Numerical manifold method for thermo-mechanical coupling simulation of fractured rock mass

As a calculation method based on the Galerkin variation,the numerical manifold method(NMM)adopts a double covering system,which can easily deal with discontinuous deformation problems and has a high calculation accuracy.Aiming at the thermo-mechanical(TM)coupling problem of fractured rock masses,this study uses the NMM to simulate the processes of crack initiation and propagation in a rock mass under the influence of temperature field,deduces related system equations,and proposes a penalty function method to deal with boundary conditions.Numerical examples are employed to confirm the effectiveness and high accuracy of this method.By the thermal stress analysis of a thick-walled cylinder(TWC),the simulation of cracking in the TWC under heating and cooling conditions,and the simulation of thermal cracking of the SwedishÄspöPillar Stability Experiment(APSE)rock column,the thermal stress,and TM coupling are obtained.The numerical simulation results are in good agreement with the test data and other numerical results,thus verifying the effectiveness of the NMM in dealing with thermal stress and crack propagation problems of fractured rock masses.

A method to interpret fracture aperture of rock slope using adaptive shape and unmanned aerial vehicle multi-angle nap-of-the-object photogrammetry

The aperture of natural rock fractures significantly affects the deformation and strength properties of rock masses,as well as the hydrodynamic properties of fractured rock masses.The conventional measurement methods are inadequate for collecting data on high-steep rock slopes in complex mountainous regions.This study establishes a high-resolution three-dimensional model of a rock slope using unmanned aerial vehicle(UAV)multi-angle nap-of-the-object photogrammetry to obtain edge feature points of fractures.Fracture opening morphology is characterized using coordinate projection and transformation.Fracture central axis is determined using vertical measuring lines,allowing for the interpretation of aperture of adaptive fracture shape.The feasibility and reliability of the new method are verified at a construction site of a railway in southeast Tibet,China.The study shows that the fracture aperture has a significant interval effect and size effect.The optimal sampling length for fractures is approximately 0.5e1 m,and the optimal aperture interpretation results can be achieved when the measuring line spacing is 1%of the sampling length.Tensile fractures in the study area generally have larger apertures than shear fractures,and their tendency to increase with slope height is also greater than that of shear fractures.The aperture of tensile fractures is generally positively correlated with their trace length,while the correlation between the aperture of shear fractures and their trace length appears to be weak.Fractures of different orientations exhibit certain differences in their distribution of aperture,but generally follow the forms of normal,log-normal,and gamma distributions.This study provides essential data support for rock and slope stability evaluation,which is of significant practical importance.

Discontinuity development patterns and the challenges for 3D discrete fracture network modeling on complicated exposed rock surfaces

Natural slopes usually display complicated exposed rock surfaces that are characterized by complex and substantial terrain undulation and ubiquitous undesirable phenomena such as vegetation cover and rockfalls.This study presents a systematic outcrop research of fracture pattern variations in a complicated rock slope,and the qualitative and quantitative study of the complex phenomena impact on threedimensional(3D)discrete fracture network(DFN)modeling.As the studies of the outcrop fracture pattern have been so far focused on local variations,thus,we put forward a statistical analysis of global variations.The entire outcrop is partitioned into several subzones,and the subzone-scale variability of fracture geometric properties is analyzed(including the orientation,the density,and the trace length).The results reveal significant variations in fracture characteristics(such as the concentrative degree,the average orientation,the density,and the trace length)among different subzones.Moreover,the density of fracture sets,which is approximately parallel to the slope surface,exhibits a notably higher value compared to other fracture sets across all subzones.To improve the accuracy of the DFN modeling,the effects of three common phenomena resulting from vegetation and rockfalls are qualitatively analyzed and the corresponding quantitative data processing solutions are proposed.Subsequently,the 3D fracture geometric parameters are determined for different areas of the high-steep rock slope in terms of the subzone dimensions.The results show significant variations in the same set of 3D fracture parameters across different regions with density differing by up to tenfold and mean trace length exhibiting differences of 3e4 times.The study results present precise geological structural information,improve modeling accuracy,and provide practical solutions for addressing complex outcrop issues.

