PROKR1-CREB-NR4A2 axis for oxidative muscle fiber specification and improvement of metabolic function.

Metabolic disorders are characterized by an imbalance in muscle fiber composition, and a potential therapeutic approach involves increasing the proportion of oxidative muscle fibers. Prokineticin receptor 1 (PROKR1) is a G protein-coupled receptor that plays a role in various metabolic functions, but its specific involvement in oxidative fiber specification is not fully understood. Here, we investigated the functions of PROKR1 in muscle development to address metabolic disorders and muscular diseases. A meta-analysis revealed that the activation of PROKR1 upregulated exercise-responsive genes, particularly nuclear receptor subfamily 4 group A member 2 (NR4A2). Further investigations using ChIP-PCR, luciferase assays, and pharmacological interventions demonstrated that PROKR1 signaling enhanced NR4A2 expression by Gs-mediated phosphorylation of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) in both mouse and human myotubes. Genetic and pharmacological interventions showed that the PROKR1-NR4A2 axis promotes the specification of oxidative muscle fibers in both myocytes by promoting mitochondrial biogenesis and metabolic function. Prokr1-deficient mice displayed unfavorable metabolic phenotypes, such as lower lean mass, enlarged muscle fibers, impaired glucose, and insulin tolerance. These mice also exhibited reduced energy expenditure and exercise performance. The deletion of Prokr1 resulted in decreased oxidative muscle fiber composition and reduced activity in the Prokr1-CREB-Nr4a2 pathway, which were restored by AAV-mediated Prokr1 rescue. In summary, our findings highlight the activation of the PROKR1-CREB-NR4A2 axis as a mechanism for increasing the oxidative muscle fiber composition, which positively impacts overall metabolic function. This study lays an important scientific foundation for the development of effective muscular-metabolic therapeutics with unique mechanisms of action.

Regarding reference state to identify priority areas for ecological restoration in a karst region.

Widespread degradation of natural ecosystems around the globe has resulted in several ecological problems. Ecological restoration is considered a global priority as an important means of mitigating ecosystem degradation and enhancing ecosystem services provision. Regarding ecosystem reference state is a prerequisite for ecological restoration. However, there were few studies focusing on how to regard reference state for ecological restoration, especially under a changing climate. Taking Guizhou Province, a typical karst region in China, as a case study area, in this study we firstly assessed ecosystem services under homogeneous climate conditions. Secondly, we defined the optimal ecosystem services as ecosystem reference state, and then evaluated restoration suitability under a comprehensive framework. Finally, ecological restoration priority areas (EPRAs), which included ecological reconstruction areas, assisted regeneration areas and conservation priority areas needing restoration, were identified by integrating restoration suitability and conservation priority areas. The results showed that the services of water conservation and habitat maintenance only increased less than 10% from 2001 to 2018. Identified ecological reconstruction areas and assisted regeneration areas covered 1078 km2 and 1159 km2 respectively. Additionally, 15 conservation priority areas with the total area of 18,507 km2 were identified as conservation priority areas needing restoration. Accounting for 11.78% of the total area, ERPAs were mostly located in the eastern part of Guizhou, including Qiandongnan, Tongren, and Zunyi. The approach proposed here for regarding ecosystem reference state after controlling climate variables and the framework for identifying ERPAs can provide a scientific reference for large-scale ecological restoration planning.

High-throughput and trace analysis of diazepam in plasma using DART-MS/MS and its pharmacokinetic application.

A high-throughput quantitative analytical method based on Direct Analysis in Real Time tandem mass spectrometry (DART-MS/MS) has been developed and validated for the determination of diazepam in rat plasma, whereby analyzing of each sample needs merely 25 µL plasma, simple solid phase extraction sample preparation and 15 s acquisition time. The multiple reaction monitoring (MRM) transitions at m/z 285.2 â†’ 193.1 and 316.0 â†’ 270.0 were selected for the monitoring of diazepam and its internal standard clonazepam respectively. A good linearity within the range of 10-2000 ng/mL, an intra- and inter-day precisions within <7.78% as to an accuracy ranging from 1.04% to 7.92% have been achieved. The method has been successfully applied to the pharmacokinetic study of diazepam in rats' plasma after a single intragastric administration at a dose of 10 mg/kg. The results indicate that this method fulfills the requirements of the bioanalysis in sensitivity and accuracy. It shows considerable promise for application of DART-MS to the quantitative investigation of other drugs.

