Isolation of Leptospirillum ferriphilum by single-layered solid medium

According to physiological and biochemical characteristics of Leptospirillum ferriphilum, a strain of object bacteria was isolated successfully. Bacteria were enriched by selective liquid medium and plated on designed single-layered agar solid medium. Colony was cultured and bacteria were collected. The morphologies of the object bacteria were observed using crystal violet staining, scanning electron microscope（SEM） and transmission electron microscope （TEM）. The result of 16S rDNA identification shows that this bacterium belongs to Leptospirillum ferriphilum and it is named as Leptospirillum ferriphilum strain D1. These results indicate that this new single-layered agar solid medium is efficient and physiological-biochemical characteristics show that the optimum simple for isolation of Leptospirillum ferriphilum. Additionally, initial pH value and its growth temperature are 1.68 and 40℃.

Dental stem cell-conditioned medium for tissue regeneration: Optimization of production and storage

BACKGROUND Mesenchymal stem cells(MSC)effects on tissue regeneration are mainly mediated by their secreted substances(secretome),inducing their paracrine activity.This Conditioned medium(CM),including soluble factors(proteins,nucleic acids,lipids)and extracellular vesicles is emerging as a potential alternative to cell therapy.However,the manufacturing of CM suffers from variable procedures and protocols leading to varying results between studies.Besides,there is no welldefined optimized procedure targeting specific applications in regenerative medicine.AIM To focus on conditioned medium produced from dental MSC(DMSC-CM),we reviewed the current parameters and manufacturing protocols,in order to propose a standardization and optimization of these manufacturing procedures.METHODS We have selected all publications investigating the effects of dental MSC secretome in in vitro and in vivo models of tissue regeneration,in accordance with the PRISMA guidelines.RESULTS A total of 351 results were identified.And based on the inclusion criteria described above,118 unique articles were included in the systematic review.DMSC-CM production was considered at three stages:before CM recovery(cell sources for CM),during CM production(culture conditions)and after production(CM treatment).CONCLUSION No clear consensus could be recovered as evidence-based methods,but we were able to describe the most commonly used protocols:donors under 30 years of age,dental pulp stem cells and exfoliated deciduous tooth stem cells with cell passage between 1 and 5,at a confluence of 70%to 80%.CM were often collected during 48 h,and stored at-80°C.It is important to point out that the preconditioning environment had a significant impact on DMSCCM content and efficiency.

Isolation and Screening of Oenococcus oeni and Its Medium Optimization

[Objectives] This study was conducted to further improve the application of Oenococcus oeni in the wine brewing industry. [Methods] O. oeni was isolated, screened, and identified, and medium optimization was performed to determine the best medium, temperature and oxygen conditions. [Results] O. oeni grew better in mFT80 medium, and the culture conditions were 25 ℃ and an anaerobic condition. [Conclusions] This study provides a reference for the development of China’s wine industry.

Response of Two Sweet Potato （Ipomoea batatas L. Lam） Varieties Regenerated on Low Cost Tissue Culture Medium

Plant tissue culture continues to be of great interest within the realms of molecular biology, plant breeding and plant health However, different plant cultivars have different culture efficiencies to tissue culture. In this research, the response of two Kenyan sweet potato varieties, KEMB 36 and Tainurey, cultured on a low cost tissue culture medium was evaluated. The low cost medium contained plant nutrients that were obtained from locally available fertilizers. Each conventional Murashige and Skoog （MS） macronutrient was individually substituted with a locally available fertilizer. The conventional source of micronutrients was substituted with Stanes~ Iodized Microfood while sucrose was obtained from table sugar. Performance of the two cultivars was monitored over a period of six weeks. KEMB 36 had a better performance than Tainurey with an average of eight nodes, seven leaves, three roots and height of four centimeters per plantlet indicating genotype-dependent response.

Modification to a Semi-selective Medium for the Isolation and Quantification of the Nematophagous Fungus of Pochonia chlamydosporia

Semi-selective media are valuable tools for detection and isolation of viable fungal propagules. After testing the in vitro sensitivity of Pochonia chlamydosporia to 9 different fungicidal compounds, boscalid and thiophanate-methyl were chosen for a modification of Kerry’s semi-selective agar. Rifampicin replaced tetracyclin to provide superior suppression of bacterial growth. The Pc medium contained 17 g/L corn meal agar, 17.5 g/L (NaCl), 75 mg/L Rose Bengal, 50 mg/L boscalid, 30 mg/L thiophanate-methyl, 50 mg/L rifampicin, 50 mg/L streptomycin sulphate, 50 mg/L chloramphenicol, 3 mL/L Triton X-100. Dilution plating of soil extracts from various California field soils amended with chlamydospores of P. chlamydosporia showed the modified medium was equivalent to Kerry’s agar in enumerating viable propagules of the fungus. While P. chlamydosporia colonies grew slower on the Pc medium, the fungal colonies appeared more uniform than on the original semi-selective agar.

