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Compared with the seismic wave fields, the influence of porosity and permeability to seismoelectric coupling may be more significant. The seismoelectric effect mainly forms the coseismic signals (CS) and interface response (IR). The influences of the porosity and permeability on the two types of seismoelectric field are still not clear. In this paper, we numerically analyze the response characteristics of the CS and IR with the dependency of porosity and permeability, respectively. The differences in the sensitivity of the CS and the IR fields to porosity and permeability are also determined. The results show that the change of permeability is mainly sensed by the IR field, while the CS has almost no obvious response to it: both the CS and IR fields are sensitive to the change of porosity, but the sensitivity of the IR field is higher and with the increase of porosity, the discrepancy between the sensitivity of the CS and IR fields to porosity changes increases. The signal amplitude anomaly will be caused in both P-wave fields and seismoelectric fields (IR and CS) at the geological body where the porosity changes, but the change of permeability has little impact on the P-wave and only induces an obvious IR amplitude anomaly in seismoelectric fields. This indicates that the seismoelectric effect can better reveal the connectivity characteristics of the geological body than the seismic wave. This investigation can help to distinguish the sensitivity of seismoelectric response to porosity and permeability more clearly.

The 1000 MW Francis turbine unit at the Baihetan hydropower station is the maximum capacity unit in the world at present, and it has adopted the runner type with long and short blades. For this ultra-high output Francis turbine, especially with the breakthrough runner structure, the hydraulic excitation phenomenon caused by internal dynamic and static interference is the key factor for the stability of the unit. In this study, the 1000 MW Francis turbine unit is taken as the research object, and the rated output conditions with different guide vane openings are selected for comparative analysis. The flow field structure and the pressure pulsation characteristics inside the guide vane and runner under different openings are obtained. The distribution characteristics and evolution law of the vortex in the runner under different guide vane openings are analyzed. The results show that the dynamic and static interference between the runner and the guide vane induces the local high-speed flow to appear in the vaneless area, and the larger the guide vane opening, the smaller the dynamic and static interference between the runner and the guide vane; the vortex in the runner mainly develops and evolves from the inlet to the outlet and is mainly distributed near the blade wall surface. The pressure pulsation inside the runner is mainly due to the action of dynamic and static interference. The pressure pulsation induced by the dynamic and static interferences shows a decreasing law from the runner inlet to the runner outlet.

Transient processes that occur in pumped storage power plants can cause high-pressure conditions, which in turn can result in vibrations in the pump–turbine structure and even damage to structural components. It is therefore crucial to research the transient process of the large pump–turbine units and the flow-induced vibrations of the structural components. The three-dimensional flow field and structural field models of a high-head prototype pump–turbine were constructed to study its flow characteristics and structural characteristics under the turbine start-up. Calculations and analyses were performed on the pressure variation and the flow-induced stress concentrations of the pump–turbine during start-up in turbine mode. The simulated pressure distributions during the turbine start-up were mapped onto the finite element calculation model of the structures of the pump–turbine to calculate the flow-induced stress concentrations. This study provides a reference to improve the design and operation of high-head prototype pump–turbines based on the findings of the flow and structural characteristics.

It is not uncommon for pump-turbine units in pumped storage power plants to experience load rejections due to the sudden disconnection of the generator from the power grid. Load rejection can suddenly increase the rotating speed of the pump-turbine and cause strong pressure fluctuations in the flow passage of the pump-turbine unit. During load rejection, the strong pressure fluctuations caused by the water hammer effect can cause strong structural vibrations, high stresses and even damage to the turbine runner, head cover, stay ring, bottom ring, head cover bolts and bottom ring bolts. In order to study, in detail, the flow-induced stress characteristics of the prototype pump-turbine unit, and the pressure variations during load rejection in a high-head pumped storage power plant were measured first. Then the measured data were used to set up computational fluid dynamics (CFD) simulations in the entire flow passage of the prototype pump-turbine and to calibrate the simulation results. The calculated pressure distributions in the flow passage during load rejection were exported and mapped on the finite element model of the stationary structures of the pump-turbine unit so that the flow-induced stresses on the head cover, stay ring, bottom ring, head cover bolts and bottom ring bolts can be calculated. The results of the analysis show that the maximum stresses in the head cover bolts and bottom ring bolts are located on the rounded corner of the bolt near the stay ring and that the stresses in the bolts vary with time during load rejection. The maximum stresses of the head cover bolts are higher than the maximum stresses of the bottom ring bolts, and the maximum stresses of the bolts are above two-thirds of the yield strength of the bolt material. It is recommended to use larger nominal diameter bolts to avoid damage to the connecting bolts of the pump-turbine unit.

SUMMARY The monoclinic medium with a horizontal symmetry plane is gradually being studied for seismic anisotropy characterization. The principle goal of this paper is to investigate the effect of fracture parameters (azimuth angle, density, aspect ratio, scale) on the exact and approximate monoclinic anisotropy parameters. We derive the monoclinic porous media based on the Chapman model which accounts for the wave-induced fluid flow and give the expressions of the Thomsen-style anisotropy parameters (nine orthorhombic anisotropy parameters: VP0, VS0, ϵ1, ϵ2, γ1, γ2, δ1, δ2, δ3, three exact monoclinic parameters: ζ1, ζ2, ζ3 and three approximate monoclinic parameters: $\widetilde{\zeta _{1}}, \widetilde{\zeta _{2}}, \widetilde{\zeta _{3}}$). The dependence of Thomsen-style anisotropy parameters associated with azimuth angle between two fracture sets is analysed. The orthorhombic anisotropy parameters and monoclinic anisotropy parameters have the same period (π) on the azimuth angle between two fracture sets. The exact and approximate monoclinic anisotropy parameters responsible for the rotation of the P-wave NMO ellipse have a similar trend versus the azimuth angle, while those responsible for the rotation of the S1- and S2-wave NMO ellipses have significant discriminations. The influence of fracture density, aspect ratio, and scale on the monoclinic parameters are also analysed. The monoclinic anisotropy parameters responsible for the rotation of the P-wave NMO ellipse decrease with fracture density and aspect ratio increasing from 0 to 0.1, while those responsible for the rotation of S1- and S2-wave NMO ellipses increase with the fracture parameters. The fracture density has a bigger influence on the monoclinic anisotropy parameters than the fracture aspect ratio. When saturated with different fluids (water and CO2), the monoclinic parameters have a similar behaviour versus the azimuth angle between two fracture sets.

