Association of SLC22A4 Gene Polymorphism with Rheumatoid Arthritis in the Chinese Population

Rheumatoid arthritis (RA) is a chronic inflammatory disease with complex genetic factors. Single‐nucleotide polymorphisms (SNPs) in the SLC22A4 gene have been previously reported to be associated with RA in Japanese but not European populations. This study further investigated the association of SLC22A4 polymorphisms, in particular slc2F1/slc2F2, with RA in the Chinese population, the largest Asian population. A total of 160 human subjects with 95 RA patients and 65 healthy controls were genotyped for slc2F1‐G/A and slc2F2‐C/T polymorphisms. The results showed that there was a significant difference in the genotype distribution of these two polymorphisms between the two groups. In addition, the presence of slc2F1 A allele and slc2F2 T allele carries a 1.93‐fold and 2.14‐fold increased risk for anticyclic citrullinated peptide (CCP) positivity, respectively. Overall, this study provided evidence that SLC22A4 gene polymorphisms played important roles in the etiology of RA in the largest Asian population, the Chinese population.     

野生和栽培防风根系的比较形态解剖学研究

研究了野生和栽培防风根系的外部形态及内部结构,研究结果表明:在外部形态上,栽培防风根系颜色较浅,侧根较多。在内部结构上,二者大致相同。二年生栽培防风主根与野生多年生防风主根中油室数目、韧皮部/木质部相近,建议生产上适当采收二年生植株根系。     

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.     

SARS病毒核蛋白基因的克隆及其表达研究

从一例输入性传染性非典型性肺炎病人血清中提取病毒RNA,通过RT-PCR方法扩增出SARS病毒核蛋白基因片段,克隆入质粒载体pUCm-T后,进行核苷酸序列的测定及分析,与已公布的SARS病毒基因序列进行比较,证实为SARS冠状病毒核蛋白基因.为了解该病毒核蛋白的抗原特性,将核蛋白基因插入表达载体,构建重组质粒pET28a-SN,转导大肠杆菌BL21(DE3)后,加IPTG诱导表达.产物经SDS-PAGE电泳分析,表达出相对分子量约为50kDa的蛋白,占整个菌体的45%左右.Western-blot分析表明,表达产物仅与SARS阳性病人血清起反应,而与正常血清不起反应.间接ELISA免疫检测,抗原滴度达112500.表明表达的核蛋白为SARS特异性抗原,这为SARS病毒的诊断试剂的研制提供了方便而安全的抗原来源.     

含霍乱肠毒素B亚单位基因植物表达载体的构建策略

采用高保真PCR方法调出CTB基因,经测序证实核酸序列正确后,再亚克隆到含植物表达调控原件的载体上,采用冻融法和电击法,将含CTB的植物表达载体转入根癌农杆菌中,通过一系列分子克隆的方法获得含CTB基因的植物及元素达载体pBI-CTB和pBI-CTBK,并经酶切证实,为下一步的植物转基因研究打下坚实的基础。     

Structural and functional characterization of a novel acidophilic 7α‐hydroxysteroid dehydrogenase

7α‐Hydroxysteroid dehydrogenase (7α‐HSDH) is an NAD(P)H‐dependent oxidoreductase belonging to the short‐chain dehydrogenases/reductases. In vitro, 7α‐HSDH is involved in the efficient biotransformation of taurochenodeoxycholic acid (TCDCA) to tauroursodeoxycholic acid (TUDCA). In this study, a gene encoding novel 7α‐HSDH (named as St‐2‐1) from fecal samples of black bear was cloned and heterologously expressed in Escherichia coli. The protein has subunits of 28.3 kDa and a native size of 56.6 kDa, which suggested a homodimer. We studied the relevant properties of the enzyme, including the optimum pH, optimum temperature, thermal stability, activators, and inhibitors. Interestingly, the data showed that St‐2‐1 differs from the 7α‐HSDHs reported in the literature, as it functions under acidic conditions. The enzyme displayed its optimal activity at pH 5.5 (TCDCA). The acidophilic nature of 7α‐HSDH expands its application environment and the natural enzyme bank of HSDHs, providing a promising candidate enzyme for the biosynthesis of TUDCA or other related chemical entities.     

In Situ Electrochemical Synthesis of MXenes without Acid/Alkali Usage in/for an Aqueous Zinc Ion Battery

The traditional method to fabricate a MXene based energy storage device starts from etching MAX phase particles with dangerous acid/alkali etchants to MXenes, followed by device assembly. This is a multistep protocol and is not environmentally friendly. Herein, an all‐in‐one protocol is proposed to integrate synthesis and battery fabrication of MXene. By choosing a special F‐rich electrolyte, MAX V₂AlC is directly exfoliated inside a battery and the obtained V₂CT_X MXene is in situ used to achieve an excellent battery performance. This is a one‐step process with all reactions inside the cell, avoiding any contamination to external environments. Through the lifetime, the device experiences three stages of exfoliation, electrode oxidation, and redox of V₂O₅. While the electrode is changing, the device can always be used as a battery and the performance is continuously enhanced. The resulting aqueous zinc ion battery achieves outstanding cycling stability (4000 cycles) and rate performance (97.5 mAh g^?1 at 64 A g^?1), distinct from all reported aqueous MXene‐based counterparts with pseudo‐capacitive properties, and outperforming most vanadium‐based zinc ion batteries with high capacity. This work sheds light on the green synthesis of MXenes, provides an all‐in‐one protocol for MXene devices, and extends MXenes’ application in the aqueous energy storage field.     