Isolation, Purification, and Identification of a Novel HX-Producing Strain and Optimization of Its Fermentation Medium

Hexanoic acid (HX) is a crucial flavor compound and precursor of ethyl caproate (EA), which determines the quality of Chinese Luzhou-flavor liquor (CLFL). The isolation, purification, identification, and optimization of fermentation conditions of HX-producing bacteria are essential for industrial CLFL production. In this study, one strain of HX-producing bacterium was isolated from six candidate bacterial strains and identified as Clostridium sartagoneforme. Then, the growth characteristics and HX production of C. sartagoneforme were investigated. Sodium acetate medium was identified as the optimal fermentation medium from four candidate media. C. sartagoneforme yielded 800.85 ± 12.87 mg/100mL HX in sodium acetate medium. Then, to further optimize the formula of the fermentation medium, the carbon and nitrogen sources and inorganic salt component of the fermentation medium were investigated using HX yields as an optimization index. Optimization was performed with a single-factor experiment and the Taguchi design method. The single-factor experiment showed that the highest HX outputs were obtained when the sodium acetate medium contained 2.5 g/L yeast extract, 1.8 g/L KCl, 20 g/L sodium acetate, 15 mL/L ethanol, and 1.5 g/L glucose. In the orthogonal experiment designed using the Taguchi design method, HX yields reached 2018.29 ± 46.37 mg/100mL in sodium acetate medium that contained 3.5 g/L yeast extract, 1.8 g/L KCl, 25 g/L sodium acetate, and 15 mL/L ethanol.

Medium-Chain Triglyceride Activated Brown Adipose Tissue and Induced Reduction of Fat Mass in C57BL/6J Mice Fed High-fat Diet

Objective To investigate activation of brown adipose tissue （BAT） stimulated by medium-chain triglyceride （MCT）. Methods 30 Male C57BL/6J obese mice induced by fed high fat diet （HFD） were divided into 2 groups, and fed another HFD with 2% MCT or long-chain triglyceride （LCT） respectively for 12 weeks. Body weight, blood biochemical variables, interscapular brown fat tissue （IBAT） mass, expressions of mRNA and protein of beta 3-adrenergic receptors （β3-AR）, uncoupling protein-1 （UCP1）, hormone sensitive lipase （HSL）, protein kinase A （PKA）, and adipose triglyceride lipase （ATGL） in IBAT were measured. Results Significant decrease in body weight and body fat mass was observed in MCT group as compared with LCT group （P〈O.05） after 12 weeks. Greater increases in IBAT mass was observed in MCT group than in LCT group （P〈O.05）. Blood TG, TC, LDL-C in MCT group were decreased significantly, meanwhile blood HDL-C, ratio of HDL-C/LDL-C and norepinephrine were increased markedly. Expressions of mRNA and protein of β3-AR, UCP1, PKA, HSL, ATGL in BAT were greater in MCT group than in LCT group （P〈O.05）. Conclusion Our results suggest that MCT stimulated the activation of BAT, possible via norepinephrine pathway, which might partially contribute to reduction of the body fat mass in obese mice fed high fat diet.

Shoot tissue culture of Robinia pseudoacacia f. decaisneana

Robinia pseudoacacia f. decaisneana is a transfiguration of Robinia pseudoacacia. For enhancing propagation coefficient of the species, the experiment of shoot tissue culture of Robinia pseudoacacia f. decaisneana was conducted in Forestry College of Shenyang Agricultural University from July 1999 to July 2001. The experiment included medium selection of explant induction survival, initial culture, subculture as well as rooting culture, and forming seedling with callus. The results showed that shoot segment in vitro survive rate is larger in spring than in autumn, and green dense callus could form plantlet. The best medium for initial culture was SH+0.5mg/L BA+0.05 mg/L NAA, with a propagation coefficient of 4.1 (per micro-cutting in a month), and for subculture it was B5+0.5 mg/L BA+0.05 mg/L NAA+ 10 mg/L Glu., with a propagation coefficient of 4.7. The best rooting medium was 1/2MS+0.5 mg/L NAA+10 mg/L Glu., with a rooting rate of 84.4%. These results provide reference data for reproduction of superior individuals of Robinia pseudoacacia f. decaisneana.

