GAS6-AS1调节miR-370-3p/SPATA2轴对卵巢癌细胞增殖、迁移、侵袭、凋亡和EMT的影响

目的:探讨长链非编码RNA GAS6反义RNA1(long non-coding RNA GAS6 antisense RNA1, lncRNA GAS6-AS1)调节miR-370-3p/精子发生相关蛋白2 (spermatogenesis-associated protein 2, SPATA2)轴对卵巢癌细胞增殖、迁移、侵袭、凋亡和上皮间质转化(epithelial mesenchymal transformation, EMT)的影响。方法:qRT-PCR、Western blot分别检测癌旁组织、卵巢癌组织、人正常卵巢上皮细胞IOSE80及卵巢癌细胞系HO-8910、SKOV3、A2780中GAS6-AS1、miR-370-3p及SPATA2蛋白表达。将SKOV3细胞分为:对照组(NC组)、 si-NC组、si-GAS6-AS1组、mimic NC组、miR-370-3p mimic组、si-GAS6-AS1+inhibitor NC组、si-GAS6-AS1+miR-370-3p inhibitor组,qRT-PCR检测细胞中GAS6-AS1、miR-370-3p表达;CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡;划痕愈合实验检测细胞迁移;Transwell实验检测细胞侵袭;Western blot检测SPATA2、细胞周期素D1(CyclinD1)、Bcl-2相关X蛋白(Bcl-2-associated X,Bax)、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、神经钙黏蛋白(N-cadherin)表达;双荧光素酶报告基因实验检测GAS6-AS1与miR-370-3p、 miR-370-3p与SPATA2的关系。结果:在卵巢癌组织和细胞中GAS6-AS1、SPATA2蛋白高表达,miR-370-3p低表达,且在SKOV3细胞中GAS6-AS1、SPATA2蛋白表达量最高,miR-370-3p表达水平最低,因此,选择SKOV3细胞为后续研究对象。与NC组、si-NC组比较,si-GAS6-AS1组GAS6-AS1、OD450值(24 h、48 h、72 h)、划痕愈合率、侵袭细胞数、SPATA2、CyclinD1、Vimentin、N-cadherin蛋白表达降低,miR-370-3p表达、细胞凋亡率、Bax、E-cadherin蛋白表达升高(P<0.05);与NC组、mimic NC组比较,miR-370-3p mimic组OD450值(24 h、48 h、72 h)、划痕愈合率、侵袭细胞数、SPATA2、CyclinD1、Vimentin、N-cadherin蛋白表达降低,miR-370-3p表达、细胞凋亡率、Bax、E-cadherin蛋白表达升高(P<0.05);miR-370-3p inhibitor减弱了沉默GAS6-AS1对SKOV3细胞增殖、迁移、侵袭及EMT的抑制及对细胞凋亡的促进作用。GAS6-AS1与miR-370-3p、miR-370-3p与SPATA2存在靶向调控关系。结论:沉默GAS6-AS1通过上调miR-370-3p来抑制SPATA2表达,从而抑制SKOV3细胞增殖、迁移、侵袭及EMT,并促进细胞凋亡。

Geochemistry and origins of hydrogen-containing natural gases in deep Songliao Basin,China:Insights from continental scientific drilling

The different reservoirs in deep Songliao Basin have non-homogeneous lithologies and include multiple layers with a high content of hydrogen gas.The gas composition and stable isotope characteristics vary significantly,but the origin analysis of different gas types has previously been weak.Based on the geochemical parameters of gas samples from different depths and the analysis of geological settings,this research covers the diverse origins of natural gas in different strata.The gas components are mainly methane with a small amount of C_(2+),and non-hydrocarbon gases,including nitrogen(N_(2)),hydrogen(H_(2)),carbon dioxide(CO_(2)),and helium(He).At greater depth,the carbon isotope of methane becomes heavier,and the hydrogen isotope points to a lacustrine sedimentary environment.With increasing depth,the origins of N_(2)and CO_(2)change gradually from a mixture of organic and inorganic to inorganic.The origins of hydrogen gas are complex and include organic sources,water radiolysis,water-rock(Fe^(2+)-containing minerals)reactions,and mantle-derived.The shales of Denglouku and Shahezi Formations,as source rocks,provide the premise for generation and occurrence of organic gas.Furthermore,the deep faults and fluid activities in Basement Formation control the generation and migration of mantle-derived gas.The discovery of a high content of H_(2)in study area not only reveals the organic and inorganic association of natural-gas generation,but also provides a scientific basis for the exploration of deep hydrogen-rich gas.