Upregulation of Tubulointerstitial nephritis antigen like 1 promotes gastric cancer growth and metastasis by regulating multiple matrix metallopeptidase expression.

BACKGROUND AND AIM: Tubulointerstitial nephritis antigen-like 1 (TINAGL1), as a novel matricellular protein, has been demonstrated to participate in cancer progression, whereas the potential function of TINAGL1 in gastric cancer (GC) remains unknown. METHODS: The expression pattern of TINAGL1 in GC was examined by immunohistochemistry, ELISA, real-time polymerase chain reaction, and Western blot. Correlation between TINAGL1 and matrix metalloproteinases (MMPs) was analyzed by the GEPIA website and Kaplan-Meier plots database. The lentivirus-based TINAGL1 knockdown, CCK-8, and transwell assays were used to test the function of TINAGL1 in vitro. The role of TINAGL1 was confirmed by subcutaneous xenograft, abdominal dissemination, and lung metastasis model. Microarray experiments, ELISA, real-time polymerase chain reaction, and Western blot were used to identify molecular mechanism. RESULTS: TINAGL1 was increased in GC tumor tissues and associated with poor patient survival. Moreover, TINAGL1 significantly promoted GC cell proliferation and migration in vitro as well as facilitated GC tumor growth and metastasis in vivo. TINAGL1 expression in GC cells was accompanied with increasing MMPs including MMP2, MMP9, MMP11, MMP14, and MMP16. GEPIA database revealed that these MMPs were correlated with TINAGL1 in GC tumors and that the most highly expressed MMP was MMP2. Mechanically, TINAGL1 regulated MMP2 through the JNK signaling pathway activation. CONCLUSIONS: Our data highlight that TINAGL1 promotes GC growth and metastasis and regulates MMP2 expression, indicating that TINAGL1 may serve as a therapeutic target for GC.

Development and Application of an MSALL-Based Approach for the Quantitative Analysis of Linear Polyethylene Glycols in Rat Plasma by Liquid Chromatography Triple-Quadrupole/Time-of-Flight Mass Spectrometry.

Polyethylene glycols (PEGs) are synthetic polymers composed of repeating ethylene oxide subunits. They display excellent biocompatibility and are widely used as pharmaceutical excipients. To fully understand the biological fate of PEGs requires accurate and sensitive analytical methods for their quantitation. Application of conventional liquid chromatography-tandem mass spectrometry (LC-MS/MS) is difficult because PEGs have polydisperse molecular weights (MWs) and tend to produce multicharged ions in-source resulting in innumerable precursor ions. As a result, multiple reaction monitoring (MRM) fails to scan all ion pairs so that information on the fate of unselected ions is missed. This Article addresses this problem by application of liquid chromatography-triple-quadrupole/time-of-flight mass spectrometry (LC-Q-TOF MS) based on the MSALL technique. This technique performs information-independent acquisition by allowing all PEG precursor ions to enter the collision cell (Q2). In-quadrupole collision-induced dissociation (CID) in Q2 then effectively generates several fragments from all PEGs due to the high collision energy (CE). A particular PEG product ion (m/z 133.08592) was found to be common to all linear PEGs and allowed their total quantitation in rat plasma with high sensitivity, excellent linearity and reproducibility. Assay validation showed the method was linear for all linear PEGs over the concentration range 0.05-5.0 µg/mL. The assay was successfully applied to the pharmacokinetic study in rat involving intravenous administration of linear PEG 600, PEG 4000, and PEG 20000. It is anticipated the method will have wide ranging applications and stimulate the development of assays for other pharmaceutical polymers in the future.

Interleukin 6 induces M2 macrophage differentiation by STAT3 activation that correlates with gastric cancer progression.

