Search

Summary: The USP7 deubiquitinase regulates proteins involved in the cell cycle, DNA repair, and epigenetics and has been implicated in cancer progression. USP7 inhibition has been pursued for the development of anti-cancer therapies. Here, we describe the discovery of potent and specific USP7 inhibitors exemplified by FX1-5303. FX1-5303 was used as a chemical probe to study the USP7-mediated regulation of p53 signaling in cells. It demonstrates mechanistic differences compared to MDM2 antagonists, a related class of anti-tumor agents that act along the same pathway. FX1-5303 synergizes with the clinically approved BCL2 inhibitor venetoclax in acute myeloid leukemia (AML) cell lines and ex vivo patient samples and leads to strong tumor growth inhibition in in vivo mouse xenograft models of multiple myeloma and AML. This work introduces new USP7 inhibitors, differentiates their mechanism of action from MDM2 inhibition, and identifies specific opportunities for their use in the treatment of AML.

The rhizosphere is one of the key determinants of plant health and productivity. Mixtures of pesticides are commonly used in intensified agriculture. However, the combined mechanisms underlying their impacts on soil microbiota remain unknown. The present study revealed that the rhizosphere microbiota was more sensitive to azoxystrobin and oxytetracycline, two commonly used pesticides, than was the microbiota present in bulk soil. Moreover, the rhizosphere microbiota enhanced network complexity and stability and increased carbohydrate metabolism and xenobiotic biodegradation as well as the expression of metabolic genes involved in defence against pesticide stress. Co-exposure to azoxystrobin and oxytetracycline had antagonistic effects on Arabidopsis thaliana growth and soil microbial variation by recruiting organic-degrading bacteria and regulating ABC transporters to reduce pesticide uptake. Our study explored the composition and function of soil microorganisms through amplicon sequencing and metagenomic approaches, providing comprehensive insights into the synergistic effect of plants and rhizosphere microbiota on pesticides and contributing to our understanding of the ecological risks associated with pesticide use.

Objective: To investigate the differences between meditation and resting states using infrared thermal imaging (IRTI) to determine facial temperature distribution features during meditation and annotate the patterns of facial temperature changes during meditation from the perspective of traditional Chinese medicine facial diagnosis. Methods: Each participant performed 10 min meditation and 10 min resting but in different sequences. A concentration test was set as the task load, followed by a meditation/resting or resting/meditation session, during which the participants' facial temperatures were observed using IRTI. Participants were scored on the Big Five Inventory (BFI) and Mindful Attention Awareness Scale (MAAS). Results: Forehead temperatures decreased more during meditation than during the resting state. The chin temperature increased only during meditation (P

Background: Esophageal squamous cell carcinoma (ESCC) is the most common pathological type of esophageal cancer in China, accounting for more than 90 %. Most patients were diagnosed with advanced-stage ESCC, for whom new adjuvant therapy is recommended. Therefore, it is urgent to explore new therapeutic targets for ESCC. Ferroptosis, a newly discovered iron-dependent programmed cell death, has been shown to play an important role in carcinogenesis by many studies. This study explored the effect of Polo like kinase 1 (PLK1) on chemoradiotherapy sensitivity of ESCC through ferroptosis Methods: In this study, we knocked out the expression of PLK1 (PLK1-KO) in ESCC cell lines (KYSE150 and ECA109) with CRISPR/CAS9. The effects of PLK1-knock out on G6PD, the rate-limiting enzyme of pentose phosphate pathway (PPP), and downstream NADPH and GSH were explored. The lipid peroxidation was observed by flow cytometry, and the changes in mitochondria were observed by transmission electron microscopy. Next, through the CCK-8 assay and clone formation assay, the sensitivity to cobalt 60 rays, paclitaxel, and cisplatin were assessed after PLK1-knock out, and the nude mouse tumorigenesis experiment further verified it. The regulation of transcription factor YY1 on PLK1 was evaluated by dual luciferase reporter assay. The expression and correlation of PLK1 and YY1, and their impact on prognosis were analyzed in more than 300 ESCC cases from the GEO database and our center. Finally, the above results were further proved by single-cell sequencing. Results: After PLK1 knockout, the expression of G6PD dimer and the level of NADPH and GSH in KYSE150 and ECA109 cells significantly decreased. Accordingly, lipid peroxidation increased, mitochondria became smaller, membrane density increased, and ferroptosis was more likely to occur. However, with the stimulation of exogenous GSH (10 mM), there was no significant difference in lipid peroxidation and ferroptosis between the PLK1-KO group and the control group. After ionizing radiation, the PLK1-KO group had higher lipid peroxidation ratio, more cell death, and was more sensitive to radiation, while exogenous GSH (10 mM) could eliminate this difference. Similar results could also be observed when receiving paclitaxel combined with cisplatin and chemoradiotherapy. The expression of PLK1, G6PD dimer, and the level of NADPH and GSH in KYSE150, ECA109, and 293 T cells stably transfected with YY1-shRNAs significantly decreased, and the cells were more sensitive to radiotherapy and chemotherapy. ESCC patients from the GEO database and our center, YY1 and PLK1 expression were significantly positively-correlated, and the survival of patients with high expression of PLK1 was significantly shorter. Further analysis of single-cell sequencing specimens of ESCC in our center confirmed the above results. Conclusion: In ESCC, down-regulation of PLK1 can inhibit PPP, and reduce the level of NADPH and GSH, thereby promoting ferroptosis and improving their sensitivity to radiotherapy and chemotherapy. Transcription factor YY1 has a positive regulatory effect on PLK1, and their expressions were positively correlated. PLK1 may be a target for predicting and enhancing the chemoradiotherapy sensitivity of ESCC.

Chlorpyrifos is a widely used organophosphorus insecticide because of its high efficiency and overall effectiveness, and it is commonly detected in aquatic ecosystems. However, at present, the impact of chlorpyrifos on the aquatic micro-ecological environment is still poorly understood. Here, we established aquatic microcosm systems treated with 0.2 and 2.0 µg/L chlorpyrifos, and employed omics biotechnology, including metagenomics and 16S rRNA gene sequencing, to investigate the effect of chlorpyrifos on the composition and functional potential of the aquatic and zebrafish intestinal microbiomes after 7 d and 14 d chlorpyrifos treatment. After 14 d chlorpyrifos treatment, the aquatic microbial community was adversely affected in terms of its composition, structure, and stability, while its diversity showed only a slight impact. Most functions, especially capacities for environmental information processing and metabolism, were destroyed by chlorpyrifos treatment for 14 d. We observed that chlorpyrifos increased the number of risky antibiotic resistance genes and aggravated the growth of human pathogens. Although no clear effects on the structure of the zebrafish intestinal microbial community were observed, chlorpyrifos treatment did alter the metabolic capacity of the zebrafish. Our study highlights the ecological risk of chlorpyrifos to the aquatic environment and provides a theoretical basis for the rational use of pesticides in agricultural production.