Borehole stability in naturally fractured rocks with drilling mud intrusion and associated fracture strength weakening:A coupled DFN-DEM approach

Borehole instability in naturally fractured rocks poses significant challenges to drilling.Drilling mud invades the surrounding formations through natural fractures under the difference between the wellbore pressure(P w)and pore pressure(P p)during drilling,which may cause wellbore instability.However,the weakening of fracture strength due to mud intrusion is not considered in most existing borehole stability analyses,which may yield significant errors and misleading predictions.In addition,only limited factors were analyzed,and the fracture distribution was oversimplified.In this paper,the impacts of mud intrusion and associated fracture strength weakening on borehole stability in fractured rocks under both isotropic and anisotropic stress states are investigated using a coupled DEM(distinct element method)and DFN(discrete fracture network)method.It provides estimates of the effect of fracture strength weakening,wellbore pressure,in situ stresses,and sealing efficiency on borehole stability.The results show that mud intrusion and weakening of fracture strength can damage the borehole.This is demonstrated by the large displacement around the borehole,shear displacement on natural fractures,and the generation of fracture at shear limit.Mud intrusion reduces the shear strength of the fracture surface and leads to shear failure,which explains that the increase in mud weight may worsen borehole stability during overbalanced drilling in fractured formations.A higher in situ stress anisotropy exerts a significant influence on the mechanism of shear failure distribution around the wellbore.Moreover,the effect of sealing natural fractures on maintaining borehole stability is verified in this study,and the increase in sealing efficiency reduces the radial invasion distance of drilling mud.This study provides a directly quantitative prediction method of borehole instability in naturally fractured formations,which can consider the discrete fracture network,mud intrusion,and associated weakening of fracture strength.The information provided by the numerical approach(e.g.displacement around the borehole,shear displacement on fracture,and fracture at shear limit)is helpful for managing wellbore stability and designing wellbore-strengthening operations.

Creep constitutive model considering nonlinear creep degradation of fractured rock

Stability analysis of underground constructions requires a model study of rock masses’ long-term performance. Creep tests under different stress conditions was conducted on intact granite and granite samples fractured at 30° and 45° angles. The experimental results indicate that the steady creep strain rates of intact and fractured rock present an exponential increase trend with the increase of stress level. A nonlinear creep model is developed based on the experimental results, in which the initial damage caused by fracture together with the damage caused by constant load have been taken into consideration. The fitting analysis results indicated that the model proposed is more accurate at identifying the full creep regions in fractured granite, especially the accelerated stage of creep deformation. The least-square fit error of the proposed creep model is significantly lower than that of Nishihara model by almost an order of magnitude. An analysis of the effects of elastic modulus, viscosity coefficient, and damage factors on fractured rock strain rate and creep strain is conducted. If no consideration is given to the effects of the damage, the proposed nonlinear creep model can degenerate into to the classical Nishihara model.

Sub-Homogeneous Peridynamic Model for Fracture and Failure Analysis of Roadway Surrounding Rock

The surrounding rock of roadways exhibits intricate characteristics of discontinuity and heterogeneity.To address these complexities,this study employs non-local Peridynamics(PD)theory and reconstructs the kernel function to represent accurately the spatial decline of long-range force.Additionally,modifications to the traditional bondbased PD model are made.By considering the micro-structure of coal-rock materials within a uniform discrete model,heterogeneity characterized by bond random pre-breaking is introduced.This approach facilitates the proposal of a novel model capable of handling the random distribution characteristics of material heterogeneity,rendering the PD model suitable for analyzing the deformation and failure of heterogeneous layered coal-rock mass structures.The established numerical model and simulation method,termed the sub-homogeneous PD model,not only incorporates the support effect but also captures accurately the random heterogeneous micro-structure of roadway surrounding rock.The simulation results obtained using this model show good agreement with field measurements from the Fucun coal mine,effectively validating the model’s capability in accurately reproducing the deformation and failure mode of surrounding rock under bolt-supported(anchor cable).The proposed subhomogeneous PD model presents a valuable and effective simulation tool for studying the deformation and failure of roadway surrounding rock in coal mines,offering new insights and potential advancements.

