Prognositic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma

Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor prognosis.Anchorage-dependent cell death(anoikis)has been proven to be indispensable in tumor metastasis,regulating the migration and adhesion of tumor cells at the primary site.However,as a type of programmed cell death,anoikis is rarely studied in osteosarcoma,especially in the tumor immune microenvironment.This study aims to clarify prognostic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma.Methods:Anoikis-related genes(ANRGs)were obtained from GeneCards.Clinical information and ANRGs expression profiles of osteosarcoma patients were sourced from the therapeutically applicable research to generate effective therapies and Gene Expression Omnibus(GEO)databases.ANRGs highly associated with tumor immune microenvironment were identified by the estimate package and the weighted gene coexpression network analysis(WGCNA)algorithm.Machine learning algorithms were performed to construct long-term survival predictive strategy,each sample was divided into high-risk and low-risk subgroups,which was further verified in the GEO cohort.Finally,based on single-cell RNA-seq from the GEO database,analysis was done on the function of signature genes in the osteosarcoma tumor microenvironment.Results:A total of 51 hub ANRGs closely associated with the tumor microenvironment were identified,from which 3 genes(MERTK,BNIP3,S100A8)were selected to construct the prognostic model.Significant differences in immune cell activation and immune-related signaling pathways were observed between the high-risk and low-risk groups based on tumor microenvironment analysis(all P<0.05).Additionally,characteristic genes within the osteosarcoma microenvironment were identified in regulation of intercellular crosstalk through the GAS6-MERTK signaling pathway.Conclusion:The prognostic model based on ANRGs and tumor microenvironment demonstrate good predictive power and provide more personalized treatment options for patients with osteosarcoma.

Variations of phyllosphere microorganisms in asymptomatic and tobacco brown spot leaves before and after spraying 12% difenoconazole + fluxapyroxad SC

12%difenoconazole+fluxapyroxad SC(commercial name:Jiangong)was first released by BASF in China in 2016.It has been registered to control many diseases,including pear scab,apple Alternaria leaf spot,tomato early blight,cucumber powdery mildew,etc.This study evaluated the bioactivity of Jiangong against Alternaria alternata and explored variations of phyllosphere microorganisms in both asymptomatic and tobacco brown spot leaves at different persistence periods(0,5,10,and 15 days post-fungicide application)using high-throughput sequencing technology.The results indicated that Jiangong effectively inhibited mycelial growth(average EC_(50) value of 0.51μg/mL),conidia germination(average EC_(50) value of 3.47μg/mL),and the carbon metabolism of A.alternata.Both asymptomatic and symptomatic leaves presented complex microbial communities.Higher fungal diversity was noted in asymptomatic leaves,while higher bacterial diversity was found in symptomatic leaves.After application,the diversity and abundance of microbial community structures in both types of leaves changed over time.Fungal microbiome communities showed greater sensitivity than bacterial groups,with the microbiome communities of asymptomatic leaves being more affected than those of symptomatic leaves.Fungal community diversity decreased for both symptomatic and asymptomatic leaves after 5 days of application,while the diversity of fungal community in symptomatic leaves showed an upward trend after 10 days of application.Meanwhile,bacterial community diversity increased in both symptomatic and asymptomatic leaves after 5 days of application but then declined in asymptomatic leaves after 15 days.The abundance of the dominant function group of phyllosphere bacteria(metabolism,genetic information processing,environmental information processing)was not affected by the application of Jiangong.However,the abundance of the dominant function group of phyllosphere fungi(animal pathogen-endophyte-wood saprotroph,endophyte-plant pathogen,plant pathogen-undefined saprotroph)was significantly affected by the application of Jiangong,and high variation was found in symptomatic leaves than that of asymptomatic leaves.The application of Jiangong-induced alterations in the community structure of the tobacco phyllosphere microbiome provides a basis for future tobacco brown spot control strategies based on phyllospheric microecology.