Tissue Culture of Toona ciliata

In this study, a screening experiment was carried out among MS, WPM, DCR, MT, B5 and N6. The results showed that for proliferation culture of Toona ciliate, the optimum basic medium was WPM, and the optimum medium was WPM ＋ 6-BA 0.5 mg/L ＋ IBA 0.05 mg/L. The proliferation coefficient reached 4.29. The optimum rooting culture medium was 1/2 MS ＋ IBA 0.5 mg/L ＋ NAA 0.5 mg/L with rooting rate of 100%. In the optimum rooting ＇culture medium, the roots of T. ciliate developed neatly, and the seedlings grew well.

富血小板血浆来源的血小板细胞外囊泡分离技术与应用

背景:血小板细胞外囊泡是循环中最丰富的囊泡,富含丰富的生物活性分子、遗传物质及蛋白质等信息分子,参与细胞通讯和物质交换,不仅具有良好的促凝活性,还具有促进组织修复和再生的巨大潜力,在再生医学具有广泛的应用前景。目的:从血小板细胞外囊泡的分泌机制、在再生医学领域的应用及临床转化的限制因素进行阐述,为血小板细胞外囊泡的临床转化提供一些理论支撑。方法:应用计算机检索2005年1月至2023年8月PubMed数据库与血小板细胞外囊泡有关的文章,英文检索词为“platelet-derived,plateletrich plasma,extracellular vesicles,isolated,microvesicles exosomes,applications”,最后纳入符合主题标准的文献62篇进行综述分析。结果与结论:(1)活化的血小板细胞外囊泡可产生血小板微囊泡和血小板外泌体两种类型的囊泡,前者的分泌可能与肌动蛋白细胞骨架的不对称性相关,后者的分泌可能与H^(+)-ATP酶的调节相关。(2)血小板细胞外囊泡是血小板浓缩物和血小板本身的潜在效应物,可能通过促进血管生成、影响细胞行为、促凝与止血以及发挥炎症作用等几方面影响组织再生。(3)血小板细胞外囊泡在组织损伤、肌肉再生、软骨再生、骨关节炎等再生医学领域已有临床前报道,作为伤口愈合的潜在治疗方法已有临床试验数据,但血小板细胞外囊泡的分离方法、样本来源以及激活剂的种类等因素限制了血小板细胞外囊泡在再生医学领域的临床转化。(4)未来血小板细胞外囊泡可能成为再生医学中替代富血小板血浆的无细胞疗法,其临床转化还需要积极寻找鉴别血小板细胞外囊泡和血小板外泌体的特异性标志物,在激活剂刺激血小板细胞外囊泡产生的机制以及血小板细胞外囊泡的最佳收集方式、最佳储存方法、保质期限、临床应用的推荐剂量和最佳临床适应证还需要继续深入研究。

Study on Tissue Culture and Adventitious Bud Induction of Eucheuma striatum

This study aimed to investigate the aseptic treatment and disinfection of E. striatum explants and the effects of three liquid culture medium （ PES, PESI, f/2） and plant hormones on adventitious bud induction from explants. The results showed that the vast majority of explants disinfected by combined treatment exhib- ited good growth and normal color with an average mortality rate of only 9%. E. striatum explants exhibited the best growth in PES liquid medium and the survival rate reached 66.67% at 40 d after culture. Under different concentrations （0.5, 2, 5 mg/L） of 6-BA, the induction rate ofE. striatum explants was significantly higher than that in control group, but no remarkable differences were found in the induction of adventitious buds from E. striatum explants. In PES liquid medium containing 6-BA and 2,4-D, the adventitious bud induction rate was 90% and the average number of induced adventitious buds was 6.63.

Posterior quadrantic disconnection maintains the activity of isolated temporal-parietal-occipital nerve tissue: neuroprotective measures in the surgical treatment of epilepsy

Extensive lesions involving the posterior quadrant of the cerebral hemisphere （temporal, parietal, and occipital lobes） induce intractable epilepsy. These patients are potential candidates for surgical treatmenttu. Maintenance of isolated nerve tissue activity after surgery plays a crucial role in the neuroprotective effects of neurosurgery treatment. Disconnection surgery of the posterior quadrant is used to completely isolate nerve fibers, while blood supply at the isolated lobes is maintained. Subsequently, cavities caused by cystic or necrotic nerve tissues should be reduced as much as possible,

Study on Tissue Culture and Rapid Propagation of Tilia amurensis

To explore the establishment of a tissue culture and rapid propagation system of Tilia amurensis,the effects of basic medium and concentrations and ratios of plant growth regulators on tissue culture and rapid propagation of T.amurensis were studied.The results showed that 1/2 MS medium was the most suitable proliferation medium,and the proliferation coefficient could reach 13.5 after adding 0.05 mg/L 6-BA and 0.03 mg/L IBA;MS medium was the most suitable medium for strong plantlets and rooting,and the best medium for strong plantlets was MS+0.1 mg/L 6-BA+0.1 mg/L IBA+0.03 mg/L GA_(3),with which the average plantlet height reached 5.15 cm;and the best rooting medium was MS+1.0 mg/L6-BA+0.05 mg/L NAA,with which the rooting rate was 93.3%and the number of roots was 5.7 roots.

