CD39hi identifies an exhausted tumor-reactive CD8+ T cell population associated with tumor progression in human gastric cancer.

The ectonucleotidase CD39 has been regarded as a promising immune checkpoint in solid tumors. However, the expression of CD39 by tumor-infiltrating CD8+ T cells as well as their potential roles and clinical implications in human gastric cancer (GC) remain largely unknown. Here, we found that GC-infiltrating CD8+ T cells contained a fraction of CD39hi cells that constituted about 6.6% of total CD8+ T cells in tumors. These CD39hi cells enriched for GC-infiltrating CD8+ T cells with features of exhaustion in transcriptional, phenotypic, metabolic and functional profiles. Additionally, GC-infiltrating CD39hiCD8+ T cells were also identified for tumor-reactive T cells, as these cells expanded in vitro were able to recognize autologous tumor organoids and induced more tumor cell apoptosis than those of expanded their CD39int and CD39-CD8+ counterparts. Furthermore, CD39 enzymatic activity controlled GC-infiltrating CD39hiCD8+ T cell effector function, and blockade of CD39 efficiently enhanced their production of cytokines IFN-γ and TNF-α. Finally, high percentages of GC-infiltrating CD39hiCD8+ T cells correlated with tumor progression and independently predicted patients' poor overall survival. These findings provide novel insights into the association of CD39 expression level on CD8+ T cells with their features and potential clinical implications in GC, and empowering those exhausted tumor-reactive CD39hiCD8+ T cells through CD39 inhibition to circumvent the suppressor program may be an attractive therapeutic strategy against GC.

Palladium-Catalyzed Sequential Cross-Coupling/Annulation of ortho-Vinyl Bromobenzene with Aryl Bromide: Bimetallic Pathway versus Pd(II)-Pd(IV) Pathway: A DFT Investigation.

The palladium-catalyzed sequential cross-coupling/annulation of ortho-vinyl bromobenzenes with aryl bromides generating phenanthrenes was characterized by density functional theory (DFT). The Pd(II)-Pd(IV) pathway (Path V) is shown to be less probable than the bimetallic pathway (Path I), the latter proceeding via the following six steps: oxidative addition, vinyl-C(sp2)-H activation, Pd(II)-Pd(II) transmetalation, C-C coupling, aryl-C(sp2)-H activation, and reductive elimination. The aryl-C(sp2)-H activation process acts as the rate-determining step (RDS) of the entire chemical transformation, with an activation free energy barrier of ca. 27.4-28.8 kcal·mol-1, in good agreement with the corresponding experimental data (phenanthrenes' yields of ca. 65-90% at 130 °C after 5 h of reaction). The K2CO3 additive effectively reduces the activation free energy barrier of the RDS through direct participation in the reaction while preferentially modulating the charge distributions and increasing the stability of corresponding intermediates and complexes along the reaction path. Furthermore, bonding and electronic structure analyses of the key structures indicate that the chemo- and regioselectivities of the reaction are strongly influenced by both electronic effects and steric hindrance.

Acteoside ameliorates learning and memory impairment in APP/PS1 transgenic mice by increasing Aß degradation and inhibiting tau hyperphosphorylation.

Alzheimer's disease (AD) is a neurodegenerative disease. Senile plaques and intracellular neurofibrillary tangles are pathological hallmarks of AD. Recent studies have described the improved cognitive and neuroprotective functions of acteoside (AS). This study aimed to investigate whether the improved cognition of AS was mediated by Aß degradation and tau phosphorylation in APP/PS1 mice. The open field, Y maze, and novel object recognition tests were used to assess cognitive behavioral changes. We evaluated the levels of Aß40 and Aß42 in serum, cortex, and hippocampus, and Aß-related scavenging enzymes, phosphorylated GSK3ß and hyperphosphorylated tau in the cortex and hippocampus of APP/PS1 mice by western blotting. Our results revealed that AS treatment ameliorated anxious behaviors, spatial learning, and memory impairment in APP/PS1 mice and significantly reduced Aß deposition in their serum, cortex, and hippocampus. AS significantly increased Aß degradation, inhibited the hyperphosphorylation of tau, and significantly decreased the activity of GSK3ß, which is involved in tau phosphorylation. Altogether, these findings indicated that the beneficial effects of AS on AD-associated anxious behaviors and cognitive impairments could be attributed to promoting Aß degradation and inhibiting tau hyperphosphorylation, which might be partly mediated by GSK3ß.

