青虾“软壳综合症”病原及其特性

从患软壳综合症的濒死青虾体内分离到一株细菌QXL0711B, 经人工感染试验, 其对青虾的半数致死浓度(LC50)为1.47×106 CFU/mL, 具有较强毒力。API 32E系统鉴定及16S rRNA序列分析, 该病原菌为维罗纳气单胞菌温和生物变种(Aeromonas veronii biovar sobria, 登录号: FJ808727)。其系统发育分析表明, 菌株QXL0711B与维罗纳气单胞菌(登录号: X71120)和维罗纳气单胞菌温和生物变种(登录号: AY987729)的亲缘关系最近,     

5-Azacytidine facilitates osteogenic gene expression and differentiation of mesenchymal stem cells by alteration in DNA methylation

Mesenchymal stem cells (MSCs) are considered to be one of the most promising therapeutic cell sources as they encompass a plasticity of multiple cell lineages. The challenge in using these cells lies in developing well-defined protocols for directing cellular differentiation to generate a desired lineage. In this study, we investigated the effect of 5-azacytidine, a DNA demethylating agent, on osteogenic differentiation of MSCs. The cells were exposed to 5-azacytidine in culture medium for 24 h prior to osteogenic induction. Osteogenic differentiation was determined by several the appearance of a number of osteogenesis characteristics, including gene expression, ALP activity, and calcium mineralization. Pretreatment of MSCs with 5-azacytidine significantly facilitated osteogenic differentiation and was accompanied by hypomethylation of genomic DNA and increased osteogenic gene expression. Taking dlx5 as a representative, methylation alterations of the “CpG island shore” in the promoter caused by 5-azacytidine appeared to contribute to osteogenic differentiation.     

固定化细胞拆分(R,S)-α-乙基-2-氧-1-吡咯烷乙酸甲酯的研究

【目的】筛选鉴定一株产酯酶用于选择性拆分(R,S)-α-乙基-2-氧-1-吡咯烷乙酸甲酯的菌株,利用该菌株固定化细胞催化拆分外消旋底物。【方法】通过富集培养、罗丹明B平板初筛及复筛培养获得一株选择性拆分(R,S)-α-乙基-2-氧-1-吡咯烷乙酸甲酯的菌株,通过对其形态、生理生化特征及16S r DNA序列分析,确立该菌株系统发育地位。优化了利用硅藻土-戊二醛吸附交联法对该菌体细胞固定化的条件,研究固定化细胞催化性质及操作稳定性。【结果】该菌为革兰氏阴性菌,鉴定其为甲基球状菌属(Methylopila)。固定化体系最优条件:聚乙烯亚胺0.15%(V/V),戊二醛0.2%(V/V),硅藻土6 g/L,菌体质量浓度100 g/L。与游离细胞相比,固定化细胞最适p H由8.0变为8.5,最适温度由35°C变为40°C,p H稳定性和温度稳定性都有所提高。Cu~(2+)、Mn~(2+)、Ca~(2+)能促进酶活,Zn~(2+)、Fe~(2+)抑制酶活。固定化细胞的有机溶剂耐受性较游离细胞有所提高。动力学分析细胞固定化后Km值变大,底物亲和力降低。利用固定化细胞水解(R,S)-α-乙基-2-氧-1-吡咯烷乙酸甲酯,底物浓度200 g/L,反应20 h,保留构型为S型,得率47.8%,对映体过量值ees为99.4%,重复使用12次后仍保留初始酶活的80%以上。【结论】开发了利用Methylopila sp.cxzy-L013固定化细胞择性拆分(R,S)-α-乙基-2-氧-1-吡咯烷乙酸甲酯的工艺,该工艺具有良好的工业应用前景。     

