四川大头茶净光合作用生理生态的初步研究

为了探讨重要生理生态因子在自然条件下对四川大头茶净光合作用(p_n)的影响,本文采用改良半叶法测定净光合作用的日变化和年变化。结果表明:春夏秋三季晴天p_n日变化呈双峰曲线型,即上,下午各出现一次P_n高峰,蜂谷则是P_n中午降低现象。冬季晴天P_n呈单峰曲线型。5月至11月P_n平均值为9.00mg·dm~(-2)·h~(-1)最高值23.44mg·dm~(-2)·h~(-1)出现在8月底。 一定程度上,P_n年变化曲线与温度年变化曲线相似,可以明显地区分为三部分。第一部分是从1月至5月,P_n随着温度的逐步上升而逐步升高。5月至10月为第二部分,此间P_n波动较大。10月至12月为第三部分,P_n随着逐步下降的温度而逐步降低。P_n季节变化显著,与植物生长、发育节奏相关。 温度和光照强度是影响P_n最重要的生态因子,相对湿度,叶绿素含量和SLW对P_n影响不大。温度与P_n呈一不对称钟形曲线,P_n的最低温度是4—5℃,最适温度是25—30℃,或稍高于30℃,最高温度则大于30℃。P_n的光饱和点在70,000lx左右,光补偿点约为3,000lx.在最适温度(25—30℃)范围内,P_n主要由光强所控制,并与光强呈正相关,一旦低于或超过最适温度,P_n则主要由温度所决定,与温度呈正相关。 遮阴实验表明,四川大头茶是具相当耐阴能力的阳性植物。     

PVP-coated graphene oxide for selective determination of ochratoxin A via quenching fluorescence of free aptamer

In this paper, we developed a simple method to detect fungi toxin (ochratoxin A) produced by Aspergillus Ochraceus and Penicillium verrucosumm, utilizing graphene oxide as quencher which can quench the fluorescence of FAM (carboxyfluorescein) attached to toxin-specific aptamer. By optimizing the experimental conditions, we obtained the detection limit of our sensing platform based on bare graphene oxide to be 1.9 μM with a linear detection range from 2 μM to 35 μM. Selectivity of this sensing platform has been carefully investigated; the results showed that this sensor specifically responded to ochratoxin A without interference from other structure analogues (N-acetyl-l-phenylalanine and warfarin) and with only limited interference from ochratoxin B. Experimental data showed that ochratoxin A as well as other structure analogues could adsorb onto the graphene oxide. As compared to the non-protected graphene oxide based biosensor, PVP-protected graphene oxide reveals much lower detection limit (21.8 nM) by two orders of magnitude under the optimized ratio of graphene oxide to PVP concentration. This sensor has also been challenged by testing 1% red wine containing buffer solution spiked with a series of concentration of ochratoxin A.     

The role of microvesicles and its active molecules in regulating cellular biology

Cell‐derived microvesicles are membrane vesicles produced by the outward budding of the plasma membrane and released by almost all types of cells. These have been considered as another mechanism of intercellular communication, because they carry active molecules, such as proteins, lipids and nucleic acids. Furthermore, these are present in circulating fluids, such as blood and urine, and are closely correlated to the progression of pathophysiological conditions in many diseases. Recent studies have revealed that microvesicles have a dual effect of damage and protection of receptor cells. However, the nature of the active molecules involved in this effect remains unclear. The present study mainly emphasized the mechanism of microvesicles and the active molecules mediating the different biological effects of receptor cells by affecting autophagy, apoptosis and inflammation pathways. The effective ways of blocking microvesicles and its active molecules in mediating cell damage when microvesicles exert harmful effects were also discussed.     

The Investigation of Protein Diffusion via H-Cell Microfluidics

In this study, we developed a microfluidics method, using a so-called H-cell microfluidics device, for the determination of protein diffusion coefficients at different concentrations, pHs, ionic strengths, and solvent viscosities. Protein transfer takes place in the H-cell channels between two laminarly flowing streams with each containing a different initial protein concentration. The protein diffusion coefficients are calculated based on the measured protein mass transfer, the channel dimensions, and the contact time between the two streams. The diffusion rates of lysozyme, cytochrome c, myoglobin, ovalbumin, bovine serum albumin, and etanercept were investigated. The accuracy of the presented methodology was demonstrated by comparing the measured diffusion coefficients with literature values measured under similar solvent conditions using other techniques. At low pH and ionic strength, the measured lysozyme diffusion coefficient increased with the protein concentration gradient, suggesting stronger and more frequent intermolecular interactions. At comparable concentration gradients, the measured lysozyme diffusion coefficient decreased drastically as a function of increasing ionic strength (from zero onwards) and increasing medium viscosity. Additionally, a particle tracing numerical simulation was performed to achieve a better understanding of the macromolecular displacement in the H-cell microchannels. It was found that particle transfer between the two channels tends to speed up at low ionic strength and high concentration gradient. This confirms the corresponding experimental observation of protein diffusion measured via the H-cell microfluidics.     