Background: Recent studies demonstrate that N6-methyladenosine (m6A) methylation plays a crucial role in colorectal cancer (CRC). Therefore, we conducted a comprehensive analysis to assess the m6A modification patterns and identify m6A-modified genes in patients with CRC recurrence.Methods: The m6A modification patterns were comprehensively evaluated by the NMF algorithm based on the levels of 27 m6A regulators, and tumor microenvironment (TME) cell-infiltrating characteristics of these modification patterns were systematically assessed by ssGSEA and CIBERSORT algorithms. The principal component analysis algorithm based on the m6A scoring scheme was used to explore the m6A modification patterns of individual tumors with immune responses. The weighted correlation network analysis and univariable and multivariable Cox regression analyses were applied to identify m6A-modified gene signatures. The single-cell expression dataset of CRC samples was used to explore the tumor microenvironment affected by these signatures.Results: Three distinct m6A modification patterns with significant recurrence-free survival (RFS) were identified in 804 CRC patients. The TME characterization revealed that the m6A modification pattern with longer RFS exhibited robust immune responses. CRC patients were divided into high- and low-score subgroups according to the m6A score individually, which was obtained from the m6A-related signature genes. The patients with low m6A scores had both longer RFS and overall survival (OS) with altered immune cell infiltration. Notably, m6A-modified genes showed significant differences related to the prognosis of CRC patients in the meta-GEO cohort and TCGA cohort. Single-cell expression indicated that ALVRL1 was centrally distributed in endothelial tip cells and stromal cells.Conclusion: The m6A modification plays an indispensable role in the formation of TME diversity and complexity. Importantly, the signatures (TOP2A, LRRC58, HAUS6, SMC4, ACVRL1, and KPNB1) were identified as m6A-modified genes associated with CRC recurrence, thereby serving as a promising predictive biomarker or therapeutic target for patients with CRC recurrence.

The Yellow River has a high sand content, and sand deposition in the pipelines behind the pumping station occurs from time to time. It is of great significance to reasonably predict the critical velocity of the small-angled V-inclined water transportation pipes. In this study, a Eulerian multiphase model was employed to simulate the solid–liquid two-phase flow. Based on the conservation of the sand transport rate, the critical velocity of the V-inclined pipe was predicted. The effects of simulated pipeline length, pipe inclination and particle size were investigated. The results show that when the simulated pipeline length reached a certain value, it did not affect the prediction of the critical velocity of the overall pipeline. The ±2∘ pipe inclination had a negligible effect on the critical velocity for transporting small-sized particles, but it led to the nonuniform and asymmetrical distribution of liquid velocity and sand deposition at the different cross-sections. As the particle size increased, the critical velocity also increased. However, the influence of particle size on the critical velocity is currently complicated, resulting in a large difference between numerical simulation and empirical formulas when transporting large-sized particles. Accurate prediction of critical velocity is important for long-distance water transportation pipelines to prevent sand deposition and reduce costs.

Adelphocoris suturalis Jakovlev (Hemiptera: Miridae) is an omnivorous agricultural pest that has severe economic impacts on a diverse range of agricultural crops. Although the targeted disruption of reproductive development among insects has been proposed as a novel control strategy for pest species, the current understanding of the physiology and molecular mechanisms of A. suturalis reproduction is very limited. In this study, we isolated a putative A. suturalisaspartic protease (AsAP) gene that is highly expressed in the fat body and ovaries of sexually mature females. The double-stranded RNA (dsRNA)-mediated knockdown of AsAP suppressed ovarian development and negatively impacted female fertility, which suggested that it plays an essential role in A. suturalis reproduction. The results of this study could help to expand our understanding of A. suturalis reproductive development and have the potential to facilitate the development of effective strategies for the better control of this pest species.

In recent years, large-capacity, high-head pump–turbine units have been developed for pumped storage power plants to effectively utilise water energy and store large amounts of electricity. Compared with the traditional Francis turbine unit, the radial distance between the trailing edge of the guide vanes and the leading edge of runner blades of high-head pump–turbine unit is smaller, so the rotor–stator interaction and the corresponding pressure fluctuations in the vaneless space of pumped storage units are more intense. The pressure fluctuations with high amplitudes and high frequencies induced by rotor–stator interaction (RSI) become the main hydraulic excitation source for the structures of the unit and may cause violent vibration and fatigue damage to structural components, and seriously affect the safe operation of the units. In this paper, the RSI of a high-head pump–turbine in turbine mode of operation is studied in detail by means of site measurement and full three-dimensional unsteady simulations. The results of RSI-induced pressure fluctuations in turbine mode are analysed experimentally and numerically. The accuracy of the numerical calculations is verified by comparing with the measured results, and the variation law of RSI is deeply analysed. The results show that the pressure fluctuations in the vaneless space are affected by the wake of the guide vane, the rotating excitation of the runner, the low-frequency excitation of the draft tube, and the asymmetric characteristics of the incoming flow of the spiral case, and shows significant differences in spatial position. The findings of the investigation are an important and valuable reference for the design and safe operation of the pumped storage power station. It is recommended to design the runner with inclined inlets to reduce the amplitudes of RSI-induced pressure fluctuations and to avoid operating the pump–turbine units under partial load for long periods of time to reduce the risk of pressure fluctuation induced severe vibration on the structures.

The thrust bearing, as the only part bearing the axial load, is extremely important in vertical hydroelectric generating units. Its working condition directly affects the safe and reliable operation of the hydroelectric generating unit. However, during operation, the oil film is easily damaged under the influence of complex factors. Installation deviation is a key point that can cause temperature and pressure changes in the oil film, affecting the force on the bearing. This article takes the thrust bearing model of the 1000 MW Francis turbine unit of the Baihetan Power Station as the research object. Based on the fluid–solid coupling theory and CFD technology, the oil film characteristics of thrust bearings are analyzed, and the influence of inclination angles and installation deviation on the oil film flow performance of thrust bearings is discussed. The results show that as the inclination angle changes from 0.0030° to 0.0048°, the axial force changes from 856 t to 368 t, and there is an approximate linear correlation between them. The radial installation deviation has an effect on the axial force. Under the optimal working condition of an inclination angle of 0.0039°, when the radial deviation of the pad changes from 0 mm to 1 mm, the axial force changes from 1573 t to 1275 t. In the process of unit installation, it is necessary to pay attention to the installation deviation of the pad. The results provide guidance for the installation of the bearing, which helps to ensure the safe and stable operation of the station.