Smart Target-Initiated Catalytic DNA Junction Circuit Amplification Strategy for the Ultrasensitive Electrochemiluminescence Detection of MicroRNA.

One of the highly attractive research directions in the electrochemiluminescence (ECL) field is how to regulate and improve ECL efficiency. Quantum dots (QDs) are highly promising ECL materials due to their adjustable luminescence size and strong luminous efficiency. MoS2 NSs@QDs, an ECL emitter, is synthesized via hydrothermal methods, and its ECL mechanism is investigated using cyclic voltammetry and ECL-potential curves. Then, a stable and vertical attachment of a triplex DNA (tsDNA) probe to the MoS2 nanosheets (NSs) is applied to the electrode. Next, an innovative ECL sensor is courageously empoldered for precise and ultrasensitive detection of target miRNA-199a through the agency of ECL-resonance energy transfer (RET) strategy and a dextrous target-initiated catalytic three-arm DNA junction assembly (CTDJA) based on a toehold strand displacement reaction (TSDR) signal amplification approach. Impressively, the ingenious system not only precisely regulates the distance between energy donor-acceptor pairs leave energy less loss and more ECL-RET efficiency, but also simplifies the operational procedure and verifies the feasibility of this self-assembly process without human intervention. This study can expand MoS2 NSs@QDs utilization in ECL biosensing applications, and the proposed nucleic acid amplification strategy can become a miracle cure for ultrasensitive detecting diverse biomarkers, which helps researchers to better study the tumor mechanism, thereby unambiguously increasing cancer cure rates and reducing the risk of recurrence.

Fusion data from FT-IR and MALDI-TOF MS result in more accurate classification of specific microbiota.

Microbes are usually present as a specific microbiota, and their classification remains a challenge. MALDI-TOF MS is particularly successful in library-based microbial identification at the species level as it analyzes the molecular weight of peptides and ribosomal proteins. FT-IR allows more accurate classification of bacteria at the subspecies level due to the high sensitivity, specificity and repeatability of FT-IR signals from bacteria, which is not achievable with MALDI-TOF MS. Previous studies have shown that more accurate identification results can be obtained by the fusion of FT-IR and MALDI-TOF MS spectral data. Here, we constructed 20 groups of model microbiota samples and used FT-IR, MALDI-TOF MS, and their fusion data to classify them. Hierarchical clustering analysis (HCA) showed that the classification accuracy of FT-IR, MALDI-TOF MS, and the fusion data was 85%, 90%, and 100%, respectively. These results indicate that both FT-IR and MALDI-TOF MS can effectively classify specific microbiota, and the fusion of their spectral data could improve the classification accuracy. The FT-IR and MALDI-TOF MS data fusion strategy may be a promising technology for specific microbiota classification.

Fabrication of MIL-101(Cr)/silver nanocomposites as SERS substrate for sensitive determination of malachite green and crystal violet in tilapia.

Nanocomposites with multiple functions have attracted much attention in designing novel SERS substrates. In this report, the enrichment ability of MIL-101(Cr) and the local surface plasma resonance (LSPR) of silver nanoparticles are combined to fabricate a SERS substrate denoted as MIL-101-MA@Ag, which can simultaneously produce high-density and uniformly distributed hot spots. Moreover, the enrichment ability of MIL-101(Cr) can further improve the sensitivity by concentrating and transferring the analytes in the vicinity of hot spots. Under optimal conditions, MIL-101-MA@Ag showed good SERS activity for malachite green (MG) and crystal violet (CV), with detection limits as low as 9.5×10-11 M and 9.2×10-12 M at 1616 cm-1, respectively. The prepared substrate has been successfully applied to detect MG and CV in tilapia, the recovery rate of fish tissue extract was 86.4~102%, and the relative standard deviation (RSD) was 8.9~15%. The results demonstrate that MOF-based nanocomposites are expected to be useful SERS substrates and have a universal applicability for the detection of other hazardous molecules.

