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Abstract Oocytes play a crucial role in transmitting maternal mitochondrial DNA (mtDNA), essential for the continuation of species. However, the effects of mitochondrial reactive oxygen species (ROS) on mammalian oocyte maturation and mtDNA maintenance remain unclear. We investigated this by conditionally knocking out the Sod2 gene in primordial follicles, elevating mitochondrial matrix ROS levels from early oocyte stages. Our data indicates that reproductive aging in Sod2 conditional knockout females begins at 6 months, with oxidative stress impairing oocyte quality, particularly affecting OXPHOS complex II and mtDNA-encoded mRNA levels. Despite unchanged mtDNA mutation load, mtDNA copy numbers exhibited significant variations. Strikingly, reducing mtDNA copy numbers by reducing mtSSB protein, crucial for mtDNA replication, accelerated reproductive aging onset to three months, underscoring the critical role of mtDNA copy number maintenance under oxidative stress conditions. This research provides new insights into the relationship among mitochondrial ROS, mtDNA, and reproductive aging, offering potential strategies for delaying aging-related fertility decline.

Abstract “Candidatus Liberibacter asiaticus” (CLas) is a phloem-limited alpha-proteobacteria causing Citrus Huanglongbing, the destructive disease currently threatening global citrus industry. Genomic analyses of CLas provide insights into its evolution and biology. Here, we sequenced and assembled whole genomes of 135 CLas strains originally from 20 citrus cultivars collected at ten citrus-growing provinces in China. The resulting dataset comprised 135 CLas genomes ranging from 1,221,309 bp to 1,308,521 bp, with an average coverage of 675X. Prophage typing showed that 44 strains contained Type 1 prophage, 89 strains contained Type 2 prophage, 44 strains contained Type 3 prophage, and 34 of them contained more than one type of prophage/phage. The SNP calling identified a total of 5,090 SNPs. Genome-based phylogenetic analysis revealed two major clades among CLas strains, with Clade I dominated by CLas strains containing Type 1 prophage (79/95) and Clade II dominated by CLas strains containing Type 1 or Type 3 prophage (80/95). This CLas genome dataset provides valuable resources for studying genetic diversity and evolutionary pattern of CLas strains.

IntroductionCitrus is one of the most important fruit crops worldwide, and the root-associated microbiota can have a profound impact on tree health and growth.MethodsIn a collaborative effort, the International Citrus Microbiome Consortium investigated the global citrus root microbiota with samples collected from nine citrus-producing countries across six continents. We analyzed 16S rDNA and ITS2 amplicon sequencing data to identify predominant prokaryotic and fungal taxa in citrus root samples. Comparative analyses were conducted between root-associated microbial communities and those from the corresponding rhizosphere and bulk soil samples. Additionally, genotype-based group-wise comparisons were performed to assess the impact of citrus genotype on root microbiota composition.ResultsTen predominant prokaryotic phyla, containing nine bacterial phyla including Proteobacteria, Actinobacteria, Acidobacteria, and Bacteroidetes and one archaeal phylum (Thaumarchaeota), and multiple fungal phyla including Ascomycota and Basidiomycota were identified in the citrus root samples. Compared with the microbial communities from the corresponding rhizosphere and bulk soil samples from the same trees, the prokaryotic and fungal communities in the roots exhibited lower diversity and complexity but greater modularity compared to those in the rhizosphere. In total, 30 root-enriched and 150 root-depleted genera in bacterial community were identified, whereas 21 fungal genera were enriched, and 147 fungal genera were depleted in the root niche compared with the rhizosphere. The citrus genotype significantly affected the root prokaryotic and fungal communities. In addition, we have identified the core root prokaryotic genera comprising Acidibacter, Allorhizobium, Bradyrhizobium, Chitinophaga, Cupriavidus, Devosia, Dongia, Niastella, Pseudomonas, Sphingobium, Steroidobacter and Streptomyces, and the core fungal genera including Acrocalymma, Cladosporium, Fusarium, Gibberella, Mortierella, Neocosmospora and Volutella. The potential functions of these core genera of root microbiota were predicted.ConclusionOverall, this study provides new insights into the assembly of microbial communities and identifies core members of citrus root microbiota across a wide geographic range. The findings offer valuable information for manipulating root microbiota to enhance plant growth and health.

Abstract Objective Vitamins and homocysteine (Hcy) are involved in liver metabolism and related to the pathogenesis of autoimmune liver disease (AILD), but consensus is lacking. This study aims to systematically summarize relevant evidence to clarify the association of serum vitamins and Hcy levels with AILD. Methods The English and Chinese literature was searched until August 29, 2023. Studies were included if they were observational studies of investigating serum vitamins and Hcy levels in patients with AILD and their healthy comparisons. Quality assessment was performed by using the Newcastle–Ottawa Scale, and a meta‐analysis was conducted using ReviewManager 5.3. The protocol was registered in the international prospective register of systematic reviews (PROSPERO), with registration number CRD42023455367. Results A total of 25 case–control studies comprising 3487 patients (1673 patients and 1814 healthy controls) were included for analysis. There were 548 autoimmune hepatitis (AIH) cases, 1106 primary biliary cholangitis (PBC) cases, and 19 primary sclerosing cholangitis (PSC) cases. We found that serum A and E were decreased in both AIH and PBC/PSC; but vitamin C was reduced only in patients with PBC, not AIH. In addition, decreased content of 25(OH)D3 was found in both AIH and PBC. However, levels of 25(OH)D did not differ between the patients and controls, and were independent of disease types and the country. Only one study that met the inclusion criteria reported vitamin B6, B9, B12, and Hcy changes, and found that vitamin B6 and B9 were significantly decreased in patients with PBC, while serum vitamin B12 and Hcy levels were significantly elevated in them. One eligible study each confirmed a reduction in plasma vitamin K1 and 1,25(OH)2D3 in patients with PBC. Conclusion Most vitamins are deficient in AILD, so appropriate vitamin supplementation should be necessary. Further studies with larger sample sizes are needed to validate these findings.

ABSTRACT“Candidatus Liberibacter asiaticus” (CLas), the causal agent of citrus Huanglongbing (HLB), is able to multiply to a high abundance in citrus fruit pith. However, little is known about the biological processes and phytochemical substances that are vital for CLas colonization and growth in fruit pith. In this study, CLas-infected fruit pith of three citrus cultivars (“Shatangju” mandarin, “Guanxi” pomelo, and “Shatian” pomelo) exhibiting different tolerance to CLas were collected and used for dual RNA-Seq and untargeted metabolome analysis. Comparative transcriptome analysis found that the activation of the CLas noncyclic TCA pathway and pathogenic-related effectors could contribute to the colonization and growth of CLas in fruit pith. The pre-established Type 2 prophage in the CLas genome and the induction of its CRISPR/cas system could enhance the phage resistance of CLas and, in turn, facilitate CLas population growth in fruit pith. CLas infection caused the accumulation of amino acids that were correlated with tolerance to CLas. The accumulation of most sugars and organic acids in CLas-infected fruit pith, which could be due to the phloem blockage caused by CLas infection, was thought to be beneficial for CLas growth in localized phloem tissue. The higher levels of flavonoids and terpenoids in the fruit pith of CLas-tolerant cultivars, particularly those known for their antimicrobial properties, could hinder the growth of CLas. This study advances our understanding of CLas multiplication in fruit pith and offers novel insight into metabolites that could be responsible for tolerance to CLas or essential to CLas population growth.IMPORTANCECitrus Huanglongbing (HLB, also called citrus greening disease) is a highly destructive disease currently threatening citrus production worldwide. HLB is caused by an unculturable bacterial pathogen, “Candidatus Liberibacter asiaticus” (CLas). However, the mechanism of CLas colonization and growth in citrus hosts is poorly understood. In this study, we utilized the fruit pith tissue, which was able to maintain the CLas at a high abundance, as the materials for dual RNA-Seq and untargeted metabolome analysis, aiming to reveal the biological processes and phytochemical substances that are vital for CLas colonization and growth. We provided a genome-wide CLas transcriptome landscape in the fruit pith of three citrus cultivars with different tolerance and identified the important genes/pathways that contribute to CLas colonization and growth in the fruit pith. Metabolome profiling identified the key metabolites, which were mainly affected by CLas infection and influenced the population dynamic of CLas in fruit pith.