A NEW METHOD OF ISOLATING KUPFFER CELLS FROM BIOPSY TISSUE OF RAT LIVER

The isolation of a high yield and purity of Kupffer cells has been reported in detail.1 This paper reports into the research about isolation Kupffer cells from biopsy tissue of liver. This method includes 5 important steps: (1) take fresh liver tissue, and mince with scissors. (2) spin at low speed to wash off red blood cells. (3) digest in collagenase for suitable time. (4) isolate Kupffer cells on a percoll density gradient. (5) cell charaterization was observed by N.S.E stain and peroxidatic activity with lumino-meter measurement and phagocytosis with latex beads.2.3

The Tissue Reactions and Changes of a Surface of Various Metal Implants after Their Introduction in a Bone Tissue in Experiment

Screw metal implants (3S, Israel) with rough or smooth polished surface were introduced in a tibial proximal condyle of not purebred rabbits. The condition of surrounding tissues in 2 and 6 months after implantation was compared by light microscopy and X-ray methods. Within 6 months after operation the considerable distinctions of radiological and morphological data were revealed not. 2 months later after introduction of implants with a rough surface the effort enclosed for its twisting is, much more, than for removal of the polished product. However, stability of fixing of implants was practically made even at 6 months. On remote rough implants there is a set of tissue scraps whereas on products with a smooth surface the tissue remains were much less. Surrounding tissues strongly join a rough surface, grow into cavities, and during removal of such products there is a considerable trauma of tissues round an implantation place. Smooth implants have the smaller area of contact with organism tissues, they are fixed due to bicortical implantation, during removal easily get out and don’t break off surrounding tissues. The signs of inflammation and formation of merged multinuclear macrophages were not found at all cases, which give evidence to the inertness of material of the mentioned articles for living organism. In some observations however and by implantation of the rough article and by introduction of polished implants, metal particles were found, but after use of the foreign body with grit-blasted treatment of surface metal was found more frequently, and its fragments had larger volume.

Simplified isolation and spheroidal aggregate culture of rat hepatocytes

IM To explore a simplified method for isolation of hepatocytes and establish a method of primary hepatocyte culture with more aggregates and longer persisence.METHODS Wistar rat hepatocytes were isolated by a single extracorporeal twostep perfusion method, and the cells were seeded on polyHEMA coated flasks and cultured with hormonally defined medium and gentle shaking at regular intervals.RESULTS The total yield of isolating hepatocytes amounted to 108 cells for each rat liver with the viability of more than 90% in all isolations. Under the nonadherent environments, the cells were found to attach to each other and form multicellular aggregates rapidly, and the aggregates became spheroidal shape after two days in culture. The morphologic characteristics and albumin synthetic function of the multicellular spheroidal aggregates can be maintained for one month.CONCLUSION The simple and reliable isolation as well as large scale and longer time culture of hepatocytes can be used for experiments in liver cell transplantation and bioartificial liver support system.

粤东上升流区不同水层可培养细菌多样性

上升流通过增强扩散和独特的环境驱动着近海生态系统中的物质循环和能量流动,直接影响环境中生物的生长、繁殖和分布,蕴藏了大量尚未培养和难培养的微生物和新基因资源。本研究从粤东上升流区表层、中层和底层共采集27份海水样品,探究细菌多样性和分布模式及其与深度的相关性。采用纯培养方法共分离获得1928株细菌,通过16S rRNA基因序列分析,结果显示这些细菌分布在5门、9纲、32目、54科、121属中的262种。其中,变形菌门(Proteobacteria)为优势门,占46.9%;放线菌纲(Actinomycetia)为优势纲,占44.8%;Microbacteriales为优势目级类群,占21.2%。总体来说,中层海水分离得到的菌株多样性明显低于表层和底层海水环境,但三个层级环境分离获得的优势种群基本相同,最优类群均为Microbacteriales目。从不同水层海水中分离得到的潜在新菌数排序为:底层>表层>中层。从不同培养基分离得到的潜在新菌率排序为2216E培养基>R2A培养基>ISP2培养基>MSM培养基>IMK培养基。通过细菌多样性与环境因子的典范对应分析(canonical correspondence analysis,CCA)表明,能显著影响细菌多样性变化的环境因子是叶绿素含量(chlorophyll content,Chl)、活性硅酸盐(active silicate salts,ASS)、活性磷酸盐(active phosphate,AP)、氨及部分氨基酸(ammonia,Amm)和生化需氧量(biochemical oxygen demand,BOD)。本研究系统调查了粤东上升流区可培养海洋细菌多样性及其分布情况,其结果为近海上升流区不同水层可培养细菌多样性研究积累了关键的资料和数据,也为近海典型上升流环境生物物种和基因资源的发掘和利用提供了重要参考。