高、低核酸含量原核生物应对资源可利用性降低和原生动物选择性摄食的适应性策略

原核生物在全球海洋生物地球化学循环中发挥重要作用,然而对它们不同生态亚群(即:高核酸(HNA)和低核酸(LNA)含量亚群)的丰度和代谢活性,以及它们的调控因素如何随着海洋环境条件的变化而变化,仍知之甚少.我们在117,000升的大体积培养体系中进行了73天的培养实验,深入研究HNA和LNA亚群对资源可利用性降低和原生动物选择性摄食随时间变化的动态响应.研究结果显示,当培养环境资源充足时, HNA亚群的代谢活性高于LNA亚群,但随着资源日益稀缺, HNA亚群代谢下降速度比LNA亚群快,从而使LNA亚群对原核生物活性的贡献稳步增加.与此同时,随着培养体系资源可利用性逐步降低,原生动物的摄食偏好从HNA亚群转向LNA亚群,使得LNA亚群对培养体系的碳流贡献增加.这些结果突显了LNA亚群在资源有限环境中的韧性,阐明了原生动物选择性摄食在资源条件变化时平衡不同亚群中的关键作用,并展示了在不同环境背景下原生动物和原核生物之间复杂的相互适应性.

富营养化对海洋生态系统的影响及其围隔实验研究

陆源的营养盐输入引起海洋中的富营养化，提高了水体中营养盐含量，改变了水体中营养盐比例，引起浮游植物水华甚至赤潮。水华发生时由于沿传统食物链的能流在浮游植物环节阻塞，能量过多地分配到微生物环，细菌大量繁殖分解有机物，导致水体缺氧，造成鱼虾贝类窒息死亡。围隔系统是物质相对守恒的系统，围隔与自然海水没有水交换，利用现场围隔实验可以定量地、系统地研究海洋生态系统对富营养化的响应。围隔实验现已成为全球海洋生态系统动力学研究的重要手段。

注水井腐蚀原因分析及防护对策

胜利油田注水井腐蚀严重,结合注水水质,分析了腐蚀严重原因:注水井的腐蚀主要是回注污水中Cl-含量高、细菌滋生严重、含较高的HCO3-及硫化物造成,且随着注水井温度的升高,腐蚀因素在不断变化。主要分析了东辛采油厂污水中高矿化度、高游离CO2对注水井造成的腐蚀;在现河采油厂注水井油套环空腐蚀主要是由于细菌滋生、繁殖造成的腐蚀。结合注水井实际现状,实施相对应的防护措施:在注水井管柱上应用渗氮油管,延长了管线使用寿命;在回注污水过程中有选择性地投加缓蚀剂、杀菌剂;在油套环空中实施环空保护技术,实施后取得较好的效果,对延长注水井有效寿命具有重要意义。

The Super-population Bridges the Habitat Diversity and Biodiversity

There is another way for new species to invade former settlers＇ habitats and successfully settle there; it is demonstrated that the su- per-population, which plays a key role in changing the world-wide environment, can create many new microhabitats in the old habitat, it is these new microhabitats that make new species easily come into the old habitat and settle there without fierce competition with former settlers. In other words, the super-population lead to the differentiation of the worldwide environment, lead to the environmental diversity in the biosphere. Through the differentiation of the environment the super-population produces many new microhabitats for new species, the appearance of many new microhabitats make it possible for new species to coexist with former settlers in the biosphere. The cqexistence of new species with former settlers or with new species＇ ＂ancestors＂ results in the increaseing in the biodiversity of the biosphere. The super-population is the founder of many new environments on the earth, it bridges the habitat diversity and the biodiversity. Now It is easy to explain the phenomenon that new species sometimes coexist with former settlers and even depend on former settlers in the biosphere rather than fierce competition between them.

