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A ratiometric fluorescent probe based on a “dual-key-and-lock” design was reported for the imaging of vicinal dithiol proteins in living cells.

Reproductive competence in male mammals depends on testicular function. Testicular development and spermatogenesis in goats involve highly complex physiological processes. In this study, six testes were, respectively, obtained from each age group, immature (1 month), sexually mature (6 months) and physically mature (12 months old) Qianbei Ma goats. RNA-Seq was performed to assess testicular mRNA expression in Qianbei Ma goats at different developmental stages. Totally, 18 libraries were constructed to screen genes and pathways involved in testis development and spermatogenesis. Totally, 9724 upregulated and 4153 downregulated DEGs were found between immature (I) and sexually mature (S) samples; 7 upregulated and 3 downregulated DEGs were found between sexually mature (S) and physically mature (P) samples, and about 4% of the DEGs underwent alternative splicing events between I and S. Select genes were assessed by qRT-PCR, corroborating RNA-Seq findings. The detected genes have key roles in multiple developmental stages of goat testicular development and spermatogenesis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to determine differentially expressed genes (DEGs). GO analysis revealed DEGs between S and P contributed to “reproduction process”, “channel activity” and “cell periphery part” between I and S, and in “ion transport process”, “channel activity” and “transporter complex part”. KEGG analysis suggested the involvement of “glycerolipid metabolism”, “steroid hormone biosynthesis” and “MAPK signaling pathway” in testis development and spermatogenesis. Genes including IGF1, TGFB1, TGFBR1 and EGFR may control the development of the testis from immature to sexually mature, which might be important candidate genes for the development of goat testis. The current study provides novel insights into goat testicular development and spermatogenesis.

Citrus Huanglongbing (HLB) is one of the most destructive diseases in the citrus industry. At present, Candidatus Liberibacter asiaticus (CLas) cannot be cultured in vitro, and there is a lack of rapid methods to test antibacterial activity, which greatly hinders the discovery of new antibacterial agents against HLB. To establish a rapid screening method for antibacterial agents against HLB with simple operation, a short cycle, and a large number of tests, the CLas contents in leaves from different citrus branches, different leaves from the same citrus branch, and two halves of the same citrus leaf were detected. Compared with the leaves on different branches and different leaves on the same branch, the difference in CLas content of the left and right halves of the same leaf was small; the difference was basically between 0.7 and 1.3. A rapid and efficient method for primary screening agents against HLB termed the “half-leaf method” was established through our long-term optimization and improvement. To verify the stability and reliability of the activity data measured using this method, 6-chloropurine riboside, which is highly soluble in water, was used as the test agent, and its antibacterial activity against HLB was tested 45 times. The results of the antibacterial activity test showed little difference in the mean values of each data group, indicating that this method could be used as a rapid method for screening agents against HLB. We used this method to test the antibacterial activity of compounds synthesized by our research group against HLB and found that some of the compounds showed good activity.

This study wasintended to investigate the protective effects of dexmedetomidine (DEX) on renal injury caused by acute stress in rats and to explore the protective pathways of DEX on rat kidney from the perspective of oxidative stress.An acute restraint stress modelwas used in this study in which the rats were restrained for 3h after swimming for 15min.In this experiment, biochemical tests and histopathological sections were performed to assess renal function,followed by measurement of oxidative stress and related pathway proteins.Meanwhile,this study used NRK-52E cells for in vitro experiments and dexamethasone for stress-induced changes.The NOX4 inhibitor GLX351322 was also used to verify the inhibitory effect of DEX on NOX4.The open-field experiments confirmed that the acute stress model had been successfully established.Acute stress-induced renal injury increased NOX4 protein expression and decreased Nrf2/HO-1/NQO1 expression levels.Significant reduction in renal NOX4 expression after DEX treatment.The DEX-treated group showed significantly normalized renal biochemical results and significantly less damage observed in pathological sections compared to the acute stress group.The experimental results suggest that DEX treatment of acute stress can affect NOX4/Nrf2/HO-1/NQO1 signaling pathway and inhibit oxidative stress.Therefore,acute stress-induced kidney injury can be prevented by DEX.At the same time,DEX has potential clinical applications in stress syndromes.