ABSTRACTCRISPR-Cas technology has widely been applied to detect single-nucleotide mutation and is considered as the next generation of molecular diagnostics. We previously reported the combination of nucleic acid amplification (NAA) and CRISPR-Cas12a system to distinguish major severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants. However, the mixture of NAA and CRISPR-Cas12a reagents in one tube could interfere with the efficiency of NAA and CRISPR-Cas12a cleavage, which in turn affects the detection sensitivity. In the current study, we employed a novel photoactivated CRISPR-Cas12a strategy integrated with recombinase polymerase amplification (RPA) to develop one-pot RPA/CRISPR-Cas12a genotyping assay for detecting SARS-CoV-2 Omicron sub-lineages. The new system overcomes the potential inhibition of RPA due to early CRISPR-Cas12a activation and cleavage of the target template in traditional one-pot assay using photocleavable p-RNA, a complementary single-stranded RNA to specifically bind crRNA and precisely block Cas12a activation. The detection can be finished in one tube at 39℃ within 1 h and exhibits a low limit of detection of 30 copies per reaction. Our results demonstrated that the photocontrolled one-pot RPA/CRISPR-Cas12a assay could effectively identify three signature mutations in the spike gene of SARS-CoV-2 Omicron variant, namely, R346T, F486V, and 49X, and distinguish Omicron BA.1, BA.5.2, and BF.7 sub-lineages. Furthermore, the assay achieved a sensitivity of 97.3% and a specificity of 100.0% and showed a concordance of 98.3% with Sanger sequencing results.IMPORTANCEWe successfully developed one-pot recombinase polymerase amplification/CRISPR-Cas12a genotyping assay by adapting photocontrolled CRISPR-Cas technology to optimize the conditions of nucleic acid amplification and CRISPR-Cas12a-mediated detection. This innovative approach was able to quickly distinguish severe acute respiratory syndrome coronavirus 2 Omicron variants and can be readily modified for detecting any nucleic acid mutations. The assay system demonstrates excellent clinical performance, including rapid detection, user-friendly operations, and minimized risk of contamination, which highlights its promising potential as a point-of-care testing for wide applications in resource-limiting settings.

Abstract Objectives “Candidatus Liberibacter asiaticus” (CLas) is an un-culturable α-proteobacterium that caused citrus Huanglongbing (HLB), a destructive disease threatening citrus production worldwide. In China, the presence of HLB was first reported in Chaoshan region of Guangdong province, China around a century ago. Thus, whole genome information of CLas strains from Chaoshan area become the most important resource to understand the population diversity and evaluation of CLas in China. Data description CLas strain GDCZ was originally from Chaozhou city (Chaoshan area) and sequenced using PacBio Sequel long-read sequencing and Illumina short-read sequencing. The genome of strain GDCZ comprised of 1,230,507 bp with an average G + C content of 36.4%, along with a circular CLasMV1 phage: CLasMV1_GDCZ (8,869 bp). The CLas strain GDCZ contained a Type 2 prophage (37,452 bp) and encoded a total of 1,057 open reading frames and 53 RNA genes. The whole genome sequence of CLas strain GDCZ from the historical HLB endemic region in China will serve as a useful resource for further analyses of CLas evolution and HLB epidemiology in China and world.

The utilization of the large pretrained model (LPM) based on Transformer has emerged as a prominent research area in various fields, owing to its robust computational capabilities. However, there remains a need to explore how LPM can be effectively employed in the agricultural domain. This research aims to enhance agricultural pest detection with limited samples by leveraging the strong generalization performance of the LPM. Through extensive research, this study has revealed that in tasks involving the counting of a small number of samples, complex agricultural scenes with varying lighting and environmental conditions can significantly impede the accuracy of pest counting. Consequently, accurately counting pests in diverse lighting and environmental conditions with limited samples remains a challenging task. To address this issue, the present research suggests a unique approach that integrates the outstanding performance of the segment anything model in class-agnostic segmentation with the counting network. Moreover, by intelligently utilizing a straightforward TopK matching algorithm to propagate accurate labels, and drawing inspiration from the GPT model while incorporating the forgetting mechanism, a more robust model can be achieved. This approach transforms the problem of matching instances in different scenarios into a problem of matching similar instances within a single image. Experimental results demonstrate that our method enhances the accuracy of the FamNet baseline model by 69.17% on this dataset. Exploring the synergy between large models and agricultural scenes warrants further discussion and consideration.

Abstract Objective The COVID-19 pandemic has been the most serious public health emergency encountered in modern assisted reproductive technology (ART) development. In order to identify lessons learned, this study reviews the effect of the pandemic on ART institutions and human sperm banks in China, and summarizes the experiences and reflections of Chinese scholars post-pandemic era. Methods This review is based on multiple consensus statements on the COVID-19 pandemic issued by Chinese experts as well as current national regulations and principles in ART institutions and human sperm banks to document the current situation of ART services in China, describe the impact of the pandemic on these services, and offer Chinese reflections on worrying issues in the post-pandemic era. Results China reached one million ART cycles in 2016, and there are currently 540 ART medical institutions and 27 human sperm banks, with 540 licensed for AIH, 91 for AID, 415 for conventional IVF and ICSI and 85 for PGT. Of these, only 4 institutions carry out 10,000 cycles or more annually, and the proportion of institutions with less than 1,000 cycles has reached 66%, which means that a considerable number of ART institutions are still not saturated. As a consequence of the COVID-19 pandemic, 63.6% of ART providers and 95.5% of human sperm banks suspended operations. By the end of May 2020, China, as an early country affected by the pandemic achieved a national resumption rate of ART medical services of 99.2% and that of human sperm banks of 100.0%. Reports from the first and largest human sperm bank in China showed that qualification, semen concentration and sperm viability rates measured at primary screening have significantly decreased post-pandemic. Much like in other countries, Chinese experts developed a consensus on prevention and control measures during the pandemic. In principle, all ART activities should be suspended during acute phases of infection spread. Chinese scholars highlight that attention should be paid to young patients with fertility requirements during and after COVID-19, and emphasize the importance of fertility evaluation and clinical intervention. In addition, couples should be reminded that during ART treatment, disinfectants should not be used excessively to minimize risks of damaging the reproductive system, gametes and zygotes. At the same time, timely and reasonable guidance for tackling negative emotions from stress response is needed to provide reassurance and to avoid irrational fear and excessive stress. Seminal parameters should be re-examined 2 months after SARS-CoV-2 vaccination, and ART treatments recommenced if no abnormalities are detected. Conclusions Given the growing frequency of outbreaks of global infectious diseases in recent years, ART institutions and human sperm banks should pay attention to improving their prevention and control capabilities. To a certain extent, decisions and measures adopted in China during COVID-19 pandemic are worthy of recognition and acceptance. Chinese scholars have discussed, proactively responded to and understand the key issues surrounding ART development during the pandemic with the aim of contributing to the substantial progress and healthy development of ART services in the world.

This study aimed to compare the sensitivity of two-phase separation and the filtration method using a mixed cellulose ester (MCE) membrane to detect enteroviruses in sewage samples. From December 2015 to July 2016, four domestic sewage samples (1 L/sample) were collected monthly from the Guangzhou Liede Sewage Treatment Plant, and each sewage sample was divided into two aliquots (500 mL). The sewage sample was concentrated using the two-phase separation and the filtration method using an MCE membrane, and the treated solutions were inoculated into cells for enterovirus isolation. Polymerase chain reaction amplification, VP1 sequencing, and enterovirus molecular typing were performed on the positive isolates. The detection rates of poliovirus (PV) and non-polio enterovirus (NPEV) obtained using the filtration method using an MCE membrane were higher than those using the two-phase separation method. McNemar's test showed that the detection rates of PV, NPEV, type 1 Sabin-like (SL1), type 2 Sabin-like (SL2), and type 3 Sabin-like (SL3) strain were not statistically significant (P > 0.05). In Guangdong Province, China, the detection rates for PV and NPEV were 53.13% and 62.50% (20/32), respectively. Twenty-seven PVs were isolated, three highly variable strains of the type 1 vaccine, with seven nucleotide substitutions in the VP1 region, compared with the type 1 Sabin strains. Eighty-seven strains of NPEV were isolated and nine serotypes were identified, among which coxsackievirus B3 (CVB3), echovirus 6(E6), and echovirus 11(E11) were the dominant strains. The filtration method using an MCE membrane is more sensitive than two-phase separation and can be used as a robust, sensitive, and cost-effective method to isolate enteroviruses from sewage.

