Hydraulic fracturing behaviors of shale under coupled stress and temperature conditions simulating different burial depths

Fracture propagation in shale under in situ conditions is a critical but poorly understood mechanical process in hydraulic fracturing for deep shale gas reservoirs. To address this, hydraulic fracturing experiments were conducted on hollow double-wing crack specimens of the Longmaxi shale under conditions simulating the ground surface(confining pressure σ_(cp)=0, room temperature(Tr)) and at depths of 1600 m(σ_(cp)=40 MPa, Ti=70 ℃) and 3300 m(σ_(cp)=80 MPa, high temperature Ti=110 ℃) in the study area.High in situ stress was found to significantly increase fracture toughness through constrained microcracking and particle frictional bridging mechanisms. Increasing the temperature enhances rather than weakens the fracture resistance because it increases the grain debonding length, which dissipates more plastic energy and enlarges grains to close microdefects and generate compressive stress to inhibit microcracking. Interestingly, the fracture toughness anisotropy in the shale was found to be nearly constant across burial depths, despite reported variations with increasing confining pressure. Heated water was not found to be as important as the in situ environment in influencing shale fracture. These findings emphasize the need to test the fracture toughness of deep shales under coupled in situ stress and temperature conditions rather than focusing on either in situ stress or temperature alone.

Application of graph neural network and feature information enhancement in relation inference of sparse knowledge graph

At present,knowledge embedding methods are widely used in the field of knowledge graph(KG)reasoning,and have been successfully applied to those with large entities and relationships.However,in research and production environments,there are a large number of KGs with a small number of entities and relations,which are called sparse KGs.Limited by the performance of knowledge extraction methods or some other reasons(some common-sense information does not appear in the natural corpus),the relation between entities is often incomplete.To solve this problem,a method of the graph neural network and information enhancement is proposed.The improved method increases the mean reciprocal rank(MRR)and Hit@3 by 1.6%and 1.7%,respectively,when the sparsity of the FB15K-237 dataset is 10%.When the sparsity is 50%,the evaluation indexes MRR and Hit@10 are increased by 0.8%and 1.8%,respectively.

Edge modes in finite-size systems with different edge terminals

We investigate the behavior of edge modes in the presence of different edge terminations and long-range(LR)hopping.Here,we mainly focus on such model crystals with two different types of structures(type I:“…-P-Q-P-Q-…”and type II:“…=P-Q=P-Q=…”),where P and Q represent crystal lines(CLs),while the symbols“-”and“=”denote the distance between the nearest neighbor(NN)CLs.Based on the lattice model Hamiltonian with LR hopping,the existence of edge modes is determined analytically by using the transfer matrix method(TMM)when different edge terminals are taken into consideration.Our findings are consistent with the numerical results obtained by the exact diagonalization method.We also notice that edge modes can exhibit different behaviors under different edge terminals.Our result is helpful in solving novel edge modes in honeycomb crystalline graphene and transition metal dichalcogenides with different edge terminals.

不同荷载作用下五边形石墨烯的变形破坏机理研究

五边形石墨烯是一种完全由碳五元环组成的准二维的亚稳态碳结构,在不同载荷作用下其变形破坏机理仍需进一步研究.本文基于ReaxFF反应力场采用分子动力学方法模拟研究了五边形石墨烯的拉伸、剪切和纳米压痕破坏过程,得到了五边形石墨烯的拉伸和剪切应力-应变曲线以及压入载荷-位移曲线,系统分析了五边形石墨烯的变形破坏机理,并验证了五边形石墨烯在不同载荷作用下是否表现出塑性变形特征的一致性问题.研究结果表明,通过拉伸或纳米压入等不同加载方式均可准确测出五边形石墨烯的本征力学性能参数,其杨氏模量265.4~285.1 N/m,与第一性原理计算结果一致.研究还发现,五边形石墨烯在拉伸、剪切和纳米压痕过程中均会出现不可逆的塑性变形特征,而引起塑性变形的原因是由于不可逆的碳五元环向碳多元环结构的转变.以上研究结果可为基于五边形石墨烯的微/纳米机电系统的实际应用提供重要指导.