Lung cancer is the main reason for cancer-associated death globally, and lung adenocarcinoma (LUAD) is the most prevalent subtype of lung cancer. Recently, AGRN is considered playing an vital role in the development of some cancers. However, the regulatory effects and mechanisms of AGRN in LUAD remain elusive. In this study, we clarified the significant upregulation of AGRN expression in LUAD by single-cell RNA sequencing combined with immunohistochemistry. Besides, we confirmed that LUAD patients with high AGRN expression are more susceptible to lymph node metastases and have a worse prognosis by a retrospective study of 120 LUAD patients. Next, we demonstrated that AGRN directly interact with NOTCH1, which results in the release of the intracellular structural domain of NOTCH1 and the subsequent activation of the NOTCH pathway. Moreover, we also found that AGRN promotes proliferation, migration, invasion, EMT and tumorigenesis of LUAD cells in vitro and in vivo, and that these effects are reversed by blocking the NOTCH pathway. Furthermore, we prepared several antibodies targeting AGRN, and clarify that Anti-AGRN antibody treatment could significantly inhibit proliferation and promote apoptosis of tumor cells. Our study highlights the important role and regulatory mechanism of AGRN in LUAD development and progression, and suggests that antibodies targeting AGRN have therapeutic potential for LUAD. We also provide theoretical and experimental evidence for further development of monoclonal antibodies targeting AGRN.

Lung adenocarcinoma (LUAD) is one of the most common malignancies worldwide. Combination chemotherapy with cisplatin (CDDP) plus pemetrexed (PEM) remains the predominant therapeutic regimen; however, chemoresistance greatly limits its curative potential. Here, through CRISPR-Cas9 screening, we identified miR-6077 as a key driver of CDDP/PEM resistance in LUAD. Functional experiments verified that ectopic overexpression of miR-6077 desensitized LUAD cells to CDDP/PEM in both cell lines and patient-derived xenograft models. Through RNA sequencing in cells and single-cell sequencing of samples from patients with CDDP/PEM treatments, we observed CDDP/PEM-induced upregulation of CDKN1A and KEAP1, which in turn activated cell-cycle arrest and ferroptosis, respectively, thus leading to cell death. Through miRNA pull-down, we identified and validated that miR-6077 targets CDKN1A and KEAP1. Furthermore, we demonstrated that miR-6077 protects LUAD cells from cell death induced by CDDP/PEM via CDKN1A-CDK1-mediated cell-cycle arrest and KEAP1-NRF2-SLC7A11/NQO1-mediated ferroptosis, thus resulting in chemoresistance in multiple LUAD cells both in vitro and in vivo. Moreover, we found that GMDS-AS1 and LINC01128 sensitized LUAD cells to CDDP/PEM by sponging miR-6077. Collectively, these results imply the critical role of miR-6077 in LUAD’s sensitivity to CDDP/PEM, thus providing a novel therapeutic strategy for overcoming chemoresistance in clinical practice.

The Tibetan Plateau (TP) is a hotspot for biodiversity conservation in the world. Understanding the impacts of land-use/cover change (LUCC) on natural habitat quality (NHQ) in terms of both historical and future urbanization and ecological restoration is important, but existing studies have only focused on the total impacts of historical LUCC on NHQ on the TP. This study aims to assess both historical and future impacts by taking the Lhasa River Basin (LRB) as an example. We quantified the LUCC from 2000 to 2020 based on remote sensing data, and simulated the LUCC during 2020–2030 under the scenarios of business-as-usual (BAU) and cropland conservation. Finally, we used the habitat quality model to assess the impacts of LUCC on NHQ. We found that the improved NHQ due to ecological restoration exceeded the deteriorated NHQ due to urban expansion, resulting in the total NHQ showing an increasing trend in the LRB during 2000–2020. Under the BAU scenario, NHQ was projected to remain stable. However, under the cropland protection scenario, NHQ was projected to deteriorate due to the large-scale occupation of natural habitats by urban expansion, which will be prominent in Chengguan District (NHQ will decrease by 22.52%). To protect both cropland and natural habitats, limiting urban sprawl and implementing ecological protection and restoration are needed.

Summary: A model was previously proposed that DA neurons provide SHH ligand to striatal interneurons, which in turn support the survival of DA neurons through the release of trophic factors such as Glial cell-derived neurotrophic factor (GDNF). However, some key clinical observations do not support this proposed model, and a recent independent study shows that striatal cholinergic neuron survival does not rely on intact DA neuron projections. To resolve this discrepancy, we generated several independent mouse lines to examine the exact role of DA neuron-derived Shh signaling in the maintenance of the basal ganglia circuit and to identify the Shh-producing cells in the adult brain. Our data suggest that the deletion of Shh in DA neurons does not affect DA neuron survival or locomotive function in cKO mice during aging, nor does it affect the long-term survival of cholinergic or FS PV + interneurons in the striatum (STR).

Biotreatment of acidic rare earth mining wastewater via acidophilic living organisms is a promising approach owing to their high tolerance to high concentrations of rare earth elements (REEs); however, simultaneous removal of both REEs and ammonium is generally hindered since most acidophilic organisms are positively charged. Accordingly, immobilization of acidophilic Galdieria sulphuraria (G. sulphuraria) by calcium alginate to improve its affinity to positively charged REEs has been used for simultaneous bioremoval of REEs and ammonium. The results indicate that 97.19%, 96.19%, and 98.87% of La, Y, and Sm, respectively, are removed by G. sulphuraria beads (GS-BDs). The adsorption of REEs by calcium alginate beads (BDs) and GS-BDs is well fitted by both pseudo first-order (PFO) and pseudo second-order (PSO) kinetic models, implying that adsorption of REEs involves both physical adsorption caused by affinity of functional groups such as –COO– and –OH and chemical adsorption based on ion exchange of Ca2+ with REEs. Notably, GS-BDs exhibit high tolerance to La, Y, and Sm with maximum removal efficiencies of 97.9%, 96.6%, and 99.1%, respectively. Furthermore, the ammonium removal efficiency of GS-BDs is higher than that of free G. sulphuraria cells at an initial ammonium concentration of 100 mg L−1, while the efficiency decreases when initial concentration of ammonium is higher than 150 mg L−1. Last, small size of GS-BDs favors ammonium removal because of their lower mass transfer resistance. This study achieves simultaneous removal of REEs and ammonium from acidic mining drainage, providing a potential strategy for biotreatment of REE tailing wastewater.