Interleukin 6 (IL-6) was abundant in the tumor microenvironment and played potential roles in tumor progression. In our study, the expression of IL-6 in tumor tissues from 36 gastric cancer (GC) patients was significantly higher than in non-tumor tissues. Moreover, the number of CD163+CD206+ M2 macrophages that infiltrated in tumor tissues was significantly greater than those infiltrated in non-tumor tissues. The frequencies of M2 macrophages were positively correlated with the IL-6 expression in GC tumors. We also found that IL-6 could induce normal macrophages to differentiate into M2 macrophages with higher IL-10 and TGF-ß expression, and lower IL-12 expression, via activating STAT3 phosphorylation. Accordingly, knocking down STAT3 using small interfering RNA decreased the expression of M2 macrophages-related cytokines (IL-10 and TGF-ß). Furthermore, supernatants from IL-6-induced M2 macrophages promote GC cell proliferation and migration. Moreover, IL-6 production and CD163+CD206+ M2 macrophage infiltration in tumors were associated with disease progression and reduced GC patient survival. In conclusion, our data indicate that IL-6 induces M2 macrophage differentiation (IL-10highTGF-ßhighIL-12 p35low ) by activating STAT3 phosphorylation, and the IL-6-induced M2 macrophages exert a pro-tumor function by promoting GC cell proliferation and migration.

MSAll strategy for comprehensive quantitative analysis of PEGylated-doxorubicin, PEG and doxorubicin by LC-high resolution q-q-TOF mass spectrometry coupled with all window acquisition of all fragment ion spectra.

The covalent attachment of polyethylene glycol (PEG) to therapeutic compounds (known as PEGylation) is one of the most promising techniques to improve the biological efficacy of small molecular weight drugs. After administration, PEGylated prodrugs can be metabolized into pharmacologically active compounds so that PEGylated drug, free drug and released PEG are present simultaneously in the body. Understanding the pharmacokinetic behavior of these three compounds is needed to guide the development of pegylated theranostic agents. However, PEGs are polydisperse molecules with a wide range of molecular weights, so that the simultaneous quantitation of PEGs and PEGylated molecules in biological matrices is very challenging. This article reports the application of a data-independent acquisition method (MSAll) based on liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (LC-q-q-TOF-MS) in the positive ion mode to the simultaneous determination of methoxyPEG2000-doxorubicin (mPEG2K-Dox) and its breakdown products in rat blood. Using the MSAll technique, precursor ions of all molecules are generated in q1, fragmented to product ions in q2 (collision cell), and subjected to TOF separation before precursor and product ions are recorded using low and high collision energies (CE) respectively in different experiments for a single sample injection. In this study, dissociation in q2 generated a series of high resolution PEG-related product ions at m/z 89.0611, 133.0869, 177.1102, 221.1366, 265.1622, 309.1878, and 353.2108 corresponding to fragments containing various numbers of ethylene oxide subunits, Dox-related product ions at m/z 321.0838 and 361.0785, and an mPEG2K-Dox specific product ion at m/z 365.0735. Detection of mPEGs and mPEG2K-Dox was based on high resolution extracted ions of mPEG and the specific compound. The method was successfully applied to a pharmacokinetic study of doxorubicin, mPEG2K (methylated polyethylene glycol 2K), and mPEG2K-doxorubicin in rats after a single intravenous injection of mPEG2K-doxorubicin. To the best of our knowledge, this is the first assay that simultaneously determines mPEG, Dox, and mPEG2K-Dox in a biological matrix. We believe the MSAll technique as applied in this study can be potentially extended to the determination of other PEGylated small molecules or polymeric compounds.

A validated UPLC-MS/MS method coupled with protein precipitation and ion exchange solid phase extraction for the quantitation of porcine relaxin B29 in dog plasma and its application to a pharmacokinetic study.

Porcine relaxin is a 6 kDa peptide hormone of pregnancy with important physiological and pharmacological effects. It contains a number of analogs of which porcine relaxin B29 is one of the most important. To support the development of porcine relaxin B29 as a new drug, we established an UPLC-MS/MS method for its quantitation in dog plasma. Sample preparation by protein precipitation and ion exchange solid phase extraction was followed by UPLC on an XBridge™ BEH300 C18 column at 40 °C in a run time of only 5.5 min. Detection was performed on a Qtrap 6500 mass spectrometer using ESI in the positive ion mode with MRM of the transitions at m/z 831.7 [M+7H]7+ â†’ 505.4 and m/z 1162.4 [M+5H]5+ â†’ 226 for pRLX B29 and internal standard (recombinant human insulin), respectively. The method was linear over the concentration range 30-2000 ng/mL with no matrix effects. Intra- and inter-day precisions were < 15% with accuracies in the range 98.8-100.6%. The method was successfully applied to a pharmacokinetic study in beagle dogs after administration of a 0.15 mg/kg intravenous dose. Graphical abstract Sample preparation and detection procedure.

Determination of rabeprazole enantiomers in dog plasma by supercritical fluid chromatography tandem mass spectrometry and its application to a pharmacokinetic study.