Accurately extracting the Region of Interest (ROI) of a palm print was crucial for subsequent palm print recognition. However, under unconstrained environmental conditions, the user's palm posture and angle, as well as the background and lighting of the environment, were not controlled, making the extraction of the ROI of palm print a major challenge. In existing research methods, traditional ROI extraction methods relied on image segmentation and were difficult to apply to multiple datasets simultaneously under the aforementioned interference. However, deep learning-based methods typically did not consider the computational cost of the model and were difficult to apply to embedded devices. This article proposed a palm print ROI extraction method based on lightweight networks. Firstly, the YOLOv5-lite network was used to detect and preliminarily locate the palm, in order to eliminate most of the interference from complex backgrounds. Then, an improved UNet was used for keypoints detection. This network model reduced the number of parameters compared to the original UNet model, improved network performance, and accelerated network convergence. The output of this model combined Gaussian heatmap regression and direct regression and proposed a joint loss function based on JS loss and L2 loss for supervision. During the experiment, a mixed database consisting of 5 databases was used to meet the needs of practical applications. The results showed that the proposed method achieved an accuracy of 98.3% on the database, with an average detection time of only 28ms on the GPU, which was superior to other mainstream lightweight networks, and the model size was only 831k. In the open-set test, with a success rate of 93.4%, an average detection time of 5.95ms on the GPU, it was far ahead of the latest palm print ROI extraction algorithm and could be applied in practice.

The extraction of ROI (region of interest) was a key step in noncontact palm vein recognition, which was crucial for the subsequent feature extraction and feature matching. A noncontact palm vein ROI extraction algorithm based on the improved HRnet for keypoints localization was proposed for dealing with hand gesture irregularities, translation, scaling, and rotation in complex backgrounds. To reduce the computation time and model size for ultimate deploying in low-cost embedded systems, this improved HRnet was designed to be lightweight by reconstructing the residual block structure and adopting depth-separable convolution, which greatly reduced the model size and improved the inference speed of network forward propagation. Next, the palm vein ROI localization and palm vein recognition are processed in self-built dataset and two public datasets (CASIA and TJU-PV). The proposed improved HRnet algorithm achieved 97.36% accuracy for keypoints detection on self-built palm vein dataset and 98.23% and 98.74% accuracy for keypoints detection on two public palm vein datasets (CASIA and TJU-PV), respectively. The model size was only 0.45 M, and on a CPU with a clock speed of 3 GHz, the average running time of ROI extraction for one image was 0.029 s. Based on the keypoints and corresponding ROI extraction, the equal error rate (EER) of palm vein recognition was 0.000362%, 0.014541%, and 0.005951% and the false nonmatch rate was 0.000001%, 11.034725%, and 4.613714% (false match rate: 0.01%) in the self-built dataset, TJU-PV, and CASIA, respectively. The experimental result showed that the proposed algorithm was feasible and effective and provided a reliable experimental basis for the research of palm vein recognition technology.

Background: In addition to being secreted into the intercellular spaces by exocytosis, insulin-like growth factor binding protein 5 (IGFBP5) may also remain in the cytosol or be transported to the nucleus. Depending on the different cellular context and subcellular distribution, IGFBP5 can act as a tumor suppressor or promoter through insulin-like growth factor -dependent or -independent mechanisms. Yet, little is known about the impacts of IGFBP5 on acute myeloid leukemia (AML) and its underlying mechanism. Methods: Here we investigated the roles of IGFBP5 in human AML by using recombinant human IGFBP5 (rhIGFBP5) protein and U937 and THP1 cell lines which stably and ectopically expressed IGFBP5 or mutant IGFBP5 (mtIGFBP5) with the lack of secretory signal peptide. Cell counting kit-8 and flow cytometry assay were conducted to assess the cell viability, cell apoptosis and cell cycle distribution. Cytotoxicity assay was used to detect the chemosensitivity. Leukemia xenograft model and hematoxylin-eosin staining were performed to evaluate AML progression and extramedullary infiltration in vivo. Results: In silico analysis demonstrated a positive association between IGFBP5 expression and overall survival of the AML patients. Both IGFBP5 overexpression and extrinsic rhIGFBP5 suppressed the growth of THP1 and U937 cells by inducing cell apoptosis and arresting G1/S transition and promoted the chemosensitivity of U937 and THP1 cells to daunorubicin and cytarabine. However, overexpression of mtIGFBP5 failed to demonstrate these properties. An in vivo xenograft mouse model of U937 cells also indicated that overexpression of IGFBP5 rather than mtIGFBP5 alleviated AML progression and extramedullary infiltration. Mechanistically, these biological consequences depended on the inactivation of insulin-like growth factor 1 receptor -mediated phosphatidylinositol-3-kinase/protein kinase B pathway. Conclusions: Our findings revealed secreted rather than intracellular IGFBP5 as a tumor-suppressor and chemosensitizer in AML. Upregulation of serum IGFBP5 by overexpression or addition of extrinsic rhIGFBP5 may serve as a suitable therapeutic approach for AML.

Objective: The purpose of the study was to assess the clinical efficacy and safety of a combined perfusion suction platform with pressure feedback control function and an ureteroscopic suction sheath that can measure the ureteropelvic pressure in implementing lithotripsies. Methods: Fifty-two patients who underwent lithotripsy under intelligent monitoring of ureteral intraluminal pressure from June 2016 to January 2018 were retrospectively recruited. The inclusion standard was stone diameter >1.5 cm but

Transient receptor potential vanilloid-6 (TRPV6) is a cation channel belonging to the TRP superfamily, specifically the vanilloid subfamily, and is the sixth member of this subfamily. Its presence in the body is primarily limited to the skin, ovaries, kidney, testes, and digestive tract epithelium. The body maintains calcium homeostasis using the TRPV6 channel, which has a greater calcium selectivity than the other TRP channels. Several pieces of evidence suggest that it is upregulated in the advanced stages of thyroid, ovarian, breast, colon, and prostate cancers. The function of TRPV6 in regulating calcium signaling in cancer will be covered in this review, along with its potential applications as a cancer treatment target.

Citri Reticulatae Pericarpium, especially the pericarp of Citrus reticulata Blanco cv. Chachiensis (PCRC), is an important edible and medicinal ingredient for health and pharmacological properties. Citrus Huanglongbing, a devastating disease that currently threatens the citrus industry worldwide, is caused by a phloem-limited alpha-proteobacterium, “Candidatus Liberibacter asiaticus” (CLas). The industry of cultivar Chachiensis has been suffering from HLB. Although HLB affected the quality of citrus fruit, whether the quality of PCRC was affected by HLB remains unclear. In this study, we compared the metabolite profiles between HLB-affected and healthy PCRC from three sources: fresh, 6-month-old, and 9-year-old PCRC, through the untargeted LC–MS method. Compared to healthy controls, various types of bioactive compounds, mainly flavonoids, terpenoids, alkaloids, coumarins, polysaccharides, and phenolic acids, accumulated in HLB-affected PCRC, especially in the HLB-affected 9-year PCRC. In particular, isorhamnetin, isoliquiritigenin, luteolin 7-O-beta-D-glucoside, limonin, geniposide, pyrimidodiazepine, scoparone, chitobiose, m-coumaric acid, malonate, and pantothenic acid, which contributed to the pharmacological activity and health care effects of PCRC, were highly accumulated in HLB-affected 9-year-old PCRC compared to the healthy control. Multibioassay analyses revealed that HLB-affected 9-year-old PCRC had a higher content of total flavonoids and total polyphenols and exhibited similar antioxidant capacity as compared to healthy controls. The results of this study provided detailed information on the quality of HLB-affected PCRC.

“Candidatus Liberibacter asiaticus” (CLas) is a notorious agent that causes Citrus Huanglongbing (HLB), which is transmitted by Diaphorina citri (D. citri). We recently found that the acquisition and transmission of CLas by D. citri was facilitated by Citrus tristeza virus (CTV), a widely distributed virus in the field. In this study, we further studied whether different CTV strains manipulate the host preference of D. citri, and whether endosymbionts variation is related to CTV strains in D. citri. The results showed that the non-viruliferous D. citri preferred to select the shoots infected with CTV, without strain differences was observed in the selection. However, the viruliferous D. citri prefered to select the mixed strain that is similar to the field’s. Furthermore, D. citri effectively acquired the CTV within 2–12 h depending on the strains of the virus. The persistence period of CTV in D. citri was longer than 24 days, without reduction of the CTV titers being observed. These results provide a foundation for understanding the transmission mode of D. citri on CTV. During the process of CTV acquisition and persistence, the titers of main endosymbionts in D. citri showed similar variation trend, but their relative titers were different at different time points. The titers of the “Candidatus Profftella armatura” and CTV tended to be positively correlated, and the titers of Wolbachia and “Candidatus Carsonella ruddii” were mostly negatively related with titers of CT31. These results showed the relationship among D. citri, endosymbionts, and CTV and provided useful information for further research on the interactions between D. citri and CLas, which may benefit the development of approaches for the prevention of CLas transmission and control of citrus HLB.