Long non-coding RNAs (lncRNAs), microRNAs (miRNAs or miRs), and genes are emerging players in cancer progression. In the present study, we explored the roles and interactions of oncogenic lncRNA small nucleolar RNA host gene 1 (SNHG1), miR-376, forkhead box protein K1 (FOXK1), and Snail in hepatocellular carcinoma (HCC). Expression of SNHG1, miR-376, and FOXK1 in HCC was characterized in clinical HCC tissues of 75 patients with HCC. The interactions between SNHG1 and miR-376 and between miR-376 and FOXK1 were predicted and confirmed by dual-luciferase reporter gene and RNA immunoprecipitation assays. Overexpression and knockdown experiments were performed in HCC cells to examine the effects of the SNHG1/miR-376/FOXK1/Snail axis on viability, apoptosis, invasiveness, and migrating abilities. Their effects on tumor growth and metastasis were validated in nude mouse models. SNHG1 and FOXK1 were upregulated, and miR-376a was downregulated in HCC. SNHG1 knockdown contributed to suppression of HCC cell viability, invasion, and migration properties and promotion of apoptosis. SNHG1 could competitively bind to miR-376a to upregulate its target gene FOXK1, which upregulated Snail. SNHG1 knockdown delayed cancer progression both in vitro and in vivo by upregulating miR-376a and downregulating FOXK1 and Snail. SNHG1 knockdown exerts anti-tumor activity in HCC, suggesting a therapeutic target.

Background: Stroke could cause long-term disability, even mortality around the world. Recently, Sodium tanshinone IIA sulfonate (STS), identified from Salvia miltiorrhiza Bunge and was found to have unique efficiency in clinical practice as a potential therapeutic agent for ischemic cerebral infarction. However, systematic investigation about the biological mechanism is still lacking. Herein, we utilized high-throughput proteomics approach to identify the underlying targets for the treatment of STS in stroke. Methods: We investigated the effect of STS on stroke outcomes on rat model of the Middle Cerebral Artery Occlusion and Reperfusion (MCAO/R), assessing by Z-Longa score, infarct volume and HE staining. Pharmacoproteomic profiling of ischemic penumbra in cortical (IPC) was performed using DIA-based label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique. Bioinformatics analysis was processed for further investigation. The expression of core proteins was semi-quantified by DIA, and the major protein correlating with stroke was examined using parallel reaction monitoring (PRM). Results: Rats in the MCAO/R group showed neurological function deterioration, which was improved by STS. There were 423 differentially expressed proteins (DEPs) in IPC being detected and quantified in both the sham group and the MCAO/R group. Meanwhile, 285 proteins were significantly changed in the STS treated group, compared to the MCAO/R model. Protein-protein interaction (PPI) network, pathway and biological function enrichment were processed for the DEPs across each two groups, the results of which were integrated for analysis. Alb, mTOR, Dync1h1, Stxbp1, Cltc, and Sptan1 were contained as the core proteins. Altered molecules were discovered to be enriched in 18 signal pathways such as phosphatidylinositol signaling system, PI3K/AKT signal pathway and HIF-1 signal pathway. The results also showed the correlation with sleep disturbances and depression post-stroke. Conclusions: We concluded that STS could prevent penumbra from progressively ongoing damage and improve neurological deficits in MCAO/R model rats. The intersected pathways and protein networks predicted by proteomics might provide much more detailed information for the therapeutic mechanisms of STS in the treatment of CIS.

Clinical studies have shown that renal injury in Corona Virus Disease 2019 (COVID-19) patients has been a real concern, which is associated with high mortality and an inflammation/apoptosis-related causality. Effective target therapy for renal injury has yet been developed. Besides, potential anti-COVID-19 medicines have also been reported to cause adverse side effects to kidney. Chinese Herbal Medicine (CHM), however, has rich experience in treating renal injury and has successfully applied in China in the battle of COVID-19. Nevertheless, the molecular mechanisms of CHM treatment are still unclear. In this study, we searched prescriptions in the treatment of renal injury extensively and the potential mechanisms to treat COVID-19 related renal injury were investigated. The association rules analysis showed that the core herbs includes Huang Qi, Fu Ling, Bai Zhu, Di Huang, Shan Yao. TCM herbs regulate core pathways, such as AGE-RAGE, PI3K-AKT, TNF and apoptosis pathway, etc. The ingredients (quercetin, formononetin, kaempferol, etc.,) from core herbs could modulate targets (PTGS2 (COX2), PTGS1 (COX1), IL6, CASP3, NOS2, and TNF, etc.), and thereby prevent the pharmacological and non-pharmacological renal injury comparable to that from COVID-19 infection. This study provides therapeutic potentials of CHM to combat COVID-19 related renal injury to reduce complications and mortality.

Objective: Although Follistatin-like protein 1 (FSTL1), as an “adipokine”, is highly expressed in preadipocytes, the detail role of FSTL1 in adipogenesis and obesity remains not fully understood. Methods: In vitro differentiation of both Fstl1−/− murine embryonic fibroblasts (MEFs) and stromal vascular fraction (SVF) were measured to assess the specific role of FSTL1 in adipose differentiation. Fstl1 adipocyte-specific knockout mice were generated to evaluate its role in obesity development. Gene expression analysis and phosphorylation patterns were performed to check out the molecular mechanism of the biological function of FSTL1. Results: FSTL1 deficiency inhibited preadipocytes differentiation in vitro and obesity development in vivo. Glycosylation at N142 site was pivotal for the biological effect of FSTL1 during adipogenesis; the conversion between PPARγ and p-PPARγ was the key factor for the function of FSTL1. Molecular mechanism studies showed that FSTL1 functions through the integrin/FAK/ERK signaling pathway. Conclusions: Our results suggest that FSTL1 promotes adipogenesis by inhibiting the conversion of PPARγ to p-PPARγ through the integrin/FAK/ERK signaling pathway. Glycosylated modification at N142 of FSTL1 is the key site to exert its biological effect.

Accumulating studies have implicated the role of long non-coding RNAs (lncRNAs) in the pathogenesis of hepatocellular carcinoma (HCC) through the regulating transcription and mRNA stability. A recent report has linked Rac GTPase-activating protein 1 (RACGAP1) to the early recurrence of HCC. The current study aimed to ascertain whether MAGI2 antisense RNA 3 (MAGI2-AS3) influences the development of HCC by regulating RACGAP1. MAGI2-AS3 expression was initially quantified in both the HCC tissues and cell lines. In order to elucidate the role of MAGI2-AS3 in the development of HCC, MAGI2-AS3 was overexpressed or silenced in HCC cells after which cell proliferation, apoptosis, invasion, and migration were evaluated. Chromatin immunoprecipitation (ChIP), RNA immunoprecipitation (RIP), and biotin-labeled RNA pull-down assays were conducted to determine the interactions among MAGI2-AS3, KDM1A, and RACGAP1. Finally, the effects of MAGI2-AS3 and RACGAP1 on the tumorigenesis of transplanted HCC cells in nude mice were evaluated. MAGI2-AS3 was found to be under-expressed in HCC tissues and cell lines. The restoration of MAGI2-AS3 was identified to markedly inhibit HCC cell growth, migrating ability, and invasiveness, and promote cell apoptosis. Interaction between MAGI2-AS3 and KDM1A was identified. KDM1A recruited by MAGI2-AS3 was found to promote H3K4me2 demethylation at the RACGAP1 promoter, which ultimately decreased the expression of RACGAP1. We also identified that RACGAP1 knockdown eliminated the stimulatory effects of MAGI2-AS3 silencing on the malignant phenotypes of HCC cells. Additionally, the expression of MAGI2-AS3 reduced tumor weight and size in HCC transplanted nude mice. Taken together, the key observations of the current study demonstrate the potential of MAGI2-AS3 as a tumor suppressor and a promising target for HCC treatment. Keywords: hepatocellular carcinoma, MAGI2-AS3, KDM1A, RACGAP1, demethylation, H3K4me2