Ligustilide protects PC12 cells from oxygen-glucose deprivation/reoxygenation-induced apoptosis via the LKB1-AMPK-mTOR signaling pathway

Autophagy has been shown to have a protective effect against brain damage.Ligustilide(LIG)is a bioactive substance isolated from Ligusticum chuanxiong,a traditional Chinese medicine.LIG has a neuroprotective effect;however,it is unclear whether this neuroprotective effect involves autophagy.In this study,PC12 cells were treated with 1×10^-5–1×10^-9 M LIG for 0,3,12 or 24 hours,and cell proliferation was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS)assay.Treatment with 1×10^-6 M LIG for 3 hours had the greatest effect on cell proliferation,and was therefore used for subsequent experiments.PC12 cells were pre-treated with 1×10^-6 M LIG for 3 hours,cultured in 95%N2/5%CO2 in Dulbecco’s modified Eagle’s medium without glucose or serum for 4 hours,and then cultured normally for 16 hours,to simulate oxygen-glucose deprivation/reoxygenation(OGD/R).Cell proliferation was assessed with the MTS assay.Apoptosis was detected by flow cytometry.The expression levels of apoptosis-related proteins,Bcl-2 and Bax,autophagy-related proteins,Beclin 1 and microtubule-associated protein l light chain 3B(LC3-II),and liver kinase B1(LKB1)-5′-adenosine monophosphate-activated protein kinase(AMPK)-mammalian target of rapamycin(mTOR)signaling pathway-related proteins were assessed by western blot assay.Immunofluorescence staining was used to detect LC3-II expression.Autophagosome formation was observed by electron microscopy.LIG significantly decreased apoptosis,increased Bcl-2,Beclin 1 and LC3-II expression,decreased Bax expression,increased LC3-II immunoreactivity and the number of autophagosomes,and activated the LKB1-AMPK-mTOR signaling pathway in PC12 cells exposed to OGD/R.The addition of the autophagy inhibitor 3-methyladenine or dorsomorphin before OGD/R attenuated the activation of the LKB1-AMPK-mTOR signaling pathway in cells treated with LIG.Taken together,our findings show that LIG promotes autophagy and protects PC12 cells from apoptosis induced by OGD/R via the LKB1-AMPK-mTOR signaling pathway.

Robot-assisted one-stage resection of rectal cancer with liver and lung metastases

The Da Vinci Surgical System may help to overcome some of the difficulties of laparoscopy for complicated abdominal surgery.The authors of this article present a case of robot-assisted, one-stage radical resection of three tumors, including robotic anterior resection for rectal cancer, segmental hepatectomy for liver metastasis, and wedge-shaped excision for lung metastasis.A 59-year-old man with primary rectal cancer and liver and lung metastases was operated upon with a one-stage radical resection approach using the Da Vinci Surgical System.Resection and anastomosis of rectal cancer were performed extracorporeally afterundocking the robot.The procedure was successfully completed in 500 min.No surgical complications occurred during the intervention and postoperative period, and no conversion to laparotomy or additional trocars were required.To the best of our knowledge, this is the first case of simultaneous resection for rectal cancer with liver and lung metastases using the Da Vinci Surgery System to be reported.The procedure is feasible and safe and its main advantages for patient are avoiding repeated operation, reducing surgical trauma, shortening recovery time, and early implementation of postoperative adjuvant therapy.