In this paper, a 1000 MW Francis turbine was used to study the influences of uneven clearance distribution caused by various radial installation deviations on the hydraulic thrust of a runner under the rated operating condition with steady-state CFD analysis. Then, the influences of radial installation deviation on the pressure pulsations of the runner and the fluid domains most affected by the deviation were investigated via unsteady CFD calculation. The results show that the radial hydraulic force on the chambers increased linearly with the increase in the radial installation deviation. Additionally, the high-pressure zone was not located in the same position as the radial deviation. With increasing values of the radial installation deviation, the high-pressure zone rotated along the opposite direction of the rotating direction of the runner. This study also found that the flow in the upper crown chamber was most affected by radial installation deviation and that the percentage of high frequencies of the pressure pulsations increased with the flow in the clearances.

BackgroundColorectal cancer (CRC) is one of the most common malignancies and its incidence and mortality are increasing yearly. 5-Fluorouracil (5-FU) has long been used as a standard first-line treatment for CRC patients. Although 5-FU-based chemotherapy is effective for advanced CRC, the consequent resistance remains a key problem and causes the poor prognosis of CRC patients. Thus, there is an urgent need to identify new biomarkers to predict the response to 5-FU-based chemotherapy.MethodsCRC samples were retrieved from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). The immune-related genes were retrieved from the ImmPort database. Single-cell sequencing results from colorectal cancer were obtained by the ArrayExpress database. 5-FU resistance-related genes were filtered and validated by R packages. ESTIMATE algorithms were used to assess the tumor microenvironment (TME). KEGG and GO analysis were performed to explore the biological signaling pathway for resistant-response patients and sensitive-response patients in the tumor microenvironment. pRRophetic algorithms were used to predict 5-FU sensitivity. GSEA and GSVA analysis was performed to excavate the biological signaling pathway of the RBP7 gene.ResultsNine immune-related genes were identified to be associated with 5-FU resistance and poor disease-free survival (DFS) of CRC patients and the signature of these genes was developed in a DFS-prognostic model. Four immune-related genes were determined to be associated with 5-FU resistance and overall survival (OS) of CRC patients. The signature of these genes was developed an OS-prognostic model. ESTIMATE scores showed a significant difference between 5-FU resistant and 5-FU sensitive CRC patients. Resistant-response patients and sensitive-response patients to 5-FU based chemotherapy showed different GO and KEGG enrichment on the tumor microenvironment. RBP7, as a tumor immune microenvironment (TIME) related gene, was found to have the potential of predicting chemotherapy resistance and poor prognosis of CRC patients. GSEA analysis showed multiple signaling differences between the high and low expression of RBP7 in CRC patients. Hypoxia and TNFα signaling via NFκB gene sets were significantly different between chemotherapy resistant (RBP7High) and chemotherapy sensitive (RBP7Low) patients. Single-cell RNA-seq suggested RBP7 was centrally distributed in endothelial stalk cells, endothelial tip cells, and myeloid cells.ConclusionsImmune-related genes will hopefully be potential prognostic biomarkers to predict chemotherapy resistance for CRC. RBP7 may function as a tumor microenvironment regulator to induce 5-FU resistance, thereby affecting the prognosis of CRC patients.

During the load rejection transient process of the prototype pump turbine units, the pressure fluctuations of the entire flow passage change drastically due to the rapid closing of guide vanes. The extremely unsteady pressure distribution in the flow domains including the crown chamber and the band chamber may cause a strong vibration on the stationary structures of the unit and result in large dynamic stress on the head cover, stay ring and bottom ring. In this paper, the numerical fluid dynamic analysis of the entire flow passage of a reversible prototype pump turbine during load rejection was performed. The flow characteristics in the runner passage, crown chamber, band chamber, seal labyrinths and balance tubes are analysed. The corresponding unsteady flow-induced dynamic behaviour of the head cover, stay vanes and bottom ring was investigated in detail. The analysed results show that the total deformation of the inner edge of the head cover closed to the main shaft is larger than that of other stationary structures of the unit during the load rejection. The maximum stress of the stay ring is larger than that of the head cover and the bottom ring and the maximum equivalent stress is located at the fillet of the stay vane trailing edge. The fluid–structure coupling calculation method and the analysed results can provide guidance for the design of stationary components of hydraulic machinery such as pump turbines, Francis turbines and centrifugal pumps with different heads.

The 1000 MW hydro-generator unit has huge geometrical dimensions and requires high installation accuracy. The control of its installation deviation will directly affect the installation quality and operational performance of the unit and the rotor eccentricity is one of the core problems faced in the installation and operation of the giant hydro-generator unit. This paper presents a method to calculate the asymmetric magnetic field by means of the transient finite element calculation to reveal the effect of the generator rotor radial deviation on the unbalanced magnetic pull of the unit. The effects of rotor dynamic eccentricity on the magnetic field are analyzed in the time and frequency domains, respectively. The results show that both the DC component and the key harmonic component of the unbalanced magnetic pull increase with the increase of the eccentricity value, and this variation relationship is nearly linear. when the eccentricity is no more than 0.15 mm, the unbalanced magnetic pull caused by the rotor eccentricity is calculated to be small, which is acceptable. The conclusions of this investigation can help to straighten out the requirements and standards for the installation deviation control of the large hydro-generator units.

To study the influence of the axial installation deviation of the runner on the hydraulic axial force of the 1000 MW Francis turbine unit, geometric models of the full flow passage of the Francis turbine with the runner sinking in the axial direction by 0, 0.5, 1, 1.5, 2.5, 4, and 5.5 mm were established. The geometric models of the upper crown clearance, lower band clearance, and pressure balance pipes were also built. The SST turbulence model was used in the CFD setup to numerically simulate the flow in the Francis turbine with different runner installation sinking values. The results show that the hydraulic axial force on the inner surface of the runner remains stable when the runner is lowered. The hydraulic axial force on the entire runner surface and the outer surface of the lower band decreases, and the hydraulic axial force on the outer surface of the upper crown clearance increases. All of these hydraulic axial forces gradually tend to stabilize as the amount descending from the runner increases. To study the reasons for the changes in hydraulic axial forces, the streamlines and fluid fields of different sections in the flow passage were analyzed in detail. It was found that periodic changes of vortices were generated in the clearance due to the influences of the geometric shape and wall rotation. These vortices affect the distribution of velocity and pressure and, thus, determine the hydraulic axial forces. The runner axial installation deviation has little influence on the streamlines, pressure, and velocity distribution in each flow passage, and only changes the velocity and pressure in the upper crown clearance and lower band clearance. Therefore, the axial installation deviation of the runner has a great effect on the hydraulic axial force on the outer surface of the upper crown and lower band and has a smaller impact on the runner passage and the hydraulic axial force on the inner surface of the runner. The conclusions in this study can be adopted as references for the installation accuracy control of other hydraulic Francis turbine units.

