敲低PFKP联合肉碱棕榈酰基转移酶抑制剂etomoxir协同对肾透明细胞癌细胞Caki-1的抗肿瘤作用

目的:研究敲低P型磷酸果糖激酶(phosphofructokinase,PFKP)联合肉碱棕榈酰基转移酶抑制剂etomoxir对肾透明细胞癌Caki-1细胞的影响,并进一步探究其作用机制。方法:利用Western blot验证对照及PFKP shRNA敲低肾透明细胞癌细胞中PFKP的敲低效率,分别检测对照组(shCtrl)、PFKP敲低组、etomoxir组(shCtrl+etomoxir)、PFKP敲低联合etomoxir组的增殖曲线。使用Annexin-V/PI染色并用流式细胞检测对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的细胞死亡,研究PFKP敲低联合etomoxir对细胞存活的影响。分别检测对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的ATP水平与脂肪酸变化。结果:Western blot结果验证了PFKP的敲低效率。流式细胞检测显示,对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的平均细胞死亡率分别为1.1、1.9、13.9、31.3%。PFKP敲低联合etomoxir组Caki-1细胞的死亡率显著高于单纯PFKP敲低与etomoxir组(P0.05)。PFKP敲低联合etomoxir组Caki-1细胞的ATP水平显著低于单纯PFKP敲低与etomoxir组(P0.05)。Etomoxir的加入抑制了PFKP敲低引起的游离脂肪酸下降(P0.05)。结论:PFKP敲低联合etomoxir对Caki-1细胞呈现协同的细胞毒抗肿瘤作用。     

64排螺旋CT对粗隆间骨折Evans分型的影响研究

目的:探讨64排螺旋CT对粗隆间骨折Evans分型的影响,为临床使用提供参考依据。方法:2015年3月至2017年3月,三甲医院高年资创伤骨科主任医师2名,医师1、医师2分别按照术前X线、术前64排螺旋CT平扫和三位重建结果对128例新鲜闭合单侧粗隆间骨折患者进行Evans分型,分别记为X线分型、CT分型。本院术者依据围术期X线、CT及术中所见骨折情况进行Evans分型(逆粗隆间骨折定义为Ⅴ型)作为最终分型。记录分型结果,计算并对比准确率、误诊率。结果:(1)剔除5例,90.09%(123/128)的患者完成研究。(2)分型结果:X线分型中,3例(最终分型Ⅲ型2例,Ⅳ型1例)无法定型;Ⅰ型正确1例,改为Ⅱ型1例;Ⅱ型正确18例,改为Ⅰ型2例,改为Ⅲ型3例,Ⅳ型2例;Ⅲ型正确45例,改为Ⅱ型7例,改为Ⅳ型1例;Ⅳ型正确19例,改为Ⅱ型3例,改为Ⅲ型15例。CT分型中,Ⅰ型正确3例,Ⅱ型正确29例,Ⅲ型正确64例,改为Ⅳ型1例,Ⅳ型正确22例,Ⅴ型正确3例。(3)CT分型的总准确率、总误诊率优于X线分型(99.19%vs67.48%、0.81%vs30.08%,P0.05)。(4)Ⅰ型、Ⅱ型、Ⅲ型、Ⅳ型骨折进行CT分型,准确率高于X线分型(P0.05),误诊率低于X线分型(P0.05);Ⅴ型骨折,两种分型准确率、误诊率相等。结论:64排螺旋CT平扫及三维重建是粗隆间骨折Evans分型较为可靠的辅助检查,可考虑推广运用。     

广西红壤土壤微生物溶磷基因的克隆与表达

本研究对广西红壤土壤微生物溶磷基因进行克隆,并在大肠杆菌中诱导表达。从广西红壤中提取宏基因组DNA,克隆了GabY基因,构建大肠杆菌原核表达载体p ETDuet-1,转化大肠杆菌DH5α得到重组工程菌。通过酶切验证重组体。重组体在诱导条件下24 h对难溶矿物质磷Ca_3(PO_4)_2的溶磷量为(40.73±1.32)μg/mL,而对照组仅为(5.11±0.08)μg/mL。重组体和对照随着诱导时间的增加,培养基pH值均有下降趋势,但重组体培养基pH值变化显著大于对照组,产酸能力显著优于对照组。证明成功克隆得到土壤微生物溶磷基因,并在大肠杆菌中得到表达。     

木薯SWEETs基因家族生物信息学及表达特性研究

SWEET (sugars will eventually be exported transporters)是植物中新发现的一类编码糖转运蛋白的基因,它在植物生长发育及糖代谢过程中发挥重要作用。该基因家族在木薯(Manihot esculenta)中尚未有详细的报道。本研究从Phytozome数据库获得了28个木薯SWEET候选基因并对其进行生物信息学分析,在华南124的木薯苗中通过荧光定量实验检测SWEET基因在旱胁迫下的表达水平。结果发现木薯SWEET基因被分为4簇,主要分布在第6条和第14条染色体上,编码234 aa与302 aa之间的氨基酸序列;木薯SWEET基因家族的表达在旱胁迫条件下发生了变化,其中明显上调的基因有9个,包括MeSWEET1b、MeSWEET2a、MeSWEET6、MeSWEET9a、MeSWEET9b、MeSWEET12、MeSWEET15a、MeSWEET15b和MeSWEET16c;而表达量明显下调的基因也有9个,为MeSWEET2b、MeSWEET3b、MeSWEET4、MeSWEET7、MeSWEET11、MeSWEET16a、MeSWEET16b、MeSWEET17a和MeSWEET17c。这些结果为进一步阐明SWEET基因家族在木薯中的功能提供理论依据。     