Cognitive deficits and Alzheimer-like neuropathological impairments during adolescence in a rat model of type 2 diabetes mellitus

Numerous studies have shown that many patients who suffer from type 2 diabetes mellitus exhibit cognitive dysfunction and neuronal synaptic impairments. Therefore, growing evidence suggests that type 2 diabetes mellitus has a close relationship with occurrence and progression of neurodegeneration and neural impairment in Alzheimer＇s disease. However, the relationship between metabolic disorders caused by type 2 diabetes mellitus and neurodegeneration and neural impairments in Alzheimer＇s disease is still not fully determined. Thus, in this study, we replicated a type 2 diabetic animal model by subcutaneous injection of newborn Sprague-Dawley rats with monosodium glutamate during the neonatal period. At 3 months old, the Barnes maze assay was performed to evaluate spatial memory function. Microelectrodes were used to measure electrophysiological function in the hippocampal CA1 region. Western blot assay was used to determine expression levels of glutamate ionotropic receptor NMDA type subunit 2 A（GluN2A） and GluN2B in the hippocampus. Enzyme-linked immunosorbent assay was used to determine levels of interleukin-1β, tumor necrosis factor α, and interleukin-6 in the hippocampus and cerebral cortex, as well as hippocampal amyloid beta（Aβ）1-40 and Aβ_（1-42） levels. Our results showed that in the rat model of type 2 diabetes mellitus caused by monosodium glutamate exposure during the neonatal period, latency was prolonged and the number of errors increased in the Barnes maze. Further, latency was increased and time in the escape platform quadrant shortened. Number of times crossing the platform was also reduced in the Morris water maze. After high frequency stimulation of the hippocampus, synaptic transmission was inhibited, expression of GluN2A and GluN2B were decreased in the hippocampus, expression of interleukin 1β, interleukin 6, and tumor necrosis factor α was increased in the hippocampus and cortex, and levels of Aβ_（1-40） and Aβ_（1-42） were increased in the hippocampus. These findings confirm that type 2 diabetes mellitus induced by neonatal monosodium glutamate exposure results in Alzheimer-like neuropathological changes and further causes cognitive deficits and neurodegeneration in young adulthood.

Nanostructuring of Mg-Based Hydrogen Storage Materials:Recent Advances for Promoting Key Applications

With the depletion of fossil fuels and global warming,there is an urgent demand to seek green,low-cost,and high-efficiency energy resources.Hydrogen has been considered as a potential candidate to replace fossil fuels,due to its high gravimetric energy density(142 MJ kg^(-1)),high abundance(H_(2)O),and environmentalfriendliness.However,due to its low volume density,effective and safe hydrogen storage techniques are now becoming the bottleneck for the"hydrogen economy".Under such a circumstance,Mg-based hydrogen storage materials garnered tremendous interests due to their high hydrogen storage capacity(~7.6 wt%for MgH_(2)),low cost,and excellent reversibility.However,the high thermodynamic stability(ΔH=-74.7 kJ mol^(-1)H_(2))and sluggish kinetics result in a relatively high desorption temperature(>300℃),which severely restricts widespread applications of MgH_(2).Nano-structuring has been proven to be an effective strategy that can simultaneously enhance the ab/de-sorption thermodynamic and kinetic properties of MgH_(2),possibly meeting the demand for rapid hydrogen desorption,economic viability,and effective thermal management in practical applications.Herein,the fundamental theories,recent advances,and practical applications of the nanostructured Mg-based hydrogen storage materials are discussed.The synthetic strategies are classified into four categories:free-standing nano-sized Mg/MgH_(2)through electrochemical/vapor-transport/ultrasonic methods,nanostructured Mg-based composites via mechanical milling methods,construction of core-shell nano-structured Mg-based composites by chemical reduction approaches,and multi-dimensional nano-sized Mg-based heterostructure by nanoconfinement strategy.Through applying these strategies,near room temperature ab/de-sorption(<100℃)with considerable high capacity(>6 wt%)has been achieved in nano Mg/MgH_(2)systems.Some perspectives on the future research and development of nanostructured hydrogen storage materials are also provided.