Abstract Abnormal activation of Wnt/β-catenin-mediated transcription is closely associated with the malignancy of pancreatic cancer. Family with sequence similarity 83 member A (FAM83A) was shown recently to have oncogenic effects in a variety of cancer types, but the biological roles and molecular mechanisms of FAM83A in pancreatic cancer need further investigation. Here, we newly discovered that FAM83A binds directly to β-catenin and inhibits the assembly of the cytoplasmic destruction complex thus inhibiting the subsequent phosphorylation and degradation. FAM83A is mainly phosphorylated by the SRC non-receptor kinase family member BLK (B-lymphoid tyrosine kinase) at tyrosine 138 residue within the DUF1669 domain that mediates the FAM83A-β-catenin interaction. Moreover, FAM83A tyrosine 138 phosphorylation enhances oncogenic Wnt/β-catenin-mediated transcription through promoting β-catenin-TCF4 interaction and showed an elevated nucleus translocation, which inhibits the recruitment of histone deacetylases by TCF4. We also showed that FAM83A is a direct downstream target of Wnt/β-catenin signaling and correlates with the levels of Wnt target genes in human clinical pancreatic cancer tissues. Notably, the inhibitory peptides that target the FAM83A-β-catenin interaction significantly suppressed pancreatic cancer growth and metastasis in vitro and in vivo. Our results revealed that blocking the FAM83A cascade signaling defines a therapeutic target in human pancreatic cancer.

Xanthomonas citri pv. citri (Xcc), a gram-negative bacterium, is the causal agent of citrus canker, one of the most devastating diseases threatening the citrus industry worldwide. Understanding the diversity and population structure of Xcc is a prerequisite for disease epidemiological monitoring and effective disease management. Recent characterization of the clustered regularly interspaced short palindromic repeats (CRISPR)/cas (CRISPR-associated proteins genes) system with a highly variable repeat number among species provides a new molecular typing method for bacterial genetic analysis. In this study, we performed systematic in silico analyses of 28 Xcc genomes and identified a credible CRISPR/cas in Xcc strains. Further analysis of CRISPR polymorphisms (repeat number and spacer types) in 129 XccA strains collected from six provinces in China identified 15 types of CRISPR arrays with 25 spacers. Phylogenetic analysis of Xcc strains based on the CRISPR locus produced a more reliable and accurate typing result compared to the commonly used loci. In addition, seven associated cas genes—cas1, cas2, cas3, cas4, cas5, cas7 (csd2), and cas8 (csd1)—were found located adjacent to the CRISPR array. BLAST results showed > 99% similarity of seven cas genes among Xcc strains. Homology analysis of spacer sequences showed that six spacers had possible phage/prophage origin. The characterization of the CRISPR/cas system among Xcc strains provided an updated strain typing method for Xcc diversity analysis and yielded a panoramic view of CRISPR evolution for further studies of Xcc-phage interactions.

Abstract Currently, the majority of the global population has been vaccinated with the COVID-19 vaccine, and characterization studies of antibodies in vivo from Omicron breakthrough infection and naive infection populations are urgently needed to provide pivotal clues about accurate diagnosis, treatment, and next-generation vaccine design against SARS-CoV-2 infection. We showed that after infection with Omicron-BA.2, the antibody levels of specific IgM against the Wuhan strain and specific IgG against Omicron were not significantly elevated within 27 days of onset. Interestingly, in this study, the levels of humoral immunity against Omicron-specific IgM were significantly increased after breakthrough infection, suggesting that the detection of Omicron-specific IgM antibodies can be used as a test criterion of Omicron breakthrough infection. In addition, we observed that serums from unvaccinated individuals and the majority of vaccinated infections possessed only low or no neutralizing activity against Omicron at the onset of Omicron breakthrough infections, and at the later stage of Omicron-BA.2 breakthrough infection, levels of neutralization antibody against the Wuhan and Omicron strains were elevated in infected individuals. The findings of this study provide important clues for the diagnosis of Omicron breakthrough infections, antibody characterization studies and vaccine design against COVID-19.

ABSTRACT “Candidatus Liberibacter asiaticus” is one of the putative causal agents of citrus Huanglongbing (HLB), a highly destructive disease threatening the global citrus industry. Several types of phages had been identified in “Ca. Liberibacter asiaticus” strains and found to affect the biology of “Ca. Liberibacter asiaticus.” However, little is known about the influence of phages in “Ca. Liberibacter asiaticus” pathogenicity. In this study, two “Ca. Liberibacter asiaticus” strains, PYN and PGD, harboring different types of phages were collected and used for pathogenicity analysis in periwinkle (Catharanthus roseus). Strain PYN carries a type 1 phage (P-YN-1), and PGD harbors a type 2 phage (P-GD-2). Compared to strain PYN, strain PGD exhibited a faster reproduction rate and higher virulence in periwinkle: leaf symptoms appeared earlier, and there was a stronger inhibition in the growth of new flush. Estimation of phage copy numbers by type-specific PCR indicated that there are multiple copies of phage P-YN-1 in strain PYN, while strain PGD carries only a single copy of phage P-GD-2. Genome-wide gene expression profiling revealed the lytic activity of P-YN-1 phage, as evidenced by the unique expression of genes involved in lytic cycle, which may limit the propagation of strain PYN and lead to a delayed infection in periwinkle. However, the activation of genes involved in lysogenic conversion of phage P-GD-1 indicated it could reside within the “Ca. Liberibacter asiaticus” genome as a prophage form in strain PGD. Comparative transcriptome analysis showed that the significant differences in expression of virulence factor genes, including genes associated with pathogenic effectors, transcriptional factors, the Znu transport system, and the heme biosynthesis pathway, could be another major determinant of virulence variation between two “Ca. Liberibacter asiaticus” strains. This study expanded our knowledge of “Ca. Liberibacter asiaticus” pathogenicity and provided new insights into the differences in pathogenicity between “Ca. Liberibacter asiaticus” strains. IMPORTANCE Citrus Huanglongbing (HLB), also called citrus greening disease, is a highly destructive disease threatening citrus production worldwide. “Candidatus Liberibacter asiaticus” is one of the most common putative causal agents of HLB. Phages of “Ca. Liberibacter asiaticus” have recently been identified and found to affect “Ca. Liberibacter asiaticus” biology. Here, we found that “Ca. Liberibacter asiaticus” strains harboring different types of phages (type 1 or type 2) showed different levels of pathogenicity and multiplication patterns in the periwinkle plant (Catharanthus roseus). Transcriptome analysis revealed the possible lytic activity of type 1 phage in a “Ca. Liberibacter asiaticus” strain, which could limit the propagation of “Ca. Liberibacter asiaticus” and lead to the delayed infection in periwinkle. The heterogeneity in the transcriptome profiles, particularly the significant differences in expression of virulence factors genes, could be another major determinant of difference in virulence observed between the two “Ca. Liberibacter asiaticus” strains. These findings improved our understanding of “Ca. Liberibacter asiaticus”-phage interaction and provided insight into “Ca. Liberibacter asiaticus” pathogenicity.

ABSTRACT Continuous surveillance of enteroviruses (EVs) in urban domestic sewage can timely reflect the circulation of EVs in the environment and crowds, and play a predictive and early warning role in EV-related diseases. To better understand the long-term epidemiological trends of circulating EVs and EV-related diseases, we conducted a 9-year (2013 to 2021) surveillance study of non-polio EVs (NPEVs) in urban sewage in Guangzhou city, China. After concentrating and isolating the viruses from the sewage samples, NPEVs were detected and molecular typing was performed. Twenty-one different NPEV serotypes were identified. The most isolated EVs were echovirus 11 (E11), followed by coxsackievirus (CV) B5, E6, and CVB3. EV species B prevailed in sewage samples, but variations in the annual frequency of different serotypes were also observed in different seasons, due to spatial and temporal factors. E11 and E6 were detected continuously before 2017, and the number of isolates was relatively stable during the surveillance period. However, after their explosive growth in 2018 and 2019, their numbers suddenly decreased significantly. CVB3 and CVB5 had alternating trends; CVB5 was most frequently detected in 2013 to 2014 and 2017 to 2018, while CVB3 was most frequently detected in 2015 to 2016 and 2020 to 2021. Phylogenetic analysis showed that at least two different transmission chains of CVB3 and CVB5 were prevalent in Guangzhou City. Our results show that in the absence of a comprehensive and systematic EV-related disease surveillance system in China, environmental surveillance is a powerful and effective tool to strengthen and further investigate the invisible transmission of EVs in the population. IMPORTANCE This study surveilled urban sewage samples from north China for 9 years to monitor enteroviruses. Samples were collected, processed, and viral identification and molecular typing were performed. We detected 21 different non-polio enteroviruses (NPEVs) with yearly variations in prevalence and peak seasons. In addition, this study is very important for understanding the epidemiology of EVs during the COVID-19 pandemic, as the detection frequency and serotypes of EVs in sewage changed considerably around 2020. We believe that our study makes a significant contribution to the literature because our results strongly suggest that environmental surveillance is an exceptionally important tool, which can be employed to detect and monitor organisms of public health concern, which would otherwise be missed and under-reported by case-based surveillance systems alone.