高寒地区湖泊浮游病毒时空分布特征

湖泊中浮游病毒的丰度在10~5~10~8mL^(-1),是浮游微生物中丰度较高的组分,在湖泊水生态系统中扮演着重要角色。病毒对宿主的侵染和裂解可以调节微生物群落的丰度、生产力、组成和种群多样性,同时还可以将宿主细胞的大量物质释放到湖泊中,改变碳循环和营养物质流通,进而影响生物地球化学循环。高寒地区湖泊水生态系统结构简单,主要以微生物为主,浮游病毒对微生物具有重要的调节作用,但目前研究较少。综述了近几年国内外对高寒地区湖泊病毒生态学的相关研究进展,主要包括病毒的时空分布以及与生物和非生物因子的关系、病毒与宿主的相互作用关系以及病毒对湖泊碳循环和微生物食物环的作用,发现高寒湖泊浮游病毒分布较广、多样性较高以及溶原性是病毒重要的生活策略,同时也得出由病毒裂解作用释放的有机碳对微生物活性和微生物食物环具有重要的作用。在此基础上,对今后我国青藏高原湖泊浮游病毒的研究提出了展望。

Soil Microbial Characteristics Under Long-Term Heavy Metal Stress:A Case Study in Zhangshi Wastewater Irrigation Area,Shenyang

Soil samples were collected from Zhangshi Wastewater Irrigation Area in the suburb of Shenyang City,China,an area with a 30-year irrigation history with heavy metal-containing wastewater.The chemical properties and microbial characteristics of the soils were examined to evaluate the present situation of heavy metal pollution and to assess the soil microbial characteristics under long-term heavy metal stress.In light of the National Environmental Quality Standards of China,the soil in the test area was heavily polluted by Cd and to a lesser degree by Zn and Cu,even though wastewater irrigation ceased in 1993.Soil metabolic quotient (qCO_2) had a significant positive correlation,while soil microbial quotient (qM) had a negative correlation with content of soil heavy metals.Soil microbial biomass carbon (MBC) had significantly negative correlation with Cd,but soil substrate-induced respiration (SIR),dehydrogenase activity (DHA),cellulase activity, and culturable microbial populations had no persistent correlations with soil heavy metal content.Soil nutrients,except for phosphorous,showed positive effects on soil microbial characteristics,which to a certain degree obscured the adverse effects of soil heavy metals.Soil Cd contributed more to the soil microbial characteristics,but qM and qCO_2 were more sensitive and showed persistent responses to heavy metals stress.It could be concluded that qM and qCO_2 can be used as bioindicators of heavy metal pollution in soils.

Mutagenesis and selection of high efficiency hydrogen-producing mutants by ultraviolet radiation

Hydrogen is an ideal, clean and sustainable energy source for the future because of its high conversion and nonpolluting nature. Biohydrogen production by dark-fermentation appears to have a great potential to be developed for practical application. However, one limiting factor affecting the development of hydrogen-production industrialization is that the hydrogen-producing capacity of bacteria is lower, so how to increase bacteria＇ s hydrogen-producing ability will be an urgent issue. In this experiment, 2 mutants, namely UV3 and UV7, were obtained by ultra-violet radiation. They grew and produced hydrogen efficiently on iron-containing medium. The hydrogen evolution of UV3 and UV7 were 2 356. 68 ml/L and 2 219. 62 ml/L at a glucose concentration of 10 g/L, respectively. With wild parent strain Ethanoligenens sp. ZGX4, the hydrogen evolution was 1 806. 02 ml/L under the same conditions. Mutants＇ hydrogen-producing capacities were about 29. 71% and 22.22% higher than that of wild parent strain ZGX4. The maximum H2 production rate by mutants UV3 and UV7 were estimated to be 32.57 mmol H2/g cell h and 31.19 mmol H2/g cell h, respectively, which were 38. 18% and 34. 78% higher than the control （23.57 mmol H：/g cell h）. The abundant products of UV3 and UV7 were ethanol and acetic, which accounted for 95% - 98% of total soluble microbial products. In each case, mutant strains UV3 and UV7 evolved hydrogen at a higher rate than the wild type, showing a possible potential for commercial hydrogen production. Another mutant named UV20＇ was also gained whose main end metabolites were butyric acid and acetic acid. This would provide researched material for a discussion of metabolic pathways of hydrogen-producing bacteria.