Oxygen Vacancy-Rich 2D TiO_(2) Nanosheets:A Bridge Toward High Stability and Rapid Hydrogen Storage Kinetics of Nano-Confined MgH_(2)

MgH_(2) has attracted intensive interests as one of the most promising hydrogen storage materials.Nevertheless,the high desorption temperature,sluggish kinetics,and rapid capacity decay hamper its commercial application.Herein,2D TiO_(2) nanosheets with abundant oxygen vacancies are used to fabricate a flower-like MgH_(2)/TiO_(2) heterostructure with enhanced hydrogen storage performances.Particularly,the onset hydrogen desorption temperature of the MgH_(2)/TiO_(2) heterostructure is lowered down to 180℃(295℃ for blank MgH_(2)).The initial desorption rate of MgH_(2)/TiO_(2) reaches 2.116 wt% min^(-1) at 300℃,35 times of the blank MgH_(2) under the same conditions.Moreover,the capacity retention is as high as 98.5% after 100 cycles at 300℃,remarkably higher than those of the previously reported MgH_(2)-TiO_(2) composites.Both in situ HRTEM observations and ex situ XPS analyses confirm that the synergistic effects from multi-valance of Ti species,accelerated electron transportation caused by oxygen vacancies,formation of catalytic Mg-Ti oxides,and stabilized MgH_(2) NPs confined by TiO_(2) nanosheets contribute to the high stability and kinetically accelerated hydrogen storage performances of the composite.The strategy of using 2D substrates with abundant defects to support nano-sized energy storage materials to build heterostructure is therefore promising for the design of high-performance energy materials.

Rockburst process and strength-weakening effect of the high-stress circular tunnel under internal unloading

To investigate the influence of unloading effect of a circular tunnel face on rockburst process,by innovatively combining rock drilling unloading devices and triaxial systems,the strain rockburst simulation under the entire stress path of“high initial stressþinternal unloadingþstress adjustment”(HUS test)was realized for the intact cubic red sandstone samples(100 mm×100 mm×100 mm).Comparative tests were conducted on cubic red sandstone samples with prefabricated circular holes(425 mm)under the stress path of“prefabricated circular hole+þhigh initial stress+stress adjustment”(PHS test),thereby highlighting the influence of internal unloading on rockburst failure.The test results revealed that with an increase in vertical stress,the sidewalls in both the HUS and PHS tests suffered strain rockburst failure.Compared with the PHS test,the initial failure stress in the HUS test is lower,and it is easier to induce sidewall rockbursts.This indicates that the internal unloading influences the sidewall failure,causing an obvious strength-weakening effect,which becomes more significant with an increase in buried depth.The strain rockburst failure was more severe in the HUS test owing to the influence of internal unloading.V-shaped rockburst pits were formed in the HUS tests,whereas in the PHS test,arcshaped rockburst pits were produced.It was also found that strain rockburst failure may occur only when the rock has a certain degree of rockburst proneness.

Modeling Hydrothermal Transfer Processes in Permafrost Regions of Qinghai-Tibet Plateau in China

Hydrothermal processes are key components in permafrost dynamics; these processes are integral to global wanning. In this study the coupled heat and mass transfer model for （CoupModel） the soil-plant-atmosphere-system is applied in high-altitude permafrost regions and to model hydrothermal transfer processes in freeze-thaw cycles. Measured meteorological forcing and soil and vegetation properties are used in the CoupModel for the period from January 1, 2009 to December 31, 2012 at the Tanggula observation site in the Qinghai-Tibet Plateau. A 24-h time step is used in the model simulation. The results show that the simulated soil temperature and water content, as well as the frozen depth compare well with the measured data. The coefficient of determination （R2） is 0.97 for the mean soil temperature and 0.73 for the mean soil water content, respectively. The simulated soil heat flux at a depth of 0-20 cm is also consistent with the monitored data. An analysis is performed on the simulated hydrothermal transfer processes from the deep soil layer to the upper one during the freezing and thawing period. At the beginning of the freezing period, the water in the deep soil layer moves upward to the freezing front and releases heat during the freezing process. When the soil layer is completely frozen, there are no vertical water ex- changes between the soil layers, and the heat exchange process is controlled by the vertical soil temperature gradient. During the thaw- ing period, the downward heat process becomes more active due to increased incoming shortwave radiation at the ground surface. The melt water is quickly dissolved in the soil, and the soil water movement only changes in the shallow soil layer. Subsequently, the model was used to provide an evaluation of the potential response of the active layer to different scenarios of initial water content and climate warming at the Tanggula site. The results reveal that the soil water content and the organic layer provide protection against active layer deepening in summer, so climate warming will cause the permafrost active layer to become deeoer and permafrost degradation.