Theoretical investigation of reactivities of amines in the <Emphasis Type="Italic">N</Emphasis>-nitrosation reactions by N<Subscript>2</Subscript>O<Subscript>3</Subscript>

N-nitrosamine is a class of carcinogenic, mutagenic, and teratogenic compounds, which can be produced from N-nitrosation of amine by nitrosating agents. N-nitrosation of 19 amines (eight acyclic amines, five heterocyclic amines, and six amines with unsaturated groups) by N₂O₃ was investigated at the CBS-QB3 level of theory. The results indicate that generally the heterocyclic amines have the highest reactivities among the three kinds of amines, whereas the reactivities of the amines with unsaturated and electron-withdrawing groups are relatively low. Frontier molecular orbital analysis indicates that the energy gap between the HOMO of an amine and the LUMO of N₂O₃ has a close connection with the reactivity of an amine. A structure-reactivity relationship of amines in the N-nitrosation reactions by N₂O₃ was established using the stepwise multivariate linear regression. The results indicate that the reactivity of an amine has a definite relationship (R_adj² = 0.947) with the heterolytic bond dissociation energy of R₁R₂N-H bond, energy of HOMO, NBO occupancy of the natural lone pair orbital of N atom, the NBO charge of the N atom, and the pyramidalization angle of an amine. This work will be helpful to gain more insight into the N-nitrosation reactions.     

蛋白质代谢、折叠、运输、定位、装配相关基因在大鼠肝再生中表达变化

为在基因转录水平了解蛋白质代谢、折叠、运输、定位、装配相关基因在大鼠肝再生中表达情况和作用,本文用搜集网站资料和查阅相关论文等方法获得上述基因,用Rat Genome 2302.0芯片检测它们在大鼠再生肝中表达情况,用真、假手术比较方法确定肝再生相关基因。初步证实上述基因中1147个基因与肝再生相关。其中,参与蛋白质代谢、折叠、运输、定位和装配的基因以上调表达为主;参与蛋白质代谢的基因主要在部分肝切除（partial hepatectomy,PH）后0.5-1h和16-30h起始表达;0.5-12h表达的促进蛋白降解基因数多于促进蛋白积累基因数,而16-48h表达的促进蛋白质积累基因数显著多于促进蛋白质降解基因数;蛋白质合成相关基因在肝再生的16、24、42和66h表达上调较多,在42h最多;几乎在整个肝再生中蛋白质降解相关基因表达上调,在早、前期较多,在后期较少;蛋白质折叠相关基因在2、16-24、42、66、72和168h表达上调较多,在66h最多;蛋白质运输和定位相关基因在整个肝再生中表达上调,在66h表达上调最多;蛋白质装配相关基因在96h前均表达上调,其中,12h表达上调基因最多。根据上述结果推测,在肝再生中期蛋白质合成旺盛,几乎整个肝再生中蛋白质降解、折叠、运输定位和装配活动活跃。     

干旱和臭氧浓度升高对元宝枫早生和晚生叶片色素和脱落酸含量的影响

臭氧和干旱是威胁我国北方城市植物生长的两大重要因素。于2012年利用开顶式气室,通过设置4个处理(AW-大气环境和水分充足;AW+60-大气增加60 nL/L臭氧+水分充足;AD-大气+干旱处理;AD+60-大气增加60 nL/L臭氧+干旱处理),开展了大气臭氧浓度升高(以下简称"臭氧")和干旱对元宝枫秋季变色期主要色素含量及脱落酸(ABA)含量的影响研究。结果表明:(1)早生叶在臭氧处理后,总叶绿素和类胡萝卜素分别下降了21%和29.6%、花青苷和类黄酮相对含量显著升高了34.1%和7.3%、脱落酸含量增加了19.8%。干旱处理后,早生叶总叶绿素显著下降了18.7%、花青苷和类黄酮相对含量分别显著升高了37%和7.4%、脱落酸含量显著升高了13%。叶片的上述生理变化将会导致叶片提前变红、叶片早衰和提前脱落。(2)晚生叶在干旱处理后总叶绿素含量减少了18.8%,脱落酸含量增加了33.4%,臭氧以及与干旱共同处理未对晚生叶产生显著影响。(3)臭氧和干旱共同处理后,早生叶总叶绿素含量、花青苷和叶片脱落酸含量存在显著交互作用,交互作用缓解了叶片总叶绿素的下降和花青苷的上升,但未缓解叶片脱落酸含量的增加。综上,早生叶和晚生叶对臭氧和干旱处理的响应不同,早生叶对臭氧处理响应大于晚生叶,而晚生叶对干旱更敏感。臭氧和干旱处理均加速了叶片衰老,二者共同处理后叶片脱落风险增加。