Tolerance of Grasses to Heavy Metals and Microbial Functional Diversity in Soils Contaminated with Copper Mine Tailings

Copper （Cu） mine tailings, because of their high content of heavy metals, are usually hostile to plant colonization. A pot experiment was conducted to determine the tolerance of four forage grasses to heavy metals in Cu mine tailings and to examine the variation in the microbial functional diversity of soils from the tailing sites in southern China. All the four grass species survived on Cu mine tailings and Cu mine tailing-soil mixture. However, on pure mine tailings, the growth was minimal, whereas the growth was maximum for the control without mine tailings. The tolerance of grasses to heavy metals followed the sequence： Paspalum notatum 〉 Festuea arundinaeea 〉 Lolium perenne 〉 Cynodon daetylon. The planting of forage grasses enhanced the soil microbial biomass. The Biolog data indicated that the soil microbial metabolic profile values （average well color development, community richness, and Shannon index） of the four forage grasses also followed the sequence： P. notatum 〉 F. arundinaeea 〉 L. perenne 〉 C. daetylon. Thus, P. notatum, under the experimental conditions of this study, may be considered as the preferred plant species for revegetation of Cu mine tailing areas.

Responses of soil microorganisms to elevated CO2 in experiment sites of Pinus sylvestriformis and Pinus koraiensis

Responses of soil microbial activities to elevated CO, in experiment sites of Pinus sylvestriformis and Pinus koratensts seecllmgs were studied in summer in 2003. The results indicated the number of bacteria decreased significantly （p 〈 0.05） under elevated CO, for Pinus syivestriformis and Pinups koraiensis. Amylase and invertase activities in soil increased for Pinus syivestriformis and decreased for Pinus koraiensis with CO2 enrichment compared with those at ambient （350 pmol·mol^-1）. The size of microbial biomass C also decreased significantly at 700 μmol- mol^-1 CO2. Bacterial community structure had some evident changes under elevated CO, by DGGE （Denaturing Gradient Gel Electrophoresis） analysis of bacterial 16S rDNA gene fragments amplified by PCR from DNA extracted directly from soil. The results suggested that responses of soil microorganisms to elevated CO2 would be related to plant species exposed to elevated CO2.

Assessment of different mcy genes for detecting the toxic to non-toxic Microcystis ratio in the field by multiplex qPCR

Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis and non-microcystin-producing Microcystis are key to predicting and treating Microcystis blooms. Multiplex qPCR is a useful tool to assess such issues. In this study, we developed multiplex qPCR methods with newly-designed probes and primers for the microcystin-synthesis related genes mcyA and mcyE. We used seven toxic Microcystis strains and four non-toxic Microcystis strains to compare the differences in the ratios of toxic and non-toxic Microcystis in mixed cultures, which were calculated using abundances of the genes mcyA, mcyB, mcyD, mcy E and phycocyanin( PC). We also compared traditional cell counting and multiplex qPCR. Hierarchical clustering and principal component analysis indicated that mcyD was the most suitable mcy gene for quantification in laboratory experiments. mcyB abundances were always higher; we suggest that the amount of toxic Microcystis measured using mcyB might overestimate the actual percentages.

Simulation of coupled pelagic-benthic ecosystem of the Yellow Sea Cold Water Mass

A one-dimensional coupled pelagic-benthic box model for the Yellow Sea Cold Water Mass (YSCWM) is developed. The model is divided into three boxes vertically according to the depths of thermocline and euphotic layer. It simulates well the oligotrophic shelf ecosystem of the YSCWM considering effects of nu- trients deposition and microbial loop. Main features of vertical structure of various variables in ecosystem of the YSCWM were captured and seasonal variability of the ecosystem was well reconstructed. Calculation shows that the contribution of microbial loop to the zooplankton can reach up to 60%. Besides, input of inorganic nutrients from atmospheric deposition is an important mechanism of production in upper layer of the YSCWM when stratified.