Free-standing N-doped carbon nanofiber membranes have been obtained from cellulose acetate (CA) and soy protein isolate (SPI). As an environmental friendliness, inexpensive and abundant polysaccharide in nature, cellulose and its derivatives have shown great potentials in the preparation of energy storage materials. This work successfully fabricated cellulose-based carbon nanofiber membranes through electrospinning, regeneration process and carbonization. Furthermore, this paper has intensively studied the electrospinning parameters of CA (main component) and SPI composite fiber membranes and explored the influence of carbonization temperature on their microstructure and electrochemical properties. The resultant CA-SPI derived carbon nanofiber membranes are applied in supercapacitors (SCs) for the first time, and display good electrochemical performance because of their unique structures and pseudo-capacitance contribution from heteroatoms. The as-received CA-SPI-800 exhibits large specific capacitance of 219.3 F g−1 at 0.2 A g−1 in 6 M KOH. CA-SPI-800 shows high cycling stability with 98.9% capacitance retention after 50,000 cycles at 20 A g−1. The symmetrical supercapacitor possesses 99% Columbic efficiency and outstanding cycling stability of 94.1% retention after 40,000 charging-discharging processes at 20 A g−1. It is expected that the design of CA-SPI nanofiber membranes will provide a successful route for the preparation of cellulose derived nanofibers, and broaden their applications in low-consumption and green energy storage devices.

Large size sandwich structure is involved in a wide range of applications, such as aerospace and ship. However, the weak connection between panel and lattice is the bottleneck problem restricting their application. To solve this problem, this work demonstrates a novel circular scanning connection method based on pulse hot-wire arc additive manufacturing (PHWAAM) to connect the centimeter-scale and meter-scale panel. This method exhibits great connection quality (without unmelted holes) and high efficiency (about 60 s for each connection). The optimized process is studied, based on systematical analysis of the microstructure, the macro defects, and the compression testing results. By comparing and analyzing three different connection modes, the optimized circular scanning process has the advantages of low heat input and high deposition efficiency. It can be observed from the microstructure that there are equiaxed crystals and short columnar crystals in the connection area. The width of acicular α phase is finer than that of other processes, which is conducive to improving the mechanical properties. The connection area of the circular scanning process has a better matching of strength and plasticity through the compression test. Finally, the proposed connection method is also applicable to fabricate multi-cell lattice sandwich structure.

Although microfiber (MF) loaded polypeptides have been widely studied in the field of medicine, load control and slow release remain significant challenges. Herein, polybutylene adipate terephthalate (PBAT) hollow porous MFs (HPMFs) were prepared by improving and regulating the groups and structures of the MFs to address these challenges. The capacity of the HPMFs for the loading of polypeptide can be improved. Measurements involving the use of X-ray photoelectron, energy-dispersive, and ultraviolet (UV) spectroscopies in conjunction with scanning electron microscopy measurements showed that the amount of polypeptide (arginylglycylaspartic acid, RGD) loaded on the HPMFs was significantly higher than that loaded on the MFs. Animal cell experiments revealed that PBAT grafted MAH has good bioactivity. By loading RGD onto HPMFs and MFs, RGD@HPMFs and RGD@MFs were obtained. A study on HeLa and A549 cancer cells showed that the inhibition rates of RGD@HPMFs were higher than that of RGD@MFs by 14.1% and 6.9%, respectively. The results obtained herein show that HPMF scaffold preparation by improving the material groups and regulating the structure of MFs can address the challenges associated with control of the load and sustained release of polypeptides and other drugs.

Background: Esophageal squamous cell carcinoma (ESCC) is among the most prevalent causes of cancer-related death in adults. Tumor microenvironment (TME) has been associated with therapeutic failure and lethal outcomes for patients. However, published reports on the heterogeneity and TME in ESCC are scanty. Methods: Five tumor samples and five corresponding non-malignant samples were subjected to scRNA-seq analysis. Bulk RNA sequencing data were retrieved in publicly available databases. Findings: From the scRNA-seq data, a total of 128,688 cells were enrolled for subsequent analyses. Gene expression and CNV status exhibited high heterogeneity of tumor cells. We further identified a list of tumor-specific genes and four malignant signatures, which are potential new markers for ESCC. Metabolic analysis revealed that energy supply-related pathways are pivotal in cancer metabolic reprogramming. Moreover, significant differences were found in stromal and immune cells between the esophagus normal and tumor tissues, which promoted carcinogenesis at both cellular and molecular levels in ESCC. Immune checkpoints, regarded as potential targets for immunotherapy in ESCC were significantly highly expressed in ESCC, including LAG3 and HAVCR2. Eventually, we constructed a cell-to-cell communication atlas based on cancer cells and immune cells and performed the flow cytometry, qRT-PCR, immunofluorescence, and immunohistochemistry analyses to validate the results. Interpretation: This study demonstrates a widespread reprogramming across multiple cellular elements within the TME in ESCC, particularly in transcriptional states, cellular functions, and cell-to-cell interactions. The findings offer an insight into the exploration of TME and heterogeneity in the ESCC and provide new therapeutic targets for its clinical management in the future.

Ciprofloxacin (CIP), one of the most frequently detected antibiotics in water systems, has become an aquatic contaminant because of improper disposal and excretion by humans and animals. It is still unknown how trace amounts of CIP affect the aquatic microbial community diversity and function. We therefore investigated the effects of CIP on the structure and function of freshwater microbial communities via 16S/18S rRNA gene sequencing and metatranscriptomic analyses. CIP treatment (7 μg/L) did not significantly alter the physical and chemical condition of the water body as well as the composition of the main species in the community, but slightly increased the relative abundance of cyanobacteria and decreased the relative abundance of eukaryotes. Metatranscriptomic results showed that bacteria enhanced their phosphorus transport and photosynthesis after CIP exposure. The replication, transcription, translation and cell proliferation were all suppressed in eukaryotes, while the bacteria were not affected in any of these aspects. This interesting phenomenon was the exact opposite to both the antibacterial property of CIP and its safety for eukaryotes. We hypothesize that reciprocal and antagonistic interactions in the microcosm both contribute to this result: cyanobacteria may enhance their tolerance to CIP through benefiting from cross-feeding and some secreted substances that withstand bacterial CIP stress would also affect eukaryotic growth. The present study thus indicates that a detailed assessment of the aquatic ecotoxicity of CIP is essential, as the effects of CIP are much more complicated in microbial communities than in monocultures. CIP will continue to be an environmental contaminant due to its wide usage and production and more attention should be given to the negative effects of antibiotics as well as other bioactive pollutants on aquatic environments. Keywords: Ciprofloxacin, Cyanobacteria, Meta-transcriptomic analysis, High-throughput sequencing, Freshwater microbiome