Accumulating evidence suggests that chronic inflammation may play a critical role in various malignancies, including bladder cancer. This hypothesis stems in part from inflammatory cells observed in the urethral microenvironment. Chronic inflammation may drive neoplastic transformation and the progression of bladder cancer by activating a series of inflammatory molecules and signals. Recently, it has been shown that the microbiome also plays an important role in the development and progression of bladder cancer, which can be mediated through the stimulation of chronic inflammation. In effect, the urinary microbiome can play a role in establishing the inflammatory urethral microenvironment that may facilitate the development and progression of bladder cancer. In other words, chronic inflammation caused by the urinary microbiome may promote the initiation and progression of bladder cancer. Here, we provide a detailed and comprehensive account of the link between chronic inflammation, the microbiome and bladder cancer. Finally, we highlight that targeting the urinary microbiome might enable the development of strategies for bladder cancer prevention and personalized treatment.

The pharmacological mechanism of curcumin against drug resistance in non-small cell lung cancer (NSCLC) remains unclear. This study aims to summarize the genes and pathways associated with curcumin action as an adjuvant therapy in NSCLC using network pharmacology, drug-likeness, pharmacokinetics, functional enrichment, protein-protein interaction (PPI) analysis, and molecular docking. Prognostic genes were identified from the curcumin-NSCLC intersection gene set for the following drug sensitivity analysis. Immunotherapy, chemotherapy, and targeted therapy sensitivity analyses were performed using external cohorts (GSE126044 and IMvigor210) and the CellMiner database. 94 curcumin-lung adenocarcinoma (LUAD) hub targets and 41 curcumin-lung squamous cell carcinoma (LUSC) hub targets were identified as prognostic genes. The anticancer effect of curcumin was observed in KEGG pathways involved with lung cancer, cancer therapy, and other cancers. Among the prognostic curcumin-NSCLC intersection genes, 20 LUAD and 8 LUSC genes were correlated with immunotherapy sensitivity in the GSE126044 NSCLC cohort; 30 LUAD and 13 LUSC genes were associated with immunotherapy sensitivity in the IMvigor210 cohort; and 12 LUAD and 13 LUSC genes were related to chemosensitivity in the CellMiner database. Moreover, 3 LUAD and 5 LUSC genes were involved in the response to targeted therapy in the CellMiner database. Curcumin regulates drug sensitivity in NSCLC by interacting with cell cycle, NF-kappa B, MAPK, Th17 cell differentiation signaling pathways, etc. Curcumin in combination with immunotherapy, chemotherapy, or targeted drugs has the potential to be effective for drug-resistant NSCLC. The findings of our study reveal the relevant key signaling pathways and targets of curcumin as an adjuvant therapy in the treatment of NSCLC, thus providing pharmacological evidence for further experimental research.

Objective: In this study, we aimed to develop an amino-terminal fragment (ATF) peptide-targeted liposome carrying β-elemene (ATF24-PEG-Lipo-β-E) for targeted delivery into urokinase plasminogen activator receptor-overexpressing bladder cancer cells combined with cisplatin (DDP) for bladder cancer treatment. Methods: The liposomes were prepared by ethanol injection and high-pressure microjet homogenization. The liposomes were characterized, and the drug content, entrapment efficiency, and in vitro release were studied. The targeting efficiency was investigated using confocal microscopy, ultra-fast liquid chromatography, and an orthotopic bladder cancer model. The effects of ATF24-PEG-Lipo-β-E combined with DDP on cell viability and proliferation were evaluated by a Cell Counting Kit-8 (CCK-8) assay, a colony formation assay, and cell apoptosis and cell cycle analyses. The anticancer effects were evaluated in a KU-19-19 bladder cancer xenograft model. Results: ATF24-PEG-Lipo-β-E had small and uniform sizes (˜79 nm), high drug loading capacity (˜5.24 mg/mL), high entrapment efficiency (98.37 ± 0.95%), and exhibited sustained drug release behavior. ATF24-PEG-Lipo-β-E had better targeting efficiency and higher cytotoxicity than polyethylene glycol (PEG)ylated β-elemene liposomes (PEG-Lipo-β-E). DDP, combined with ATF24-PEG-Lipo-β-E, exerted a synergistic effect on cellular apoptosis and cell arrest at the G2/M phase, and these effects were dependent on the caspase-dependent pathway and Cdc25C/Cdc2/cyclin B1 pathways. Furthermore, the in vivo antitumor activity showed that the targeted liposomes effectively inhibited the growth of tumors, using the combined strategy. Conclusions: The present study provided an effective strategy for the targeted delivery of β-elemene (β-E) to bladder cancer, and a combined strategy for bladder cancer treatment.

Cancer is a common and complex disease with various genetic and environmental risks. Chemotherapy and targeted therapy are one of the main therapeutic approaches for the treatment of advanced cancer patients. However, prolonged patient treatment often provokes drug resistance phenomena that render the therapeutic treatment highly ineffective. Therefore, the development of novel and effective treatment strategies to overcome drug resistance is urgent for cancer therapy. A variety of factors contribute to drug resistance, including tumor cell heterogeneity, drug efflux, drug inactivation, changes in tumor microenvironment, DNA damage repair disorders, epigenetic alterations in cancer cells and so on. However, how to circumvent drug resistance to improve anticancer efficacy remains to be determined. Currently, the most important approach for overcoming drug resistance is drug combination therapeutic regimen, however, it is easy to induce new drug resistance or even multi-drug resistance (MDR) which is defined as the resistance of cancer cells to multiple anticancer drugs with different structures and mechanisms of action. Whereas, the “multi-components and multi-targets” manifestations of Traditional Chinese Medicine (TCM) make them an attractive direction for developing new adjuvant therapeutics to reverse drug resistance. In recent years, several compounds derived from TCM have been reported to augment sensitivity or overcome resistance of anticancer therapeutic drugs, including elemene, bufalin, curcumin, magnolol, honokiol, resveratrol, quercetin, and so on. Thus, the combination of TCM and conventional anticancer drugs will hopefully provide a novel strategy for enhancing the effects of anticancer strategies and overcoming their resistance in the treatment of cancer patients. In the current paper, we review the molecular mechanisms of drug resistance and the therapeutic strategies. We also discuss the current basic research and clinical studies in overcoming drug resistance from Traditional Chinese Medicine. Finally, we focus the integrated traditional Chinese and Western medicine as a promising new therapeutic approach to enhance the effects of anticancer strategies or overcome their resistance in the treatment of cancer patients.