Citrus Huanglongbing (HLB), caused by “Candidatus Liberibacter asiaticus” (CLas), is the most destructive disease threatening global citrus industry. Most commercial cultivars were susceptible to HLB, although some showed tolerant to HLB phenotypically. Identifying tolerant citrus genotypes and understanding the mechanism correlated with tolerance to HLB is essential for breeding citrus variety tolerance/resistance to HLB. In this study, the graft assay with CLas-infected bud were performed in four citrus genotypes, including Citrus reticulata Blanco, C. sinensis, C. limon, and C. maxima. HLB tolerance was observed in C. limon and C. maxima, while C. Blanco and C. sinensis were susceptible to HLB. The time-course transcriptomic analysis revealed a significant variation in differentially expressed genes (DEGs) related to HLB between susceptible and tolerant cultivar group at early and late infection stage. Functional analysis of DEGs indicated that the activation of genes involved in SA-mediated defense response, PTI, cell wall associated immunity, endochitinase, phenylpropanoid and alpha-linolenic/linoleic lipid metabolism played an important in the tolerance of C. limon and C. maxima to HLB at early infection stage. In addition, the overactive plant defense combined with the stronger antibacterial activity (antibacterial secondary and lipid metabolism) and the suppression of pectinesterase were contributed to the long-term tolerance to HLB in C. limon and C. maxima at late infection stage. Particularly, the activation of ROS scavenging genes (catalases and ascorbate peroxidases) could help to reduce HLB symptoms in tolerant cultivars. In contrast, the overexpression of genes involved in oxidative burst and ethylene metabolism, as well as the late inducing of defense related genes could lead to the early HLB symptom development in susceptible cultivars at early infection stage. The weak defense response and antibacterial secondary metabolism, and the induce of pectinesterase were responsible for sensitivity to HLB in C. reticulata Blanco and C. sinensis at late infection stage. This study provided new insights into the tolerance/sensitivity mechanism against HLB and valuable guidance for breeding of HLB-tolerant/resistant cultivars.

Grafting is a commonly used method for citrus propagation and transmitting 'Candidatus Liberibacter asiaticus' (CLas), the putative causing agent of citrus Huanglongbing (HLB). Optimization of the grafting inoculation methods facilitates the material preparation in HLB research. Citrus buds with CLas were grafted onto healthy sour tangerine (Citrus sunki Hort. ex Tanaka) seedlings by different methods such as top grafting ('T' grafting and 'V' grafting) and side grafting (abdominal grafting). Along with the symptom observation, titers of CLas in the leaves were detected by RT-qPCR monthly. The correlation between the growth status of buds or different grafting methods and the success rate of HLB transmission were analyzed. Our results suggest that sufficient DNA could be extracted to accurately detect the CLas from even only 0.0125 g leaf midrib or branch bark. The probability of CLas transmission was higher in plants inoculated with buds in better growth conditions. The success rate of 'T' grafting was significantly higher than that of side grafting and 'V' grafting. Additionally, in terms of HLB transmission efficiency, the two-bud grafting scheme was superior to the single-bud and three-bud grafting schemes. In conclusion, the grafting combinations with the highest HLB transmission efficiency were screened to provide a methodological reference for the practice or research of grafting to obtain plant material.

The SARS-CoV-2 Delta variant has spread rapidly worldwide. Here, the authors characterise a single chain of transmission of Delta in China, and find evidence that it is more infectious and replicates faster during early infection compared to early pandemic lineages.

In this article, the finite difference parabolic equation (FDPE) method is presented to calculate the propagation loss (PL) for electromagnetic waves (EWs) in the forest environment. The FDPE method is more efficient and convenient than the empirical models and has more advantages on compatibility and accuracy for long-range EWs prediction. The Debye–Cole dual dispersion model is used to simulate the effective permittivity of vegetation. The results of the FDPE model are compared with those of the advanced refractive effects prediction system and measurement results, and a good agreement is observed. Research found that PL for EWs varies with the effective permittivity. Also, the effective permittivity is a function of radio frequency, weight moisture content, and volume content of vegetation. Thus, it is necessary to establish a statistical model to determine some relations between the PL and plant biophysical parameters. The polynomial fitting method is adopted to process a large amount of PL data for obtaining a linear function. Then, the volume content and moisture content of vegetation can be determined according to the polynomial fitting function. It provides a novel and efficient method for forestry remote telemetry, which is specifically suitable for large-scale inaccessible region with serious environment.

Till now, the medicines approved for acute myeloid leukemia with internal tandem duplication mutation of FMS-like tyrosine kinase 3 (FLT3-ITD) display not ideal efficacy. This study aimed to evaluate the effects of 15,16-dihydrotanshinone I on FLT3-ITD acute myeloid leukemia cells. The inhibitory effect of this compound against MV4-11 was determined using CCK-8 assay. Western blotting detecting caspase-3, PARP, and annexin V-APC/7-AAD was carried out. Activation of FLT3, STAT5, and Mcl-1 expression was analyzed by western blotting. The results showed that MV4-11 was sensitive toward dihydrotanshinone I in a dose-dependent manner (p

Visual Question Answering (VQA) about diseases is an essential feature of intelligent management in smart agriculture. Currently, research on fruit tree diseases using deep learning mainly uses single-source data information, such as visible images or spectral data, yielding classification and identification results that cannot be directly used in practical agricultural decision-making. In this study, a VQA model for fruit tree diseases based on multimodal feature fusion was designed. Fusing images and Q&A knowledge of disease management, the model obtains the decision-making answer by querying questions about fruit tree disease images to find relevant disease image regions. The main contributions of this study were as follows: (1) a multimodal bilinear factorized pooling model using Tucker decomposition was proposed to fuse the image features with question features: (2) a deep modular co-attention architecture was explored to simultaneously learn the image and question attention to obtain richer graphical features and interactivity. The experiments showed that the proposed unified model combining the bilinear model and co-attentive learning in a new network architecture obtained 86.36% accuracy in decision-making under the condition of limited data (8,450 images and 4,560k Q&A pairs of data), outperforming existing multimodal methods. The data augmentation is adopted on the training set to avoid overfitting. Ten runs of 10-fold cross-validation are used to report the unbiased performance. The proposed multimodal fusion model achieved friendly interaction and fine-grained identification and decision-making performance. Thus, the model can be widely deployed in intelligent agriculture.

BackgroundHuanglongbing (HLB, yellow shoot disease) is a highly destructive citrus disease associated with a nonculturable bacterium, “Candidatus Liberibacter asiaticus” (CLas), which is transmitted by Asian citrus psyllid (ACP, Diaphorina citri). In Mexico, HLB was first reported in Tizimin, Yucatán, in 2009 and is now endemic in 351 municipalities of 25 states. Understanding the population diversity of CLas is critical for HLB management. Current CLas diversity research is exclusively based on analysis of the bacterial genome, which composed two regions, chromosome (> 1,000 genes) and prophage (about 40 genes).Methods and resultsIn this study, 40 CLas-infected ACP samples from 20 states in Mexico were collected. CLas was detected and confirmed by PCR assays. A prophage gene(terL)-based typing system (TTS) divided the Mexican CLas strains into two groups: Term-G including four strains from Yucatán and Chiapas, as well as strain psy62 from Florida, USA, and Term-A included all other 36 Mexican strains, as well as strain AHCA1 from California, USA. CLas diversity was further evaluated to include all chromosomal and prophage genes assisted by using machine learning (ML) tools to resolve multidimensional data handling issues. A Term-G strain (YTMX) and a Term-A strain (BCSMX) were sequenced and analyzed. The two Mexican genome sequences along with the CLas genome sequences available in GenBank were studied. An unsupervised ML was implemented through principal component analysis (PCA) on average nucleotide identities (ANIs) of CLas whole genome sequences; And a supervised ML was implemented through sparse partial least squares discriminant analysis (sPLS-DA) on single nucleotide polymorphisms (SNPs) of coding genes of CLas guided by the TTS. Two CLas Geno-groups, Geno-group 1 that extended Term-A and Geno-group 2 that extended Term-G, were established.ConclusionsThis study concluded that: 1) there were at least two different introductions of CLas into Mexico; 2) CLas strains between Mexico and USA are closely related; and 3) The two Geno-groups provide the basis for future CLas subspecies research.