Identifying flow regime in the aquifer of fractured rock system in Germi Chai Basin,Iran

Considering the importance of fractured rock aquifers in the hydrogeologic process,this research aimed to analyze the flow regime,internal degree of karstification,and estimate storage volume in fractured rock aquifers of the Germi Chai Basin in northwest Iran,which is attributed to its active tectonics,erosion,and the lithological diversity.Given the geological setting,the hypothesis is that this basin is characterized by a high degree of karstification and diffuse or intermediate flow regime leading to variation in discharge flow rate.The hydrodynamic and hadrochemical analysis was conducted on 9 well distributed springs across the basin from 2019 to 2020.The maximum flow rate in most of the springs appeared in the early wet season despite their different levels of fluctuations on the monthly discharge time series.Analyzing the spring recession curve form revealed an aquifer containing multiple micro-regimes withαrecession coefficients and a degree of karstification ranging between 0.001 to 0.06 and 0.55 to 2.61,respectively.These findings indicated a dominant diffuse and intermediate flow system resulting from the development of a high density of fractures in this area.The electrical conductivity of the spring changes inversely proportional to the change in flow discharge,indicating the reasonable hydrological response of the aquifer to rainfall events.Hydrograph analysis revealed that the delay time of spring discharge after rainfall events mostly varies between 10 to 30 days.The total dynamic storage volume of the spring for a given period(2019-2020)was estimated to be approximately 1324 million cubic meters reflecting the long-term drainage potential and high perdurability of dynamic storage.Estimating the maximum and minimum ratio revealed that the springs recharging system in Germi Chai Basin comes under the slow aquifers category.This finding provides valuable insight into the hydrogeological properties of fractured rock aquifers contributing to effective water management strategy.

Development and application of novel high‐efficiency composite ultrafine cement grouts for roadway in fractured surrounding rocks

The fractured surrounding rocks of roadways pose major challenges to safe mining.Grouting has often been used to reinforce the surrounding rocks to mitigate the safety risks associated with fractured rocks.The aim of this study is to develop highly efficient composite ultrafine cement(CUC)grouts to reinforce the roadway in fractured surrounding rocks.The materials used are ultrafine cement(UC),ultrafine fly ash(UF),ultrafine slag(US),and additives(superplasticizer[SUP],aluminate ultrafine expansion agent[AUA],gypsum,and retarder).The fluidity,bleeding,shrinkage,setting time,chemical composition,microstructure,degree of hydration,and mechanical property of grouting materials were evaluated in this study.Also,a suitable and effective CUC grout mixture was used to reinforce the roadway in the fractured surrounding rock.The results have shown that the addition of UF and US reduces the plastic viscosity of CUC,and the best fluidity can be obtained by adding 40%UF and 10%US.Since UC and UF particles are small,the pozzolanic effect of UF promotes the hydration reaction,which is conductive to the stability of CUC grouts.In addition,fine particles of UC,UF,and US can effectively fill the pores,while the volumetric expansion of AUA and gypsum decreases the pores and thus affects the microstructure of the solidified grout.The compressive test results have shown that the addition of specific amounts of UF and US can ameliorate the mechanical properties of CUC grouts.Finally,the CUC22‐8 grout was used to reinforce the No.20322 belt roadway.The results of numerical simulation and field monitoring have indicated that grouting can efficaciously reinforce the surrounding rock of the roadway.In this research,high‐performance CUC grouts were developed for surrounding rock reinforcement of underground engineering by utilizing UC and some additives.

Impact of effective stress on permeability for carbonate fractured-vuggy rocks

To gain insight into the flow mechanisms and stress sensitivity for fractured-vuggy reservoirs,several core models with different structural characteristics were designed and fabricated to investigate the impact of effective stress on permeability for carbonate fractured-vuggy rocks(CFVR).It shows that the permeability performance curves under different pore and confining pressures(i.e.altered stress conditions)for the fractured core models and the vuggy core models have similar change patterns.The ranges of permeability variation are significantly wider at high pore pressures,indicating that permeability reduction is the most significant during the early stage of development for fractured-vuggy reservoirs.Since each obtained effective stress coefficient for permeability(ESCP)varies with the changes in confining pressure and pore pressure,the effective stresses for permeability of four representative CFVR show obvious nonlinear characteristics,and the variation ranges of ESCP are all between 0 and 1.Meanwhile,a comprehensive ESCP mathematical model considering triple media,including matrix pores,fractures,and dissolved vugs,was proposed.It is proved theoretically that the ESCP of CFVR generally varies between 0 and 1.Additionally,the regression results showed that the power model ranked highest among the four empirical models mainly applied in stress sensitivity characterization,followed by the logarithmic model,exponential model,and binomial model.The concept of“permeability decline rate”was introduced to better evaluate the stress sensitivity performance for CFVR,in which the one-fracture rock is the strongest,followed by the fracture-vug rock and two-horizontalfracture rock;the through-hole rock is the weakest.In general,this study provides a theoretical basis to guide the design of development and adjustment programs for carbonate fractured-vuggy reservoirs.