Rabeprazole is a novel benzimidazole proton pump inhibitor used for the treatment of gastrointestinal disorders. It is a chiral molecule that gives rise to the possibility of stereoselective pharmacokinetics. To investigate this phenomenon, a rapid and sensitive chiral assay based on supercritical fluid chromatography tandem mass spectrometry was developed and applied to the determination of (R)-rabeprazole and (S)-rabeprazole in dog plasma. Sample preparation involved protein precipitation with acetonitrile after the addition of (R)-lansoprazole as internal standard. Baseline separation of enantiomers in 4.5 min was achieved on an Acquity UPC2 system using an ACQUITY UPC2 Trefoil CEL2 column maintained at 60°C and a mobile phase consisting of methanol/CO2 (30:70, v/v) delivered at 2.5 mL/min. Detection was achieved by multiple reaction monitoring of the transitions at m/z 360.0→242.2 (rabeprazole) and 370.3→252.0 (internal standard) in the positive ion mode. The assay was linear in the range of 1-1000 ng/mL and free of matrix effects. Intra- and interday precisions were less than 10.0% with accuracy in the range of -2.6 to 3.1%. The method was successfully applied to a pharmacokinetic study of rabeprazole enantiomers after administration of a single oral dose of 10 mg racemate to beagle dogs.

Single administration of Selective Internal Radiation Therapy versus continuous treatment with sorafeNIB in locally advanced hepatocellular carcinoma (SIRveNIB): study protocol for a phase iii randomized controlled trial.

BACKGROUND: Approximately 20 % of hepatocellular carcinoma (HCC) patients diagnosed in the early stages may benefit from potentially curative ablative therapies such as surgical resection, transplantation or radiofrequency ablation. For patients not eligible for such options, prognosis is poor. Sorafenib and Selective Internal Radiation Therapy (SIRT) are clinically proven treatment options in patients with unresectable HCC, and this study aims to assess overall survival following either SIRT or Sorafenib therapy for locally advanced HCC patients. METHODS: This investigator-initiated, multi-centre, open-label, randomized, controlled trial will enrol 360 patients with locally advanced HCC, as defined by Barcelona Clinic Liver Cancer stage B or stage C, without distant metastases, and which is not amenable to immediate curative treatment. Exclusion criteria include previous systemic therapy, metastatic disease, complete occlusion of the main portal vein, or a Child-Pugh score of >7. Eligible patients will be randomised 1:1 and stratified by centre and presence or absence of portal vein thrombosis to receive either a single administration of SIRT using yttrium-90 resin microspheres (SIR-Spheres®, Sirtex Medical Limited, Sydney, Australia) targeted at HCC in the liver by the trans-arterial route or continuous oral Sorafenib (Nexavar®, Bayer Pharma AG, Berlin, Germany) at a dose of 400 mg twice daily until disease progression, no further response, complete regression or unacceptable toxicity. Patients for both the Sorafenib and SIRT arms will be followed-up every 4 weeks for the first 3 months and 12 weekly thereafter. Overall survival is the primary endpoint, assessed for the intention-to-treat population. Secondary endpoints are tumour response rate, time-to-tumour progression, progression free survival, quality of life and down-staging to receive potentially curative therapy. DISCUSSION: Definitive data comparing these two therapies will help to determine clinical practice in the large group of patients with locally advanced HCC and improve outcomes for such patients. TRIAL REGISTRATION: ClinicalTrials.gov identifier, NCT01135056 , first received 24, May 2010.

Significant Improvement of Metabolic Characteristics and Bioactivities of Clopidogrel and Analogs by Selective Deuteration.

In the search for prodrug analogs of clopidogrel with improved metabolic characteristics and antiplatelet bioactivity, a group of clopidogrel and vicagrel analogs selectively deuterated at the benzylic methyl ester group were synthesized, characterized, and evaluated. The compounds included clopidogrel-d3 (8), 2-oxoclopidogrel-d3 (9), vicagrel-d3 (10a), and 12 vicagrel-d3 analogs (10b-10m) with different alkyl groups in the thiophene ester moiety. The D3C-O bond length in 10a was shown by X-ray single crystal diffraction to be shorter than the H3C-O bond length in clopidogrel, consistent with the slower rate of hydrolysis of 8 than of clopidogrel in rat whole blood in vitro. A study of the ability of the compounds to inhibit ADP-induced platelet aggregation in fresh rat whole blood collected 2 h after oral dosing of rats with the compounds (7.8 µmol/kg) showed that deuteration increased the activity of clopidogrel and that increasing the size of the alkyl group in the thiophene ester moiety reduced activity. A preliminary pharmacokinetic study comparing 10a with vicagrel administered simultaneously as single oral doses (72 µmol/kg of each drug) to male Wistar rats showed 10a generated more of its active metabolite than vicagrel. These results suggest that 10a is a potentially superior antiplatelet agent with improved metabolic characteristics and bioactivity, and less dose-related toxicity.

