O139霍乱弧菌检测方法的研究─诊断血清的制备

本文以Ｏ１３９死菌免疫健康家兔,经吸收去除非特异性凝集素制成的特异性诊断血清,专供诊断霍乱弧菌Ｏ１３９之用。采用玻片凝集试验对５株Ｏ１３９菌株及１２６株肠道菌进行验证,在敏感性和特异性方面均获得满意的结果。     

Repellent,Insecticidal and Antimicrobial Activities of Leaf Essential Oils from Three Eucalyptus Species

Developing effective and eco‐friendly antimicrobials and pesticides has become a highly important issue. The repellent, insecticidal and antimicrobial activity of essential oils (EOs) isolated by hydrodistillation from dried leaves of the three Eucalyptus species (E. cloeziana, E. umbellata and E. benthamii) were investigated. During GC/MS analysis, α‐pinene (47.36 %), 1,8‐cineol (38.53 %) and α‐pinene (35.31 %) were identified as major components of E. cloeziana, E. umbellata and E. benthamii, respectively. The EOs from E. cloeziana exhibited the longest effective protection time (465 min, at 50.0 % w/w) for humans among the EOs studied. The effective protection time was 30 min and 300 min at concentrations of 12.5 % (w/w) and 25.0 % (w/w), respectively. Fumigating insecticidal activity of EOs from three Eucalyptus species was tested by airtight fumigation in conical flask, which indicated that essential oils had a highly and rapidly insecticidal activity on Culex pipiens quinquefasciatus. The antimicrobial activity of EOs was evaluated by using disc diffusion and agar dilution methods. There was no significant difference in the antibacterial activity of EOs from E. cloeziana and E. umbellate and they had the same MICs (20 mL/L) on Staphylococcus aureus, Salmonella typhi, Bacillus subtilis and Escherichia coli. E. benthamii had the worst microbial inhibitory effect among the three Eucalyptus essential oils and the MIC value for the test species is 40 mL/L except for Rhodotorula Harrison (10 mL/L).     

Streptococcus pneumoniae aminopeptidase N contributes to bacterial virulence and elicits a strong innate immune response through MAPK and PI3K/AKT signaling

Streptococcus pneumoniae is a Gram-positive pathogen with high morbidity and mortality globally but some of its pathogenesis remains unknown. Previous research has provided evidence that aminopeptidase N (PepN) is most likely a virulence factor of S. pneumoniae. However, its role in S. pneumoniae virulence and its interaction with the host remains to be confirmed. We generated a pepN gene deficient mutant strain and found that its virulence for mice was significantly attenuated as were in vitro adhesion and invasion of host cells. The PepN protein could induce a strong innate immune response in vivo and in vitro and induced secretion of IL-6 and TNF-α by primary peritoneal macrophages via the rapid phosphorylation of MAPK and PI3K/AKT signaling pathways and this was confirmed using specific pathway inhibitors. In conclusion, PepN is a novel virulence factor that is essential for the virulence of S. pneumoniae and induces host innate immunity via MAPK and PI3K/AKT signaling.

     

Knowledge Mapping of Opuntia Milpa Alta Since 1998: A Scientometric Analysis

Opuntia Milpa Alta is a cactus cultivated, domesticated, hybridized and selected from the plant Opuntia ficus-indica by Mexican agricultural experts, which can be used as fruit and vegetable. Opuntia Milpa Alta leaves and fruit are superior to wild varieties and suitable for storage and transportation. In 1998, Opuntia Milpa Alta was introduced to China from Mexico by the Quality Product Development Center of the Ministry of Agriculture of China. Up to now, the Opuntia Milpa Alta has been cultivated on a certain scale in China. This study aims to identify the research progress and development trends of Opuntia Milpa Alta in China. Papers published between 1998 to 2019 from two major Chinese academic databases (CNKI and Wangfang) with a topic search related to Opuntia Milpa Alta were collected. The research progress and development trends were analyzed based on CiteSpace software of text mining and visualization. The analysis found that Opuntia Milpa Alta has gone through three obvious research phases after being introduced to China. In the first phase, the researchers paid attention to its cultivation method. Subsequently, researchers began to use extraction methods to extract some of its components, such as polysaccharides and flavonoids. Finally, these extracted ingredients began to be used in some biomedical research.     