Constraints on Characteristics and Distribution of Gas Hydrate and Free Gas Using Broad-Band Processing of Three-Dimensional Seismic Data

Gas hydrate drilling expeditions in the Pearl River Mouth Basin,South China Sea,have identified concentrated gas hydrates with variable thickness.Moreover,free gas and the coexistence of gas hydrate and free gas have been confirmed by logging,coring,and production tests in the foraminifera-rich silty sediments with complex bottom-simulating reflectors(BSRs).The broad-band processing is conducted on conventional three-dimensional(3D)seismic data to improve the image and detection accuracy of gas hydratebearing layers and delineate the saturation and thickness of gas hydrate-and free gas-bearing sediments.Several geophysical attributes extracted along the base of the gas hydrate stability zone are used to demonstrate the variable distribution and the controlling factors for the differential enrichment of gas hydrate.The inverted gas hydrate saturation at the production zone is over 40% with a thickness of 90 m,showing the interbedded distribution with different boundaries between gas hydrate-and free gas-bearing layers.However,the gas hydrate saturation value at the adjacent canyon is 70%,with 30-m-thick patches and linear features.The lithological and fault controls on gas hydrate and free gas distributions are demonstrated by tracing each gas hydrate-bearing layer.Moreover,the BSR depths based on broad-band reprocessed 3D seismic data not only exhibit variations due to small-scale topographic changes caused by seafloor sedimentation and erosion but also show the upward shift of BSR and the blocky distribution of the coexistence of gas hydrate and free gas in the Pearl River Mouth Basin.

基于GAS算法的卵砾石粒径自动识别应用研究

粒径和级配是表征床面组成的重要指标,基于GAS粒径自动识别技术可自动识别粗粒床面粒径并生成级配曲线,能够大幅提高现场采样和分析的效率。为了验证GAS的分割效果,采用GAS提供的默认参数进行分割,同时应用ImageJ软件手动分割进行验证。结果表明:GAS级配曲线的相对误差为5.7%,相关系数为0.992。另外,采用单参数和多参数敏感性分析法来标准化参数调整方案,gre、can1和can2对GAS提取的级配曲线和特征粒径有显著影响,其中gre起主导作用,而can1和can2控制着砾石边界的检测完整性。

Deep-large faults controlling on the distribution of the venting gas hydrate system in the middle of the Qiongdongnan Basin, South China Sea

Many locations with concentrated hydrates at vents have confirmed the presence of abundant thermogenic gas in the middle of the Qiongdongnan Basin(QDNB).However,the impact of deep structures on gasbearing fluids migration and gas hydrates distribution in tectonically inactive regions is still unclear.In this study,the authors apply high-resolution 3D seismic and logging while drilling(LWD)data from the middle of the QDNB to investigate the influence of deep-large faults on gas chimneys and preferred gasescape pipes.The findings reveal the following:(1)Two significant deep-large faults,F1 and F2,developed on the edge of the Songnan Low Uplift,control the dominant migration of thermogenic hydrocarbons and determine the initial locations of gas chimneys.(2)The formation of gas chimneys is likely related to fault activation and reactivation.Gas chimney 1 is primarily arises from convergent fluid migration resulting from the intersection of the two faults,while the gas chimney 2 benefits from a steeper fault plane and shorter migration distance of fault F2.(3)Most gas-escape pipes are situated near the apex of the two faults.Their reactivations facilitate free gas flow into the GHSZ and contribute to the formation of fracture‐filling hydrates.

Inter-layer interference for multi-layered tight gas reservoir in the absence and presence of movable water