Real-time arrival picking of rock microfracture signals based on convolutional-recurrent neural network and its engineering application

Accurately picking P-and S-wave arrivals of microseismic(MS)signals in real-time directly influences the early warning of rock mass failure.A common contradiction between accuracy and computation exists in the current arrival picking methods.Thus,a real-time arrival picking method of MS signals is constructed based on a convolutional-recurrent neural network(CRNN).This method fully utilizes the advantages of convolutional layers and gated recurrent units(GRU)in extracting short-and long-term features,in order to create a precise and lightweight arrival picking structure.Then,the synthetic signals with field noises are used to evaluate the hyperparameters of the CRNN model and obtain an optimal CRNN model.The actual operation on various devices indicates that compared with the U-Net method,the CRNN method achieves faster arrival picking with less performance consumption.An application of large underground caverns in the Yebatan hydropower station(YBT)project shows that compared with the short-term average/long-term average(STA/LTA),Akaike information criterion(AIC)and U-Net methods,the CRNN method has the highest accuracy within four sampling points,which is 87.44%for P-wave and 91.29%for S-wave,respectively.The sum of mean absolute errors(MAESUM)of the CRNN method is 4.22 sampling points,which is lower than that of the other methods.Among the four methods,the MS sources location calculated based on the CRNN method shows the best consistency with the actual failure,which occurs at the junction of the shaft and the second gallery.Thus,the proposed method can pick up P-and S-arrival accurately and rapidly,providing a reference for rock failure analysis and evaluation in engineering applications.

Dynamic response and failure process of horizontal-layered fractured structure rock slope under strong earthquake

Rock slope with horizontal-layered fractured structure(HLFS)has high stability in its natural state.However,a strong earthquake can induce rock fissure expansion,ultimately leading to slope failure.In this study,the dynamic response,failure mode,and spectral characteristics of rock slope with HLFS under strong earthquake conditions were investigated based on the large-scale shaking table model test.On this basis,multiple sets of numerical calculation models were further established by UDEC discrete element program.Five influencing factors were considered in the parametric study of numerical simulations,including slope height,slope angle,bedding-plane spacing and secondary joint spacing as well as bedrock dip angle.The results showed that the failure process of rock slope with HLFS under earthquake action is mainly divided into four phases,i.e.,the tensile crack of the slope shoulder joints and shear dislocation at the top bedding plane,the extension of vertical joint cracks and increase of shear displacement,the formation of step-through sliding surfaces and the instability,and finally collapse of fractured rock mass.The acceleration response of slopes exhibits elevation amplification effect and surface effect.Numerical simulations indicate that the seismic stability of slopes with HLFS exhibits a negative correlation with slope height and angle,but a positive correlation with bedding-plane spacing,joint spacing,and bedrock dip angle.The results of this study can provide a reference for seismic stability evaluation of weathered rock slopes.

Failure characteristics and fracture mechanism of overburden rock induced by mining:A case study in China

This study employs similar simulation testing and discrete element simulation coupling to analyze the failure and deformation processes of a model coal seam's roof.The caving area of the overburden rock is divided into three zones:the delamination fracture zone,broken fracture zone,and compaction zone.The caving and fracture zones'heights are approximately 110 m above the coal seam,with a maximum subsidence of 11 m.The delamination fracture zone's porosity range is between 0.2 and 0.3,while the remainder of the roof predominantly exhibits a porosity of less than 0.1.In addition,the numerical model's stress analysis revealed that the overburden rock's displacement zone forms an'arch-beam'structure starting from 160 m,with the maximum and minimum stress values decreasing as the distance of advancement increases.In the stress beam interval of the overburden rock,the maximum value changes periodically as the advancement distance increases.Based on a comparative analysis between observable data from on-site work and numerical simulation results,the stress data from the numerical simulation are essentially consistent with the actual results detected on-site,indicating the validity of the numerical simulation results.