Primary colonic melanoma presenting as ileocecal intussusception: Case report and literature review

Primary malignant melanoma originating in the colon is an extremely rare disease.Herein,we report a case of primary melanoma of the ascending colon.The patient was a 57-year-old male who was admitted to our hospital for persistent abdominal pain and episodes of bloody stool,nausea and vomiting.A computed tomography scan revealed lower intestinal intussusception and enlarged lymph nodes in the abdominal cavity and retroperitoneum.During laparoscopic operation,multiple enlarged lymph nodes were found.Several segments of the proximal small intestine were incarcerated into the distal small intestine,forming an internal hernia and obstruction.The necrotic terminal ileum was invaginated into the ascending cecum.Subsequently,adhesive internal hernia reduction and palliative right hemicolectomy were performed.Pathologic examination of the excised specimen revealed a polypoid mass in the ascending colon.Histological examination showed epithelioid and spindle tumor cells with obvious cytoplasmic melanin deposition.Immunohistochemical staining revealed that the tumor cells were positive for S-100,HmB-45 and vimentin,confirming the diagnosis of melanoma.The patient history and a thorough postoperative investigation excluded the preexistence or coexistence of a primary lesion elsewhere in the skin,anus or oculus or at other sites.Thus,we consider our case to represent an aggressive primary colon melanoma presenting as ileocecal intussusception and intestinal obstruction.

2017 Chinese expert consensus on the clinical application ofserum marker for thyroid cancer

In recent years, the clinical incidence of thyroid cancer has been increasing year by year, and its risk assessment and clinical management methods have also been accordingly modified and constantly improved. There are great differences between the clinical diagnostic and therapeutic modes and disease management of thyroid cancer employed by various medical institutions in China, particularly with regard to the clinical application of serum marker of thyroid cancer. To this end, the China Anti-Cancer Association Thyroid Cancer Specialized Committee Chinese Association of Thyroid Oncology organized this compilation of ExpertConsensus on Clinical Application of Serum Marker of Thyroid Cancer to help and impel relevant clinical institutions and professionals to standardize clinical diagnosis, treatment, and long-term management of thyroid cancer, and to properly utilize the serum marker for scientific auxiliary clinical diagnosis and assessment of thyroid cancer before and after operation.

Epithelial-mesenchymal transition in colorectal cancer tissue of patients with Lynch syndrome

AIM: To explore the epithelial-mesenchymal transition (EMT) in tissue from patients with Lynch syndrome, and to interpret biological behaviour of Lynch syndrome.

Individualized Treatment and Palliative Care for A90-Year-Old Patient with Primary Gastric Diffuse Large-B Cell Lymphoma:4 Year Follow-up and Inspiration

A 90-year-old man was diagnosed with primary gastric diffuse large B-cell lymphoma(PGDLBL)by PET/CT examination,gastroscopy,biopsy and histopathological analysis at a regular physical check in April,2016.The patient received R-CO chemotherapy(rituximab,cyclophosphamide,and vincristine)and radiotherapy subsequently,with enteral nutritional treatment through 3-cavity nasogastric tube due to development of pyloric obstruction.To satisfy patient's strong desire of eating by himself,we performed surgery of exploratory laparotomy and Roux-en-Y gastric bypass(RGB)to relieve pylorus obstruction.Postoperatively,the patient resumed oral feeding,supplemented by nasogastric tube feeding at 1350-1550 Kcal daily.He is now 94 years old with fairly well nutrition and normal communication.The outcome of 4 year follow-up suggests that nutritional treatment and palliative medicine are important for improving prognosis and life-quality of very elderly patients with end-stage tumors apart from the effective chemotherapy,radiotherapy,and surgery.

