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Abstract A combined model was developed using contrast-enhanced CT-based radiomics features and clinical characteristics to predict liver fibrosis stages in patients with chronic liver disease (CLD). We retrospectively analyzed multiphase CT scans and biopsy-confirmed liver fibrosis. 160 CLD patients were randomly divided into 7:3 training/validation ratio. Clinical laboratory indicators associated with liver fibrosis were identified using Spearman's correlation and multivariate logistic regression correlation. Radiomic features were extracted after segmenting the entire liver from multiphase CT images. Feature dimensionality reduction was performed using RF-RFE, LASSO, and mRMR methods. Six radiomics-based models were developed in the training cohort of 112 patients. Internal validation was conducted on 48 randomly assigned patients. Receptor Operating Characteristic (ROC) curves and confusion matrices were constructed to evaluate model performance. The radiomics model exhibited robust performance, with AUC values of 0.810 to 1.000 for significant fibrosis, advanced fibrosis, and cirrhosis. The integrated clinical-radiomics model had superior diagnostic efficacy in the validation cohort, with AUC values of 0.836 to 0.997. Moreover, these models outperformed established biomarkers such as the aspartate aminotransferase to platelet ratio index (APRI) and the fibrosis 4 score (FIB-4), as well as the gamma glutamyl transpeptidase to platelet ratio (GPR), in predicting the fibrotic stages. The clinical-radiomics model holds considerable promise as a non-invasive diagnostic tool for the assessment and staging of liver fibrosis in the patients with CLD, potentially leading to better patient management and outcomes.

Abstract Polygonatum kingianum Collett & Hemsl., is one of the most important traditional Chinese medicines in China. The purpose of this study is to investigate the relationship between herb quality and microbial-soil variables, while also examining the composition and structure of the rhizosphere microbial community in Polygonatum kingianum, the ultimate goal is to provide a scientific approach to enhancing the quality of P. kingianum. Illumina NovaSeq technology unlocks comprehensive genetic variation and biological functionality through high-throughput sequencing. And in this study it was used to analyze the rhizosphere microbial communities in the soils of five P. kingianum planting areas. Conventional techniques were used to measure the organic elements, pH, and organic matter content. The active ingredient content of P. kingianum was identified by High Performance Liquid Chromatography (HPLC) and Colorimetry. A total of 12,715 bacterial and 5487 fungal Operational Taxonomic Units (OTU) were obtained and taxonomically categorized into 81 and 7 different phyla. Proteobacteria, Bacteroidetes, and Acidobacteriae were the dominant bacterial phyla Ascomycota and Basidiomycota were the dominat fungal phyla. The key predictors for bacterial community structure included hydrolysable nitrogen and available potassium, while for altering fungal community structure, soil organic carbon content (OCC), total nitrogen content (TNC), and total potassium content (TPOC) were the main influencing factors. Bryobacter and Candidatus Solibacter may indirectly increase the polysaccharide content of P. kingianum, and can be developed as potential Plant Growth Promoting Rhizobacteria (PGPR). This study has confirmed the differences in the soil and microorganisms of different origins of P. kingianum, and their close association with its active ingredients. And it also broadens the idea of studying the link between plants and microorganisms.

ABSTRACTNeurensin-2 (NRSN2) performs a pro-carcinogenic function in multiple cancers. However, the function of NRSN2 in HPV-infected laryngeal carcinoma (LC) remains unclear. HPV transfection was performed in LC cells. The mRNA and protein levels were monitored using RT-qPCR, immunoblotting, and IF. Cell viability and proliferation were found using the CCK-8 assay and Edu staining. Cell invasion, migration, and apoptosis were probed using the Transwell, wound healing, and flow cytometry, respectively. The autophagosome was observed using TEM. NRSN2 was overexpressed in HPV-transfected LC cells. Inhibition of NRSN2 restrained the autophagy and malignant behavior of HPV-transfected LC cells. Meanwhile, the inhibition of AMPK/ULK1 pathway limited the increased autophagy of HPV-transfected LC cells caused by NRSN2 overexpression. Furthermore, NRSN2 knockdown inhibits autophagy by suppressing AMPK/ULK1 pathway, thereby restraining the malignant behavior of HPV-transfected LC cells. Our research confirmed that HPV transfection increased the autophagy and malignant behavior of LC cells by regulating the NRSN2-mediated activation of the AMPK/ULK1 pathway, offering a new target for cure of LC.