Application of next-generation sequencing to screen for pathogenic mutations in 123 unrelated Chinese patients with Marfan syndrome or a related disease

Marfan syndrome(MFS) is a systemic connective tissue disease principally affecting the ocular, skeletal and cardiovascular systems. This autosomal dominant disorder carries a prevalence of 1:3,000 to 1:5,000. This study aims to define the mutational spectrum of MFS related genes in Chinese patients and to establish genotype-phenotype correlations in MFS. Panel-based targeted next-generation sequencing was used to analyze the FBN1, TGFBR1 and TGFBR2 genes in 123 unrelated Chinese individuals with MFS or a related disease. Genotype-phenotype correlation analyses were performed in mutation-positive patients. The results showed that 97 cases/families(78.9%; 97/123) harbor at least one(likely) pathogenic mutation, most of which were in FBN1; four patients had TGFBR1/2 mutations; and one patient harbored a SMAD3 mutation. Three patients had two FBN1 mutations, and all patients showed classical MFS phenotypes. Patients with a dominant negative-FBN1 mutation had a higher prevalence of ectopia lentis(EL). Patients carrying a haploinsufficiency-FBN1 mutation tended to have aortic dissection without EL. This study extends the spectrum of genetic backgrounds of MFS and enriches our knowledge of genotype-phenotype correlations.     

Biogenic production of DMSP and its degradation to DMS—their roles in the global sulfur cycle

Dimethyl sulfide(DMS) is the most abundant form of volatile sulfur in Earth's oceans, and is mainly produced by the enzymatic clevage of dimethylsulfoniopropionate(DMSP). DMS and DMSP play important roles in driving the global sulfur cycle and may affect climate. DMSP is proposed to serve as an osmolyte, a grazing deterrent, a signaling molecule, an antioxidant, a cryoprotectant and/or as a sink for excess sulfur. It was long believed that only marine eukaryotes such as phytoplankton produce DMSP. However, we recently discovered that marine heterotrophic bacteria can also produce DMSP, making them a potentially important source of DMSP. At present, one prokaryotic and two eukaryotic DMSP synthesis enzymes have been identified.Marine heterotrophic bacteria are likely the major degraders of DMSP, using two known pathways: demethylation and cleavage.Many phytoplankton and some fungi can also cleave DMSP. So far seven different prokaryotic and one eukaryotic DMSP lyases have been identified. This review describes the global distribution pattern of DMSP and DMS, the known genes for biosynthesis and cleavage of DMSP, and the physiological and ecological functions of these important organosulfur molecules, which will improve understanding of the mechanisms of DMSP and DMS production and their roles in the environment.     

Novel hepacivirus in Asian house shrew,China

<正>Dear Editor,Hepatitis C virus (HCV) is a leading global cause of various liver diseases, including chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. The genome of HCV is monopartite, single-stranded, positive RNA, about 10 kb in size.HCV is the prototype species of the Hepacivirus genus,which contains 14 species according to the update from the International Committee on Taxonomy of Viruses (Smith et al., 2016). Prior to 2005, humans were thought to be the only     

氰酸盐诱导氧化应激促进肾小管上皮细胞上皮–间充质转化

该研究探讨氰酸盐(cyanate)诱导肾小管上皮细胞氧化应激损伤和促进肾纤维化的作用。氰酸盐作用HK-2肾小管上皮细胞后, CCK8法检测其对细胞活力的影响;倒置显微镜观察细胞形态的改变; DCFH-DA法检测细胞ROS水平;细胞免疫荧光和Western blot分别检测E-cadherin、Fibronectin、α-SMA的表达; Western blot检测TGF-β的表达水平。结果显示, 2 mmol/L氰酸盐明显下调HK-2细胞的活力(P<0.05),细胞形态变为长梭形。氰酸盐作用24 h后, HK-2细胞内ROS水平呈浓度依赖性升高。免疫荧光和Western blot结果均显示,氰酸盐作用24 h后, HK-2的Fibronectin、α-SMA表达升高, E-cadherin表达下降; TGF-β的表达水平随氰酸盐浓度升高而上调(P<0.05)。以上结果表明,氰酸盐诱导肾小管上皮细胞产生过量ROS,上调TGF-β水平促进细胞上皮–间充质细胞转化(epithelia-mesenchymal transition, EMT)。     

细胞自噬研究进展

细胞自噬是真核生物在进化过程中高度保守、基于溶酶体的一种胞内降解途径,对维持细胞和生物体的稳态平衡有重要作用。研究表明,自噬参与生物体发育、免疫反应、代谢调节、细胞凋亡和衰老等多种过程。自噬功能异常与神经退行性疾病、肿瘤等的发生发展密切相关。近30年,我们对细胞自噬的认识无论是在分子机制上还是生理功能方面都有了长足的发展。为进一步加深对细胞自噬的认识,该文主要对细胞自噬的概念、自噬核心机器的组成及调控机制、自噬类型、生理功能及与疾病的关系作一简单综述。