How does environmental policy stringency influence green innovation for environmental managements?

In the struggle to limit global climate change and rising temperatures, the United Nations Climate Change Conference (COP27) was held in Egypt last November. Bringing together nations to recognize climate change as a global concern and to create new "building blocks" to enhance the implementation of the Paris Agreement through actions that can move the world toward a greener, and carbon free future. This study examines a panel of high-income economies from the Organization for Economic Cooperation and Development (OECD) to investigate the empirical linkage between Green Innovations (GI), Disaggregated trade (exports and imports), Environmental policy stringency (EPS), and Consumption-based carbon dioxide emissions from 1990 to 2020. We proceed with the panel cointegration check based on the results of the diagnostic tests. The method of moment quantile regressions (MMQR) is used to investigate the relationships between CCO2 and various variables in different quantiles. The data show that GI, export, imports, and EPS are major contributors in explaining the substantial variance in CCO2 emissions in the chosen panel. Specifically, severe environmental rules boost the benefits of green technologies through the use of environmentally friendly technology. Imports, on the other hand, have been determined to be harmful to environmental quality. As a result, member economies should reform their environmental policies to include consumption-based emissions objectives and discourage people' desire for carbon-intensive items from developing countries. This will eventually result in a decrease in consumption-based carbon emissions, assisting in the achievement of true emissions reduction goals and COP27 targets.

Ameliorative effects of silybin against avermectin-triggered carp spleen mitochondrial dysfunction and apoptosis through inhibition of PERK-ATF4-CHOP signaling pathway.

The aim of this study was to investigate the splenic tissue damage of environmental biological drug avermectin to freshwater cultured carp and to evaluate the effect of silybin on the splenic tissue damage of carp induced by avermectin. A total of 60 carp were divided into 4 groups with 15 carp in each group, including the control group fed with basic diet, experimental group fed with basal diet and exposed to avermectin (avermectin group), experimental group fed with basal diet supplement silybin (silybin group), and experimental group fed with basal diet supplement silybin and exposed to avermectin (silybin + avermectin group). The whole test period lasted for 30 days, and spleen tissue was collected for analysis. In this study, H&E staining, mitochondrial purification and membrane potential detection, ATP detection, DHE staining, biochemical tests, qPCR, immunohistochemistry, and apoptosis staining were used to evaluate the biological processes of spleen tissue injury, mitochondrial function, oxidative stress, apoptosis, and endoplasmic reticulum stress. The results show that silybin protected carp splenic tissue damage caused by chronic avermectin exposure, decreased mitochondrial membrane potential, decreased ATP content, ROS accumulation, oxidative stress, apoptosis, and endoplasmic reticulum stress. Silybin may ameliorate the splenic tissue damage of cultured freshwater carp caused by environmental biopesticide avermectin by alleviating mitochondrial dysfunction and inhibiting PERK-ATF4-CHOP-driven mitochondrial apoptosis. Adding silybin into the diet becomes a feasible strategy to resist the pollution of avermectin and provides a theoretical basis for creating a good living environment for freshwater carp.

Cytokine profiles of macular neovascularization in the elderly based on a classification from a pachychoroid/drusen perspective.