Effects of thawing-induced softening on fracture behaviors of frozen rock

Due to the presence of ice and unfrozen water in pores of frozen rock,the rock fracture behaviors are susceptible to temperature.In this study,the potential thawing-induced softening effects on the fracture behaviors of frozen rock is evaluated by testing the tension fracture toughness(KIC)of frozen rock at different temperatures(i.e.-20℃,-15℃,-12℃,-10℃,-8℃,-6℃,-4℃,-2℃,and 0℃).Acoustic emission(AE)and digital image correlation(DIC)methods are utilized to analyze the microcrack propagation during fracturing.The melting of pore ice is measured using nuclear magnetic resonance(NMR)method.The results indicate that:(1)The KIC of frozen rock decreases moderately between-20℃ and-4℃,and rapidly between-4℃ and 0℃.(2)At-20℃ to-4℃,the fracturing process,deduced from the DIC results at the notch tip,exhibits three stages:elastic deformation,microcrack propagation and microcrack coalescence.However,at-4℃e0℃,only the latter two stages are observed.(3)At-4℃e0℃,the AE activities during fracturing are less than that at-20℃ to-4℃,while more small events are reported.(4)The NMR results demonstrate a reverse variation trend in pore ice content with increasing temperature,that is,a moderate decrease is followed by a sharp decrease and-4℃ is exactly the critical temperature.Next,we interpret the thawing-induced softening effect by linking the evolution in microscopic structure of frozen rock with its macroscopic fracture behaviors as follow:from-20℃ to-4℃,the thickening of the unfrozen water film diminishes the cementation strength between ice and rock skeleton,leading to the decrease in fracture parameters.From-4℃ to 0℃,the cementation effect of ice almost vanishes,and the filling effect of pore ice is reduced significantly,which facilitates microcrack propagation and thus the easier fracture of frozen rocks.

P-and SV-wave dispersion and attenuation in saturated microcracked porous rock with aligned penny-shaped fractures

P-and SV-wave dispersion and attenuation have been extensively investigated in saturated poroelastic media with aligned fractures.However,there are few existing models that incorporate the multiple wave attenuation mechanisms from the microscopic scale to the macroscopic scale.Hence,in this work,we developed a unified model to incorporate the wave attenuation mechanisms at different scales,which includes the microscopic squirt flow between the microcracks and pores,the mesoscopic wave-induced fluid flow between fractures and background(FB-WIFF),and the macroscopic Biot's global flow and elastic scattering(ES)from the fractures.Using Tang's modified Biot's theory and the mixed-boundary conditions,we derived the exact frequency-dependent solutions of the scattering problem for a single penny-shaped fracture with oblique incident P-and SV-waves.We then developed theoretical models for a set of aligned fractures and randomly oriented fractures using the Foldy approximation.The results indicated that microcrack squirt flow considerably influences the dispersion and attenuation of P-and SV-wave velocities.The coupling effects of microcrack squirt flow with the FB-WIFF and ES of fractures cause much higher velocity dispersion and attenuation for P waves than for SV waves.Randomly oriented fractures substantially reduce the attenuation caused by the FB-WIFF and ES,particularly for the ES attenuation of SV waves.Through a comparison with existing models in the limiting cases and previous experimental measurements,we validated our model.

Numerical investigation of geostress influence on the grouting reinforcement effectiveness of tunnel surrounding rock mass in fault fracture zones

Grouting is a widely used approach to reinforce broken surrounding rock mass during the construction of underground tunnels in fault fracture zones,and its reinforcement effectiveness is highly affected by geostress.In this study,a numerical manifold method(NMM)based simulator has been developed to examine the impact of geostress conditions on grouting reinforcement during tunnel excavation.To develop this simulator,a detection technique for identifying slurry migration channels and an improved fluid-solid coupling(FeS)framework,which considers the influence of fracture properties and geostress states,is developed and incorporated into a zero-thickness cohesive element(ZE)based NMM(Co-NMM)for simulating tunnel excavation.Additionally,to simulate coagulation of injected slurry,a bonding repair algorithm is further proposed based on the ZE model.To verify the accuracy of the proposed simulator,a series of simulations about slurry migration in single fractures and fracture networks are numerically reproduced,and the results align well with analytical and laboratory test results.Furthermore,these numerical results show that neglecting the influence of geostress condition can lead to a serious over-estimation of slurry migration range and reinforcement effectiveness.After validations,a series of simulations about tunnel grouting reinforcement and tunnel excavation in fault fracture zones with varying fracture densities under different geostress conditions are conducted.Based on these simula-tions,the influence of geostress conditions and the optimization of grouting schemes are discussed.