Performance of BAC process for treatment of micro-polluted water

Biological activated carbon (BAC) has been developed on the granular activated carbon by immobilization of selected and acclimated species of bacteria to treat the micro-polluted water. The BAC removal efficiencies for nitrobenzene, permanganate index, turbidity and ammonia were investigated. Effects of shock loading and SEM (Scanning Electron Microscope) observation on BAC were studied. Backwashing and its intensity of BAC were also discussed. The results showed that BAC took short time to start up and recover to the normal condition after shock loading. The shock loading studies showed that the removal efficiency of BAC was not completely inhibited even at high concentration of nitrobenzene. Backwashing performed once every 10-20 d, or an average of 15 d. Backwashing intensity was 12-14 L/(s·m2) with air and 3-4 L/(s·m2) with water.

Distribution of Microbial Populations and Their Relationship with Environmental Variables in the North Yellow Sea,China

In order to understand the large-scale spatial distribution characteristics of picoplankton,nanophytoplankton and virio-plankton and their relationship with environmental variables in coastal and offshore waters,flow cytometry(FCM) was used to ana-lyze microbial abundance of samples collected in summer from four depths at 36 stations in the North Yellow Sea(NYS).The data revealed spatial heterogeneity in microbial populations in the offshore and near-shore waters of the NYS during the summer.For the surface layer,picoeukaryotes were abundant in the near-shore waters,Synechococcus was abundant in the offshore areas,and bacte-rial and viral abundances were high in the near-shore waters around the Liaodong peninsula.In the near-shore waters,no significant vertical variation of picophytoplankton(0.2-2μm) abundance was found.However,the nanophytoplankton abundance was higher in the upper layers(from the surface to 10 m depth) than in the bottom layer.For the offshore waters,both pico-and nanophytoplankton(2-20μm) abundance decreased sharply with depth in the North Yellow Sea Cold Water Mass(NYSCWM).But,for the vertical dis-tribution of virus and bacteria abundance,no significant variation was observed in both near-shore and offshore waters.Autotrophic microbes were more sensitive to environmental change than heterotrophic microbes and viruses.Viruses showed a positive correla-tion with bacterial abundance,suggesting that the bacteriophage might be prominent for virioplankton(about 0.45μm) in summer in the NYS and that viral abundance might play an important role in microbial loop functions.

Nitrogen Biological Cycle Characteristics of Seepweed(Suaeda salsa) Wetland in Intertidal Zone of Huanghe(Yellow) River Estuary

From April 2008 to November 2009, the nitrogen （N） cycle of plant-soil system in seepweed （Suaeda salsa） wetland in the intertidal zone of the Huanghe （Yellow） River estuary was studied. Results showed that soil N had sig- nificant seasonal fluctuations and vertical distribution, and the net N mineralization rates in topsoil were significantly different in growing season （p 〈 0.01）. The N/P ratio （9.87 ±1.23） of S. salsa was less than 14, indicating that plant growth was limited by N. The N accumulated in S. salsa litter at all times during decomposition, which was ascribed to the N immobilization by microbes from the environment. Soil organic N was the main N stock of plant-soil system, accounting for 97.35% of the total N stock. The N absorption and utilization coefficients of S. salsa were very low （0.0145 and 0.3844, respectively）, while the N cycle coefficient was high （0.7108）. The results of the N turnovers among compartments of S. salsa wetland showed that the N uptake amount of aboveground part and root were 7.764 g/m2and 4.332 g/m2, respectively. The N translocation amounts from aboveground part to root and from root to soil were 3.881 g/m2 and 0.626 g/m2, respectively. The N translocation amount from aboveground living body to litter was 3.883 g/m2, the annual N return amount from litter to soil was more than 0.125（-） g/m2 （minus represented immobili- zation）, and the net N mineralization amount in topsoil （0-15 cm） in growing season was 1.190 g/m2. The assessment of N biological cycle status orS. salsa wetland indicated that N was a very important limiting factor and the ecosystem was situated in unstable and vulnerable status. The S. salsa was seemingly well adapted to the low-nutrient status and vulnerable habitat, and the N quantitative relationships determined in the compartment model might provide scientific base for us to reveal the special adaptive strategy orS. salsa to the vulnerable habitat in the following studies.