PURPOSE: To classify macular neovascularization (MNV) based on pachychoroid and drusen features and to examine the aqueous humor cytokine signatures of each group. METHODS: In total, 106 consecutive eyes with treatment-naïve MNV and 104 control eyes were examined. The aqueous humor concentrations of 15 cytokines were compared among the MNV groups classified based on the presence of drusen and/or pachychoroid features. Multidimensional scaling analysis was used to visualize the similarity level of the MNV subtypes according to their cytokine profiles. RESULTS: Thirty-one, 18, 43, and 10 eyes were classified into the pachychoroid-associated, drusen-associated, pachychoroid/drusen-associated, and non-drusen/non-pachychoroid MNV groups, respectively. Compared with the control group, cytokines were differently upregulated among the MNV groups. CRP and CXCL12 were significantly upregulated in all MNV groups, whereas CXCL13 and IL-8 were significantly upregulated in three MNV groups, excluding the non-pachychoroid/non-drusen-associated MNV group. Ang-2 was significantly upregulated in three MNV groups except the drusen-associated MNV group. PlGF was significantly upregulated in the pachychoroid-associated and drusen-associated MNV groups. CCL-2 was significantly upregulated in the pachychoroid-associated and pachychoroid/drusen-associated MNV groups. VEGF was downregulated in the pachychoroid-associated and drusen-associated MNV groups, respectively. Multidimensional scaling analysis showed a distinct cytokine profile for each MNV group. CONCLUSION: All MNV groups showed distinct cytokine profiles. Eyes with "neovascular age-related macular degeneration with drusen and concomitant pachychoroid" may share a similar etiology to those with "pachychoroid neovasculopathy" and "choroidal neovascularization with drusen," but have a distinct etiology to those without these. These findings suggest the importance of evaluating drusen and the choroid during the diagnosis of neovascular age-related macular degeneration.

Effect of cleaner residential heating policy on air pollution: A case study in Shandong Province, China.

Coal-fired heating in winter is a primary source of air pollution in many countries. In northern China, the use of scattered coal for winter heating has led to severe environmental issues. In this study, we use a quasi-natural experiment in Shandong Province, China, to investigate the effectiveness of a cleaner heating transition policy. Specifically, we use a difference-in-differences approach to identify the effects of the cleaner heating transition policy on air pollution using high-resolution hourly data. Our findings indicate that implementation of the policy could effectively reduce air pollution by decreasing a PM2.5 by 7.32%, PM10 by 2.62%, SO2 by 3.98%, and NO2 by 4.67%. In addition, we used event study and a series of robustness checks to further support our findings. Notably, our findings indicate that implementation of the policy includes a spatial spillover effect, which differs according to the level of compulsory implementation and the distance to a city centre. Overall, our findings can help promote the application of a cleaner transitioning policy for the entire country and offer guidance for further policy development regarding the effective reduction of winter air pollution in the developing world.

Outcome of retroarticular drilling for osteochondritis dissecans of the talus in a pediatric population.

BACKGROUND: Outcomes of bone marrow stimulation for osteochondritis dissecans (OCD) of the talus in pediatric patients is not optimal. The objective was to evaluate the retroarticular drilling technique for talar OCD. METHODS: A retrospective case-series study of pediatric cases treated for talar OCD with retroarticular drilling was done. Clinical and radiological outcome scores were recorded as follows: the percentage of patients who had a successful treatment, the percentage for every category of the Berndt and Harty treatment result grading and the percentage for every radiographical outcome score were computed. RESULTS: Nineteen patients (18 girls; mean age: 14.6 ± 2.1 years) were included. The mean follow-up was 14.8 (±11.7) months. 26.3% required revision surgery. The Berndt and Harty scores were: 57.9% good, 10.5% fair, 31.6% poor. Radiological outcomes were: 21% healed, 47.4% partially healed, 31.6% no healing. The radiological outcome score was better for younger patients (P = 0.01) and those with an open physis (P = 0.001). CONCLUSION: 26.3% of patients needed revision surgery after talar OCD retroarticular drilling and 21% were healed radiographically. Skeletal immaturity and a younger age were associated to a better radiological outcome.

Structural Elucidation of Garcipaucinones A and B From Garcinia paucinervis Using Quantum Chemical Calculations.

Two tocotrienol derivatives, garcipaucinones A (1) and B (2), and a biosynthetically related known analogue (3) were isolated from the fruit of Garcinia paucinervis. Their structures including absolute configurations were unequivocally determined by spectroscopic methods complemented with electronic circular dichroism (ECD) calculations and gauge-independent atomic orbital (GIAO) NMR calculations. Compounds 1 and 2 are the first naturally occurring tocotrienol derivatives with a 3,10-dioxatricyclo-[7.3.1.02,7]tridecane skeleton incorporating an unusual γ-pyrone motif. A reasonable biosynthetic pathway for formation of the two compounds is proposed. The antiproliferative and anti-inflammatory activities of compounds 1 and 2 were also evaluated.