Introduction and objectives: We performed a systematic review and meta-analysis to evaluate the prevalence of concomitant Sjögren's syndrome (SS) with primary biliary cholangitis (PBC) in adults and quantify the impact of SS on PBC. Methods: PubMed, Web of Science and Cochrane library were searched using subject terms and predefined inclusion and exclusion criteria. Results: Seventeen articles were included. The prevalence of SS in PBC patients ranged from 3.5 to 73% (35% pooled) (95% CI: 28–41%; p

Integrin β6 (ITGB6), an epithelial-specific receptor, is downregulated in the gingival epithelium of periodontitis and is associated with inflammation response and periodontitis development. However, the transcriptional regulatory mechanism of ITGB6 downregulation in the human gingival epithelium remains unclear. Fibroblast-stimulating lipopeptide-1 (FSL-1), an oral biofilm component, promotes an epithelial cell-driven proinflammatory response in periodontitis partially by suppressing ITGB6 expression. The aim of the current study was to investigate the transcriptional regulatory mechanism of ITGB6 inhibition by FSL-1 in human epithelial cells (HaCaT and primary human gingival epithelial cells), and to delineate the transcriptional mechanism of ITGB6 suppression in periodontitis. We found that FSL-1 inhibited ITGB6 transcription through increasing forkhead box protein O1 (FOXO1) expression and inhibiting signal transducer and activator of transcription 3 (STAT3) activation. Furthermore, FOXO1 bound to STAT3 directly, leading to decreased STAT3 phosphorylation induced by FSL-1. Consequently, the binding of phosphorylated STAT3 to the ITGB6 promoter was decreased, and ITGB6 transcription was therefore downregulated following FSL-1 stimulation. The reciprocal action of STAT3 and FOXO1 on ITGB6 downregulation was also confirmed by the immunostaining of the inflammatory epithelium associated with periodontitis. Our findings suggest that the interaction of FOXO1–STAT3 may be a useful signal target for the treatment of periodontitis.

BackgroundBurkholderia pseudomallei (B. pseudomallei) is a highly infectious agent and causes melioidosis, in both humans and animals, which is endemic in Southeast Asia and Northern Australia.ObjectivesThis study aims to determine the molecular epidemiology, resistant determinants, and genomic diversity of the clinical isolates of B. pseudomallei to further elucidate the phylogenetic and evolutionary relationship of the strains with those in other endemic regions.MethodsIn this study, we obtained eight clinical B. pseudomallei isolates from Guangdong province from 2018 to 2019. All the isolates were sequenced using the Illumina NovaSeq platform. The draft genomes of B. pseudomallei were further used to find antibiotic-resistant genes (ARGs), virulence factors, and gene mutations. Multilocus sequence typing (MLST) and single nucleotide polymorphism (SNP) analysis were performed to characterize the diversity and epidemiology of the strains.ResultsAll isolates were susceptible to antibiotics commonly used for melioidosis treatment. Class D beta-lactamases genes OXA-57 and OXA-59, as well as various mutation factors such as amrA, amrB, omp38, gyrA, and ceoB were identified. MLST analysis of the B. pseudomallei strains identified eight different sequence types (STs): ST1774, ST1775, ST271, ST562, ST46, ST830, ST1325, and ST10. Phylogenetic analysis found that the strains used in this study showed high genetic diversity. We also report 165 virulence factors among B. pseudomallei strains responsible for different neurological disorders, pneumonia, skin lesions, and abscesses. All strains recovered in this study were susceptible to commonly used antibiotics. However, high genetic diversity exists among the isolates. The surveillance, diagnosis, and clinical features of melioidosis varied in different geographical locations. These regional differences in the clinical manifestations have implications for the practical management of the disease.ConclusionThe present study reports the identification of different mutation and virulence factors among B. pseudomallei strains responsible for different neurological disorders, pneumonia, skin lesions, and abscesses.

Abstract Background Avian pathogenic Escherichia coli (APEC) strains cause infectious diseases in poultry. Resveratrol is extracted from Polygonum cuspidatum, Cassia tora Linn and Vitis vinifera, and displays good antimicrobial activity. The present study aimed to investigate the antibiofilm effect of resveratrol on APEC in vitro. The minimum inhibitory concentration (MIC) of resveratrol and the antibiotic florfenicol toward APEC were detected using the broth microdilution method. Then, the effect of resveratrol on swimming and swarming motility was investigated using a semisolid medium culture method. Subsequently, the minimum biofilm inhibitory concentration (MBIC) and the biofilm eradication rate were evaluated using crystal violet staining. Finally, the antibiofilm activity of resveratrol was observed using scanning electron microscopy (SEM). Meanwhile, the effects of florfenicol combined with resveratrol against biofilm formation by APEC were evaluated using optical microscopy (OM) and a confocal laser scanning microscopy (CLSM). Results The MICs of resveratrol and florfenicol toward APEC were 128 μg/mL and 64 μg/mL, respectively. The swimming and swarming motility abilities of APEC were inhibited in a resveratrol dose-dependent manner. Furthermore, resveratrol showed a significant inhibitory activity against APEC biofilm formation at concentrations above 1 μg/mL (p

The holoparasitic dodder (Cuscuta spp.) is able to transfer mRNA and certain plant pathogens (e.g., viruses and bacteria) from the host plant. “Candidatus Liberibacter asiaticus,” the phloem-limited causative agent of citrus Huanglongbing, can be transferred from citrus to periwinkle (Catharanthus roseus) mediated by dodder. However, characterization of mRNA transport between dodder and citrus/periwinkle remains unclear. In this study, we sequenced transcriptomes of dodder and its parasitizing host, sweet orange (Citrus sinensis “Newhall”) and periwinkle (Catharanthus roseus), to identify and characterize mRNA transfer between dodder and the host plant during parasitism. The mRNA transfer between dodder and citrus/periwinkle was bidirectional and most of the transfer events occurred in the interface tissue. Compared with the citrus–dodder system, mRNA transfer in the periwinkle–dodder system was more frequent. Function classification revealed that a large number of mRNAs transferred between dodder and citrus/periwinkle were involved in secondary metabolism and stress response. Dodder transcripts encoding proteins associated with microtubule-based processes and cell wall biogenesis were transferred to host tissues. In addition, transcripts involved in translational elongation, plasmodesmata, and the auxin-activated signaling pathway were transmitted between dodder and citrus/periwinkle. In particular, transcripts involved in shoot system development and flower development were transferred between the host and dodder in both directions. The high abundance of dodder-origin transcripts, encoding MIP aquaporin protein, and S-adenosylmethionine synthetase 1 protein, in citrus and periwinkle tissues indicated they could play an important biological role in dodder–host interaction. In addition, the uptake of host mRNAs by dodder, especially those involved in seed germination and flower development, could be beneficial for the reproduction of dodder. The results of this study provide new insights into the RNA-based interaction between dodder and host plants.

Coronavirus disease 2019 (COVID-19) pandemic caused by SARS-CoV-2 variants is a new and unsolved threat; therefore, it is an urgent and unmet need to develop a simple and rapid method for detecting and tracking SARS-CoV-2 variants. The spike gene of SARS-CoV-2 was amplified by isothermal recombinase-aided amplification (RAA) followed by the cleavage of CRISPR-Cas12a in which five allele-specific crRNAs and two Omicron-specific crRNAs were designed to detect and distinguish major SARS-CoV-2 variants of concerns (VOCs), including alpha, beta, delta variants, and Omicron sublineages BA.1 and BA.2. The whole reaction can be carried out in one tube at 39°C within 1.5–2 h, and the results can be read out by a fluorescence meter or naked eyes. Our results show that the RAA/CRISPR-Cas12a-based assay could readily distinguish the signature mutations, i.e., K417N, T478K, E484K, N501Y, and D614G, with a sensitivity of 100.0% and a specificity of 94.9–100.0%, respectively. The assay had a low limit of detection (LOD) of 104 copies/reaction and a concordance of 92.59% with Sanger sequencing results when detecting 54 SARS-CoV-2 positive clinical samples. The two Omicron-specific crRNAs can readily and correctly distinguish Omicron BA.1 and BA.2 sublineages with a LOD of as low as 20 copies/reaction. Furthermore, no cross-reaction was observed for all crRNAs analyzed when detecting clinical samples infected with 11 common respiratory pathogens. The combination of isothermal amplification and CRISPR-Cas12a-mediated assay is suitable for rapid detection of major SARS-CoV-2 variants in point-of-care testing and in resource-limiting settings. This simple assay could be quickly updated for emerging variants and implemented to routinely monitor and track the spread of SARS-CoV-2 variants.