Effect of dynamic loading orientation on fracture properties of surrounding rocks in twin tunnels

For expedited transportation,vehicular tunnels are often designed as two adjacent tunnels,which frequently experience dynamic stress waves from various orientations during blasting excavation.To analyze the impact of dynamic loading orientation on the stability of the twin-tunnel,a split Hopkinson pressure bar(SHPB)apparatus was used to conduct a dynamic test on the twin-tunnel specimens.The two tunnels were rotated around the specimen’s center to consider the effect of dynamic loading orientation.LS-DYNA software was used for numerical simulation to reveal the failure properties and stress wave propagation law of the twin-tunnel specimens.The findings indicate that,for a twin-tunnel exposed to a dynamic load from different orientations,the crack initiation position appears most often at the tunnel corner,tunnel spandrel,and tunnel floor.As the impact direction is created by a certain angle(30°,45°,60°,120°,135°,and 150°),the fractures are produced in the middle of the line between the left tunnel corner and the right tunnel spandrel.As the impact loading angle(a)is 90°,the tunnel sustains minimal damage,and only tensile fractures form in the surrounding rocks.The orientation of the impact load could change the stress distribution in the twin-tunnel,and major fractures are more likely to form in areas where the tensile stress is concentrated.

贝母素乙调节RhoA/ROCK信号通路对膝骨关节炎大鼠软骨损伤的影响

目的 探讨贝母素乙调节Ras同源基因家族成员A(RhoA)/Rho相关的卷曲螺旋激酶(ROCK)信号通路对膝骨关节炎(knee osteoarthritis, KOA)大鼠软骨损伤的影响。方法 以碘乙酸钠(MIA)法制备KOA大鼠模型,随机分为模型组、贝母素乙(3.5 mg/kg)组、贝母素乙(3.5 mg/kg)+LPA(RhoA激活剂,1 mg/kg)组,每组10只,另选10只大鼠为对照组。机械刺激法和热辐射法检测大鼠膝关节疼痛症状;检测大鼠关节肿胀度、膝关节宽度并以步态评分评测其关节功能;透射电镜检测大鼠膝关节软骨细胞超微结构;TUNEL染色检测大鼠关节软骨细胞凋亡比;酶联免疫吸附(ELISA)法检测大鼠血清和关节液促炎因子白细胞介素(IL)-18、单核细胞趋化蛋白1(MCP-1)水平;免疫印迹检测各组大鼠关节软骨组织凋亡(Cleaved caspase-3、Bax)和RhoA/ROCK通路相关蛋白表达。结果 贝母素乙可降低KOA大鼠关节肿胀度、膝关节宽度、步态评分、软骨细胞凋亡比、血清及关节液IL-18、MCP-1水平、软骨组织Cleaved caspase-3、Bax、RhoA、ROCK1、ROCK1蛋白表达,升高机械痛阈值、热痛阈值(P<0.05);LPA可减弱贝母素乙对KOA大鼠关节软骨损伤的改善作用。结论 贝母素乙可通过抑制RhoA/ROCK信号激活而抑制KOA大鼠关节炎症反应和关节软骨细胞凋亡,减轻软骨损伤。

桃叶珊瑚苷调节RhoA/ROCK信号通路对胶质母细胞瘤细胞活力和上皮间质转化的影响

[目的]研究桃叶珊瑚苷(AU)对胶质母细胞瘤(GBM)细胞活力和上皮间质转化(EMT)的影响,并对其作用机制进行探讨。[方法]将U87细胞随机分为对照组、AU低浓度组、AU中浓度组、AU高浓度组、Y-27632组、AU高浓度+Y-27632组。细胞计数器试剂盒(CCK-8)法检测细胞活力,流式细胞术检测细胞凋亡,Transwell小室实验检测细胞迁移和侵袭,蛋白免疫印迹法(Western Blot)检测基质金属蛋白酶(MMP)2、MMP9、波形蛋白(Vimentin)、上皮钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)、Ras同源基因家族成员A(RhoA)、Rho相关卷曲螺旋蛋白激酶(ROCK)1、ROCK2表达。构建GBM裸鼠模型,随机分为裸鼠对照组、AU组、Y-27632组、AU+Y-27632组,测量肿瘤质量与体积,免疫组化法检测移植瘤组织RhoA、ROCK1、ROCK2蛋白表达。[结果]GBM细胞活力随着AU浓度的升高而逐渐降低(P<0.05),选择U87作为后续实验细胞,选择10、25、50μmol/L浓度作为AU后续实验浓度。与对照组比较,AU低、中、高浓度组和Y-27632组细胞活力、迁移和侵袭细胞数、MMP2、MMP9、N-cadherin、Vimentin、RhoA、ROCK1、ROCK2表达显著下降,凋亡率、E-cadherin表达显著升高(P<0.05),其中高浓度AU和Y-27632组共同处理的细胞变化更显著(P<0.05)。AU和Y-27632均能抑制移植瘤质量和体积,降低RhoA/ROCK信号通路蛋白表达(P<0.05)。[结论]AU能抑制GBM细胞活力、迁移侵袭和EMT,促进细胞凋亡,其作用机制可能与抑制RhoA/ROCK信号通路有关。

