)和TNM分期(P=0.009)相关;RPN2高表达的患者总体生存期(OS)和无复发生存期(RFS)较RPN2低表达的患者短(OS:P=0.027;RFS:P=0.036)。肝癌HepG2细胞转染RPN2小干扰RNA后,细胞生长能力显著受抑制。结论:RPN2在肝癌中表达显著升高,RPN2的表达与肝癌的恶性进展有关,RPN2显著促进肝癌细胞生长。     

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。     

水椰八角铁甲不同日龄和性别成虫表皮碳氢化合物的种类及动态

水椰八角铁甲Octodonta nipae(Maulik)是一种严重为害棕榈科植物的外来入侵害虫。为了明确日龄和性别等个体因素对水椰八角铁甲成虫表皮碳氢化合物的影响,本研究采用化学萃取法和气相色谱-质谱联用技术,分析了不同日龄的水椰八角铁甲雌、雄成虫表皮碳氢化合物的组成和含量。结果表明,水椰八角铁甲成虫的表皮碳氢化合物由21种C_(18)-C_(30)的碳氢化合物构成,包括11种正构烷烃、2种单甲基烷烃、1种多甲基烷烃、3种正构烯烃、1种单甲基烯烃和3种环烷烃。不同日龄和性别成虫的表皮碳氢化合物的组成种类相同,但是一些组分的含量有明显变化,例如1 d、5 d、10 d和15 d雌、雄成虫之间正二十四烷、正二十六烷、2-甲基十九烷和1-二十烯的含量存在显著差异。水椰八角铁甲成虫表皮碳氢化合物具有性二型,如性成熟雄虫的表皮的正二十烷、正二十一烷、正二十四烷、正二十六烷、2-甲基十九烷、10-甲基二十烷、1-二十烯和1-二十二烯的含量显著高于性成熟雌虫。本文的研究结果可为检验检疫过程中水椰八角铁甲近缘种,以及日龄、性别和性成熟的快速鉴定提供支持。     

中国斑叶蝉族昆虫地理分布格局聚类分析

分析了斑叶蝉族昆虫在中国以及贵州动物地理区的分布情况,探讨其分布格局形成、起源和演化原因。根据中国和贵州斑叶蝉族的地理分布数据,运用MEGA 6.0、SPSS 22.0和ArcGIS 10.2等软件,对斑叶蝉族昆虫的区及亚区分布进行支序聚类分析,结果表明我国斑叶蝉族现代分布中心为西部山地高原亚区、华南区的台湾亚区及滇南山地亚区,分布热点地区为西双版纳地区、海南地区和台湾地区。贵州斑叶蝉现代分布中心为黔东低山丘陵省、黔北中山峡谷省和黔南低山河谷省,分布热点地区为铜仁北部的沿河地区、遵义的务川地区及贵州黔东南州的榕江、雷山地区。斑叶蝉族昆虫中国分布区形成的顺序先是东北区,其次是青藏区和蒙新区,最后是西南区、华北区、华南区和华中区。贵州分布区形成的顺序先是黔西高原中山省和黔中山原丘陵省,其次是黔南低山河谷省,最后是黔北中山峡谷省和黔东低山丘陵省。中国分布区中,B21和B22聚类群属级阶元的相似性最高,区间关联性最强。在贵州分布区中黔东低山丘陵省和黔北中山峡谷省属级阶元的相似性最高,物种交流最为频繁。目前,斑叶蝉族昆虫的地理分布格局主要是历史气候变化、当前气候条件以及植被覆盖等生态环境共同作用的结果;区系起源和演化主要受地质构造运动作用;斑叶蝉在各区的分布相似性可能与气候变化引起的物种由南向北扩散有关。     

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.     

绵马贯众多糖超声提取工艺优化及抗氧化活性分析

绵马贯众是中国传统常用中药,本研究以温度、时间、超声功率、液料比为影响因子,多糖得率为评价指标,通过响应面法优化超声辅助提取绵马贯众多糖的工艺条件,同时测定其基本理化性质及抗氧化活性。研究结果表明,绵马贯众多糖的最佳提取工艺条件为:温度64℃、时间60 min、超声功率210 W、液料比27 mL/g。此时多糖得率为9.57%,与预测值接近。理化性质分析表明绵马贯众多糖为含少量蛋白的酸性多糖。体外抗氧化研究表明绵马贯众多糖具有很强的DPPH自由基清除活性,IC50值为0.29 mg/mL;较好的羟基自由基清除活性,其IC50值为1.10 mg/mL;对DNA的氧化损伤有显著的保护作用。绵马贯众多糖可以作为一种潜在的抗氧化剂应用于食品和化妆品等领域。     

Microbial assemblages associated with the rhizosphere and endosphere of an herbage,Leymus chinensis

Root-associated microbiomes play significant roles in plant productivity, health and ecological services. However, our current understanding of the microbial assemblages in the rhizosphere and endosphere of herbage is still limited. To gain insights into these microbial assemblages, Illumina MiSeq high-throughput sequencing was performed to investigate the characteristics of microbial communities of an herbage, Leymus chinensis. Hierarchical clustering analysis and principal coordinate analysis (PCoA) results showed that microbial communities of the rhizosphere and endosphere samples were clearly distinguished. Rhizosphere soil communities showed a greater sensitivity than root endosphere communities using linear discriminant analysis (LDA) effect size (LEfSe). Rhizosphere and endosphere communities performed their respective functions in the soil as a cohesive collective, and Rhizobiales were observed to function as generalists. Redundancy analysis (RDA) and variance partitioning analysis (VPA) results revealed that the contribution of the interaction between soil physicochemical parameters and soil enzymes was greater than their individual contributions. In summary, this study is the first to elucidate the microbial diversity and community structure of L. chinensis and compare the diversity and composition between rhizospheric and endosphere microbiomes.     

Aging shifts mitochondrial dynamics toward fission to promote germline stem cell loss

Changes in mitochondrial dynamics (fusion and fission) are known to occur during stem cell differentiation; however, the role of this phenomenon in tissue aging remains unclear. Here, we report that mitochondrial dynamics are shifted toward fission during aging of Drosophila ovarian germline stem cells (GSCs), and this shift contributes to aging‐related GSC loss. We found that as GSCs age, mitochondrial fragmentation and expression of the mitochondrial fission regulator, Dynamin‐related protein (Drp1), are both increased, while mitochondrial membrane potential is reduced. Moreover, preventing mitochondrial fusion in GSCs results in highly fragmented depolarized mitochondria, decreased BMP stemness signaling, impaired fatty acid metabolism, and GSC loss. Conversely, forcing mitochondrial elongation promotes GSC attachment to the niche. Importantly, maintenance of aging GSCs can be enhanced by suppressing Drp1 expression to prevent mitochondrial fission or treating with rapamycin, which is known to promote autophagy via TOR inhibition. Overall, our results show that mitochondrial dynamics are altered during physiological aging, affecting stem cell homeostasis via coordinated changes in stemness signaling, niche contact, and cellular metabolism. Such effects may also be highly relevant to other stem cell types and aging‐induced tissue degeneration.