Dynamics of Microbial Activity Related to N Cycling in Cd-Contaminated Soil During Growth of Soybean

The potential influences of cadmium (Cd) on the biochemical processes of the soil nitrogen (N) cycle, along with the dynamics of ammonification, nitrification, and denitrification processes in the rhizosphere and non-rhizosphere (bulk soil), respectively, were investigated in a Cd-stressed system during an entire soybean growing season. In terms of Cd pollution at the seedling stage, the ammonifying bacteria proved to be the most sensitive microorganisms, whereas the effects of Cd on denitrification were not obvious. Following the growth of soybeans, the influences of Cd on ammonification in the bulk soil were: toxic impacts at the seedling stage, stimulatory effects during the early flowering stage, and adaptation to the pollutant during the podding and ripening stages. Although nitrification and denitrification in the bulk soil decreased throughout the entire growth cycle, positive adaptation to Cd stress was observed during the ripening stage. Moreover, during the ripening stage, denitrification in the bulk soil under high Cd treatment (20 mg kg-1) was even higher than that in the control, indicating a probable change in the ecology of the denitrifying microbes in the Cd-stressed system. Changes in the activity of microbes in the rhizosphere following plant growth were similar to those in the non-rhizosphere in Cd treatments; however, the tendency of change in the rhizosphere seemed to be more moderate. This suggested that there was some mitigation of Cd stress in the rhizosphere.

Environmental Capacity of Petroleum Hydrocarbon Pollutants in Jiaozhou Bay, China: Modeling and Calculation

An environmental capacity model for the petroleum hydrocarbon pollutions （PHs） in Jiaozhou Bay is constructed based on field surveys, mesocosm, and parallel laboratory experiments. Simulated results of PHs seasonal successions in 2003 match the field surveys of Jiaozhou Bay resaonably well with a highest value in July. The Monte Carlo analysis confirms that the variation of PHs concentration significantly correlates with the river input. The water body in the bay is reasonably subjected to self-purification processes, such as volatilization to the atmosphere, biodegradation by microorganism, and transport to the Yellow Sea by water exchange. The environmental capacity of PHs in Jiaozhou Bay is 1500 tons per year IF the seawater quality criterion （Grade Ⅰ/Ⅱ, 0.05 mgLl） in the region is to be satisfied. The contribution to self-purification by volatilization, biodegradation, and transport to the Yellow Sea accounts for 48%, 28%, and 23%, respectively, which make these three processes the main ways of PHs purification in Jiaozhou Bay.

Microbial Remediation of Heavy Metal(loid)Contaminated Soil: A Review

Because of the rapid development of industrial processes, increased urban pollution and agricultural chemicals applied in recent years, heavy metal(loid) pollution in soil has been very serious, and there is an urgent need for fast and efficient removal of heavy metal(loid) pollution. Currently, environmental microorganisms are always used to perform biological alteration or improvement of soils and sewage. Using functional microorganisms that are resistant to toxic heavy metal(loid) ions for alteration and transformation of heavy metal(loid)s in ionic form is an effective measure for microbial remediation of heavy metal(loid)contaminated soil. This paper reviewed the microbial remediation mechanism of heavy metal(loid) contaminated soils, and the approaches for breeding bacteria those can be used for highly efficient removal of heavy metal(loid)s, as well as the application examples of microbial remediation and transformation of heavy metal(loid) contaminated soil, and finally described the future trends and further research work of heavy metal(loid) contaminated soils by microbial remediation.

Security evaluation of compounded microbial flocculant

A new kind of compounded microbial flocculant (CMBF) for water and wastewater treatment has been developed through biological technology. In order to discuss its biological security, four groups of experiments, rat acute toxicity test via mouth, salmonella assay in vitro, mouse micronucleus in vivo test and teratogenesis test were conducted to evaluate its general toxicity, genotoxicity and generative toxicity. The experimental results showed that this type of compounded microbial flocculant was a substantial non-toxic substance based on the fact that LD50 value was over 10 mg/kg. The results from salmonella in vivo test and mouse micronucleus in vivo test revealed that the compounded microbial flocculant is a genetically non-toxic substance. Furthermore, compounded microbial flocculant has little effect on the growth of all the rats, and any morphologic abnormal phenomenon hasn’t appeared.