Summary ‘Candidatus Liberibacter asiaticus’ (CLas) is a phloem‐limited non‐culturable α‐proteobacterium associated with citrus Huanglongbing, a highly destructive disease threatening global citrus industry. Research on CLas is challenging due to the current inability to culture CLas in vitro and the low CLas titre in citrus plant. Here, we develop a CLas enrichment system using the holoparasitic dodder plant (Cuscuta campestris) as an amenable host to acquire and enrich CLas from CLas‐infected citrus shoots maintained hydroponically. Forty‐eight out of fifty‐five (87%) dodder plants successfully parasitized CLas‐infected citrus shoots with detectable CLas by PCR. Among 48 dodders cultures, 30 showed two‐ to 419‐fold CLas titre increase as compared to the corresponding citrus hosts. The CLas population rapidly increased and reached the highest level in dodder tendrils at 15 days after parasitizing citrus shoot. Genome sequencing and assembly derived from CLas‐enriched dodder DNA samples generated a higher resolution than those obtained for CLas from citrus hosts. No genomic variation was detected in CLas after transmission from citrus to dodder during short‐term parasitism. Dual RNA‐Seq experiments showed similar CLas gene expression profiles in dodder and citrus samples, yet dodder samples generated a higher resolution of CLas transcriptome data. The ability of dodder to support CLas multiplication to high levels, as well as its advantage in CLas genomic and transcriptomic analyses, make it an optimal model for further studies on CLas–host interaction.

In response to the difficulty of traditional image processing methods to quickly and accurately extract regions of interest from non-contact dorsal hand vein images in complex backgrounds, this study proposes a model based on an improved U-Net for dorsal hand keypoint detection. The residual module was added to the downsampling path of the U-Net network to solve the model degradation problem and improve the feature information extraction ability of the network; the Jensen–Shannon (JS) divergence loss function was used to supervise the final feature map distribution so that the output feature map tended to Gaussian distribution and improved the feature map multi-peak problem; and Soft-argmax is used to calculate the keypoint coordinates of the final feature map to realize end-to-end training. The experimental results showed that the accuracy of the improved U-Net network model reached 98.6%, which was 1% better than the original U-Net network model; the improved U-Net network model file was only 1.16 M, which achieved a higher accuracy than the original U-Net network model with significantly reduced model parameters. Therefore, the improved U-Net model in this study can realize dorsal hand keypoint detection (for region of interest extraction) for non-contact dorsal hand vein images and is suitable for practical deployment in low-resource platforms such as edge-embedded systems.

Mangrove-associated fungi are important sources for the discovery of new bioactive natural products. Three new isocoumarins (1–3) and one new pyrone derivative (4) were isolated from the ethyl acetate extract of the fermentation broth of the mangrove endophytic fungus Phomopsis sp. DHS-11. Nuclear magnetic resonance (NMR) spectroscopy (one-dimensional and two-dimensional) and mass spectrometry were used to determine the structures of these new compounds. The absolute configurations for the new isocoumarins 1–3 were determined by comparing their experimental and calculated electronic circular dichroism (ECD) spectra, while the configuration for the new pyrone-derivative 4 was tentatively solved by comparison of its 13C NMR data with reported data. In the biological activity test, compounds 1 and 3 showed cytotoxic activity against HeLa cells with IC50 values of 11.49 ± 1.64 µM and 8.70 ± 0.94 µM, respectively. The initial structure and activity relationship (SAR) analysis revealed that the length of the side chain at C-3 for isocoumarin-type compounds 1–3 could affect the cytotoxicity against HeLa cells. Compound 4 exhibited cytotoxic activities against human hepatoma cells HepG2 with an IC50 value of 34.10 ± 2.92 µM. All compounds have no immunosuppressive activity.

Diaphorina citri Kuwayama (D. citri) is an insect vector of phloem-limited ‘Candidatus Liberibacter asiatus’ (CLas), the presumed pathogen of citrus Huanglongbing (HLB). Recently, our lab has preliminarily found it acquired and transmitted Citrus tristeza virus (CTV), which was previously suggested to be vectored by species of aphids. However, the influences of one of the pathogens on the acquisition and transmission efficiency of the other pathogen remain unknown. In this study, CLas and CTV acquisition and transmission by D. citri at different development stages under field and laboratory conditions were determined. CTV could be detected from the nymphs, adults, and honeydew of D. citri but not from the eggs and exuviates of them. CLas in plants might inhibit CTV acquisition by D. citri as lower CTV–positive rates and CTV titers were detected in D. citri collected from HLB-affected trees compared to those from CLas–free trees. D. citri were more likely to obtain CTV than CLas from host plants co-infected with the two pathogens. Intriguingly, CTV in D. citri facilitated the acquisition and transmission of CLas, but CLas carried by D. citri had no significant effect on the transmission of CTV by the same vector. Molecular detection and microscopy methods confirmed the enrichment of CTV in the midgut after a 72-h acquisition access period. Collectively, these results raise essential scientific questions for further research on the molecular mechanism of pathogen transmission by D. citri and provide new ideas for the comprehensive prevention and control of HLB and CTV.

SWI/SNF related, matrix associated, actin-dependent regulator of chromatin, subfamily a, member 4 (SMARCA4, also known as BRG1), an ATPase subunit of the switch/sucrose non-fermentable (SWI/SNF) chromatin remodeling complex, plays an important regulatory role in many cytogenetic and cytological processes during cancer development. However, the biological function and mechanism of SMARCA4 in oral squamous cell carcinoma (OSCC) remain unclear. The present study aimed to investigate the role of SMARCA4 in OSCC and its potential mechanism. Using a tissue microarray, SMARCA4 expression was found to be highly upregulated in OSCC tissues. In addition, SMARCA4 upregulate expression led to increased migration and invasion of OSCC cells in vitro, as well as tumor growth and invasion in vivo. These events were associated with the promotion of epithelial–mesenchymal transition (EMT). Bioinformatic analysis and luciferase reporter assay confirmed that SMARCA4 is a target gene of microRNA miR-199a-5p. Further mechanistic studies showed that the miR-199a-5p regulated SMARCA4 can promote the invasion and metastasis of tumor cells through EMT. These findings indicate that the miR-199a-5p- SMARCA4 axis plays a role in tumorigenesis by promoting OSCC cell invasion and metastasis through EMT regulation. Our findings provide insights into the role of SMARCA4 in OSCC and the mechanism involved, which may have important implications for therapeutic purposes.

In recent years, the occurrence of rice pests has been increasing, which has greatly affected the yield of rice in many parts of the world. The prevention and cure of rice pests is urgent. Aiming at the problems of the small appearance difference and large size change of various pests, a deep neural network named YOLO-GBS is proposed in this paper for detecting and classifying pests from digital images. Based on YOLOv5s, one more detection head is added to expand the detection scale range, the global context (GC) attention mechanism is integrated to find targets in complex backgrounds, PANet is replaced by BiFPN network to improve the feature fusion effect, and Swin Transformer is introduced to take full advantage of the self-attention mechanism of global contextual information. Results from experiments on our insect dataset containing Crambidae, Noctuidae, Ephydridae, and Delphacidae showed that the average mAP of the proposed model is up to 79.8%, which is 5.4% higher than that of YOLOv5s, and the detection effect of various complex scenes is significantly improved. In addition, the paper analyzes and discusses the generalization ability of YOLO-GBS model on a larger-scale pest data set. This research provides a more accurate and efficient intelligent detection method for rice pests and others crop pests.

Huanglongbing (HLB) is the most harmful bacterial disease in citrus production in the world, and has been seriously ravaging the citrus groves of South China since the 1930s. The surveillance of the epidemiological characteristics of HLB is of utmost priority for citrus production in this region. In order to explore the effects of disease control measures, analyses on the space–time statistical features of the HLB epidemic, from 2019 to 2021, within six orchards in the Guangdong province are presented. Overall, the number of citrus plants in the orchards usually slightly decreased year by year. The reduction was mainly related to the level of plant susceptibility, which is correlated with citrus varieties. The maximum disease severity (incidence and race increment) was correlated with the awareness of this disease and the management intensity applied by the manager. A higher disease index was found in the conventional management orchards than in the comprehensive prevention and control orchards. Proper insect-protective screen houses can effectively prevent the epidemic of HLB, without affecting the fruit quality, and can also aid with higher yields. A high correlation was found between the geometry and topography of orchards and the HLB epidemic due to the wind direction from May to September and the Asia citrus psyllid activity characteristics. For flat orchards, the incidence of HLB in the north and entrance areas was higher than that in the southwest. In the mountain area, the incidence of the windward side in the south was higher than that of the leeward side in the north. Diseased trees tended to have an edge effect in the grove, whereas the trees of the same disease scale were found clustered in their distribution. These results allow a better understanding of HLB epidemiology and provide guidance for the early warning of HLB in new groves in areas that are severely affected by this disease. Furthermore, they also provide a scientific basis for the comprehensive prevention and control of HLB in old groves.