Performance of mobile digital X-ray fluoroscopy using a novel flat panel detector for intraoperative use.

BACKGROUND: Technologies employing digital X-ray devices are developed for mobile settings. OBJECTIVE: To develop a mobile digital X-ray fluoroscopy (MDF) for intraoperative guidance, using a novel flat panel detector to focus on diagnostics in outpatient clinics, operating and emergency rooms. METHODS: An MDF for small-scale field diagnostics was configured using an X-ray source and a novel flat panel detector. The imager enabled frame rates reaching 30 fps in full resolution fluoroscopy with maximal running time of 5 minutes. Signal-to-noise (SNR), contrast-to-noise (CNR), and spatial resolution were analyzed. Stray radiation, exposure radiation dose, and effective absorption dose were measured for patients. RESULTS: The system was suitable for small-scale field diagnostics. SNR and CNR were 62.4 and 72.0. Performance at 10% of MTF was 9.6 lp/mm (53 µ m) in the no binned mode. Stray radiation at 100 cm and 150 cm from the source was below 0.2 µ Gy and 0.1 µ Gy. Exposure radiation in radiography and fluoroscopy (5 min) was 10.2 µ Gy and 82.6 mGy. The effective doses during 5-min-long fluoroscopy were 0.26 mSv (wrist), 0.28 mSv (elbow), 0.29 mSv (ankle), and 0.31 mSv (knee). CONCLUSIONS: The proposed MDF is suitable for imaging in operating rooms.

Development of a mini-mobile digital radiography system by using wireless smart devices.

The current technologies that trend in digital radiology (DR) are toward systems using portable smart mobile as patient-centered care. We aimed to develop a mini-mobile DR system by using smart devices for wireless connection into medical information systems. We developed a mini-mobile DR system consisting of an X-ray source and a Complementary Metal-Oxide Semiconductor (CMOS) sensor based on a flat panel detector for small-field diagnostics in patients. It is used instead of the systems that are difficult to perform with a fixed traditional device. We also designed a method for embedded systems in the development of portable DR systems. The external interface used the fast and stable IEEE 802.11n wireless protocol, and we adapted the device for connections with Picture Archiving and Communication System (PACS) and smart devices. The smart device could display images on an external monitor other than the monitor in the DR system. The communication modules, main control board, and external interface supporting smart devices were implemented. Further, a smart viewer based on the external interface was developed to display image files on various smart devices. In addition, the advantage of operators is to reduce radiation dose when using remote smart devices. It is integrated with smart devices that can provide X-ray imaging services anywhere. With this technology, it can permit image observation on a smart device from a remote location by connecting to the external interface. We evaluated the response time of the mini-mobile DR system to compare to mobile PACS. The experimental results show that our system outperforms conventional mobile PACS in this regard.

Overcoming the challenge of potent endogenous interferences in limaprost quantification: An innovative methodology combining differential mobility spectrometry with LC-MS/MS for ultra-high sensitivity, selectivity and significantly enhanced throughput.

Limaprost, an orally administered analogue of prostaglandin E1, possesses potent vasodilatory, antiplatelet, and cytoprotective properties. Due to its extremely low therapeutic doses and exceedingly low plasma concentrations, the pharmacokinetic and bioequivalence studies of limaprost necessitate a highly sensitive quantitative method with a sub-pg/mL level of lower limit of quantification. Moreover, the intensity of endogenous interferences can even exceed the maximum concentration level of limaprost in human plasma, presenting further challenge to the quantification of limaprost. As a result, existing methods have not yet met the necessary level of sensitivity, selectivity, and throughput needed for the quantitative analysis of limaprost in pharmacokinetic and bioequivalence investigations. This study presents a new methodology that combines differential mobility spectrometry (DMS) with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and utilizes a distinctive strategy to achieve more accurate DMS conditions. This integration yields a method that is currently the most sensitive and features the shortest analytical time, making it the sole technique capable of meeting the requirements for limaprost pharmacokinetic and bioequivalence investigations. This method demonstrates robustness and is successfully employed in a pharmacokinetic investigation of limaprost in human subjects, underscoring that the combination of DMS with LC-MS/MS serves as an efficacious strategy for overcoming the challenges inherent in analyzing biological samples afflicted by multiple interferences.