Due to the dissimilarity among different producing layers,the influences of inter-layer interference on the production performance of a multi-layer gas reservoir are possible.However,systematic studies of inter-layer interference for tight gas reservoirs are really limited,especially for those reservoirs in the presence of water.In this work,five types of possible inter-layer interferences,including both absence and presence of water,are identified for commingled production of tight gas reservoirs.Subsequently,a series of reservoir-scale and pore-scale numerical simulations are conducted to quantify the degree of influence of each type of interference.Consistent field evidence from the Yan'an tight gas reservoir(Ordos Basin,China)is found to support the simulation results.Additionally,suggestions are proposed to mitigate the potential inter-layer interferences.The results indicate that,in the absence of water,commingled production is favorable in two situations:when there is a difference in physical properties and when there is a difference in the pressure system of each layer.For reservoirs with a multi-pressure system,the backflow phenomenon,which significantly influences the production performance,only occurs under extreme conditions(such as very low production rates or well shut-in periods).When water is introduced into the multi-layer system,inter-layer interference becomes nearly inevitable.Perforating both the gas-rich layer and water-rich layer for commingled production is not desirable,as it can trigger water invasion from the water-rich layer into the gas-rich layer.The gas-rich layer might also be interfered with by water from the neighboring unperforated water-rich layer,where the water might break the barrier(eg weak joint surface,cement in fractures)between the two layers and migrate into the gas-rich layer.Additionally,the gas-rich layer could possibly be interfered with by water that accumulates at the bottom of the wellbore due to gravitational differentiation during shut-in operations.

Geochemical characteristics and exploration significance of ultra-deep Sinian oil and gas from Well Tashen 5,Tarim Basin,NW China

The Well Tashen 5(TS5),drilled and completed at a vertical depth of 9017 m in the Tabei Uplift of the Tarim Basin,NW China,is the deepest well in Asia.It has been producing both oil and gas from the Sinian at a depth of 8780e8840 m,also the deepest in Asia in terms of oil discovery.In this paper,the geochemical characteristics of Sinian oil and gas from the well were investigated and compared with those of Cambrian oil and gas discovered in the same basin.The oil samples,with Pr/Ph ratio of 0.78 and a whole oil carbon isotopic value of31.6‰,have geochemical characteristics similar to those of Ordovician oils from the No.1 fault in the North Shuntuoguole area(also named Shunbei area)and the Middle Cambrian oil from wells Zhongshen 1(ZS1)and Zhongshen 5(ZS5)of Tazhong Uplift.The maturity of light hydrocarbons,diamondoids and aromatic fractions all suggest an approximate maturity of 1.5%e1.7%Ro for the samples.The(4-+3-)methyldiamantane concentration of the samples is 113.5 mg/g,indicating intense cracking with a cracking degree of about 80%,which is consistent with the high bottom hole temperature(179℃).The Sinian gas samples are dry with a dryness coefficient of 0.97.The gas is a mixture of kerogen-cracking gas and oil-cracking gas and has Ro values ranging between 1.5%and 1.7%,and methane carbon isotopic values of41.6‰.Based on the equivalent vitrinite reflectance(R_(eqv)=1.51%e1.61%)and the thermal evolution of source rocks from the Cambrian Yu'ertusi Formation of the same well,it is proposed that the Sinian oil and gas be mainly sourced from the Cambrian Yu'ertusi Formation during the Himalayan period but probably also be joined by hydrocarbon of higher maturity that migrated from other source rocks in deeper formations.The discovery of Sinian oil and gas from Well TS5 suggests that the ancient ultra-deep strata in the northern Tarim Basin have the potential for finding volatile oil or condensate reservoirs.

Geological characteristics and exploration breakthroughs of coal rock gas in Carboniferous Benxi Formation,Ordos Basin,NW China

To explore the geological characteristics and exploration potential of the Carboniferous Benxi Formation coal rock gas in the Ordos Basin,this paper presents a systematic research on the coal rock distribution,coal rock reservoirs,coal rock quality,and coal rock gas features,resources and enrichment.Coal rock gas is a high-quality resource distinct from coalbed methane,and it has unique features in terms of burial depth,gas source,reservoir,gas content,and carbon isotopic composition.The Benxi Formation coal rocks cover an area of 16×104km^(2),with thicknesses ranging from 2 m to 25 m,primarily consisting of bright and semi-bright coals with primitive structures and low volatile and ash contents,indicating a good coal quality.The medium-to-high rank coal rocks have the total organic carbon(TOC)content ranging from 33.49%to 86.11%,averaging75.16%.They have a high degree of thermal evolution(Roof 1.2%-2.8%),and a high gas-generating capacity.They also have high stable carbon isotopic values(δ13C1of-37.6‰to-16‰;δ13C2of-21.7‰to-14.3‰).Deep coal rocks develop matrix pores such as gas bubble pores,organic pores,and inorganic mineral pores,which,together with cleats and fractures,form good reservoir spaces.The coal rock reservoirs exhibit the porosity of 0.54%-10.67%(averaging 5.42%)and the permeability of(0.001-14.600)×10^(-3)μm^(2)(averaging 2.32×10^(-3)μm^(2)).Vertically,there are five types of coal rock gas accumulation and dissipation combinations,among which the coal rock-mudstone gas accumulation combination and the coal rock-limestone gas accumulation combination are the most important,with good sealing conditions and high peak values of total hydrocarbon in gas logging.A model of coal rock gas accumulation has been constructed,which includes widespread distribution of medium-to-high rank coal rocks continually generating gas,matrix pores and cleats/fractures in coal rocks acting as large-scale reservoir spaces,tight cap rocks providing sealing,source-reservoir integration,and five types of efficient enrichment patterns(lateral pinchout complex,lenses,low-amplitude structures,nose-like structures,and lithologically self-sealing).According to the geological characteristics of coal rock gas,the Benxi Formation is divided into 8 plays,and the estimated coal rock gas resources with a buried depth of more than 2000 m are more than 12.33×10^(12)m^(3).The above understandings guide the deployment of risk exploration.Two wells drilled accordingly obtained an industrial gas flow,driving the further deployment of exploratory and appraisal wells.Substantial breakthroughs have been achieved,with the possible reserves over a trillion cubic meters and the proved reserves over a hundred billion cubic meters,which is of great significance for the reserves increase and efficient development of natural gas in China.