The use of pesticides to ensure global food security is the most important pest control strategy in modern agriculture but causes extensive soil pollution. This pollution involves potential risks to human health and ecosystems. In addition to soil animal growth, the adverse impact of pesticides on the gut microbiomes of nontarget soil fauna remains largely unknown. Here, the effect of the fungicide azoxystrobin (AZ) on soil and the gut microbiota of soil animals (Enchytraeus crypticus) was studied. The tested concentrations of AZ altered the bacterial community in the soil and E. crypticus gut and were slightly toxic with respect to E. crypticus adult mortality and reproduction. The most abundant bacterial phylum, Proteobacteria, significantly increased in response to the 2 and 5 mg/kg AZ treatments, which implied a disordered unhealthy gut bacterial community. Furthermore, bacterial community analysis between the soil and gut showed that the main effect of AZ on the gut microbiota was directly through AZ, not soil microbiota. In addition, AZ exposure significantly enhanced the number and total abundance of antibiotic resistance genes (ARGs) in the E. crypticus gut; these genes may enter the soil food web to affect higher trophic levels and cause a more serious ecological risk. Our study reported the effect of pesticides on the gut of soil animals and on the enrichment of ARGs as global emerging contaminants, revealing unknown potential impacts of fungicides on ecosystem services and sustainable food production. Keywords: Gut microbiota, Antibiotic resistance genes, Azoxystrobin, Enchytraeus, Fungicide

Summary: The molecular basis of the earliest neuronal changes that lead to Alzheimer’s disease (AD) is unclear. Here, we analyze neural cells derived from sporadic AD (SAD), APOE4 gene-edited and control induced pluripotent stem cells (iPSCs). We observe major differences in iPSC-derived neural progenitor (NP) cells and neurons in gene networks related to neuronal differentiation, neurogenesis, and synaptic transmission. The iPSC-derived neural cells from SAD patients exhibit accelerated neural differentiation and reduced progenitor cell renewal. Moreover, a similar phenotype appears in NP cells and cerebral organoids derived from APOE4 iPSCs. Impaired function of the transcriptional repressor REST is strongly implicated in the altered transcriptome and differentiation state. SAD and APOE4 expression result in reduced REST nuclear translocation and chromatin binding, and disruption of the nuclear lamina. Thus, dysregulation of neural gene networks may set in motion the pathologic cascade that leads to AD. : Meyer et al. derive neural progenitors, neurons, and cerebral organoids from sporadic Alzheimer’s disease (SAD) and APOE4 gene-edited iPSCs. SAD and APOE4 expression alter the neural transcriptome and differentiation in part through loss of function of the transcriptional repressor REST. Thus, neural gene network dysregulation may lead to Alzheimer’s disease. Keywords: neural progenitor, induced pluripotent stem cell, Alzheimer’s disease, apolipoprotein E, neural differentiation, neurogenesis, REST, polycomb, epigenetic, organoid

Stress-mediated programmed cell death (PCD) in bacteria has recently attracted attention, largely because it raises novel possibilities for controlling pathogens. How PCD in bacteria is regulated to avoid population extinction due to transient, moderate stress remains a central question. Here, we report that the YihE protein kinase is a key regulator that protects Escherichia coli from antimicrobial and environmental stressors by antagonizing the MazEF toxin-antitoxin module. YihE was linked to a reactive oxygen species (ROS) cascade, and a deficiency of yihE stimulated stress-induced PCD even after stress dissipated. YihE was partially regulated by the Cpx envelope stress-response system, which, along with MazF toxin and superoxide, has both protective and destructive roles that help bacteria make a live-or-die decision in response to stress. YihE probably acts early in the stress response to limit self-sustaining ROS production and PCD. Inhibition of YihE may provide a way of enhancing antimicrobial lethality and attenuating virulence.

Amidst global climate change and increased anthropogenic activities, the Qilian Mountains face critical ecological and environmental challenges, including vegetation degradation and declining carbon stocks. Therefore, effective mitigation strategies are critical to building China’s ecological security and enhancing carbon storage. This study constructed and optimized an ecological corridor in the Qilian Mountains using models, such as minimum spanning tree and carbon stock calculation. It combined geographic meta cellular automata and other methods to model ecological recovery in the Qilian Mountains following corridor construction and investigated the areas of particular concern. The results show that: (1) the distribution of the 118 extracted ecological source sites (2020) exhibited a pattern of “dense in the east and sparse in the west.” Compared with the results of the 2010 extraction, the source site area had shrunk by 0.87 % and number of sites had increased by 45.68 %. This implies that the shrinkage and fragmentation of source sites are still increasing, underscoring the urgent need for ecological corridors. (2) Compared to the pre-optimization period, the network node connectivity of the corridor network formed by the optimized 114 primary corridors and 131 secondary corridors has increased by 110 % and the network connectivity has increased by 112 %, which enhances the connectivity of the corridors. (3) From 2000 to 2020, the carbon stock in the Qilian Mountains decreased and then increased; with the construction of a corridor mountain range, the carbon stock can be further increased by 1.4748 million tons. Additionally, the Geographic Cellular Automata (GCA) model simulated that in 2030, the area of forest land in the Qilian Mountains, area of grassland, and carbon stock will increase by 71.75 km2, 313.87 km2, and 6.663 million tons, respectively. These results confirm that ecological protection and corridor construction promote ecological restoration and enhance carbon stocks in the Qilian Mountains, providing a scientific basis for future ecological projects and carbon stock analysis in the region.

Background: Accumulating studies have found that circular RNAs (circRNAs) have a regulatory effect in a variety of tumors. However, to date, the relationship between specific circRNAs and colorectal cancer (CRC) remains elusive. Methods: An RNA-sequencing method based on different metastatic potential of CRC cell lines was applied to evaluate the circRNA expression profile. Additionally, we conducted a series of experiments to assess the relationship between circRNAs and CRC progression. Results: Circ_0001742 was upregulated in CRC cells with high metastatic potential, and circ_0001742 overexpression was observed to facilitate proliferation, migration and metastasis while knockdown will inhibit. More importantly, we found that circ_0001742 acted as a sponge for miR-431-5p, thus affecting ALG8 levels and the development of CRC. Conclusions: This study demonstrated an essential function for the circ_0001742/miR-431-5p/ALG8 axis in CRC development, and it may be a promising therapeutic target for CRC.