Effect of lipo-prostaglandin E1 on cystatin C, ß2-microglobulin, and estimated glomerular filtration rate in patients with decompensated heart failure and renal dysfunction: a single-center, nonrandomized controlled study.

A nonrandomized controlled study was conducted to evaluate the effect of lipo-prostaglandin E1 (lipo-PGE1) on cystatin C (CysC), ß2-microglobulin (B2MG), and estimated glomerular filtration rate (eGFR) in patients with decompensated heart failure (DHF) and renal dysfunction. A total of 286 enrolled patients with DHF and renal dysfunction were nonrandomly assigned a 7-day standard treatment without (n = 146) or with (n = 140) lipo-PGE1 intervention. According to the baseline eGFR, patients were further classified into mild, moderate, and severe renal dysfunction subgroups. By the end of study period, there was no evidence of an immense improvement in B2MG, CysC, and eGFR in response to standard treatment (all P > 0.05). On the contrary, a noticeable decrease of B2MG and CysC was observed in patients receiving lipo-PGE1 intervention, as well as an increase in eGFR (all P < 0.05). Moreover, lipo-PGE1 intervention led to greater changes in renal function variables from baseline than with standard management (all P < 0.05). Most important, the favorable renal protective effects of lipo-PGE1 were maintained in three subgroups. Lipo-PGE1 intervention brought a substantial renoprotective benefit to hospitalized DHF patients as compared with standard therapy, suggesting it might offer a promising therapeutic option for the management of renal dysfunction associated with DHF.

A multilayer affective computing model with evolutionary strategies reflecting decision-makers' preferences in process control.

Many industrial control problems related to multi-objective optimization, such as controller parameters tuning, often require operators to perform multiple-step interactions without considering the changes of decision-makers' affective states and quantitative description of decision-makers' preferences during the interactive decision. Regarding this problem, we developed a multilayer affective computing model (MACM), including three factors: human personality, emotional space, and affective states, to demonstrate the iterative affective computing during the interactions. First, a concise model of affective computing-driven interactive decision-making was built before three submodules involved were described in detail. (1) An affective state recognition method based on facial expressions was presented, providing the basis for obtaining expert affective states during decision-making. (2) An identification method of affective parameters was given, providing an approach to describing personalized affective state-changing rules of different persons. (3) A definition of decision-makers' preferences in interactive decision-making was specified. In addition, a decision-makers' preferences mining method was developed by the MACM and an iterative learning control (ILC) strategy. Thus, we proposed affective computing-driven interactive decision-making method, which provided a simplified approach to converting the interactive decision problems based on decision-makers' preferences to decision issues based on incremental decision vector, along with assisting computers to learn from human experts and perform decision-making automatically in a general sense. Then, two typical process control cases-PI controller tuning for decoupling problem and manipulate vector optimization for batch processes were used to show the correctness and effectiveness of the contributions. Compared with other traditional optimization algorithms without affective state tracking and recognition (fuzzy control, ILC, reinforcement learning, and so on), experimental results indicated that the proposed method could achieve good performance. Finally, this study presented the efficiency and limitations of using this technique for a specific application.

Assessing the Cultural Ecosystem Services Value of Protected Areas Considering Stakeholders' Preferences and Trade-Offs-Taking the Xin'an River Landscape Corridor Scenic Area as an Example.