Effect of safety valve types on the gas venting behavior and thermal runaway hazard severity of large-format prismatic lithium iron phosphate batteries

The safety valve is an important component to ensure the safe operation of lithium-ion batteries(LIBs).However,the effect of safety valve type on the thermal runaway(TR)and gas venting behavior of LIBs,as well as the TR hazard severity of LIBs,are not known.In this paper,the TR and gas venting behavior of three 100 A h lithium iron phosphate(LFP)batteries with different safety valves are investigated under overheating.Compared to previous studies,the main contribution of this work is in studying and evaluating the effect of gas venting behavior and TR hazard severity of LFP batteries with three safety valve types.Two significant results are obtained:(Ⅰ)the safety valve type dominates over gas venting pressure of battery during safety venting,the maximum gas venting pressure of LFP batteries with a round safety valve is 3320 Pa,which is one order of magnitude higher than other batteries with oval or cavity safety valve;(Ⅱ)the LFP battery with oval safety valve has the lowest TR hazard as shown by the TR hazard assessment model based on gray-fuzzy analytic hierarchy process.This study reveals the effect of safety valve type on TR and gas venting,providing a clear direction for the safety valve design.

Experimental investigation on coal pore-fracture variation and fractal characteristics synergistically affected by solvents for improving clean gas extraction

Chemical solvents instead of pure water being as hydraulic fracturing fluid could effectively increase permeability and improve clean methane extraction efficiency.However,pore-fracture variation features of lean coal synergistically affected by solvents have not been fully understood.Ultrasonic testing,nuclear magnetic resonance analysis,liquid phase mass spectrometry was adopted to comprehensively analyze pore-fracture change characteristics of lean coal treated by combined solvent(NMP and CS_(2)).Meanwhile,quantitative characterization of above changing properties was conducted using geometric fractal theory.Relationship model between permeability,fractal dimension and porosity were established.Results indicate that the end face fractures of coal are well developed after CS2and combined solvent treatments,of which,end face box-counting fractal dimensions range from 1.1227 to 1.4767.Maximum decreases in ultrasonic longitudinal wave velocity of coal affected by NMP,CS_(2)and combined solvent are 2.700%,20.521%,22.454%,respectively.Solvent treatments could lead to increasing amount of both mesopores and macropores.Decrease ratio of fractal dimension Dsis 0.259%–2.159%,while permeability increases ratio of NMR ranges from 0.1904 to 6.4486.Meanwhile,combined solvent could dissolve coal polar and non-polar small molecules and expand flow space.Results could provide reference for solvent selection and parameter optimization of permeability-enhancement technology.