Cohort profile: design and methods of the Chinese colorectal, breast, lung, liver, and stomach cancer screening trial(C-BLAST)

Given the rapid changes in social structure(urbanization),economic structure(industrialization),and demographic structure(population aging)in China,cancer has become a major public health problem1.Extensive evidence has indicated that screening can decrease cancer mortality,particularly among high-risk groups,and several representative national and regional cancer screening programs have been launched in China to cope with the increasing burden of cancer.

Tuning redox activity through delithiation induced protective layer and Fe-O coordination for Li-rich cathode with improved voltage and cycle performance

Li-rich layered transition metal oxides are one of the most promising cathode materials for their high energy density.However,the cathodes usually suffer from severe potential dropping and capacity fading during cycling,which are associated with the surface oxygen release and accompanied by cation densification and structural collapse.Herein,an integrative approach of simultaneous constructing uniform 3d Fe-ion doping in the transition metal layer and Li-rich Li_(5)FeO_(4) shell to grab the oxygen and prevent interfacial side reactions is proposed.The introduction of Fe induces higher redox potential and stronger 3 d Fe-O_(2)p covalent bond,triggering reversible anionic redox via a reductive coupling mechanism.And the delithiated product of Li-rich Li_(5)FeO_(4) not only acts as a protective layer alleviating the side reactions but also enhances the surface kinetic property.With the benefit of promoted reversibility of oxygen redox and enhanced surface stability,the cathode exhibits high reversible capacity and superior cycle performance.Density function theory calculation indicates that the O_(2)p non-bonding state in the cathode incorporated with Fe sits at a lower energy band,resulting in higher energy storage voltage and improved oxygen stability.Consequently,the modified cathode exhibits a discharge specific capacity of 307 m A h g^(-1)(1 C=250 m A g^(-1)),coulombic efficiency of 82.09%in the initial cycle at 0.1 C and 88.34%capacity retention after 100 cycles at 1 C.The work illustrates a strategy that could simultaneously enhance oxygen redox reversibility and interface stability by constructing lattice bond coordination and delithiation induced protective layer to develop Li-rich materials with high reversible capacity and long lifespan.

Botulinum toxin type A plus rehabilitative training for improving the motor function of the upper limbs and activities of daily life in patients with stroke and brain injury