Huanglongbing (HLB), which is mainly associated with “Candidatus Liberibacter asiaticus” (CLas), is currently threatening citrus production around the world. This destructive century-old disease results in callose deposition in phloem plasmodesmata and starch overaccumulation in leaf chloroplasts. Heat treatment with high humidity has proven to be effective in eliminating the CLas titers in potted citrus trees. This study explored the curative mechanisms and impact of heat treatment (from 32 to 48 °C with 75% ± 5% relative humidity, 14 h in light and 10 h in dark for four days) on mandarin trees (Citrus reticulata Blanco) that were severely infected by CLas using Solexa/Illumina's digital gene expression (DGE) profiling. A total of 838 differentially expressed genes (DEGs) were identified, of which 510 were downregulated. Protein-protein interaction analysis of the DEGs indicated that the thermotherapy of HLB trees downregulated the circadian clock related genes in chloroplasts and upregulated the RNA-editing process in the mitochondria, characterized by 141 genes encoding pentatricopeptide repeat-containing proteins (PPR) being all upregulated. Furthermore, the expression of phenylpropanoid and flavonoid biosynthesis genes, and auxin-induced genes were suppressed. The defense response pathway of the thermotherapy treated diseased trees was apparently disrupted, as indicated by the downregulation of LRR receptor-like serine/threonine-protein kinase FLS2 gene. This study improves our understanding of the curative mechanisms of heat treatment on HLB trees and CLas biological regulation. Importantly, this provides new insights on the involvement of RNA modification by PPR genes and changes in the circadian clock after the heat stress treatment of CLas-infected citrus.

Magnetoelectric composites have attracted much attention due to their strong magnetoelectric coupling effect. In this paper, PbTiO3-NiFe2O4 (PTO/NFO) composite ceramics were prepared by sol-gel method. Effect of sintering temperatures on the structure, dielectric properties and multiferroic properties were studied. XRD results show that PTO/NFO ceramics have been successfully prepared. When the sintering temperature is 1050 °C, the secondary phase PbO2 exists in the ceramics. SEM results show that there were two kinds of grains with different sizes. EDS analysis suggested that the large particles are NFO and the small particles are PTO. The sintering temperature has a great influence on the dielectric and ferroelectric properties of the prepared ceramics. When the sintering temperature is 950 °C, the ceramics show the maximum dielectric constant at low frequency. When the sintering temperature is 1150 °C, the maximum residual polarization is 1.236 μC/cm2. The conductive mechanism of ceramics is dominated by Space Charge – Limited Conduction. The remanent magnetization and coercive field decrease with increasing the sintering temperature. The magnetoelectric coupling coefficient shows that ceramics have strong magnetoelectric coupling performance (5.7 V/cm·Oe).

In this paper, xPbTiO3/(1-x)NiFe2O4 (PTO/NFO, x = 0.1, 0.2, 0.3, 0.4 and 0.5) ceramic composites were prepared by mixing of sol-gel synthesized PTO and NFO powders, followed by their uniaxial pressing and sintering at 1150 °C. The effects of PTO/NFO composition on crystal structure, surface morphology, electrical, magnetic and magnetoelectric coupling properties were studied. XRD results showed that the prepared ceramics contain only PTO and NFO phases without any impurity phase. The samples were relatively dense while the grains were evenly distributed. The dielectric constant and loss varied monotonically with the frequency for different molar ratios. The hysteresis loop of the composites with high amount of high resistance NFO phase (x = 0.1, 0.2) has the shape characteristic for a material with large leakage current. The samples with x = 0.1 showed the largest saturated magnetization, while the sample with x = 0.2 had the maximum remnant magnetization. It was also shown that the polarization was affected by the external magnetic field of 1 mT and the strongest ME coupling (relative polarization change of 15.4%) was observed for the 0.5PbTiO3/0.5NiFe2O4 composite. In addition, the relevant calculation showed that the leakage mechanism may not be the main factor affecting the polarization of the obtained ceramics.

Pine wilt disease causes huge economic losses to pine wood forestry because of its destructiveness and rapid spread. This paper proposes a detection and location method of pine wood nematode disease at a large scale adopting UAV (Unmanned Aerial Vehicle) remote sensing and artificial intelligence technology. The UAV remote sensing images were enhanced by computer vision tools. A Faster-RCNN (Faster Region Convolutional Neural Networks) deep learning framework based on a RPN (Region Proposal Network) network and the ResNet residual neural network were used to train the pine wilt diseased dead tree detection model. The loss function and the anchors in the RPN of the convolutional neural network were optimized. Finally, the location of pine wood nematode dead tree was conducted, which generated the geographic information on the detection results. The results show that ResNet101 performed better than VGG16 (Visual Geometry Group 16) convolutional neural network. The detection accuracy was improved and reached to about 90% after a series of optimizations to the network, meaning that the optimization methods proposed in this paper are feasible to pine wood nematode dead tree detection.

Over the last two decades, Salmonella1, 4, [5], 12:i:-has been frequently detected in many countries or regions around the world, and nowadays it has become one of the major serotypes responsible for human infectious diarrhea. The etiology and genetic features of Salmonella1, 4, [5], 12:i:-is quite similar with Salmonella Typhimurium (1, 4, [5], 12:i:1, 2) but lacking the second phase flagellar antigen. It has evolved from Salmonella Typhimurium through multiple independent events involving different clonal groups or multiple clones. In this study, we reviews the epidemic characteristics including human infections, animal and animal-derived foods of Salmonella1, 4, [5], 12:i:-, providing support for the prevention and control of infection and scientific researches.

Citrus psyllid is the only insect vector of citrus Huanglongbing (HLB), which is the most destructive disease in the citrus industry. There is no effective treatment for HLB, so detecting citrus psyllids as soon as possible is the key prevention measure for citrus HLB. It is time-consuming and laborious to search for citrus psyllids through artificial patrol, which is inconvenient for the management of citrus orchards. With the development of artificial intelligence technology, a computer vision method instead of the artificial patrol can be adopted for orchard management to reduce the cost and time. The citrus psyllid is small in shape and gray in color, similar to the stem, stump, and withered part of the leaves, leading to difficulty for the traditional target detection algorithm to achieve a good recognition effect. In this work, in order to make the model have good generalization ability under outdoor light condition, a high-definition camera to collect data set of citrus psyllids and citrus fruit flies under natural light condition was used, a method to increase the number of small target pests in citrus based on semantic segmentation algorithm was proposed, and the cascade region-based convolution neural networks (R-CNN) (convolutional neural network) algorithm was improved to enhance the recognition effect of small target pests using multiscale training, combining CBAM attention mechanism with high-resolution feature retention network high-resoultion network (HRNet) as feature extraction network, adding sawtooth atrous spatial pyramid pooling (ASPP) structure to fully extract high-resolution features from different scales, and adding feature pyramid networks (FPN) structure for feature fusion at different scales. To mine difficult samples more deeply, an online hard sample mining strategy was adopted in the process of model sampling. The results show that the improved cascade R-CNN algorithm after training has an average recognition accuracy of 88.78% for citrus psyllids. Compared with VGG16, ResNet50, and other common networks, the improved small target recognition algorithm obtains the highest recognition performance. Experimental results also show that the improved cascade R-CNN algorithm not only performs well in citrus psylla identification but also in other small targets such as citrus fruit flies, which makes it possible and feasible to detect small target pests with a field high-definition camera.

Co0.5Zn0.5Fe2O4/Ba0.8Sr0.2TiO3 (CZFO/BST) composite ceramics with different molar ratios (1:3, 1:2, 1:1, 2:1 and 3:1) were prepared by combining chemical co-precipitation and sol-gel method. Effects of molar ratio on the microstructure, dielectric and multiferroic properties were investigated. The formation of the individual phases and the composites was confirmed by XRD results and small amount of secondary phase (Ba2Fe2Ti4O13) was observed. The grain sizes of magnetic (CZFO) and ferroelectric phase (BST), measured by SEM, were about 5 µm and 0.5 µm, respectively. The sample with molar ratio (1:2) has the largest dielectric constant, while the sample with molar ratio (3:1) shows the lowest dielectric constant. A distinct loss peak can be observed for all the samples. Both the peak position and peak intensity increase with the frequency, indicating relaxation polarization process generated by space charge or interface polarization. The ceramics with molar ratio (1:1) shows the smallest leakage current (∼ 10−7 A/cm at 1.5 kV/cm), while the leakage current (∼ 10−5 A/cm at 1.5 kV/cm) of the sample with molar ratio (3:1) is the largest. The ferroelectric hysteresis loop is not apparent due to the low Curie temperature of the ferroelectric phase, but the sample with molar ratio (2:1) shows the best ferroelectric properties. It was found that with the increase of CZFO content, the values of saturation (Ms) and remnant (Mr) magnetization increase at first and then decrease. The sample with molar ratio (3:1) has the maximum Ms value (about 50.34 emu/g), while the sample with molar ratio (1:2) shows the minimal Mr value (about 0.46 emu/g). This anomalous magnetic property is induced by the interface interaction between the two phases.