Repositioning fertilizer manufacturing subsidies for improving food security and reducing greenhouse gas emissions in China

China removed fertilizer manufacturing subsidies from 2015 to 2018 to bolster market-oriented reforms and foster environmentally sustainable practices.However,the impact of this policy reform on food security and the environment remains inadequately evaluated.Moreover,although green and low-carbon technologies offer environmental advantages,their widespread adoption is hindered by prohibitively high costs.This study analyzes the impact of removing fertilizer manufacturing subsidies and explores the potential feasibility of redirecting fertilizer manufacturing subsidies to invest in the diffusion of these technologies.Utilizing the China Agricultural University Agri-food Systems model,we analyzed the potential for achieving mutually beneficial outcomes regarding food security and environmental sustainability.The findings indicate that removing fertilizer manufacturing subsidies has reduced greenhouse gas(GHG)emissions from agricultural activities by 3.88 million metric tons,with minimal impact on food production.Redirecting fertilizer manufacturing subsidies to invest in green and low-carbon technologies,including slow and controlled-release fertilizer,organic-inorganic compound fertilizers,and machine deep placement of fertilizer,emerges as a strategy to concurrently curtail GHG emissions,ensure food security,and secure robust economic returns.Finally,we propose a comprehensive set of government interventions,including subsidies,field guidance,and improved extension systems,to promote the widespread adoption of these technologies.

Development and technology status of energy storage in depleted gas reservoirs

Utilizing energy storage in depleted oil and gas reservoirs can improve productivity while reducing power costs and is one of the best ways to achieve synergistic development of"Carbon Peak–Carbon Neutral"and"Underground Resource Utiliza-tion".Starting from the development of Compressed Air Energy Storage(CAES)technology,the site selection of CAES in depleted gas and oil reservoirs,the evolution mechanism of reservoir dynamic sealing,and the high-flow CAES and injection technology are summarized.It focuses on analyzing the characteristics,key equipment,reservoir construction,application scenarios and cost analysis of CAES projects,and sorting out the technical key points and existing difficulties.The devel-opment trend of CAES technology is proposed,and the future development path is scrutinized to provide reference for the research of CAES projects in depleted oil and gas reservoirs.

Risk assessment of oil and gas investment environment in countries along the Belt and Road Initiative

With the implementation of the Belt and Road Initiative, China is deepening its cooperation in oil and gas resources with countries along the Initiative. In order to better mitigate risks and enhance the safety of investments, it is of significant importance to research the oil and gas investment environment in these countries for China's overseas investment macro-layout. This paper proposes an indicator system including 27 indicators from 6 dimensions. On this basis, game theory models combined with global entropy method and analytic hierarchy process are applied to determine the combined weights, and the TOPSIS-GRA model is utilized to assess the risks of oil and gas investment in 76 countries along the Initiative from 2014 to 2021. Finally, the GM(1,1) model is employed to predict risk values for 2022-2025. In conclusion, oil and gas resources and political factors have the greatest impact on investment environment risk, and 12 countries with greater investment potential are selected through cluster analysis in conjunction with the predicted results. The research findings may provide scientific decisionmaking recommendations for the Chinese government and oil enterprises to strengthen oil and gas investment cooperation with countries along the Belt and Road Initiative.

Sealing capacity evaluation of underground gas storage under intricate geological conditions

Evaluating underground gas storage(UGS)sealing capacity is essential for its safe construction and operational efficiency.This involves evaluating both the static sealing capacity of traps during hydrocarbon accumulation and the dynamic sealing capacity of UGS under intensive gas injection and withdrawal,and alternating loads.This study detailed the methodology developed by Sinopec.The approach merges disciplines like geology,geomechanics,and hydrodynamics,employing both dynamic-static and qualitative-quantitative analyses.Sinopec's evaluation methods,grounded in the in situ stress analysis,include mechanistic studies,laboratory tests,geological surveys,stress analysis,and fluid-solid interactions.Through tests on the static and dynamic sealing capacity of UGS,alongside investigations into sealing mechanisms and the geological and geomechanical properties of cap rocks and faults,A geomechanics-rock damage-seepage mechanics dynamic coupling analysis method has been developed to predict in situ stress variations relative to pore pressure changes during UGS operations and evaluate fault sealing capacity and cap rock integrity,thereby setting the maximum operational pressures.Utilizing this evaluation technique,Sinopec has defined performance metrics and criteria for evaluating the sealing capacity of depleted gas reservoirs,enabling preliminary sealing capacity evaluations at UGS sites.These evaluations have significantly informed the design of UGS construction schemes and the evaluation of fault sealing capacity and cap rock integrity during UGS operations.