BACKGROUND:Botulinum toxin type A(BTX-A)is mostly to be used to treat various diseases of motor disorders,whereas its effect on muscle spasm after stroke and brain injury needs further observation.OBJECTIVE:To observe the effect of BTX-A plus rehabilitative training on treating muscle spasm after stroke and brain injury.DESIGN:A randomized controlled observation.SETTINGS:Department of Rehabilitation,Department of Neurology and Department of Neurosurgery,the Second Hospital of Hebei Medical University.PARTICIPANTS:Sixty inpatients with brain injury and stroke were selected from the Department of Rehabilitation,Department of Neurology and Department of Neurosurgery,the Second Hospital of Hebei Medical University from January 2001 to August 2006.They were all confirmed by CT and MRI,and had obvious increase of spastic muscle strength in upper limbs,their Ashworth grades were grade 2 or above.The patients were randomly divided into treatment group(n=30)and control group(n=30).METHODS:①Patients in the treatment group undertook comprehensive rehabilitative trainings,and they were administrated with domestic BTX-A,which was provided by Lanzhou Institute of Biological Products,Ministry of Health(S10970037),and the muscles of flexion spasm were selected for upper limbs,20-25 IU for each site.②Patients in the treatment group were assessed before injection and at 1 and 2 weeks,1 and 3 months after injection respectively,and those in the control group were assessed at corresponding time points.The recovery of muscle spasm was assessed by modified Ashworth scale(MAS,grade 0-Ⅳ;Grade 0 for without increase of muscle strength;GradeⅣfor rigidity at passive flexion and extension);The recovery of motor function of the upper limbs was evaluated with Fugl-Meyer Assessment(FMA,total score was 226 points,including 100 for exercise,14 for balance,24 for sense,44 for joint motion,44 for pain and 66 for upper limb);The ADL were evaluated with Barthel index,the total score was 100 points,60 for mild dysfunction,60-41 for moderate dysfunction,<40 for severe dysfunction).MAIN OUTCOME MEASURES:Changes of MAS grade,FMA scores and Barthel index before and after BTX-A injection.RESULTS:All the 60 patients with brain injury and stroke were involved in the analysis of results.①FMA scores of upper limbs:The FMA score in the treatment group at 2 weeks after treatment was higher than that before treatment[(14.98±10.14),(13.10±9.28)points,P<0.05],whereas there was no significant difference at corresponding time point in the control group.The FMA scores at 1 and 3 months in the treatment group[(23.36±10.69),(35.36±11.36)points]were higher than those in the control group[(20.55±10.22),(30.33±10.96)points,P<0.01].②MAS grades of upper limbs:There were obviously fewer cases of gradeⅢin MAS at 2 weeks after treatment than before treatment in the treatment group(0,9 cases,P<0.05),whereas there was no obvious difference in the control group.There were obviously fewer cases of gradeⅢin MAS at 2 weeks and 1 month after treatment in the treatment group(0,0 case)than the control group(5,2 cases,P<0.01).③Barthel index of upper limbs:The Barthel index at 2 weeks after treatment was higher than that before treatment in the treatment group[(30.36±22.25),(28.22±26.21)points,P<0.05],whereas there was no significant difference in the control group.The Barthel indexes at 1 and 3 months after treatment in the treatment group were obviously higher than those in the control group[(20.55±10.22),(30.33±10.96)points,P<0.01].CONCLUSION:BTX-A has obvious efficacy on decreasing muscle tension after stroke and brain injury,and relieving muscle spasm;Meanwhile,the combination with rehabilitative training can effectively ameliorate the motor function of upper limbs and ADL of the patients.

莲心碱通过抑制细胞自噬减轻体外胰腺腺泡细胞急性炎症反应

目的探讨莲心碱(liensinine,LSN)对雨蛙素诱导的急性胰腺炎细胞模型的保护作用及其分子机制。方法胰腺腺泡细胞AR42J分为对照组、模型组、20μmol/L LSN组和40μmol/L LSN组。模型组加入雨蛙素刺激造模;20μmol/L LSN组和40μmol/L LSN组加入LSN预处理12 h,再加入雨蛙素刺激造模。采用Western blot检测LC3、Beclin-1和p62蛋白表达水平,采用ELISA法检测炎症因子白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子(tumor necrosis factor-α,TNF-α)分泌水平,采用淀粉酶活性检测试剂盒检测淀粉酶含量,采用Calcein AM细胞活性检测试剂盒检测细胞存活率,采用乳酸脱氢酶(lactic dehydrogenase,LDH)细胞毒性检测试剂盒检测细胞损伤,采用TUNEL法检测细胞凋亡。结果与对照组比较,模型组LC3-Ⅱ/LC3-Ⅰ比率和Beclin-1蛋白水平显著升高(P<0.01),而p62蛋白水平显著降低(P<0.01);与模型组比较,20μmol/L LSN组和40μmol/L LSN组LC3-Ⅱ/LC3-Ⅰ比率和Beclin-1蛋白水平显著降低(P<0.05),而p62蛋白水平显著升高(P<0.05);与20μmol/L LSN组比较,40μmol/L LSN组LC3-Ⅱ/LC3-Ⅰ比率和Beclin-1蛋白水平显著减少(P<0.05),而p62蛋白水平显著增加(P<0.05)。与对照组比较,模型组IL-1β、IL-6和TNF-α分泌水平显著增加(P<0.01),淀粉酶活性显著升高(P<0.01),细胞存活率显著降低(P<0.01),LDH活性显著升高(P<0.01),细胞凋亡率显著增加(P<0.01);与模型组比较,20μmol/L LSN组和40μmol/L LSN组IL-1β、IL-6和TNF-α分泌水平显著降低(P<0.05),淀粉酶活性显著下降(P<0.05),细胞存活率显著升高(P<0.05),LDH活性显著降低(P<0.05),细胞凋亡率显著下降(P<0.05);与20μmol/L LSN组比较,40μmol/L LSN组IL-1β、IL-6和TNF-α分泌水平显著减少(P<0.05),淀粉酶活性显著降低(P<0.05),细胞存活率显著提高(P<0.05),LDH活性显著下降(P<0.05),细胞凋亡率显著降低(P<0.05)。结论莲心碱可有效保护雨蛙素诱导的胰腺腺泡细胞损伤,其作用机制可能与抑制细胞自噬相关。