Background and Purpose: RAS mutations limit the effectiveness of anti-epidermal growth factor receptor (EGFR) monoclonal antibodies in combination with chemotherapy for metastatic colorectal cancer (mCRC) patients. Therefore, new cell death forms have focused on identifying indirect targets to inhibit Ras-induced oncogenesis. Recently, emerging evidence has shown the potential of triggering ferroptosis for cancer therapy, particularly for eradicating aggressive malignancies that are resistant to traditional therapies. Methods: KRAS mutant CRC cell HCT116 and Lovo were treated with cetuximab and β-elemene, a bioactive compound isolated from Chinese herb Curcumae Rhizoma. Ferroptosis and epithelial-mesenchymal transformation (EMT) were detected in vitro and in vivo. Orthotopic CRC animal model were established and the tumor growth was monitored by IVIS bioluminescence imaging. Tumor tissues were collected to determine ferroptosis effect and the expression of EMT markers after the treatment. Results: CCK-8 assay showed that synergetic effect was obtained when 125 µg/ml β-elemene was combined with 25 µg/ml cetuximab in KRAS mutant CRC cells. AV/PI staining suggested a non-apoptotic mode of cell death after the treatment with β-elemene and cetuximab. In vitro, β-elemene in combination with cetuximab was shown to induce iron-dependent reactive oxygen species (ROS) accumulation, glutathione (GSH) depletion, lipid peroxidation, upregulation of HO-1 and transferrin, and downregulation of negative regulatory proteins for ferroptosis (GPX4, SLC7A11, FTH1, glutaminase, and SLC40A1) in KRAS mutant CRC cells. Meanwhile, combinative treatment of β-elemene and cetuximab inhibited cell migration and decreased the expression of mesenchymal markers (Vimentin, N-cadherin, Slug, Snail and MMP-9), but promoted the expression of epithelial marker E-cadherin. Moreover, ferroptosis inhibitors but not other cell death suppressors abrogated the effect of β-elemene in combination with cetuximab on KRAS mutant CRC cells. In vivo, co-treatment with β-elemene and cetuximab inhibited KRAS mutant tumor growth and lymph nodes metastases. Conclusions: Our data for the first time suggest that the natural product β-elemene is a new ferroptosis inducer and combinative treatment of β-elemene and cetuximab is sensitive to KRAS mutant CRC cells by inducing ferroptosis and inhibiting EMT, which will hopefully provide a prospective strategy for CRC patients with RAS mutations.

Background There is a lack of data on the prevalence of potentially inappropriate medications (PIMs) in community-dwelling older adults in China. Objective To assess the prevalence of potentially inappropriate medication in community dwelling older adults in China and to investigate risk factors associated with the use of such medication. Setting Ambulatory Care Clinic of Xiangya Hospital, Central South University. Method A cross-sectional retrospective review of prescriptions for older patients (aged ≥ 65 years) was performed using the 2019 American Geriatrics Society (AGS) Beers Criteria and the 2017 Chinese Criteria. We only assessed potentialli inappropriate medications independently from diagnosis and drug–drug interactions. Each patient was classified as PIM-user or non-PIM-user, according to whether a patient took at least one potentially inappropriate medication. Main outcome measure Prevalence of potentially inappropriate medication based on the two criteria. Results A total of 8477 medications among 1874 elderly patients were examined over the study period from January 1 to December 31, 2018. The Chinese Criteria detected significantly more PIM-users than the AGS Beers Criteria (50.6% vs 35.0%, P

Objective: In this study, we aimed to develop an amino-terminal fragment (ATF) peptide-targeted liposome carrying β-elemene (ATF24-PEG-Lipo-β-E) for targeted delivery into urokinase plasminogen activator receptor-overexpressing bladder cancer cells combined with cisplatin (DDP) for bladder cancer treatment. Methods: The liposomes were prepared by ethanol injection and high-pressure microjet homogenization. The liposomes were characterized, and the drug content, entrapment efficiency, and in vitro release were studied. The targeting efficiency was investigated using confocal microscopy, ultra-fast liquid chromatography, and an orthotopic bladder cancer model. The effects of ATF24-PEG-Lipo-β-E combined with DDP on cell viability and proliferation were evaluated by a Cell Counting Kit-8 (CCK-8) assay, a colony formation assay, and cell apoptosis and cell cycle analyses. The anticancer effects were evaluated in a KU-19-19 bladder cancer xenograft model. Results: ATF24-PEG-Lipo-β-E had small and uniform sizes (˜79 nm), high drug loading capacity (˜5.24 mg/mL), high entrapment efficiency (98.37 ± 0.95%), and exhibited sustained drug release behavior. ATF24-PEG-Lipo-β-E had better targeting efficiency and higher cytotoxicity than polyethylene glycol (PEG)ylated β-elemene liposomes (PEG-Lipo-β-E). DDP, combined with ATF24-PEG-Lipo-β-E, exerted a synergistic effect on cellular apoptosis and cell arrest at the G2/M phase, and these effects were dependent on the caspase-dependent pathway and Cdc25C/Cdc2/cyclin B1 pathways. Furthermore, the in vivo antitumor activity showed that the targeted liposomes effectively inhibited the growth of tumors, using the combined strategy. Conclusions: The present study provided an effective strategy for the targeted delivery of β-elemene (β-E) to bladder cancer, and a combined strategy for bladder cancer treatment.

A significant number of processing cores in any many-core systems nowadays and likely in the future have to be switched off or forced to be idle to become dark cores, in light of ever increasing power density and chip temperature. Although these dark cores cannot make direct contributions to the chip's throughput, they can still be allocated to applications currently running in the system for the sole purpose of heat dissipation enabled by the temperature gradient between the active and dark cores. However, allocating dark cores to applications tends to add extra waiting time to applications yet to be launched, which in return can have adverse implications on the overall system performance. Another big issue related to dark core allocation stems from the fact that application characteristics are prone to undergo rapid changes at runtime, making a fixed dark core allocation scheme less desirable. In this paper, a runtime dark core allocation and dynamic adjustment scheme is thus proposed. Built upon a dynamic programming network (DPN) framework, the proposed scheme attempts to optimize the performance of currently running applications and simultaneously reduce waiting times of incoming applications by taking into account both thermal issues and geometric shapes of regions formed by the active/dark cores. The experimental results show that the proposed approach achieves an average of 61% higher throughput than the two state-of-the-art thermal-aware runtime task mapping approaches, making it the runtime resource management of choice in many-core systems.