Miscibility of light oil and flue gas under thermal action

The miscibility of flue gas and different types of light oils is investigated through slender-tube miscible displacement experiment at high temperature and high pressure.Under the conditions of high temperature and high pressure,the miscible displacement of flue gas and light oil is possible.At the same temperature,there is a linear relationship between oil displacement efficiency and pressure.At the same pressure,the oil displacement efficiency increases gently and then rapidly to more than 90% to achieve miscible displacement with the increase of temperature.The rapid increase of oil displacement efficiency is closely related to the process that the light components of oil transit in phase state due to distillation with the rise of temperature.Moreover,at the same pressure,the lighter the oil,the lower the minimum miscibility temperature between flue gas and oil,which allows easier miscibility and ultimately better performance of thermal miscible flooding by air injection.The miscibility between flue gas and light oil at high temperature and high pressure is more typically characterized by phase transition at high temperature in supercritical state,and it is different from the contact extraction miscibility of CO_(2) under conventional high pressure conditions.

Discovery and inspiration of large-and medium-sized glutenite-rich oil and gas fields in the eastern South China Sea:An example from Paleogene Enping Formation in Huizhou 26 subsag,Pearl River Mouth Basin

Based on the practice of oil and gas exploration in the Huizhou Sag of the Pearl River Mouth Basin,the geochemical indexes of source rocks were measured,the reservoir development morphology was restored,the rocks and minerals were characterized microscopically,the measured trap sealing indexes were compared,the biomarker compounds of crude oil were extracted,the genesis of condensate gas was identified,and the reservoir-forming conditions were examined.On this basis,the Paleogene Enping Formation in the Huizhou 26 subsag was systematically analyzed for the potential of oil and gas resources,the development characteristics of large-scale high-quality conglomerate reservoirs,the trapping effectiveness of faults,the hydrocarbon migration and accumulation model,and the formation conditions and exploration targets of large-and medium-sized glutenite-rich oil and gas fields.The research results were obtained in four aspects.First,the Paleogene Wenchang Formation in the Huizhou 26 subsag develops extensive and thick high-quality source rocks of semi-deep to deep lacustrine subfacies,which have typical hydrocarbon expulsion characteristics of"great oil generation in the early stage and huge gas expulsion in the late stage",providing a sufficient material basis for hydrocarbon accumulation in the Enping Formation.Second,under the joint control of the steep slope zone and transition zone of the fault within the sag,the large-scale near-source glutenite reservoirs are highly heterogeneous,with the development scale dominated hierarchically by three factors(favorable facies zone,particle component,and microfracture).The(subaqueous)distributary channels near the fault system,with equal grains,a low mud content(<5%),and a high content of feldspar composition,are conducive to the development of sweet spot reservoirs.Third,the strike-slip pressurization trap covered by stable lake flooding mudstone is a necessary condition for oil and gas preservation,and the NE and nearly EW faults obliquely to the principal stress have the best control on traps.Fourth,the spatiotemporal configuration of high-quality source rocks,fault transport/sealing,and glutenite reservoirs controls the degree of hydrocarbon enrichment.From top to bottom,three hydrocarbon accumulation units,i.e.low-fill zone,transition zone,and high-fill zone,are recognized.The main area of the channel in the nearly pressurized source-connecting fault zone is favorable for large-scale hydrocarbon enrichment.The research results suggest a new direction for the exploration of large-scale glutenite-rich reservoirs in the Enping Formation of the Pearl River Mouth Basin,and present a major breakthrough in oil and gas exploration.