日本沼虾三群体线粒体16S rRNA基因片段序列的差异与系统进化

通过对日本沼虾(Macrobrachium nipponense)3个群体线粒体DNA 16S rRNA基因片段进行扩增和测定,得到长度为495bp的片段,其碱基A、T、G和C的平均含量分别为28.6%、36.1%、22.7%和12.5%,AT含量明显高于GC含量。通过对日本沼虾16SrRNA基因片段遗传特征的研究发现其种内变异很小,在3个群体中只有5个位点发生转换。另外,利用其454bp的同源序列,以中国明对虾(Fenneropenaeus chinensis)为外群探讨了沼虾属日本沼虾、罗氏沼虾(M.rosenbergii)等8种沼虾的系统进化关系。用MEGA3.1软件中的NJ法构建的分子进化树,日本沼虾3个群体先聚在一起后与海南沼虾聚在一起;另外,罗氏沼虾与马氏沼虾、短腕沼虾与贪食沼虾亲缘关系较近先聚在一起,然后再与大臂沼虾和等齿沼虾聚在一起,最后才与外群中国明对虾聚在一起。     

The sex ratio of wild Chinese alligators Alligator sinensis

The Chinese alligator Alligator sinensis is one of the most endangered crocodilian species, and typically exhibits temperature-dependent sex determination. It is extremely important to clarify the sex structure of Chinese alligators to implement recovery projects successfully. However, the sex ratio of wild Chinese alligators remains unknown. In this study, we collected 28 years of sex ratio data from Chinese alligators residing in the natural and artificial habitats of Changxing Nature Reserve, China, and examined the differences in the sex ratio dynamics between these two populations. We observed that the sex ratio of wild Chinese alligators is 1 male to 4.507 females, which was significantly lower compared to that of the captive population （1 to 2.040; P 〈 0.001）, and is significantly different to previously documented sex ratios for this species （all P 〈 0.01）. Furthermore, we documented an annually stable （P = 1.000） female-biased sex ratio for wild alligators at hatching [1 male to 4.747 females; 0.174 （0.167~）.182）], in contrast to a dramatically fluctuating sex ratio （P 〈 0.001） in captivity [1 male to 1.674 females; 0.374 （0.246-0.593）], showing a potential mechanism for adjusting the sex structure. Finally, we found that the hatchling sex ratios were similar to that of the population sex ratio （P = 0.748）, with little correlation to air temperature values in the 60-70 day incubation period during the breeding season （July and August; both P 〉 0.05）. Overall, this study indicates that the stabilized female-biased sex ratio of Changxing Chinese alligators might result from selection pressure caused by local mate competition and major inbreeding .     

482例产褥期阴道微生态调查

目的了解产褥期妇女阴道微生态状况,为临床保健提供循证依据。方法选择长兴县妇幼保健院产后门诊复查的产褥期妇女482例及来院体检的健康妇女86例,采集阴道分泌物作pH、病原体、清洁度和阴道微生态功能检查,进行阴道微生态学评价、比较。结果 482例产褥期阴道pH为(4.62±0.62);阴道清洁度Ⅲ~Ⅳ127例;微生态正常者75例,占15.56%;微生态失调者407例,占84.43%。86例健康妇女阴道pH(4.38±0.65);阴道清洁度Ⅲ~Ⅳ14例;微生态正常者34例,占39.53%;微生态失调者52例,占60.47%。差异有统计学意义。结论产褥期妇女阴道微生态发生改变,表现为pH升高,清洁度下降,微生态失调比例明显升高,应引起临床保健工作者关注。     

肿瘤抑制因子hTid1研究进展

hTid1(human tumorous imaginal disc 1)是果蝇肿瘤抑制因子Tid56的人类同源蛋白。hTid1属于DnaJ蛋白家族成员,主要定位于线粒体基质中,作为Hsp70蛋白的辅助分子伴侣发挥作用。然而,越来越多的文献报道,hTid1可以与线粒体外的许多蛋白相互作用,进而调控细胞内许多的信号通路。该文综述了近年来hTid1蛋白的最新研究进展,并主要从hTid1蛋白的结构和功能、与肿瘤的相关性、与神经系统的联系及在细胞信号通路中的作用等方面进行系统的阐述。     