Purpose: To investigate the correlation between the expression of PD-L1, SOCS3 and immune-related biomarkers CD276, CD4, CD8 in hepatocellular carcinoma (HCC) and further determine the relationship with clinicopathologic characteristics and the prognostic value of their co-expression in HCC patients. Methods: We assessed the expression of PD-L1, CD276, SOCS3, CD4 and CD8 by immunohistochemistry in tumor tissue from 74 HCC patients who underwent curative hepatectomy. Results: High expression of PD-L1 was significantly associated with high Edmondson grade (p0.05). High expression of CD8 was respectively significantly associated with worse overall survival (OS) (p=0.002). There was no significantly difference between CD4 and CD8 high-expression and overall survival (OS) (p=0.100). Both high expression of PD-L1 (p=0.003) and low expression of SOCS3 (p=0.015) was significantly associated with worse overall survival (OS). But CD276 only had a trendency (p=0.166). Additionally, multivariate Cox regression models implied that PD-L1, SOCS3, as well as both CD4 and CD8 was an independent prognostic factor for OS (p

Silver phosphate is a promising visible-light responsive photocatalyst. However, the serious photocorrosion limits its application in photocatalysis. Herein, a novel ternary graphene oxide (GO)-supported silver phosphate (APO)/cobalt ferrite (CFO) heterojunction composite was successfully synthesized via a simple ion-exchange deposition method for photocatalytic RhB degradation coupled with hydrogen generation from water-splitting under visible light irradiation. The results show that the photocatalytic degradation efficiency of the ternary APO-based composite is 2.37 times that of pure APO. Meanwhile, the photocatalytic H2 evolution reaction (HER) of the ternary APO-based catalyst was realized for the first time. This enhanced performance could be attributed to the fact that the charge recombination can be effectively suppressed and the photocorrosion effect of APO can be inhibited as the photogenerated electrons on the conduction band of APO can be injected into CFO through the GO mediator. Furthermore, the possible photocatalytic mechanism of the ternary APO-based catalyst was proposed to illustrate its excellent photocatalytic activity and stability. The present work sheds light on the design of multifunctional photocatalysts to simultaneously solve the problems of energy crisis and environmental pollution.

5-Fluorouracil (5-FU) is known as a first-line chemotherapeutic agent against colorectal cancer (CRC), but drug resistance occurs frequently and significantly limits its clinical success. Our previous study showed that the protocadherin 17 (PCDH17) gene was frequently methylated and functioned as a tumor suppressor in CRC. However, the relationship between PCDH17 and 5-FU resistance in CRC remains unclear. Here, we revealed that PCDH17 was more highly expressed in 5-FU-sensitive CRC tissues than in 5-FU-resistant CRC tissues, and high expression of PCDH17 was correlated with high BECN1 expression. Moreover, this expression profile contributed to superior prognosis and increased survival in CRC patients. Restoring PCDH17 expression augmented the 5-FU sensitivity of CRC in vitro and in vivo by promoting apoptosis and autophagic cell death. Furthermore, autophagy played a dominant role in PCDH17-induced cell death, as an autophagy inhibitor blocked cell death to a greater extent than the pancaspase inhibitor Z-VAD-FMK. PCDH17 inhibition by siRNA decreased the autophagy response and 5-FU sensitivity. Mechanistically, we showed that c-Jun NH2-terminal kinase (JNK) activation was a key determinant in PCDH17-induced autophagy. The compound SP600125, an inhibitor of JNK, suppressed autophagy and 5-FU-induced cell death in PCDH17-reexpressing CRC cells. Taken together, our findings suggest for the first time that PCDH17 increases the sensitivity of CRC to 5-FU treatment by inducing apoptosis and JNK-dependent autophagic cell death. PCDH17 may be a potential prognostic marker for predicting 5-FU sensitivity in CRC patients.

In order to ensure stable grid operatiFon and improve power quality, active or passive load rejection of pumped storage power stations (PSPS) inevitably occurs from time to time. The rapid closing of the guide vanes will cause drastic changes in pressure pulsations in the flow channel of the pump-turbine (PT) unit. The high-level pressure pulsations during load rejection transfer to the entire flow passage of the PT unit and generate strong vibrations on the head-cover and the connecting bolts. In this study, the 1D/3D joint simulation of the pipeline in a pumped storage power station and the turbine flow channels including the flow domains of the runner, crown chamber, band chamber, upper and lower labyrinths and pressure balance tubes is carried out first. Then, by applying the calculated pressure loads on the head-cover, stay vanes and bottom ring of the PT unit, the flow-induced dynamic behavior of the structures including the head-cover bolts is analyzed in detail. The results demonstrate that pressure loads on head-cover bolts change dramatically during the load rejection process. The flow-induced deformation of the inner head-cover during the load rejection is larger than that of other structures, and the flow-induced displacement and stress of different head-cover bolts are not uniform. The achieved conclusions in this study can be a useful reference for the design and operation of head-cover bolts for other PT units and high-head Francis turbine units.

Pumped storage-power plants play an extremely important role in the modern smart grid due to their irreplaceable advantages in load peak-valley regulation, frequency modulation, and phase modulation. The number of start-stops per day of pump-turbine units is therefore also increasing. During the start-up transient process in turbine mode, the complex flow in runner passage, crown and band chambers, and seal labyrinth is able to induce severe vibration of non-rotating structures such as head cover, stay-ring, and pose a threat to the safe operation of the pump-turbine unit. In this article, the flow-induced vibration of the structures of a pump-turbine unit during its start-up process in turbine mode is studied. In the first place, this investigation establishes a three-dimensional model of the full flow passage and carries out a full three-dimensional CFD calculation based on one-dimensional pipeline calculation results for the start-up transient process. In the next place, by applying the fluid–structure interaction calculation method, the finite element analysis of non-rotating components of the pump-turbine unit is carried out. The flow-induced stresses and deformations of head cover, stay-ring, etc., are obtained and analyzed. The results reveal that the maximum deformation of the non-rotating structures is located at the inner edge of the head cover while the maximum stress appears at the trailing edge fillet of a stay vane. In summary, the dynamic stress of the non-rotating structures changes largely during the start-up process. The stress is strongly related to the axial thrust caused by the fluid flow. The achieved results can provide guidance for further fatigue life assessment of non-rotating structures and contribute to the structural safety design of pump-turbine units.