太阳抵近探测计划——观测位置的新突破

太阳抵近探测计划(SCOPE)旨在突破高温和强辐射防护、长距离运载、远距离轨道控制和先进载荷等一系列极限技术,将探测器投送至距太阳中心仅有5个太阳半径的位置上,深入太阳大气层进行探测,研究太阳附近极亮、极热的深空探测“无人区”,在全新的位置上对太阳进行超近距离原位探测和遥感观测,针对磁场和等离子体结构实现空间分辨率优于0.1"的观测。探测器将穿越太阳爆发磁重联电流片,显微观测太阳爆发磁结构,原位探测磁重联电流片、日冕物质抛射与激波以及各类带电粒子,以推动揭示太阳爆发理论机理。探测器将在最靠近太阳的区域显微监测和探测日冕中尺度小于0.1"的纳耀斑、光球中尺度小于0.06"的磁场和速度场结构及其演化特征,确认日冕加热机制;探测太阳风重离子电荷态,确认太阳起源及加速机制。探测器还将在最接近太阳的区域内与原位探测日冕磁场,显微观测太阳极区磁场和等离子体结构和演化特征;揭示太阳附近尘埃分布特征,进入无尘埃区,确定太阳系尘埃盘内边界。困扰太阳物理界近百年的两个科学难题,即太阳爆发机理难题和日冕加热与太阳风加速机理难题,随着该计划的顺利实施将被破解;在最接近日心的位置处实现日冕磁场,包括太阳极区磁场原位探测“0到1”的突破也将实现。本文将介绍SCOPE的背景、科学目标和预期产出、有效载荷以及卫星平台关键技术。

中深部孤岛工作面窄煤柱沿空掘巷支护技术研究

孤岛工作面沿空掘巷存在围岩变形严重、控制难度大、成本高的问题。以某矿5103回风顺槽为研究对象,运用多种设计方法,综合确定护巷煤柱最佳宽度为6 m;提出顶板围岩浅部基础层、中部承载层和深部强化层多圈层高效支护技术,以长锚杆+柔性锚杆+长锚索为主形成多圈层承载结构。通过在5103回风顺槽进行工业试验表明,巷道围岩变形得到有效控制,取得良好的安全、经济效益,可为相似条件下孤岛工作面的开采提供借鉴。

黄唇鱼迟缓爱德华氏菌分离鉴定及药敏分析

2023年入夏以来,东莞市黄唇鱼(Bahaba taipingensis)自然保护区救护基地出现黄唇鱼发病和死亡病例,症状主要为食欲下降,体表发红、充血,眼球突出,角膜白浊溃烂,吻部溃烂;解剖后发现腹腔有大量腹水,肝肿大呈褐色。为确定其病因,针对性制定防治措施,采集死亡病鱼的心脏、肝脏、脾脏、肾脏、肠、鳃丝等组织样品。从病死鱼体内分离到一株优势菌株,命名为DG230920,通过对菌株的形态特征、生理生化特性和16S rRNA基因序列综合分析,鉴定该菌为迟缓爱德华氏菌(Edwardsiella tarda)。病理组织学检测结果显示,心脏、肝脏、脾脏、肾脏和鳃均有明显的炎症反应,肠出血、黏膜上皮细胞脱落,脾淋巴细胞稀疏、内皮细胞变性坏死,肾小管变性坏死,鳃见鳃丝脱落、出血。对分离菌株进行药敏分析,结果为对环丙沙星、左氧氟沙星高度敏感,最低抑菌质量浓度仅≤0.25 mg·mL^(-1);其次是厄他培南,最低抑菌质量浓度≤0.5 mg·mL^(-1)。结果表明,黄唇鱼发病原因可能是迟缓爱德华氏菌感染,分离菌株遗传序列稳定,对临床使用的抗菌药物均为敏感。研究提示,应加强黄唇鱼迟缓爱德华氏菌感染的防治,选用敏感药物科学治疗。