lncRNA GAS5调控miR-452-5p/Mcl-1通路对急性肺损伤大鼠肺组织细胞焦亡的影响

目的探讨长链非编码RNA生长抑制特异因子5(lncRNA GAS5)调控miR-452-5p/髓样细胞白血病序列-1(Mcl-1)通路对急性肺损伤(ALI)大鼠肺组织细胞焦亡的影响。方法取SD大鼠随机分成6组(每组10只):对照组、模型组、pUC57-lncRNA GAS5组、miR-452-5p抑制剂组、pUC57-NC+miR-452-5p-NC组、pUC57-lncRNA GAS5+miR-452-5p激活剂组。模型组和各干预组大鼠以腹腔注射脂多糖的方法构建ALI模型,对照组大鼠腹腔注射等量生理盐水,造模同时进行分组干预。然后检测大鼠肺功能,比较各组用力肺活量(FVC)、0.3秒用力呼气容积(FEV0.3)/FVC、潮气量(VT);HE染色观察大鼠肺组织病理损伤,并以Holfbauer评分评估其损伤程度;酶联免疫吸附法(ELISA)测定大鼠肺泡灌洗液(BALF)和血清炎症因子白细胞介素(IL)-18、IL-1β水平;免疫印迹法检测大鼠肺组织焦亡相关指标[消皮素D(GSDMD)、核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、天冬氨酸特异性半胱氨酸蛋白酶(Caspase)-1、Caspase-11]及Mcl-1蛋白表达;实时荧光定量PCR检测大鼠肺组织lncRNA GAS5、miR-452-5p及Mcl-1表达;双荧光素酶报告基因实验验证大鼠肺泡上皮细胞L2中lncRNA GAS5对miR-452-5p的靶向调节。结果与对照组比较,模型组大鼠FVC、FEV0.3/FVC、lncRNA GAS5表达、Mcl-1蛋白及mRNA表达降低(均P<0.05),VT、Holfbauer评分、IL-18及IL-1β水平、GSDMD、NLRP3、Caspase-1及Caspase-11蛋白表达、miR-452-5p表达升高(均P<0.05)。与模型组比较,pUC57-lncRNA GAS5组、miR-452-5p抑制剂组大鼠FVC、FEV0.3/FVC、Mcl-1蛋白及mRNA表达均升高(均P<0.05),VT、Holfbauer评分、IL-18及IL-1β水平、GSDMD、NLRP3、Caspase-1及Caspase-11蛋白表达、miR-452-5p表达均降低(均P<0.05);pUC57-NC+miR-452-5p-NC组大鼠各指标无显著差异(均P>0.05)。与pUC57-lncRNA GAS5组比较,pUC57-lncRNA GAS5+miR-452-5p激活剂组大鼠FVC、FEV0.3/FVC、Mcl-1蛋白及mRNA表达降低(均P<0.05),VT、Holfbauer评分、IL-18及IL-1β水平、GSDMD、NLRP3、Caspase-1及Caspase-11蛋白表达、miR-452-5p表达升高(均P<0.05)。结论lncRNA GAS5可通过靶向下调miR-452-5p而促进Mcl-1表达,进而抑制ALI大鼠炎症反应及肺组织细胞焦亡,最终减轻大鼠肺损伤并改善其肺功能。

LncRNA GAS5-AS1靶向miR-106a-5p调控结直肠癌细胞增殖、迁移和侵袭

背景长链非编码RNA(long non-coding RNA,LncRNA)生长停滞特异性转录本5反义RNA(growth arrest specific transcript 5 antisense RNA,GAS5-AS1)被发现在多种癌症中扮演肿瘤抑制因子的作用.但是其在结直肠癌(colorectal cancer,CRC)进展中的作用和机制尚不清楚.目的探讨LncRNA GAS5-AS1靶向miR-106a-5p在CRC进展中的作用.方法实时荧光定量聚合酶链反应分析LncRNA GAS5-AS1和miR-106a-5p在43对CRC组织和癌旁组织样本中的表达.分别转染pcDNA、pcDNA-LncRNA GAS5-AS1、anti-miR-NC、anti-miR-106a-5p、si-NC、si-LncRNA GAS5-AS1、pcDNA-LncRNA GAS5-AS1+miR-NC、pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics至HCT8细胞.细胞计数试剂盒-8法检测细胞活力;Transwell小室法检测迁移和侵袭细胞数.通过双荧光素酶报告实验确定LncRNA GAS5-AS1和miR-106a-5p的靶向关系.结果与癌旁组织比较,CRC组织中LncRNA GAS5-AS1表达降低(P<0.05),miR-106a-5p表达升高(P<0.05).与转染pcDNA比较,转染pcDNA-LncRNA GAS5-AS1后HCT8细胞活力、迁移、侵袭、miR-106a-5p表达显著降低(P<0.05).与转染anti-miR-NC比较,转染anti-miR-106a-5p后HCT8细胞活力、迁移和侵袭细胞数显著降低(P<0.05).与转染si-NC比较,转染si-LncRNA GAS5-AS1后miR-106a-5p表达水平显著升高(P<0.05).与转染miR-NC比较,转染miR-106a-5p能显著降低LncRNA GAS5-AS1-WT载体的荧光素酶活性,表明miR-106a-5p是LncRNA GAS5-AS1的直接靶点.与转染pcDNA-LncRNA GAS5-AS1+miR-NC比较,转染pcDNA-LncRNA GAS5-AS1+miR-106a-5p mimics后HCT8细胞活力、迁移和侵袭细胞数显著升高(P<0.05).结论LncRNA GAS5-AS1通过靶向miR-106a-5p来抑制CRC细胞增殖、迁移和侵袭.