In situ visible–near infrared spectroscopy holds great potential in providing information supporting field applications, decision making and management in soil science, especially combined with the information present in the archived soil spectra. However, soil moisture can drastically affect the reflectance curve and reduce prediction accuracy. The external parameter orthogonalization (EPO) can remove the moisture effect but the effect of elimination is urgently needed to be improved. Herein, firstly, we implemented EPO on 50 bootstrapped calibration datasets, which generated from the local soil spectra library, with 255 combinations of PC1 to PC8, resulting in a total of 50*255 models in PLSR and cubist, respectively. Secondly, we calculated the mean prediction results from these 255 combinations and selected the top 10 validation performance results. Thirdly, we performed correlation analysis on SOM with each segment induced by singular value decomposition on the difference of laboratory and in situ spectra to determine which PC should be removed in the modified EPO. Results revealed that top 10 prediction results with principles without PC2 and the moisture effect was mainly in PC1, and PC2 with significant correlation with SOM were removed from the EPO procedure. EPO with removing PC2 (namely Modified EPO) improved the correlation of SOM with some optional bands that directly and indirectly were associated with SOM to improve the SOM prediction accuracy. Modified EPO improved the accuracy in predicting SOM with increased R2 (9 %–44 % and 7 %–17 %) and root mean square error (1 %–9 % and 63 %–68 %) in the PLSR and Cubist model, respectively. Our study highlights the advantage of modified EPO in improving the elimination efficiency of water in spectra, and of PC analysis biplots in approximating the removed PCs.

Thiacloprid is one of the first generation of neonicotinoid insecticide with a chloropyridine structure like imidacloprid and acetamiprid. Recent studies have revealed its environmental and non-target organism toxicity, leading to restrictions on its use in many countries and regions. Despite limitations, thiacloprid has been detected in various environmental samples, food sources, and biological specimens, posing a significant threat to human health, necessitating advanced detection methods for monitoring. In this study, a highly specific monoclonal antibody against thiacloprid via a multi-immunogen strategy was prepared and a rapid and sensitive enzyme-linked immunosorbent assay for the detection of thiacloprid residues in honey and medicinal herbs was established. The half maximal inhibitory concentration (IC50) of this method was 0.38 ng/mL, improving the sensitivity by 1.2−480.6 times compared to existing reports, and the limit of detection (IC20) was 0.097 ng/mL. The method was successfully applied to the determination of thiacloprid residues in honey and medicinal herbs (Crataegi fructus, Citri reticulatae pericarpium), achieving recovery rates ranging from 87.50 % to 116.11 %. The obtained results were verified using the LC-MS/MS method. The multi-immunogen strategy proposed in this study provides an approach for the preparation of highly sensitive and specific monoclonal antibodies, and immunoassay established based on it has good application prospects in complex matrices.

Abstract Veronica anagallis-aquatica L. and Veronica undulata Wall. are widely used ethnomedicinal plants in China. The two species have different clinical efficacies, while their extremely similar morphology and unclear interspecific relationship make it difficult to accurately identify them, leading to increased instances of mixed usage. This article reports on the complete chloroplast genomes sequence of these two species and their related Veronica species to conduct a comparative genomics analysis and phylogenetic construction. The results showed that the chloroplast (cp) genomes of Veronica exhibited typical circular quadripartite structures, with total lengths of 149,386 to 152,319 base pairs (bp), and GC content of 37.9 to 38.1%, and the number of genes was between 129–134. The total number of simple sequence repeats (SSRs) in V. anagallis-aquatica and V. undulata is 37 and 36, while V. arvensis had the highest total number of 56, predominantly characterized by A/T single bases. The vast majority of long repeat sequence types are forward repeats and palindromic repeats. Selective Ka/Ks values showed that three genes were under positive selection. Sequence differences often occur in the non-coding regions of the large single-copy region (LSC) and small single-copy region (SSC), with the lowest sequence variation in the inverted repeat regions (IR). Seven highly variable regions (trnT-GGU-psbD, rps8-rpl16, trnQ-UUG, trnN-GUU-ndhF, petL, ycf3, and ycf1) were detected, which may be potential molecular markers for identifying V. anagallis-aquatica and V. undulata. The phylogenetic tree indicates that there is a close genetic relationship between the genera Veronica and Neopicrorhiza, and V. anagallis-aquatica and V. undulata are sister groups. The molecular clock analysis results indicate that the divergence time of Veronica may occur at ∼ 9.09 Ma, and the divergence time of these two species occurs at ∼ 0.48 Ma. It is speculated that climate change may be the cause of Veronica species diversity and promote the radiation of the genus. The chloroplast genome data of nine Veronica specie provides important insights into the characteristics and evolution of the chloroplast genome of this genus, as well as the phylogenetic relationships of the genus Veronica.

Abstract Background Multiple species of the genera Cytauxzoon and Hepatozoon can infect wild felines, but the diversity of these and other apicomplexan parasites in Eurasian lynx is scarcely known. The aim of this study was to detect Cytauxzoon and Hepatozoon species with molecular methods in Eurasian lynxes and their ticks in northwestern China. Methods DNA was extracted from the heart, liver, spleen, lung, and kidney samples of three Eurasian lynxes as well as from their five ixodid ticks. These DNA samples were screened with polymerase chain reactions (PCRs) for Cytauxzoon with the partial cytochrome b gene (CytB), cytochrome c oxidase subunit I gene (COI), and small subunit ribosomal RNA gene (18S rRNA), and Hepatozoon with three different fragments of small subunit ribosomal RNA gene (18S rRNA). PCR products were sequenced, aligned, and phylogenetically analyzed. Results One adult female of Eurasian lynx (#1, adult female) was co-infected with Cytauxzoon manul and Hepatozoon felis genotype I, while an adult male lynx (#2) was infected with C. manul. Interestingly, H. felis genotype I was both detected in a male cub (#3) and two out of five infesting Hyalomma asiaticum ticks. Conclusions For the first time, Cytauxzoon manul is reported here from Eurasian lynx. In addition, H. felis has not been known to occur in this host species in China and Central Asia. Thus, the findings of this study extend our knowledge on the geographical distribution and host range of these haemoprotozoan parasites. Moreover, this is also the first evidence of C. manul and H. felis co-infection in Eurasian lynx. Graphical Abstract