The peptide hormone insulin has essential roles in regulating insect metabolism, growth, and reproduction. There are, however, few studies assessing the effects of insulin signaling on reproduction in Miridae (Hemiptera). Here, we used RNA interference (RNAi)-mediated knockdown to examine the role of three critical insulin signaling pathway components (insulin receptor, InR; insulin receptor substrate 1, IRS1; and forkhead box O, FOXO) on reproductive capacity in the mirid Adelphocoris suturalis. Knockdown of AsIRS1 led to a significant reduction in egg maturation in unmated females. To further verify the role of AsIRS1, we examined several reproductive parameters following knockdown. Suppression of AsIRS1 transcript levels throughout the reproductive period resulted in reduced lifetime fecundity, egg hatch rate, and oviposition capacity as well as statistically significant reductions in female survival rate and longevity. These findings demonstrate that the insulin signaling pathway plays a key role in the reproductive development of A. suturalis, and that IRS1 is a key regulatory factor. These findings provide an important theoretical basis for the regulation of insect reproduction by insulin and introduce a new target for potential development is A. suturalis control.

Curcumenol, an effective ingredient of Wenyujin, has been reported that exerted its antitumor potential in a few cancer types. However, the effect and molecular mechanism of curcumenol in lung cancer are largely unknown. Here, we found that curcumenol induced cell death and suppressed cell proliferation in lung cancer cells. Next, we demonstrated that ferroptosis was the predominant method that contributed to curcumenol-induced cell death of lung cancer in vitro and vivo for the first time. Subsequently, using RNA sequencing, we found that the long non-coding RNA H19 (lncRNA H19) was significantly downregulated in lung cancer cells treated with curcumenol, when compared to untreated controls. Overexpression of lncRNA H19 eliminated the anticancer effect of curcumenol, while lncRNA H19 knockdown promoted ferroptosis induced by curcumenol treatment. Mechanistically, we showed that lncRNA H19 functioned as a competing endogenous RNA to bind to miR-19b-3p, thereby enhanced the transcription activity of its endogenous target, ferritin heavy chain 1 (FTH1), a marker of ferroptosis. In conclusion, our data show that the natural product curcumenol exerted its antitumor effects on lung cancer by triggering ferroptosis, and the lncRNA H19/miR-19b-3p/FTH1 axis plays an essential role in curcumenol-induced ferroptotic cell death. Therefore, our findings will hopefully provide a valuable drug for treating lung cancer patients.

Understanding the dynamics of vegetation responses to climate change can provide important information for environmental management. The Qilian Mountain Nature Reserve in Northwest China, a high-altitude mountainous region that is of critical ecological importance, has been affected by drastic climate change. However, the response of vegetation to climate change in this area has not been established. This study aims to address three unanswered questions. First, what is the relative importance of temperature and precipitation change in driving regional vegetation change? Second, how does this vegetation–climate relationship manifest itself across different timescales (e.g., seasonal)? Third, what is the spatial heterogeneity of the vegetation–climate relationship? Based on MODIS data and daily meteorological data from 41 stations within and adjacent to the reserve, we used Ensemble Empirical Mode Decomposition (EEMD), Theil-Sen median trend analysis, the Mann-Kendall nonparametric test, and Pearson correlation analysis to study the evolution of temperature, precipitation, and the normalized difference vegetation index (NDVI). The key results are as follows: (1) during 1985–2019, the temperature in the reserve has increased by an average of 0.048 ℃/a, with the largest average annual increases occurring in spring (0.069 ℃) and summer (0.056 ℃); (2) during the same period, precipitation in the reserve increased by an average of 1.17 mm/a, with the largest annual increases occurring in autumn (0.669 mm) and summer (0.545 mm); (3) during 2001–2019, the NDVI increased by an average of 0.004/a; an average annual increase was experienced in all seasons, although the largest increase occurred in summer; (4) there is a statistically significant positive correlation (0.619) between the NDVI and temperature at the annual scale; (5) in contrast, the correlation between the NDVI and precipitation is weaker and not statistically significant (0.368), which indicates that temperature is the dominant factor affecting vegetation changes in the reserve; (6) there exist seasonal differences in the vegetation–climate relationship. The positive correlation between NDVI and temperature is strongest in spring whereas the positive correlation between NDVI and precipitation is strongest in summer; (7) the vegetation–climate relations demonstrate a degree of spatial heterogeneity driven by variability in climate factors and ecosystems. The value of this research is that it analyzes the response characteristics of vegetation to climate change using multiple methods and at multiple scales, providing a useful reference for understanding vegetation changes and their response to climate change in high-altitude mountain regions.

Reasonable design of three-dimensional (3D) catalytic particle electrodes (CPEs) is crucial for achieving efficient electrocatalytic oxidation of organic pollutants. Herein, the novel Fe

Cancer progression and metastases are the leading causes of poor outcomes in patients with colon cancer. Colon cancer metastasis is a multigene, multistep, multistage complex process in which target genes, microRNAs, epithelial-stromal transformation, tumour stem cells, the tumour microenvironment, and various cell signalling pathways are implicated in the progression and metastasis of colon cancer. Although conventional therapies have made significant advances in treating the progression and metastasis of colorectal cancer, they have failed to improve survival outcomes. Natural compounds may have more significant potential in preventing and treating colon cancer. Active natural compounds exert their antitumor effects by inducing tumour cell differentiation, promoting tumour cell apoptosis, inhibiting tumour vascular growth, and regulating immunity. Natural compounds, combined with conventional therapies, can target mutant genes and various cellular signalling pathways, inhibit epithelial-stromal transformation, and improve the tumour microenvironment to inhibit tumour progression and metastasis. The synergism of natural compounds and conventional therapeutics has the potential to become a promising therapy for treating colorectal cancer progression and metastases.

Pumped-storage power stations which can be flexibly adjusted for power generation and energy storage, are widely constructed worldwide. Pumped-storage units work under different operating conditions according to the demand of the power grid, and vortex ropes in the draft tube will appear and show different shapes under the partial load conditions of the unit in generation mode. The unstable pressure pulsation caused by the eccentric vortex ropes inside the draft tube may cause the vibration of the unit during operation. So it is important to study the characteristics of draft tube vortex ropes for flow and vibration analysis. In this paper, a three-dimensional model of the unit is fully established, and the unsteady calculations of 60 %, 86.7%, and 100 % load conditions in turbine mode are carried out by numerical calculations to obtain the flow characteristics of the full flow field. The pressure and velocity distributions, shapes of vortex rope in the draft tube under different load conditions are extracted from the CFD analyses, respectively. The pressure pulsations of different monitoring points set up inside the draft tube are evaluated in the time domain and frequency domain. The results show that the vortex rope in the draft tube is more obvious under small flow discharge, and the vortex rope itself is not sensitive to the selection of Q criterion threshold, but the vortex on the draft tube wall is greatly affected by the selection of the threshold. The influence of draft tube vortex ropes on the vibration of the pumped-storage unit is also discussed.