The achievement of reproductive competence in male mammals is dependent on the testis. Goat testis’ development and spermatogenesis involve physiological events with high complexity. In the current work, 6 testes were respectively collected from immature, sexually mature and physically mature Qianbei Ma goats (1, 6 and 12 months old, respectively). RNA-Seq was carried out to reveal changes in testis mRNA expression levels in Qianbei Ma goats at various developmental stages, and gene expression profiling at different ages was established. Totally 18 libraries were established for screening genes and pathways associated with testis development and spermatogenic processes. Totally 9,724 upregulated and 4,153 downregulated genes were identified between immature (I) and sexually mature (S) testes; 7 upregulated and 3 downregulated genes were detected between sexually mature (S) and physically mature (P) testes, and approximately 4% of genes were alternately spliced between the I and S groups. Selected genes were verified by qRT-PCR, in agreement with sequencing data indicating their reliability. Those genes have critical functions in various developmental stages of goat testicular development and spermatogenesis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were carried out to evaluate differentially expressed genes (DEGs). GO analysis suggested DEGs were involved in “reproduction process”, “channel activity” and “cell periphery part” between I and S, and in “ion transport process”, “channel activity” and “transporter complex part” between S and P. KEGG analysis indicated that pathways such as “glycerolipid metabolism”, “steroid hormone biosynthesis” and “MAPK signaling pathway” may be involved in testis development and spermatogenesis. Genes including IGF1, TGFB1, TGFBR1 and EGFR may regulate the development of the testis from immature to sexually mature, which may be key candidate genes for the development of goat testis. These findings provide novel insights into goat testicular development and spermatogenesis.

Metabolic reprogramming is a feature of tumour cells and is essential to support their rapid proliferation. The glycolytic activity of liver cancer cells is significantly higher than that of normal liver cells, and the rapidly proliferating tumour cells are powered by aerobic glycolysis. Lipid metabolism reprogramming enables tumour cells to meet their needs for highly proliferative growth and is an important driving force for the development of hepatocellular carcinoma (HCC).To explore the influence of different metabolic subtypes of HCC and analyse their significance in guiding prognosis and treatment based on the molecular mechanism of glycolysis and fatty acid oxidation (FAO).By downloading related data from public databases including the Cancer Genome Atlas (TCGA), the Molecular Signatures Database, and International Cancer Genome Consortium, we utilised unsupervised consensus clustering to divide TCGA Liver Hepatocellular Carcinoma samples into four metabolic subgroups and compared single nucleotide polymorphism, copy number variation, tumour microenvironment, and Genomics of Drug Sensitivity in Cancer and Tumour Immune Dysfunction and Exclusion between different metabolites. The differences and causes of survival and the clinical characteristics between them were analysed, and a prognostic model was established based on glycolysis and FAO genes. Combined with the clinical features, a Norman diagram was created to compare the pros and cons of each model.In the four metabolic subgroups, with the increase in glycolytic expression, the median survival of patients showed the worst results, while FAO showed the best. When comparing the follow-up analysis of each group, we considered that the differences between them might be related to reactive oxygen species, somatic copy number variation of key genes, and immune microenvironment. It was also found that the FAO group and the low-risk group had better efficacy and response to immune checkpoint blockade treatment and anti-tumour drugs.There are obvious differences in genes, chromosomes, and clinical characteristics between metabolic subgroups. The establishment of a prognostic model could predict patient prognosis and guide clinical treatment.

This paper analyzes the research status of 3S technology in China's wetlands. The research of 3S technology in wetland landscape pattern and driving force analysis is increasing day by day. Most of the research uses 3S technology to calculate the landscape index, and reveals the change rule of wetland landscape pattern characteristics with natural and human factors from the type scale and landscape scale. The use of 3S technology for wetland information extraction and dynamic monitoring research is gradually increasing. In most studies, wetland information was extracted according to different classification criteria, and spatio-temporal characteristics of wetland were analyzed based on area change, spatial distribution and shape change. With the development, planning and protection of wetland resources, the advantages of 3S technology will become more prominent.