CaMKK2调控肝细胞癌化疗耐药性的作用和机制

目的 探讨钙/钙调蛋白依赖性蛋白激酶激酶2(calcium/calmodulin-dependent protein kinase kinase 2,CaMKK2)调控肝细胞癌(hepatocellular carcinoma, HCC)化疗耐药性的作用及其机制。方法 (1)为了检测CaMKK2在HCC耐药细胞株中的表达变化,将实验分为亲本组和耐药组。采用浓度梯度递增法建立奥沙利铂(oxaliplatin, OXA)耐药细胞株MHCC97H/OXA和Hep3B/OXA。采用Western blot检测CaMKK2的磷酸化和总蛋白表达水平。(2)为了检测CaMKK2对肝细胞癌化疗药性的调控作用,将实验分为对照组和CaMKK2敲除组。采用CRISPR/Cas9技术敲除MHCC97H/OXA和Hep3B/OXA细胞株中的CaMKK2基因表达,采用Western blot验证CaMKK2敲除效率。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)实验检测CaMKK2敲除对MHCC97H/OXA和Hep3B/OXA细胞株细胞存活率的影响。采用流式细胞术检测CaMKK2敲除对MHCC97H/OXA和Hep3B/OXA细胞株凋亡的影响。采用Western blot检测CaMKK2敲除对微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)、p62、腺苷酸活化蛋白激酶(adenosine 5′-monophosphate-activated protein kinase, AMPK)和UNC-51样激酶1(UNC51-like kinase 1,ULK1)蛋白表达水平的影响。(3)为了验证CaMKK2对肝细胞癌化疗药性的调控作用,将实验分为CaMKK2敲除+空载体组和CaMKK2敲除+CaMKK2载体组。采用Western blot检测CaMKK2的蛋白表达水平。采用CCK-8实验检测重新表达CaMKK2对CaMKK2敲除的细胞耐药性的影响。采用Western blot检测重新表达CaMKK2对CaMKK2敲除的细胞中AMPK、ULK1和LC3蛋白表达水平的影响。结果 (1)与亲本组相比,耐药组HCC细胞株中CaMKK2的总蛋白表达水平无显著变化(P>0.05),而CaMKK2的磷酸化水平显著升高(P<0.01)。(2)与对照组比较,CaMKK2敲除组细胞中CaMKK2表达水平显著减少(P<0.01)。与对照组比较,CaMKK2敲除组HCC耐药细胞对OXA的敏感性显著提高(P<0.05),OXA细胞凋亡率显著升高(P<0.01)。与对照组比较,CaMKK2敲除组LC3Ⅱ/LC3Ⅰ显著降低,p62蛋白水平显著升高,p-AMPK/AMPK以及p-ULK1/ULK1显著降低(均P<0.01)。(3)与CaMKK2敲除+空载体组比较,CaMKK2敲除+CaMKK2载体组HCC耐药细胞对OXA的敏感性显著降低(P<0.01),LC3Ⅱ/LC3Ⅰ、p-AMPK/AMPK和p-ULK1/ULK1显著升高(P<0.01)。结论 基因敲除CaMKK2有效逆转HCC化疗耐药性,其作用机制与调控AMPK/ULK1介导的自噬通路相关。