The Molecular Mechanisms of Phytophthora infestans in Response to Reactive Oxygen Species Stress.

Reactive oxygen species (ROSs) are critical for the growth, development, proliferation, and pathogenicity of microbial pathogens; however, excessive levels of ROSs are toxic. Little is known about the signaling cascades in response to ROS stress in oomycetes such as Phytophthora infestans, the causal agent of potato late blight. Here, P. infestans was used as a model system to investigate the mechanism underlying the response to ROS stress in oomycete pathogens. Results showed severe defects in sporangium germination, mycelium growth, appressorium formation, and virulence of P. infestans in response to H2O2 stress. Importantly, these phenotypes mimic those of P. infestans treated with rapamycin, the inhibitor of target of rapamycin (TOR, 1-phosphatidylinositol-3-kinase). Strong synergism occurred when P. infestans was treated with a combination of H2O2 and rapamycin, suggesting that a crosstalk exists between ROS stress and the TOR signaling pathway. Comprehensive analysis of transcriptome, proteome, and phosphorylation omics showed that H2O2 stress significantly induced the operation of the TOR-mediated autophagy pathway. Monodansylcadaverine staining showed that in the presence of H2O2 and rapamycin, the autophagosome level increased in a dosage-dependent manner. Furthermore, transgenic potatoes containing double-stranded RNA of TOR in P. infestans (PiTOR) displayed high resistance to P. infestans. Therefore, TOR is involved in the ROS response and is a potential target for control of oomycete diseases, because host-mediated silencing of PiTOR increases potato resistance to late blight.

A Novel Hemoglobin Variant Hb Liaobu [α107(G14)Val→Leu, HBA2: c.322G>C] Detected by Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry.

We here describe a novel hemoglobin (Hb) variant, Hb Liaobu [α107(G14)Val→Leu, HBA2: c.322G>C], in a Chinese family. The structurally abnormal α chain variant could not be detected using capillary electrophoresis (CE) and was subsequently characterized by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS), and further confirmed by reversed phase high performance liquid chromatography (HPLC). Sanger sequencing revealed a novel base mutation on the α2-globin gene and RNA analysis by reverse transcription polymerase chain reaction (RT-PCR) showed the presence of an abnormal HBA transcript. The isopropanol stability test indicated the stable state of this structural Hb variant. In conclusion, a new Hb variant, Hb Liaobu, was discovered and characterized. It was proven to be a nonpathogenic variant. Our study resolved the confusion in the clinical diagnosis of individuals with this novel Hb variant in this family.

Transcriptomic Analysis of the Photosynthetic, Respiration, and Aerenchyma Adaptation Strategies in Bermudagrass (Cynodon dactylon) under Different Submergence Stress.

Submergence impedes photosynthesis and respiration but facilitates aerenchyma formation in bermudagrass. Still, the regulatory genes underlying these physiological responses are unclear in the literature. To identify differentially expressed genes (DEGs) related to these physiological mechanisms, we studied the expression of DEGs in aboveground and underground tissues of bermudagrass after a 7 d treatment under control (CK), shallow submergence (SS), and deep submergence (DS). Results show that compared with CK, 12276 and 12559 DEGs were identified under SS and DS, respectively. Among them, the DEGs closely related to the metabolism of chlorophyll biosynthesis, light-harvesting, protein complex, and carbon fixation were down-regulated in SS and DS. Meanwhile, a large number of DEGs involved in starch and sucrose hydrolase activities, glycolysis/gluconeogenesis, tricarboxylic acid (TCA) cycle, and oxidative phosphorylation were down-regulated in aboveground tissues of bermudagrass in SS and DS. Whereas in underground tissues of bermudagrass these DEGs were all up-regulated under SS, only beta-fructofuranosidase and α-amylase related genes were up-regulated under DS. In addition, we found that DEGs associated with ethylene signaling, Ca2+-ROS signaling, and cell wall modification were also up-regulated during aerenchyma formation in underground tissues of bermudagrass under SS and DS. These results provide the basis for further exploration of the regulatory and functional genes related to the adaptability of bermudagrass to submergence.

[An anesthesia depth computing method study based on wavelet transform and artificial neural network].