腰椎间盘突出症260例临床分析

目的:总结腰椎间盘突出症的临床特点及诊治要点。方法:回顾性分析260例腰椎间盘突出症手术患者的临床资料。结果:直腿抬高与影像学检查结果符合率为100%,治疗优良率达88.08%,有效率100%。结论:腰、下肢和臀部疼痛、下肢麻木、体位改变、运动障碍、感觉障碍、肌萎缩都是腰椎间盘突出症的主要临床表现;直腿抬高试验高试验可作为早期诊断的重要参考指标,要要根据惠者体征、病程等具体情况选择适合的最佳治法。     

Metabolic engineering of Escherichia coli for efficient osmotic stress-free production of compatible solute hydroxyectoine

Compatible solutes are key for the ability of halophilic bacteria to resist high osmotic stress. They have received wide attention from researchers for their excellent osmotic protection properties. Hydroxyectoine is a particularly important compatible solute, but its production by microbes faces several challenges, including low titer/yield, the presence of the byproduct ectoine, and the requirement of high salinity. Here, we aimed to metabolically engineer Escherichia coli to efficiently produce hydroxyectoine in the absence of osmotic stress without accumulating the byproduct ectoine. First, combinatorial optimization of the expression strength of key genes in the ectoine synthesis module and hydroxyectoine synthesis module was conducted. After optimization of the expression of these genes, 12.12 g/L hydroxyectoine and 0.24 g/L ectoine were obtained at 36 h in shake-flask fermentation with the addition of the co-substrate α-ketoglutarate. Further optimization of the addition of α-ketoglutarate achieved the sole production of hydroxyectoine (i.e., no ectoine accumulation), indicating that the supply of α-ketoglutarate is critically important for sole hydroxyectoine production. Finally, quorum sensing-based auto-regulation of intracellular α-ketoglutarate pool was implemented as an alternative to α-ketoglutarate addition by coupling the expression of sucA with the esaI/esaR circuit, which led to 14.93 g/L hydroxyectoine with a unit cell yield of 1.678 g/g and no ectoine accumulation in the absence of osmotic stress. This is the highest reported titer of sole hydroxyectoine production under salinity-free fermentation to date.     

454 pyrosequencing analysis on microbial diversity of an expanded granular sludge bed reactor treating high NaCl and nitrate concentration wastewater

The treatment of high-salinity, high-nitrate wastewater was investigated in a single expanded granular sludge bed reactor. Complete denitrification was achieved when nitrate concentration was as high as 6,000 mg N/L and the salinity of influent reached 11% NaCl at liquid up-flow velocity of 3.0 m/h, hydraulic retention time of, 24 h and the C/N molar ratio of 2.0. Furthermore, 454-pyrosequencing technology was used to analyze archaea bacterial diversity under high salinity and high nitrate conditions. Results showed that the total number of effective sequences was 5749 consisting of 5678 bacterial sequences and 71 archaea sequences after denoising and filtering out chimeras, which could be affiliated to 5 phylogenetic groups, including Proteobacteria, Firmicutes, Euryarchaeota, Crenarchaeota and unclassified phylum. Although Proteobacteria was the dominant microbial population, two archaea phylogenetic groups-Crenarchaeota and Euryarchaeota were observed in this study.     

Histone lysine methyltransferases and demethylases in Plasmodium falciparum

Dynamic histone lysine methylation, regulated by methyltransferases and demethylases, plays fundamental roles in chromatin structure and gene expression in a wide range of eukaryotic organisms. A large number of SET-domain-containing proteins make up the histone lysine methyltransferase (HKMT) family, which catalyses the methylation of different lysine residues with relatively high substrate specificities. Another large family of Jumonji C (JmjC)-domain-containing histone lysine demethylases (JHDMs) reverses histone lysine methylation with both lysine site and methyl-state specificities. Through bioinformatic analysis, at least nine SET-domain-containing genes were found in the malaria parasite Plasmodium falciparum and its sibling species. Phylogenetic analysis separated these putative HKMTs into five subfamilies with different putative substrate specificities. Consistent with the phylogenetic subdivision, methyl marks were found on K4, K9 and K36 of histone H3 and K20 of histone H4 by site-specific methyl-lysine antibodies. In addition, most SET-domain genes and histone methyl-lysine marks displayed dynamic changes during the parasite asexual erythrocytic cycle, suggesting that they constitute an important epigenetic mechanism of gene regulation in malaria parasites. Furthermore, the malaria parasite and other apicomplexan genomes also encode JmjC-domain-containing proteins that may serve as histone lysine demethylases. Whereas prokaryotic expression of putative active domains of four P. falciparum SET proteins did not yield detectable HKMT activity towards recombinant P. falciparum histones, two protein domains expressed in vitro in a eukaryotic system showed HKMT activities towards H3 and H4, respectively. With the discovery of these Plasmodium SET- and JmjC-domain genes in the malaria parasite genomes, future efforts will be directed towards elucidation of their substrate specificities and functions in various cellular processes of the parasites.