Oleaginous yeasts utilize renewable resources to produce lipids, which benefits sustainable development, and it is of great interest to screen robust lipid producers. Curvibasidium sp. belongs to nonconventional yeast that are very limitedly studied. Here, two cold-adaptive strains of Curvibasidium sp., namely, Y230 and Y231, isolated from the medicinal lichen Usnea diffracta were investigated for their potential in lipid production. Genome mining of Curvibasidium sp. Y231 was performed, and the special features related to fatty acid biosynthesis were revealed. Glucose, xylose, and glycerol were tested as sole carbon sources for yeast cell growth and lipid production. The total lipid contents of Curvibasidium sp. Y230 and Y231 range from 38.43% to 54.62% of the cell dry cell weight at 20°C, and glucose is the optimal carbon source. These results indicate that the Curvibasidium sp. strains are promising for sustainable lipid production. Our study provides basis for exploration of lichen-derived strains for biotechnological applications, and also benefits utilization of other nonconventional yeasts for sustainable production based on genome-based studies.

The recent COVID-19 pandemic has once again caught the attention of people on the probable zoonotic transmission from animals to humans, but the role of companion animals in the coronavirus (CoV) epidemiology still remains unknown. The present study was aimed to investigate epidemiology and molecular characterizations of CoVs from companion animals in Chengdu city, Southwest China. 523 clinical samples from 393 animals were collected from one veterinary hospital between 2020 and 2021, and the presence of CoVs was detected by end-point PCR using pan-CoV assay targeting the RdRp gene. Partial and complete S genes were sequenced for further genotyping and genetic diversity analysis. A total of 162 (31.0%, 162/523) samples and 146 (37.2%, 146/393) animals were tested positive for CoVs. The positive rate in rectal swabs was higher than that in eye/nose/mouth swabs and ascitic fluid but was not statistically different between clinically healthy and diseased ones. Genotyping identified twenty-two feline enteric coronavirus (FCoV) I, four canine enteric coronavirus (CECoV) I, fourteen CECoV IIa, and one CECoV IIb, respectively. Eight complete S genes, including one canine respiratory coronavirus (CRCoV) strain, were successfully obtained. FCoV strains (F21071412 and F21061627) were more closely related to CECoV strains than CRCoV, and C21041821-2 showed potential recombination event. In addition, furin cleavage site between S1 and S2 was identified in two strains. The study supplemented epidemiological information and natural gene pool of CoVs from companion animals. Further understanding of other functional units of CoVs is needed, so as to contribute to the prevention and control of emerging infectious diseases.

Background. Recent evidence shows that adipogenic differentiation of orbital fibroblasts (OFs) promotes the development of thyroid-associated ophthalmopathy (TAO), an organ-specific immune disease. Furthermore, miR-96-5p has been linked to adipogenic differentiation of C2C12 myoblasts and is significantly correlated with the severity of TAO. The purpose of this study is to look into the role of miR-96-5p in the adipogenesis of OFs with TAO. Methods. The orbital tissues from TAO patients and non-TAO participants were collected, and primary OFs were isolated and cultured for further analysis. miR-96-5p expression was examined using qRT-PCR. The adipogenic differentiation of OFs was then studied. Results. Orbital fibroblasts isolated from adipose tissues of TAO patients (t-OFs) demonstrated greater adipogenic differentiation ability than OFs isolated from adipose tissues of non-TAO participants. miR-96-5p was found to be overexpressed in the orbital tissues of TAO patients and t-OFs. Further research revealed that miR-96-5p, by targeting Smad7, could exacerbate PPAR-γ/C/EBPα signaling-induced adipogenic differentiation of t-OFs. However, inhibiting miR-96-5p could block t-OFs adipogenic differentiation-mediated adipogenesis via Smad7/PPAR-γ/C/EBPα. Conclusions. miR-96-5p plays a critical regulatory role in the development of TAO by targeting Smad7 and promoting adipogenic differentiation of OFs.

Lysosomal polarity is considered a key indicator of lysosomal function due to its significant impact on membrane fluidity and enzymatic reactions in lysosomes. Monitoring lysosomal polarity can gain insight into the related physiological and pathological processes and develop new diagnostic methods. However, current fluorescent probes with lysosomal polarity response suffer from narrow linear range, photobleaching and complicated preparation. Herein, a ratiometric fluorescent probe (r-bCDs) for intracellular lysosomal polarity imaging is designed and constructed by amide bond assembly of polarity-sensitive red fluorescent carbon dots (rCDs) and referenced blue fluorescent carbon dots (bCDs). r-bCDs show a much wider linear range of polarity response (orientation polarizability Δf from 0.020 to 0.315) than other probes, and the interference of uneven distribution and instrument factors can be effectively eliminated by ratiometric fluorescent sensing. Imaging of intracellular lysosomal polarity with r-bCDs is implemented to observe the polarity variation caused by the change of cell state and the difference between cancer cells and normal cells. This work provides a promising tool for studying the related physiological and pathological processes and developing new diagnostic methods.