In the Junggar Basin plague focus, Rhombomys opimus and Meriones meridianus live together, and their parasitic fleas have a similar community structure. However, R. opimus has significantly higher positive rates of Yersinia pestis and anti-F1 antibody compared with M. meridianus. In this study, Y. pestis- and antiF1 antibody-negative R. opimus and M. penicilliger were collected in Qitai county, Fukang city and Mulei county of the eastern part of the Junggar Basin. The genomic DNA was extracted from their spleen tissues, and RT1-Db1*exon1 was amplified through PCR procedure and then sequenced. Sequence analysis was performed and molecular diversity parameters were calculated and compared. The results showed that there were significant differences in nucleotide composition, amino acid composition, number and distribution of single nucleotide polymorphism (SNP) sites and number of haplotypes between R. opimus and M. penicilliger. The nucleotide diversity (π) for R. opimus was 0.00420 ± 0.00139, the haplotype diversity (h) was 0.833 ± 0.086, and the average number of nucleotide differences (K) was 2.02564. The π for M. penicilliger was 0.06569 ± 0.02524, and the h was 1.000 ± 0.045, and the K was 10.4444. The fixation index (FST) value between R. opimus and M. penicilliger was 0.9207. Furthermore, the FST value within R. opimus (0.0275) was significantly lower than that within M. penicilliger (0.2106), indicating a greater genetic variation of M. penicilliger compared with R. opimus. In conclusion, the genetic diversity analysis based on RT1-Db1*exon1 showed that M. penicilliger had higher gene polymorphism and greater genetic differentiation compared with R. opimus in the Junggar Basin plague focus, which might be associated with the low infection rate of Y. pestis.

Nanoplastic contamination has been of intense concern by virtue of the potential threat to human and ecosystem health. Animal experiments have indicated that exposure to nanoplastics (NPs) can deposit in the liver and contribute to hepatic injury. To explore the mechanisms of hepatotoxicity induced by polystyrene-NPs (PS-NPs), mice and AML-12 hepatocytes were exposed to different dosages of 20 nm PS-NPs in this study. The results illustrated that in vitro and in vivo exposure to PS-NPs triggered excessive production of reactive oxygen species and repressed nuclear factor erythroid-derived 2-like 2 (NRF2) antioxidant pathway and its downstream antioxidase expression, thus leading to hepatic oxidative stress. Moreover, PS-NPs elevated the levels of NLRP3, IL-1β and caspase-1 expression, along with an activation of NF-κB, suggesting that PS-NPs induced hepatocellular inflammatory injury. Nevertheless, the activaton of NRF2 signaling by tert-butylhydroquinone mitigated PS-NPs-caused oxidative stress and inflammation, and inbihited NLRP3 and caspase-1 expression. Conversely, the rescuing effect of NRF2 signal activation was dramatically supressed by treatment with NRF2 inhibitor brusatol. In summary, our results demonstrated that NRF2-NLRP3 pathway is involved in PS-NPs-aroused hepatotoxicity, and the activation of NRF2 signaling can protect against PS-NPs-evoked liver injury. These results provide novel insights into the hepatotoxicity elicited by NPs exposure.

Ecosystem surface plays an important role in maintaining ecological balance and supporting economic development. Accurate and effective identification of ecosystem surface with different ecological uses allows for better categorization and management and contributes to the orderly restoration of ecosystems. In this study, the Human Footprint Index (HFI) is constructed by comprehensively considering the factors of biodiversity, human activity impacts, and geometric features, and combined with the multi-factor knowledge constraints such as changes in land cover. We carry out accurate and effective identification and analysis of ecosystem surfaces, using Changzhou City’s tree cover as a test area. The results show that: (1) The degree of tree cover affected by human activities in Changzhou City varies greatly, and Gross Domestic Product (GDP) and elevation are the main factors affecting the level of the HFI. (2) The distribution of native ecosystem surface and artificial ecosystem surface are uneven, with more than 95% of native ecosystem surfaces distributed in the peripheral areas of the city with higher elevations and farther away from the city center. Artificial ecosystem surfaces are more likely to be found in areas with high GDP and frequent human activities, mainly in urban centers with a large proportion of buildings and along roads. (3) The overall identification accuracy of the ecosystem surface is 83.20%, which is higher in the urban center area with a large proportion of buildings and slightly lower in the area with a large proportion of agricultural production land. The results of the study provide a method for the objective, accurate and effective identification of ecosystem surface, which can improve the enthusiasm of local governments in fulfilling their ecological protection duties and contribute to the efficient achievement of the goal of ecological civilization construction.

Summary: Background: Anlotinib is a new type of tyrosine kinase inhibitor that targets vascular endothelial growth factor receptors 1/2/3, platelet-derived growth factor receptors α/β, and fibroblast growth factor receptors 1–4 and c-Kit, with a broad spectrum of inhibitory effects on tumor angiogenesis and growth. It has been proven effective in HER2-negative metastatic breast cancer, but its efficacy in early-stage triple-negative breast cancer (TNBC) is unknown. This phase 2 study aims to evaluate the efficacy and safety of adding anlotinib to neoadjuvant chemotherapy in patients with TNBC. Methods: Patients with clinical stage II/III TNBC were treated with 5 cycles of anlotinib (12 mg, d1-14, q3w) plus 6 cycles of taxanes (docetaxel 75 mg/m2 ,d1, q3w or nab-paclitaxel 125 mg/m2, d1 and d8, q3w) and lobaplatin (30 mg/m2, d1, q3w), followed by surgery. The primary endpoint was pathological complete response (pCR; ypT0/is ypN0) and the secondary endpoints include breast pCR (bpCR), axillary pCR (apCR), residual cancer burden (RCB), objective response rate (ORR), survival, and safety. Exploratory endpoints were efficacy biomarkers based on Fudan University Shanghai Cancer Center Immunohistochemical (FUSCC IHC) classification for TNBC and next-generation sequencing (NGS) of DNA from tumor tissue and blood samples of patients with 425-gene panel. This trial is registered with www.chictr.org.cn (ChiCTR2100043027). Findings: From Jan 2021 to Aug 2022, 48 patients were assessed and 45 were enrolled. All patients received at least one dose of study treatment and underwent surgery. The median age was 48.5 years (SD: 8.7), 71% were nodal involved, and 20% had stage III. In the intention-to-treat population, 26 out of 45 patients achieved pCR (57.8%; 90% CI, 44.5%–70.3%), and 39 achieved residual cancer burden class 0-I (86.7%; 95% CI, 73.2%–94.9%). The bpCR and apCR rate were 64.4% (29/45) and 71.9% (23/32), respectively. No recurrence or metastasis occurred during the short-term follow-up. Based on the FUSCC IHC-based subtypes, the pCR rates were 68.8% (11/16) for immunomodulatory subtype, 58.3% (7/12) for basal-like immune-suppressed subtype and 33.3% (4/12) for luminal androgen receptor subtype, respectively. NGS revealed that the pCR were 77% (10/13) and 50% (14/28) in MYC-amplified and wild-type patients, respectively, and 78% (7/9) and 53% (17/32) in gBRCA1/2-mutated and wild-type patients, respectively. The median follow-up time of the study was 14.9 months (95% CI: 13.5–16.3 months). There was no disease progression or death during neoadjuvant therapy. No deaths occurred during postoperative follow-up. In the safety population (N = 45), Grade 3 or 4 treatment emergent adverse events occurred in 29 patients (64%), and the most common events were neutropenia (38%), leukopenia (27%), thrombocytopenia (25%), anemia (13%), and hypertension (13%), respectively. Interpretation: The addition of anlotinib to neoadjuvant chemotherapy showed manageable toxicity and encouraging antitumor activity for patients with clinical stage II/III TNBC. Funding: Chongqing Talents Project, Chongqing Key Project of Technology Innovation and Application Development and Chongqing Outstanding Youth Natural Science Foundation.