脆弱拟杆菌BF839治疗寻常型银屑病：一项单臂、开放初步临床试验

银屑病被认为是一种T细胞主导的炎症性疾病,其发病与肠道菌群失调密切相关。脆弱拟杆菌 (Bacteroides fragilis,BF) 可通过调节T细胞的细胞因子表达起抗炎作用。目前尚无脆弱拟杆菌用于治疗银屑病的相关报道,文中率先探究脆弱拟杆菌BF839对银屑病的治疗效果。选择2019年4月至2019年10月广州医科大学附属第二医院就诊的27例银屑病患者,维持原治疗不变,口服脆弱拟杆菌BF839 12周,对比治疗前后银屑病皮损面积与严重程度指数 (Psoriasis area and severity index,PASI) 评分,统计治疗12周后药物减停率。结果表明,12周试验完成率为96.3% (26/27),12周PASI30 (PASIN定义为治疗后PASI评分下降≥N%的患者比例) 为65.4%,PASI50为42.3%,PASI75为19.2%;治疗前PASI评分为9.1±5.9,治疗12周后PASI评分为5.8±4.9,具有显著统计学差异 (P<0.01);治疗12周后皮肤瘙痒程度用视觉模拟量表 (Visual analog scale,VAS) 评分有效率为42.3%,治疗前VAS评分为2.9±2.2,治疗12周后VAS评分为2.3±2.1,无显著统计学差异 (P>0.05)。患者治疗12周内不良反应率为3.8% (1/26),其中便秘1例,药物减停率为60.0%。以上结果提示脆弱拟杆菌BF839可能对银屑病治疗有一定疗效,可降低PASI评分及药物使用率,不良反应少,值得进一步研究。     

A FOXM1-Targeted Peptide Overcomes 5-Fluorouracil Resistance via Modulating ABC Transporters in Liver Cancer HepG2 Cells

Drug resistance largely limits the efficacy and efficiency of chemotherapeutics, which is a first-line treatment for liver cancer, consequently triggering a complete failure in clinical application. There are numerous attempts in exploring potential strategies for avoiding drug resistance, but none of them has effectively addressed this problem. Therefore, novel molecular targets and agents proposed for addressing drug resistance are needed. This study established 5-fluorouracil (5-Fu)-resistant HepG2 cells (HepG2/R) and showed that a FOXM1-targeted peptide, P201, reactivated 5-Fu to attenuate HepG2/R cell viability, proliferation, migration and promote apoptosis. Moreover, both pharmacological studies and RNA genomic sequencing results uncovered that combination of P201 and 5-Fu notably decreased expressions of FOXM1, MDR1 and ABCG2 compared to 5-Fu alone, indicating P201 overcame 5-Fu resistance mainly through inhibiting FOXM1 and ABC transporters. Therefore, P201 could inhibit ABC transporters by targeting FOXM1 in HepG-2/R cells, overcoming 5-Fu resistance and enhancing anti-cancer drug sensitivity. FOXM1 may be a new target for overcoming 5-Fu resistance in HepG2 cell while the combination treatment of P201 and 5-Fu may serve as a potential strategy for treating liver cancer.

     

WormFarm: a quantitative control and measurement device toward automated Caenorhabditis elegans aging analysis

Caenorhabditis elegans is a leading model organism for studying the basic mechanisms of aging. Progress has been limited, however, by the lack of an automated system for quantitative analysis of longevity and mean lifespan. To address this barrier, we developed ‘WormFarm’, an integrated microfluidic device for culturing nematodes. Cohorts of 30–50 animals are maintained throughout their lifespan in each of eight separate chambers on a single WormFarm polydimethylsiloxane chip. Design features allow for automated removal of progeny and efficient control of environmental conditions. In addition, we have developed computational algorithms for automated analysis of video footage to quantitate survival and other phenotypes, such as body size and motility. As proof‐of‐principle, we show here that WormFarm successfully recapitulates survival data obtained from a standard plate‐based assay for both RNAi‐mediated and dietary‐induced changes in lifespan. Further, using a fluorescent reporter in conjunction with WormFarm, we report an age‐associated decrease in fluorescent intensity of GFP in transgenic worms expressing GFP tagged with a mitochondrial import signal under the control of the myo‐3 promoter. This marker may therefore serve as a useful biomarker of biological age and aging rate.     