SLC9A3-AS1调控miR-148a-3p/ROCK1信号轴影响肾癌细胞生物学功能

目的检测lncRNA SLC9A3-AS1在肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)组织与肾癌细胞中的表达水平,探讨其促进肾癌细胞恶性生物学行为的机制。方法应用GEPIA2在线软件(http://gepia2.cancer-pku.cn/)分析TCGA数据库中SLC9A3-AS1在ccRCC组织中的表达水平;采用实时荧光定量聚合酶链反应(qRT-PCR)检测SLC9A3-AS1在不同肾癌细胞系、24例ccRCC组织与癌旁正常肾脏组织中的表达水平;应用细胞增殖/毒性检测试剂盒(cell counting kit-8,CCK-8)和Transwell小室迁移实验检测敲低SLC9A3-AS1表达对肾癌细胞增殖与迁移的影响;应用蛋白质免疫印迹法检测增殖与迁移相关信号通路蛋白的表达水平;采用双荧光素酶报告基因实验验证SLC9A3-AS1与miR-148a-3p/ROCK1轴的靶向调控关系。结果GEPIA2软件分析结果显示,相较正常肾脏组织,SLC9A3-AS1在肾透明细胞癌组织中表达显著上调(P<0.01)。qRT-PCR结果显示,相较癌旁正常肾脏组织,SLC9A3-AS1在24例ccRCC组织中表达显著上调(P<0.01);相较永生化肾小管上皮细胞,SLC9A3-AS1在4种肾癌细胞系中表达均显著上调,以786-O细胞最为显著(P<0.01)。干扰SLC9A3-AS1表达,可显著抑制786-O细胞的增殖与迁移能力,上调E-cadherin的蛋白表达水平,而下调N-cadherin、MMP2的蛋白表达水平(均P<0.05);过表达miR-148a-3p可显著抑制786-O细胞的增殖与迁移能力(P<0.01)。双荧光素酶报告基因检测结果表明,SLC9A3-AS1可特异性结合miR-148a-3p,后者进一步靶向结合ROCK1 mRNA的3′UTR区域;过表达miR-148a-3p可明显下调ROCK1 mRNA的表达水平,敲低miR-148a-3p表达则产生相反的效应;敲低SLC9A3-AS1表达可显著下调786-O细胞中ROCK1 mRNA与蛋白的表达水平(P<0.01);下调miR-148a-3p表达可部分逆转SLC9A3-AS1沉默对786-O细胞增殖与迁移的抑制作用(P<0.05)。结论SLC9A3-AS1在ccRCC中通过调控miR-148a-3p/ROCK1轴发挥促癌作用,有望成为ccRCC的一个新的分子标志物。

In-situ observations of damage-fracture evolution in surrounding rock upon unloading in 2400-m-deep tunnels

The damage-fracture evolution of deep rock mass has obvious particularity,which is revealed in 2400-mdeep tunnels by field tests.The evolution of the excavation damaged zone depth is consistent with that of the fractured zone depth.The ratio of the excavation damaged zone depth to the excavation fractured zone depth is greater than 2.0 in a rock mass with both high strength and good integrity,but less than1.5 in a rock mass with lower strength or poor integrity.Zonal disintegration in a rock mass with high strength and fair integrity is more likely to occur when it contains more than two groups of primary fractures in damaged zones.Fractures develop outward in zonal disintegration but are totally different from the single-zone fracture,in which the fractures develop inward,and it is the starting position of the fractured zone when the excavation surface of the middle pilot is 7–9 m close to the pre-set borehole and it stops after the excavation surface of the baseplate is 11–14 m away.The most intense evolution occurs around 2–4 m from the pre-set borehole in the sidewall expansion stage.The research results provide a reference for the monitoring scheme and support design of CJPL-Ⅲin its future construction.