Summary: In glioma molecular subtyping, existing biomarkers are limited, prompting the development of new ones. We present a multicenter study-derived consensus immune-related and prognostic gene signature (CIPS) using an optimal risk score model and 101 algorithms. CIPS, an independent risk factor, showed stable and powerful predictive performance for overall and progression-free survival, surpassing traditional clinical variables. The risk score correlated significantly with the immune microenvironment, indicating potential sensitivity to immunotherapy. High-risk groups exhibited distinct chemotherapy drug sensitivity. Seven signature genes, including IGFBP2 and TNFRSF12A, were validated by qRT-PCR, with higher expression in tumors and prognostic relevance. TNFRSF12A, upregulated in GBM, demonstrated inhibitory effects on glioma cell proliferation, migration, and invasion. CIPS emerges as a robust tool for enhancing individual glioma patient outcomes, while IGFBP2 and TNFRSF12A pose as promising tumor markers and therapeutic targets.

High levels of copper released in the soil, mainly from anthropogenic activity, can be hazardous to plants, animals, and humans. The present research aimed to estimate the suitability and effectiveness of rapeseed (Brassica napus L.) as a possible soil remediation option and to uncover underlying adaptive mechanisms A pot experiment was conducted to explore the effect of copper stress on agronomic and yield traits for 32 rapeseed genotypes. The copper-tolerant genotype H2009 and copper-sensitive genotype ZYZ16 were selected for further physiological, metabolomic, and transcriptomic analyses. The results exhibited a significant genotypic variation in copper stress tolerance in rapeseed. Specifically, the ratio of seed yield under copper stress to control ranged from 0.29 to 0.74. Furthermore, the proline content and antioxidant enzymatic activities in the roots were greater than those in the shoots. The accumulated copper in the roots accounted for about 50% of the total amount absorbed by plants; thus, the genotypes possessing high root volumes can be used for rhizofiltration to uptake and sequester copper. Additionally, the pectin and hemicellulose contents were significantly increased by 15.6% and 162%, respectively, under copper stress for the copper-tolerant genotype, allowing for greater sequestration of copper ions in the cell wall and lower oxidative stress. Comparative analysis of transcriptomes and metabolomes revealed that excessive copper enhanced the up-regulation of functional genes or metabolites related to cell wall binding, copper transportation, and chelation in the copper-tolerant genotype. Our results suggest that copper-tolerant rapeseed can thrive in heavily copper-polluted soils with a 5.85% remediation efficiency as well as produce seed and vegetable oil without exceeding food quality standards for the industry. This multi-omics comparison study provides insights into breeding copper-tolerant genotypes that can be used for the phytoremediation of heavy metal-polluted soils.

In recent years, nanoplastics (NPs) and triclosan (TCS, a pharmaceutical and personal care product) have emerged as environmental pollution issues, and their combined presence has raised widespread concern regarding potential risks to organisms. However, the combined toxicity and mechanisms of NPs and TCS remain unclear. In this study, we investigated the toxic effects of polystyrene NPs and TCS and their mechanisms on KGN cells, a human ovarian granulosa cell line. We exposed KGN cells to NPs (150 μg/mL) and TCS (15 μM) alone or together for 24 hours. Co-exposure significantly reduced cell viability. Compared with exposure to NPs or TCS alone, co-exposure increased reactive oxygen species (ROS) production. Interestingly, co-exposure to NPs and TCS produced synergistic effects. We examined the activity of superoxide dismutase (SOD) and catalase (CAT), two antioxidant enzymes; it was significantly decreased after co-exposure. We also noted an increase in the lipid oxidation product malondialdehyde (MDA) after co-exposure. Furthermore, co-exposure to NPs and TCS had a more detrimental effect on mitochondrial function than the individual treatments. Co-exposure activated the NRF2–KEAP1–HO-1 antioxidant stress pathway. Surprisingly, the expression of SESTRIN2, an antioxidant protein, was inhibited by co-exposure treatments. Co-exposure to NPs and TCS significantly increased the autophagy-related proteins LC3B-II and LC3B-Ⅰ and decreased P62. Moreover, co-exposure enhanced CASPASE-3 expression and inhibited the BCL-2/BAX ratio. In summary, our study revealed the synergistic toxic effects of NPs and TCS in vitro exposure. Our findings provide insight into the toxic mechanisms associated with co-exposure to NPs and TCS to KGN cells by inducing oxidative stress, activations of the NRF2-KEAP1-HO-1 pathway, autophagy, and apoptosis.

Background: In this study, we utilized whole genome sequencing (WGS) of clinical extremely drug-resistant tuberculosis (EDR-TB) strains collected during 2014–2020 in Beijing to detect clustered strains. Methods: A retrospective cohort study was conducted by inclusion of EDR-TB patients with positive cultures in Beijing between 2014 and 2020. Results: A total of 95 EDR-TB patients were included in our analysis. Up on the WGS based genotyping, 94 (94/95, 98.9%) out of 95 were identified as lineage 2 (East Asia). The pairwise genomic distance analysis identified 7 clusters, ranging in size from 2 to 5 isolates. The clustering rate of EDR-TB was 21.1%; while no patients had significantly higher odds of clustering. All isolates harbor rpoB RRDR mutations that confer RIF resistance and katG or inhA promoter mutations that confer INH resistance. Of 95 EDR-TB isolates, a total of 15 mutation types were recorded in the transcriptional regulator mmpR5. In vitro susceptibility testing results revealed that 14 (14/15, 93.3%) out of 15 mutation types were resistant to CFZ; whereas only 3 (3/15, 20.0%) showed resistance to BDQ. Interestingly, 12 isolates harbored mutations within rrl locus, whereas only mutations at positions 2294 and 2296 conferred CLA resistance. Favorable outcomes of EDR-TB patients were positively associated with more effective drugs in the regimes. Conclusion: WGS data demonstrate limited transmission of EDR-TB in this metropolis city. WGS-based drug susceptibility predictions will bring benefits to EDR-TB patients to formulate optimal therapeutic regimens.