Research on thermal insulation materials properties under HTHP conditions for deep oil and gas reservoir rock ITP-Coring

Deep oil and gas reservoirs are under high-temperature conditions,but traditional coring methods do not consider temperature-preserved measures and ignore the influence of temperature on rock porosity and permeability,resulting in distorted resource assessments.The development of in situ temperaturepreserved coring(ITP-Coring)technology for deep reservoir rock is urgent,and thermal insulation materials are key.Therefore,hollow glass microsphere/epoxy resin thermal insulation materials(HGM/EP materials)were proposed as thermal insulation materials.The materials properties under coupled hightemperature and high-pressure(HTHP)conditions were tested.The results indicated that high pressures led to HGM destruction and that the materials water absorption significantly increased;additionally,increasing temperature accelerated the process.High temperatures directly caused the thermal conductivity of the materials to increase;additionally,the thermal conduction and convection of water caused by high pressures led to an exponential increase in the thermal conductivity.High temperatures weakened the matrix,and high pressures destroyed the HGM,which resulted in a decrease in the tensile mechanical properties of the materials.The materials entered the high elastic state at 150℃,and the mechanical properties were weakened more obviously,while the pressure led to a significant effect when the water absorption was above 10%.Meanwhile,the tensile strength/strain were 13.62 MPa/1.3%and 6.09 MPa/0.86%at 100℃ and 100 MPa,respectively,which meet the application requirements of the self-designed coring device.Finally,K46-f40 and K46-f50 HGM/EP materials were proven to be suitable for ITP-Coring under coupled conditions below 100℃ and 100 MPa.To further improve the materials properties,the interface layer and EP matrix should be optimized.The results can provide references for the optimization and engineering application of materials and thus technical support for deep oil and gas resource development.

黄芪糖蛋白干扰lncRNA GAS5/miR-21/TLR4信号轴抑制细胞焦亡改善佐剂性关节炎大鼠心肌损伤

目的基于长链非编码RNA生长停滞特异性转录本5/微小RNA 21/Toll样受体4(lncRNA GAS5/miR-21/TLR4)信号轴探究黄芪糖蛋白(HQGP)对佐剂性关节炎(AA)大鼠心肌损伤的作用及机制。方法将SD大鼠随机分为正常对照组、模型组、甲氨蝶呤(MTX)组、HQGP组,每组6只。复制AA模型,第19天开始给药,连续4周。检测各组AA大鼠足趾肿胀度、关节炎指数(AI),HE染色观察大鼠心肌组织病理变化,透射电镜观察心肌组织超微结构的变化及焦亡小体情况,ELISA检测血清白细胞介素6(IL-6)、IL-18、IL-1β和肿瘤坏死因子α(TNF-α)水平,乳酸脱氢酶(LDH)试剂盒检测心肌组织LDH释放量,实时荧光定量PCR检测心肌组织核因子κB p65(NF-κB p65)、胱天蛋白酶1(caspase-1)、含pyrin结构域核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、消皮素D(GSDMD)的mRNA以及lncRNA GAS5/miR-21/TLR4信号轴分子的表达,Western blot法检测心肌组织TLR4、NF-κB p65、caspase-1、NLRP3和GSDMD蛋白水平。结果与正常对照组相比,模型组大鼠足趾肿胀度、AI显著升高;大鼠心肌纤维溶解、断裂,肌节结构模糊,心肌纤维排列紊乱,组织间有炎性细胞浸润,线粒体嵴明显断裂稀疏,焦亡小体数量增加;血清促炎细胞因子IL-6、IL-18、IL-1β和TNF-α表达显著升高;心肌组织中GAS5表达显著降低,miR-21表达显著升高,LDH释放量、TLR4、NF-κB p65、caspase-1、NLRP3、GSDMD的mRNA和蛋白表达水平显著升高。与模型组比较,HQGP组大鼠足趾肿胀度、AI和心肌组织病理状态明显改善;血清IL-6、IL-18、IL-1β和TNF-α表达显著降低;心肌组织中GAS5表达显著升高,miR-21表达显著降低,LDH释放量、TLR4、NF-κB p65、caspase-1、NLRP3的mRNA和蛋白表达水平显著降低,GSDMD的mRNA表达降低且蛋白水平显著降低。结论HQGP通过上调lncRNA GAS5抑制miR-21/TLR4信号传导,抑制细胞焦亡,减少促炎细胞因子表达,改善AA大鼠心肌损伤。