Elevating the discharge voltage plateau is regarded as the most effective strategy to improve the energy density of Li||CFx batteries in consideration of the finite capacity of CFx (x ∼ 1) cathodes. Here, an electrolyte, with LiBF4 in 1,3-dimethyl-2-imidazolidinone (DMI)/1,2-dimethoxyethane (DME), is developed for the first time to substantially promote the discharge voltage of CFx without compromising the available discharge capacity. DME possesses the property of low viscosity, while DMI functions to increase the voltage plateau during discharge owing to its moderate nucleophilicity and donor number, which decreases the energy barrier for breaking C-F bonds. The optimized electrolyte exhibits a significantly high average discharge voltage of 2.69 V at a current density of 10 mA g-1, which is 11.6% higher than the control electrolyte (2.41 V). In addition, a high energy density of 2099 Wh kg-1 is achieved in the optimized electrolyte (vs 1905 Wh kg-1 in the control electrolyte), showing great potential for practical applications.

Due to the inherent volume expansion effect and low conductivity of silicon monoxide (SiOx, 0

As an emerging visual task, vehicle re-identification refers to the identification of the same vehicle across multiple cameras. Herein, we propose a novel vehicle re-identification method that uses an improved ResNet-50 architecture and utilizes the topology information of a surveillance network to rerank the final results. In the training stage, we apply several data augmentation approaches to expand our training data and increase their diversity in a cost-effective manner. We reform the original RestNet-50 architecture by adding non-local blocks to implement the attention mechanism and replacing part of the batch normalization operations with instance batch normalization. After obtaining preliminary results from the proposed model, we use the reranking algorithm, whose core function is to improve the similarity scores of all images on the most likely path that the vehicle tends to appear to optimize the final results. Compared with most existing state-of-the-art methods, our method is lighter, requires less data annotation, and offers competitive performance.

Ulcerative colitis (UC) is a common chronic recurrent inflammatory bowel disease. This study attempts to reveal the improvement mechanism of floral mushroom polysaccharide (FMPS) on UC from the perspective of coordinated interaction between intestinal microbes and intestinal helper T cell 17 (Th17)/regulatory T cell (Treg) balance.Dextran sulfate sodium (DSS)-induced colitis mice model is used for the experiment. The results suggest that FMPS up-regulated the expression of occludin, ZO-1, and MUC2, and down-regulated the secretion of TNF-α, IL-1β, and IL-6 in colitis mice. Importantly, FMPS restores intestinal Th17/Treg balance. Meanwhile, FMPS can regulate intestinal microorganisms and improve the level of short-chain fatty acids (SCFAs) in colitis mice. Intestinal microbial depletion and fecal microbiota transplantation (FMT) experiments reveal that FMPS ameliorated UC is mediated by intestinal microbiome. Flow cytometry further proves that FMPS restores intestinal Th17/Treg balance in a microbial-dependent manner.These results indicate that FMPS has the potential to improve UC, and its mechanism depends on the restoration of Th17/Treg balance mediated by intestinal microorganisms. Therefore, it is suggested that FMPS dietary supplement can be potentially used to intervene UC.

Background To investigate the effects of PD-1 ligands signaling pathway on microglia and transcriptomic expression and function of neural retina. Methods PD-L1/PD-L2 deficient mice were generated by Cyagen Biosciences Inc. (Suzhou, China). We performed quantitative and qualitative analyses of retinal microglia using mouse retinal western blotting and immunofluorescence experiments. The transcriptomic sequencing of neural retina was performed and analyzed by OE Biotech Co. Ltd. (Shanghai, China). The morphology of neural retina in PD-L1/PD-L2 deficient mice was evaluated using haematoxylin and eosin (HE) staining and in vivo optical coherence tomography (OCT). The visual evoked potential (VEP) test and measuring the photo negative response (PhNR) test assist in approaching changes in visual function in mice. Results PD-L1/PD-L2 deficiency decreased the total number of retinal microglia in neural retina. The activated microglia (M2 microglia) was increased in PD-L1/PD-L2 deficient mice, accompanied by increased STAT6 pathway activation. Transcriptome sequencing and analysis revealed the activation of the PI3K-Akt and MAPK signalling pathways in neural retina of PD-L1/PD-L2 knockout mice. The PD-L1/PD-L2 deficiency did not impact on the morphology and visual function of the neural retina. Conclusions PD-L1/PD-L2 deficiency alters numbers and types of microglia and transcriptomic landscape of neural retina.