Improving the accuracy of cultural ecosystem services (CESs) value assessment and paying more attention to the preferences and trade-offs of stakeholders in the administration of CESs are of vital importance for achieving resilient ecosystem management. Combining methodologies from sociology (Q method) and economics (choice experiment), an assessment framework of CESs is introduced to examine stakeholders' preferences and willingness to pay to participate in CESs in protected areas so as to explore how the value of CESs in protected areas can be optimized. The results show that the selection of CESs by stakeholders reflects certain synergies and trade-offs. Visitors can be classified as preferring humanistic-natural recreation, aesthetic-sense of place, or environmental education according to the factor ranking of the Q method. Visitors have a higher willingness to pay for humanistic heritage and a lower willingness to pay for sense of place experience, which can be measured at $6.55 per visit and $0.96 per visit, respectively. This indicates that the local customs and characteristics should be further explored and promoted through traditional festival celebrations and farming activities in further development of protected areas, apart from protecting local cultural heritages such as Huizhou ancient villages and halls. Furthermore, it is also necessary to actively explore the synergistic development of CESs, promote social participation, raise stakeholders' awareness of available services, manage visitors and stakeholders from a demand perspective, and promote the realization of the value of ecological products in protected areas.

Controlling Non-Grain Production Based on Cultivated Land Multifunction Assessment.

The control of non-grain production (NGP) has become a great challenge for cultivated land protection in China in recent years. A control method for NGP that can coordinate the conflicts between cultivated land protection and farmers' interest is urgently needed. Taking Tongxiang City as an example, this research proposed a solution for the control and management of NGP based on cultivated land multifunctional assessment. The GIS and AHP approach were used to assess production function via a comprehensive evaluation index. The InVEST and FMSPA models were applied to assess ecological function while, the Maxent model was applied to assess recreational function, then multifunctional comprehensive zoning was conducted through natural breakpoint method and spatial overlay analysis. Five development-oriented function zones were considered, including the core area of grain production plus areas for ecological agriculture, leisure agriculture, compound agriculture, and general farmland. Differentiated control measures for NGPs in each functional subarea are proposed considering the current NGP distribution of Tongxiang city. This research can provide a reference for subsequent improvement of land management policies and can aid the achievement of sustainable agricultural development and rural revitalization.

Considering Farmers' Heterogeneity to Payment Ecosystem Services Participation: A Choice Experiment and Agent-Based Model Analysis in Xin'an River Basin, China.

The concept of watershed ecological compensation is one payment for ecosystem services (PES) program that incentivizes stakeholders undertake environmental conservation activities that improve the provision of ecosystem services. Defining the heterogeneity of farmers' willingness to participate in watershed ecological compensation is critically important for fully understanding stakeholders' demands. Accordingly, we designed a choice experiment survey to analyze the heterogeneity of policy preferences and willingness to receive compensation between upstream and midstream farmers in Xin'an River basin, China. Moreover, we simulated the impact of farmers' social capitals' heterogeneity with an agent-based model. The results show that there are significant differences in the preferences of agricultural waste recycling rate and agricultural water quality between farmers in the upstream and midstream. The total willingness of farmers in the upstream and midstream to participate in ecological compensation are RMB 149.88 (USD 22.54)/month and RMB 57.40 yuan (USD 8.63)/month, respectively. Social network size has a negative effect on farmers' willingness to participate the programs. Our findings suggest that the characteristics of farmers' influence their willingness to participate in the PES program. The results of this research can be used to improve PES management policies in the future, as well as to support sustainable environmental development and rural revitalization.

An action recognition method for manual acupuncture techniques using a tactile array finger cot.

The current measurement systems for the physical parameters (rotation frequency, and amplitude) of Traditional Chinese Medicine (TCM) manual acupuncture tend to cause disturbance and inconvenience in clinical application and do not accurately capture the tactile signals from the physician's finger during manual acupuncture operations. In addition, the literature rarely discusses classification of the four basic manual acupuncture techniques (reinforcing by twirling and rotating (RFTR), reducing by twirling and rotating (RDTR), reinforcing by lifting and thrusting (RFLT), and reducing by lifting and thrusting (RDLT)). To address this problem, we developed a multi-PVDF film-based tactile array finger cot to collect piezoelectric signals from the acupuncturist's finger-needle contact during manual acupuncture operations. In order to recognize the four typical TCM manual acupuncture techniques, we developed a method to capture piezoelectric signals in related "windows" and subsequently extract features to model acupuncture techniques. Next, we created an ensemble learning-based action classifier for manual acupuncture technique recognition. Finally, the proposed classifier was employed to recognize the four types of manual acupuncture techniques performed by 15 TCM physicians based on the piezoelectric signals collected using the tactile array finger cot. Among all the approaches, our proposed feature-based CatBoost ensemble learning model achieved the highest validation accuracy of 99.63% and the highest test accuracy of 92.45%. Moreover, we provide the efficiency and limitations of using this action recognition method.