The consortium blockchain has been widely applied in various fields such as agricultural product traceability, supply chain management, and logistics transportation. As an indispensable component of a consortium blockchain, the consensus algorithm ensures the consistency and trustworthiness of each node in the network. However, existing consensus algorithms in large-scale consortium blockchain scenarios suffer from low system throughput and high latency due to the complexity of communication processes, rendering them impractical for real-world use. To address these issues, this paper proposes a novel consensus algorithm called credit evaluation-based practical Byzantine fault tolerance (CE-PBFT). This algorithm designs a new node credit evaluation model that considers node completion rate, consensus decay, and node behavior. It effectively measures and reflects the specific reliability status of nodes during system operation, thereby enhancing system reliability and security. Additionally, the paper introduces the innovative use of decision tree algorithms to analyze network node behavior and simplifies the existing consensus protocol. Nodes are categorized as excellent, good, ordinary, or poor based on the classification results, and non-Byzantine nodes are dynamically selected accordingly. This greatly improves the overall efficiency of the system. The performance of CE-PBFT is validated through experiments and compared with PBFT, G-PBFT, RBFT, WBFT and PPoR. Experimental results demonstrate that in large-scale consortium scenarios, CE-PBFT significantly improves system throughput, effectively reduces transaction latency and communication overhead, and outperforms the compared protocols.

Pharmaceutical compounds often coexist in mixtures rather than as individual entities. However, little is known about their co-adsorption and co-mobility in soil and groundwater. In this study, we investigated the adsorption of a quinolone antibiotic (nalidixic acid, NA) and an anti-inflammatory agent (niflumic acid, NFA) onto two soils from France and Sweden in water-saturated soil columns. Despite its lower hydrophobicity, adsorption of NA is much greater than NFA, which can be ascribed to the presence of both carbonyl and carboxylic groups in NA molecule. The data suggest that chemical adsorption to soil components can mainly take place through hydrogen bonding and surface complexation mechanisms, prevailing over hydrophobic interactions. Accordingly, more sorption of NA and NFA was observed in the Swedish soil because it contains more clay content, and much greater Al and Fe contents than the French soil. Injection of NA/NFA mixture in the column did not modify the breakthrough behavior compared to single systems, although cooperative adsorption was observed under static batch conditions. Ca2+ inhibited NA adsorption by forming a soluble NA-Ca2+ complex but promoted NFA adsorption both in single and binary systems. Their mobility in soil columns was well predicted using a new transport model that accounts for both kinetics and binding reactions of NA and NFA to soil constituents. This work will help in accurately predicting the mobility of coexisting pharmaceutical compounds in soils.

Whether receptor activity-modifying proteins (RAMPs) play a key role in human cancer prognosis and immunity remains unknown. We used data from the public databases, The Cancer Genome Atlas, Therapeutically Applicable Research to Generate Effective Treatments, and the Genotype-Tissue Expression project. We utilized bioinformatics methods, R software, and a variety of online databases to analyze RAMPs. In general, RAMPs were significantly and differentially expressed in multiple tumors, and RAMP expression was closely associated with prognosis, immune checkpoints, RNA-editing genes, tumor mutational burden, microsatellite instability, ploidy, and stemness indices. In addition, the expression of RAMPs is strongly correlated with tumor-infiltrating lymphocytes in human cancers. Moreover, the RAMP co-expression network is largely involved in many immune-related biological processes. Quantitative reverse transcription polymerase chain reaction and Western blot proved that RAMP3 was highly expressed in glioma, and RAMP3 promoted tumor proliferation and migration. RAMPs exhibit potential as prognostic and immune-related biomarkers in human cancers. Moreover, RAMPs can be potentially developed as therapeutic targets or used to enhance the efficacy of immunotherapy.

The pore solution composition (OH−, Na+, K+, Ca2+ and SO42-, S2O32-, S2- concentrations) of hardened concrete materials, including binary systems of PC mixed with a single SCM and with two SCMs, was investigated. Based on database comprising more than 400 entries with more than 80 parameters, machine learning (ML) is applied to predict ion concentrations. Catboost model is the optimal model. The concentrations of OH−, Na+, K+ were predicted with high accuracy (R2 of 0.92–0.95). The prediction accuracy of S is low, could also reaches a R2 of 0.79. But the prediction accuracy of linear regression model is very low, with R2 of 0.18–0.7. PC_MgO, PC_Na2O, PC_K2O and SCM_SiO2 rank high in the characteristic importance analysis for predicting the concentration of OH−, Na+, K+. Compared with the classical pore solution prediction methods (Taylor's and NIST algorithm), the ML model is more accurate. Due to the use of more data and kinds of methods, the ML prediction results in this paper are also better than Cristhiana's ML model. These ML models can be used to predict pore solution of more than 28 d old normal PC concrete with silica fume, fly ash, slag, limestone or quartz powder, but PC or SCM with high phosphorus oxide is not suitable.

Summary: Promoters are essential tools for basic and translational neuroscience research. An ideal promoter should possess the shortest possible DNA sequence with cell-type selectivity. However, whether ultra-compact promoters can offer neuron-specific expression is unclear. Here, we report the development of an extremely short promoter that enables selective gene expression in neurons, but not glial cells, in the brain. The promoter sequence originates from the human CALM1 gene and is only 120 bp in size. The CALM1 promoter (pCALM1) embedded in an adeno-associated virus (AAV) genome directed broad reporter expression in excitatory and inhibitory neurons in mouse and monkey brains. Moreover, pCALM1, when inserted into an all-in-one AAV vector expressing SpCas9 and sgRNA, drives constitutive and conditional in vivo gene editing in neurons and elicits functional alterations. These data demonstrate the ability of pCALM1 to conduct restricted neuronal gene expression, illustrating the feasibility of ultra-miniature promoters for targeting brain-cell subtypes.

This study conducted a two-year field artificial intelligence (AI) greenhouse rice planting experiment with different canopy humidity (normal humidity, NH; high humidity, and HH) and irrigation regimes (continuous flooding, CF; drought cultivation, DC; alternate wetting-drying, AWD) to test whether high canopy humidity from heading to maturity deteriorates rice grain quality, whether appropriate water management can alleviate these adverse effects, and the related mechanisms. The results showed that compared with NH, HH significantly decreased the head rice rate while increasing the protein, amino acid, amylopectin, amylose, and chalkiness. Moreover, HH significantly decreased the peak viscosity, breakdown, and number of small starch granules, while increasing the setback, number of large starch granules, relative crystallinity, gelatinization temperature, and enthalpy. Under NH and HH, AWD treatment resulted in a higher head rice rate, peak viscosity, breakdown, key enzyme activities of starch synthesis, amylose, amylopecan, relative crystallinity, small starch granules, gelatinization temperature, and enthalpy than DC and CF treatments, while lower chalkiness, setback, protein, amino acid, and large starch granules were observed. HH increased the chalkiness by promoting the formation of large starch granules, thus reducing the milling quality. The increase in amylose and relative crystallinity further causes HH to deteriorate the cooking and eating quality. AWD could alleviate the deterioration of rice grain milling, appearance, and eating quality caused by HH by improving starch granules.