General anesthesia is an essential part of surgery to ensure the safety of patients. Electroencephalogram (EEG) has been widely used in anesthesia depth monitoring for abundant information and the ability of reflecting the brain activity. The paper proposes a method which combines wavelet transform and artificial neural network (ANN) to assess the depth of anesthesia. Discrete wavelet transform was used to decompose the EEG signal, and the approximation coefficients and detail coefficients were used to calculate the 9 characteristic parameters. Kruskal-Wallis statistical test was made to these characteristic parameters, and the test showed that the parameters were statistically significant for the differences of the four levels of anesthesia: awake, light anesthesia, moderate anesthesia and deep anesthesia ( P < 0.001). The 9 characteristic parameters were used as the input of ANN, the bispectral index (BIS) was used as the reference output, and the method was evaluated by the data of 8 patients during general anesthesia. The accuracy of the method in the classification of the four anesthesia levels of the test set in the 7:3 set-out method was 85.98%, and the correlation coefficient with the BIS was 0.977 0. The results show that this method can better distinguish four different anesthesia levels and has broad application prospects for monitoring the depth of anesthesia.

[Application of Improved Wavelet Threshold in Denoising of ECG Signals].

The ECG signal is susceptible to interference from the external environment during the acquisition process, affecting the analysis and processing of the ECG signal. After the traditional soft-hard threshold function is processed, there is a defect that the signal quality is not high and the continuity at the threshold is poor. An improved threshold function wavelet denoising is proposed, which has better regulation and continuity, and effectively solves the shortcomings of traditional soft and hard threshold functions. The Matlab simulation is carried out through a large amount of data, and various processing methods are compared. The results show that the improved threshold function can improve the denoising effect and is superior to the traditional soft and hard threshold denoising.

VdNPS, a Nonribosomal Peptide Synthetase, Is Involved in Regulating Virulence in Verticillium dahliae.

Nonribosomal peptide synthetases (NPS) are known for the biosynthesis of antibiotics, toxins, and siderophore production. They are also a virulence determinant in different phytopathogens. However, until now, the functional characterization of NPS in Verticillium dahliae has not been reported. Deletion of the NPS gene in V. dahliae led to the decrease of conidia, microsclerotia, and pathogenicity. ΔVdNPS strains were tolerant to H2O2, and the genes involved in H2O2 detoxification, iron/copper transport, and cytoskeleton were differentially expressed in ΔVdNPS. Interestingly, ΔVdNPS strains exhibited hypersensitivity to salicylic acid (SA), and the genes involved in SA hydroxylation were up-regulated in ΔVdNPS compared with wild-type V. dahliae under SA stress. Additionally, during infection, ΔVdNPS induced more pathogenesis-related gene expression, higher reactive oxygen species production, and stronger SA-mediated signaling transduction in host to overcome pathogen. Uncovering the function of VdNPS in pathogenicity could provide a reliable theoretical basis for the development of cultivars with durable resistance against V. dahliae-associated diseases.

[Development of a Wearable Wireless ECG Monitoring System with Ultra-low Power Consumption].

This study describes the development of a wireless and wearable ECG monitoring system with ultra-low power consumption. The system is mainly composed of a connection part of an ECG electrode sticker, an electrocardiogram collecting part, a data storage part, a Bluetooth main control unit, a charging module, a voltage regulator and a lithium battery. The low-power ECG acquisition chip ADS1292R and the ultra-low-power Bluetooth microcontroller nRF51822 together constitute the ECG signal acquisition and wireless data communication part. The collected ECG signals can be sent to the mobile APP through the Bluetooth function provided by the MCU, and can completly display and analysis to achieve low power system. After testing, the system power consumption is only (3.7 V×2.87 mA)10.619 mW, and if it is optimized, it can further reduce power consumption, therefore, the system design can have good applicability.

[EEG Signal Analysis Methods and Their Applications].

EEG is a weak physiological electrical signal, which has important value in clinical and laboratory research. This paper mainly introduces several common methods of EEG signal processing, including power spectrum analysis, time-frequency analysis, bispectral analysis, etc, it mainly introduces their principles and applications in EEG signal processing, and provides methods and approaches for studying EEG.