Recently more start-stops per day of the pump-turbine units are required by the power grid for load regulation. During the turbine start-up and shut-down transient processes, the flow and pressure of the prototype pump-turbine change dramatically at different guide vane opening angles. The extremely unsteady pressure fluctuation in the flow passages can induce large deformation, stress concentration, and strong vibration of the pump-turbine runner. Therefore, it is significantly important to study the unsteady flow characteristics and the corresponding flow-Induced vibration of the runner during turbine start-up and shut-down transient process. In this investigation, a 3D model of the pump-turbine unit including the flow passages of the spiral casing, stay vane, guide vane, runner, and draft tube, crown chamber, band chamber, labyrinth seals, and balance pipes are constructed. The flow characteristics of the unit during the turbine start-up and shut-down process were analysed by coupling the 1D pipeline calculation and 3D turbine flow domain calculation. By mapping the pressure distribution of the fluid domain to the runner structure domain, the flow-induced dynamic behavior of the pump-turbine runner is performed, and the large deformation and stress concentration of the runner are investigated in detail. The flow-induced vibration results achieved are able to provide meaningful suggestions for safe operation and for improving the pump-turbine runner design.

Hydropower is a major renewable clean energy and is widely used worldwide. The reversible pump-turbine unit of the pumped-storage power station is able to work in two main operating modes as required by the power grid: turbine-mode for power generation and pump-mode for power storage. In order to absorb unstable energies such as wind and solar energy and improve the quality of the electricity, reversible pump-turbines need to frequently change operating conditions, and experience more start-stops under different operating modes in a short period. The unstable flow during these transient processes will lead to high-level stresses on the structural components of the pump-turbine units. Therefore, it is of great engineering and academic significance to study the flow characteristics and structural characteristics of the unit during the transient processes. This paper has established a numerical calculation model for a prototype reversible pump-turbine unit, has carried out the CFD calculations of the pump-turbine fluid domains during the pump shutdown transient process, and has analysed the corresponding structural dynamic characteristics of the stationary components of the unit with the fluid-structure coupling method. The pressure variation trend of the spiral case outlet during pump shutdown has the same trend as that of the spiral case domain, and the guide vane flow domain. The maximum flow-induced deformation and stress of the stationary structures have a strong correlation with the axial thrust values of the head cover. The maximum deformation occurs at the inner edge of the head cover, and the maximum stress appears in the fillet of the stay vane leading edge. An increase in the number of shutdowns will result in a higher real risk of fatigue damage to the stay vanes. The conclusions obtained are of great value for safe operation, field condition monitoring, fault diagnosis, and predictive maintenance of the pump-turbine units.

The dramatic changes in the internal flow and the corresponding structural behavior during the turbine start-up transient process of pump-turbines are extremely complex. The clearances in the upper crown chamber and bottom ring chamber affect the results of the flow field and structural field of the pump-turbine runner. Most of the previous studies ignored the effects of the clearance flow field to simplify the numerical simulations. In this study, numerical calculations were performed on the entire flow passage of a high-head prototype pump-turbine during the start-up in turbine mode, and the model with and without the clearance are analysed respectively. The causes of the flow field characteristic difference and external characteristic difference caused by the existence of intermediate clearance in the model are studied in detail. The results show that the clearance flow field has a great influence on the axial forces on the runner, which is mainly due to the higher pressure of the clearance flow field compared with the flow field in the runner; At the same time, because the existence of clearance flow field only has a small effect on the distribution of internal flow field, the hydraulic torque of runner considering clearance effects is basically the same as the one without clearance.

Balancing the relationship between rural development and the protection of water resources is a challenging undertaking. This study develops a coupling coordination degree (CCD) model to examine the non-linear interaction between rural development and water environment in the 11 prefectures of Gansu, northwestern China. There are three key findings. First, economic development is the key driver of rural development, whereas social development has relatively little impact. For the water environment subsystem, improved water efficiency has been the key contributor, whereas environmental carrying capacity is secondary. Second, the CCD increased steadily in the studied period, which suggests that the relationship between rural development and water environment has gradually changed from antagonistic to mutually beneficial. However, this change is not occurring rapidly and in fact shows signs of slowing. Third, the complex spatial differences of the CCD are related to the level of economic and social development, the process of urban–rural integration, and regional natural conditions. The findings of this study have great significance for further quantitative analysis of the interaction and mutual feedback mechanism between the rural economy and the water environment in China and support evidence-based policymaking.

A propanol-tolerant neutral protease was purified and characterized from Bacillus sp. ZG20 in this study. This protease was purified to homogeneity with a specific activity of 26,655 U/mg. The reco...

Alkaline protease was purified from Bacillus sp. ZJ1502 isolated from fermented bean curd and its enzymatic properties were investigated. The final purification fold and specific activity were 18.6 and 30,230 U/mg, respectively. The molecular weight was 14 kDa by SDS-PAGE. The optimal pH and temperature were 10.0 and 40 °C, respectively. Alkaline protease showed high stability at pH 9–11. Mn2+ and Tween-80 improved its activity by 22% and 31%, respectively, while SDS, CMC and EDTA respectively inhibited its activity by 33%, 47% and 22%. Alkaline protease exhibited poor tolerance to n-butyl alcohol and ethanol, but showed resistance to H2O2. 29.8% of the original activity was still retained after 0.5 M H2O2 treatment for 3 min. The Km and Vmax values of this enzyme towards casein were 16.7 mg/ml and 14.7 µg/(min·ml), respectively. This study provides a basis for understanding enzymatic properties of Bacillus sp. ZJ1502 alkaline protease.

The compound γ-aminobutyric acid (GABA) is widely used in various fields. The combinatory strategy of the gene knockout and overexpression in a wild-type Escherichia coli K12 to improve the production of GABA was investigated. In E. coli, gabP encodes GABA permease responsible for absorbing extracellular GABA from medium. gabT and puuE encode GABA transaminases that can catalyze the reversible interconversion between GABA and succinic semialdehyde (SSA). These three genes were firstly deleted in E. coli K12 using λRed/ET system. Another three genes related to the biosynthesis and transportation of GABA, gadA, gadB and gadC, were overexpressed. The selected E. coli ZGK12-TPE with simultaneous deletion of gabT, gabP and puuE produced 0.73 g/L of GABA from 10 g/L of monosodium glutamate (MSG), much higher than the wild-type E. coli K12 with a GABA titer of only 0.11 g/L. After the transformation of the recombinant plasmid pTrc99a-gadABC in E. coli ZGK12-TPE, 6.40 g/L of GABA could be produced. Finally, 19.79 g/L of GABA was successfully obtained from fermentation medium in 1 L shake flask. This study provides new insight into the efficient biosynthesis of GABA.