长兴县合溪水库集雨区苗木地不同施肥管理模式对径流磷素流失的影响

为保护主要饮用水源合溪水库水质,长兴县将合溪水库集雨区农业种植结构向苗木种植方式调整,但缺乏优化施肥管理模式。针对该问题,选取集雨区典型苗木种植地进行径流小区定位试验,设置了撒施(对照)、地膜覆盖、秸秆覆盖、区外设置植草缓冲带、条施、穴施6个处理,研究不同施肥管理模式下苗木地径流磷素流失特征,筛选最优施肥管理模式,实现最大化的削减农田磷素流失。结果表明:不同施肥管理模式下,苗木地地表径流年总磷素径流流失通量大小顺序为:撒施(对照)地膜覆盖条施穴施区外植草缓冲带秸秆覆盖,撒施(对照)、地膜覆盖、条施、穴施、区外设置植草缓冲带和秸秆覆盖六种施肥管理模式下的年总磷素径流流失通量分别为:9.60、9.14、5.49、4.44、2.48、1.37 kg hm-2a-1,说明穴施、区外设置植草缓冲带和秸秆覆盖是较佳的施肥管理模式,若将其优化组合将显著消减合溪水库集雨区苗木地径流磷素流失及其对合溪水库水质的潜在影响。此外,不同施肥管理模式下,苗木地径流水样磷素流失的首要形态均为颗粒态磷,其次为溶解态磷,其中,撒施(对照)、地膜覆盖、秸秆覆盖、区外植草缓冲带、条施和穴施小区径流年均流失的颗粒态磷占流失总磷的比例分别是66.09%、70.69%、67.97%、71.63%、68.11%和67.87%。     

Genome analysis and signature discovery for diving and sensory properties of the endangered Chinese alligator

Crocodilians are diving reptiles that can hold their breath under water for long periods of time and are crepuscular animals with excellent sensory abilities. They comprise a sister lineage of birds and have no sex chromosome. Here we report the genome sequence of the endangered Chinese alligator (Alligator sinensis) and describe its unique features. The next-generation sequencing generated 314 Gb of raw sequence, yielding a genome size of 2.3 Gb. A total of 22 200 genes were predicted in Alligator sinensis using a de novo, homology- and RNA-based combined model. The genetic basis of long-diving behavior includes duplication of the bicarbonate-binding hemoglobin gene, co-functioning of routine phosphate-binding and special bicarbonate-binding oxygen transport, and positively selected energy metabolism, ammonium bicarbonate excretion and cardiac muscle contraction. Further, we elucidated the robust Alligator sinensis sensory system, including a significantly expanded olfactory receptor repertoire, rapidly evolving nerve-related cellular components and visual perception, and positive selection of the night vision-related opsin and sound detection-associated otopetrin. We also discovered a well-developed immune system with a considerable number of lineage-specific antigen-presentation genes for adaptive immunity as well as expansion of the tripartite motif-containing C-type lectin and butyrophilin genes for innate immunity and expression of antibacterial peptides. Multifluorescence in situ hybridization showed that alligator chromosome 3, which encodes DMRT1, exhibits significant synteny with chicken chromosome Z. Finally, population history analysis indicated population admixture 0.60-1.05 million years ago, when the Qinghai-Tibetan Plateau was uplifted.     

Interface-Induced Ag Monolayer Film for Surface-Enhanced Raman Scattering Detection of Water-Insoluble Enrofloxacin

Water-insoluble molecules usually show poor surface-enhanced Raman scattering (SERS) signals, because they are hardly adsorbed on the surface of most commonly used SERS substrates, such as aqueous Ag or Au colloids. In this work, a highly sensitive and reproducible Ag monolayer film (Ag MLF) SERS substrate prepared by self-assembly of Ag nanoparticles (Ag NPs) on water/oil interface can realize the trace SERS detection of water-insoluble enrofloxacin. The positively charged phase transfer catalyst can transfer the negatively charged Ag nanoparticles in aqueous solution to the water/oil interface. At the same time, the water-insoluble enrofloxacin can also be attracted to the interface because of its lipophilic group. The type/volume of the oil phase and phase transfer catalyst and the vortex mixing time were all optimized to maximize the SERS effect of Ag MLF. Results showed that trace water-insoluble enrofloxacin can be identified by Ag MLF and its detection sensitivity was significantly improved. The proposed novel Ag MLF can be further applied to detect other water-insoluble molecules in SERS.