Radiation therapy (RT) in combination with an immune checkpoint inhibitor (ICI) represents a promising regimen for non-small cell lung cancer (NSCLC); however, the underlying mechanisms are poorly characterized. We identified a specific dose of RT that conferred tumor regression and improved survival in NSCLC models when combined with ICIs. The immune-modulating functions of RT were ascribed to activated lung-resident Scgb1a1+ club cells. Notably, mice with club-cell-specific knockout of synaptosome-associated protein 23 failed to benefit from the combination treatment, indicating a pivotal role of club cell secretome. We identified eight club cell secretory proteins that inhibited immunosuppressive myeloid cells, reduced pro-tumor inflammation and enhanced antitumor immunity. Notably, CC10, a member of club cell secretome, was increased in plasma of patients with NSCLC responding to the combination therapy. By revealing an immunoregulatory role of club cells, our studies have the potential to guide future clinical trials of ICIs in NSCLC. Mittal and colleagues investigate the mechanisms underlying the therapeutic effects of radiation therapy in combination with checkpoint blockade, finding a role for activated lung-resident secretory club cells in modulating antitumor immune responses.

AIMS We investigated the fate of Salmonella in lettuce seeds grown in a hydroponic system and the potentials of applying photodynamic inactivation (PDI) to enhance microbial safety of hydroponic farming systems. METHODS AND RESULTS Lettuce was grown from Salmonella-contaminated seeds, and rose bengal-mediated PDI was applied. Without intervention, Salmonella could persist in plants and hydroponic farming environment throughout 6 weeks of lettuce growth. Cross-contamination from Salmonella-inoculated to noninoculated seedlings was observed. PDI significantly decreased Salmonella from 3.90 ± 0.31 log colony-forming unit (CFU) per plant to 2.77 ± 0.49 log CFU per plant without extra illumination needed (p

Enterovirus 71 (EV71) is the major pathogens of human hand, foot, and mouth disease (HFMD). EV71 efficiently escapes innate immunity responses of the host to cause infection. At present, no effective antiviral drugs for EV71 are available. Anemoside B4 (B4) is a natural saponin isolated from the roots of Pulsatilla chinensis (Bunge) Regel. P. chinensis extracts that shows a wide variety of biological activities. In this study, we investigated the antiviral activities of B4 against EV71 both in cell culture and in suckling mice. We showed that B4 (12.5–200 μM) dose dependently increased the viability of EV71-infected RD cells with an IC(50) value of 24.95 ± 0.05 μM against EV71. The antiviral activity of B4 was associated with enhanced interferon (IFN)-β response, since knockdown of IFN-β abolished its antiviral activity. We also confirmed that the enhanced IFN response was mediated via activation of retinoic acid-inducible gene I (RIG-I) like receptors (RLRs) pathway, and it was executed by upregulation of 14-3-3 protein, which disrupted the interaction between yes-associated protein (YAP) and interferon regulatory factor 3 (IRF3). By using amino acids in cell culture (SILAC)-based proteomics profiling, we identified the Hippo pathway as the top-ranking functional cluster in B4-treated EV71-infected cells. In vivo experiments were conducted in suckling mice (2-day-old) infected with EV71 and subsequently B4 (200 mg · kg(−1) · d(−1), i.p.) was administered for 16 days. We showed that B4 administration effectively suppressed EV71 replication and improved muscle inflammation and limb activity. Meanwhile, B4 administration regulated the expressions of HFMD biomarkers IL-10 and IFN-γ, attenuating complications of EV71 infection. Collectively, our results suggest that B4 could enhance the antiviral effect of IFN-β by orchestrating Hippo and RLRs pathway, and B4 would be a potential lead compound for developing an anti-EV71 drug.