Identification and fine mapping of qRBSDV-6 MH , a major QTL for resistance to rice black-streaked dwarf virus disease

Rice black streaked-dwarf virus (RBSDV) disease is recently expanding in southern China and poses a serious threat to rice crops. Few studies related to the genetics and breeding of RBSDV resistance have been reported. We have previously mapped a number of quantitative trait loci (QTLs) for RBSDV resistance by using a recombinant inbred line population of ‘Zhenshan 97’ (ZS97, susceptible)/‘Minghui 63’ (MH63, resistant) with natural infection data in two locations. In the present study, we confirmed the presence of a number of resistant QTLs on chromosomes 6, 7, and 9 from MH63 by using the same population in four different locations. We then focused on a major QTL, qRBSDV-6 ^MH , on chromosome 6 and introduced it into a highly susceptible japonica rice variety, ‘Huaidao 5’, using MH63 as the donor via marker-assisted selection, to generate seven backcross inbred lines (BILs). Natural infection and artificial inoculation-based tests revealed that all of the BILs had a significantly higher resistance to RBSDV than the recurrent parent. These results demonstrate that qRBSDV-6 ^MH is a stable major resistance QTL of high breeding value. We also constructed a set of chromosome segment substitution lines (CSSLs) specific to the qRBSDV-6 ^MH region and these used as fine mapping population. Combining the genotypes of CSSLs with the phenotypes from natural infection data in a highly RBSDV epidemic area during two different sowing seasons, we were able to precisely map qRBSDV-6 ^MH to the markers S18 and S23 at a physical distance of 627.6 kb on the Nipponbare reference genome.     

Overexpression of an Acidic Endo-β-1,3-1,4-glucanase in Transgenic Maize Seed for Direct Utilization in Animal Feed

Background

Incorporation of exogenous glucanase into animal feed is common practice to remove glucan, one of the anti-nutritional factors, for efficient nutrition absorption. The acidic endo-β-1,3-1,4-glucanase (Bgl7A) from Bispora sp. MEY-1 has excellent properties and represents a potential enzyme supplement to animal feed.

Methodology/Principal Findings

Here we successfully developed a transgenic maize producing a high level of Bgl7AM (codon modified Bgl7A) by constructing a recombinant vector driven by the embryo-specific promoter ZM-leg1A. Southern and Western blot analysis indicated the stable integration and specific expression of the transgene in maize seeds over four generations. The β-glucanase activity of the transgenic maize seeds reached up to 779,800 U/kg, about 236-fold higher than that of non-transgenic maize. The β-glucanase derived from the transgenic maize seeds had an optimal pH of 4.0 and was stable at pH 1.0–8.0, which is in agreement with the normal environment of digestive tract.

Conclusion/Significance

Our study offers a transgenic maize line that could be directly used in animal feed without any glucanase production, purification and supplementation, consequently simplifying the feed enzyme processing procedure.     

Microdissection and Chromosome Painting of the Alien Chromosome in an Addition Line of Wheat - Thinopyrum intermedium

In this study, chromosome painting was developed and used to identify alien chromosomes in TAi-27, a wheat - Thinopyrum intermedium addition line, and the chromosomes of the three different genomes of Th. Intermedium. The smallest alien chromosome of TAi-27 was microdissected and its DNA amplified by DOP-PCR was used as a probe to hybridize with metaphase chromosomes of TAi-27 and Th . intermedium . Results showed that hybridization signals were observed in all regions of a pair of the smallest alien chromosomes and the pericentromeric area of another pair of alien chromosomes in TAi-27, indicating that the probe from microdissected chromosome is species specific. In Th . intermedium , 14 chromosomes had wide and strong hybridization signals distributed mainly on the pericentromere area and 9 chromosomes with narrow and weak signals on the pericentromere area. The remaining chromosomes displayed a very weak or no signal. Sequential FISH/GISH on Th . intermedium chromosomes using the DNAs of microdissected chromosome, Pseudoroegneria spicata (St genome) and pDbH12 (a J^s genome specific probe) as the probes indicated that the microdissected chromosome belonged to the St genome, three genomes (J^s, J and St) in Th . intermedium could be distinguished, in which there is no hybridization signal on J genome that is similar to the genome of Th . bessarabicum . Our results showed that the smallest alien chromosomes may represent a truncated chromosome and the repetitive sequence distribution might be similar in different chromosomes within the St genome. However, the repetitive sequence distributions are different within the J^s genome, within a single chromosome, and among different genomes in Th . intermedium . Our results suggested that chromosome painting could be feasible in some plants and useful in detecting chromosome variation and repetitive sequence distribution in different genomes of polyploidy plants, which is helpful for understanding the evolution of different genomes in polyploid plants.