Background This study comprehensively investigates the association between the expression of nicotinamide N-methyltransferase (NNMT) and clinical outcomes of urothelial bladder cancer (UBC), as well as the molecular mechanisms by which NNMT in cancer-associated fibroblast (CAF) modulates tumor progression and immunotherapy resistance in UBC.Methods Single-cell transcriptomic analyses, immunohistochemical and immunofluorescence assays were performed on bladder cancer samples to validate the relationship between NNMT expression and clinical outcomes. A series of experiments, including chromatin immunoprecipitation assay, liquid chromatography tandem mass spectrometry assay, and CRISPR‒Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats and CRISPR-associated protein 9) knockout, together with in vivo models, have been established to determine the molecular functions of NNMT in CAFs in UBC.Results We demonstrated that elevated expression of the nicotinamide adenine dinucleotide (NAD+) metabolism enzyme NNMT in CAFs (NNMT+ CAFs) was significantly associated with non-response to programmed death-ligand 1 (PD-L1) blockade immunotherapy in patients with UBC and predicted the unfavorable prognosis of UBC in two independent large cohorts. Targeting NNMT using the inhibitor 5-Amino-1-methylquinolinium iodide significantly reduced tumor growth and enhanced the apoptotic effects of the anti-PD-L1 antibody in UBC mouse models. Mechanistically, NNMT+ CAFs recruit tumor-associated macrophages via epigenetic reprogramming of serum amyloid A (SAA) to drive tumor cell proliferation and confer resistance to programmed death-1/PD-L1 blockade immunotherapy.Conclusions NNMT+ CAFs were significantly associated with non-response to PD-L1 blockade immunotherapy in patients with UBC. Elevated NNMT, specifically in CAFs, upregulates SAA expression and enhances the recruitment and differentiation of macrophages in the tumor microenvironment, thereby directly or indirectly promoting tumor progression and conferring resistance to immunotherapies in bladder cancer.

Zearalenone (ZEN) is a mycotoxin that extensively contaminates food and feed, posing a significant threat to public health. However, the mechanisms behind ZEN-induced intestinal immunotoxicity remain unclear. In this study, Sprague-Dawley (SD) rats were exposed to ZEN at a dosage of 5 mg/kg/day b.w. for a duration of 14 days. The results demonstrated that ZEN exposure led to notable pathological alterations and immunosuppression within the intestine. Furthermore, ZEN exposure caused a significant reduction in the levels of apolipoprotein E (ApoE) and liver X receptor (LXR) (P

Abstract Background In the family Trypanosomatidae, the genus Trypanosoma contains protozoan parasites that infect a diverse range of hosts, including humans, domestic animals, and wildlife. Wild rodents, as natural reservoir hosts of various pathogens, play an important role in the evolution and emergence of Trypanosomatidae. To date, no reports are available on the trypanosomatid infection of pikas (Lagomorpha: Ochotonidae). Methods In this study, Mongolian pikas and their fleas were sampled at the China–Mongolia border, northwestern China. The samples were analyzed with polymerase chain reaction (PCR) and sequencing for the presence of Trypanosomatidae on the basis of both the 18S ribosomal RNA (18S rRNA) gene and the glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene. The morphology of trypomastigotes was also observed in peripheral blood smears by microscopy. Results Molecular and phylogenetic analyses revealed a new genotype of the Trypanosoma lewisi clade that was found both in pika blood and flea samples. This genotype, which probably represents a new species, was provisionally designated as “Trypanosoma sp. pika”. In addition, a novel genotype belonging to the genus Blechomonas of Trypanosomatidae was detected in fleas. On the basis of its molecular and phylogenetic properties, this genotype was named Blechomonas luni-like, because it was shown to be the closest related to B. luni compared with other flea-associated trypanosomatids. Conclusions To the best of our knowledge, this is the first study to report any trypanosomatid species in Mongolian pikas and their fleas. Further studies are needed to investigate the epidemiology of these protozoan parasites, as well as to evaluate their pathogenicity for humans or domestic animals. Graphical Abstract

Abstract Background Rodents play an important role in the life cycle of ixodid and argasid ticks, particularly as hosts of larvae and nymphs. The great gerbil (Rhombomys opimus), the preferred prey item of several carnivores (e.g. the red fox and marbled polecat), is the dominant rodent species in the Gurbantunggut Desert in northwestern China. The aim of this study was to investigate tick species associated with different hosts in the habitat of great gerbils, including wildlife and livestock. Methods During 2018–2023, ticks were removed from 326 great gerbils, two red foxes (Vulpes vulpes), three marbled polecats (Vormela peregusna), 35 pastured sheep (Ovis aries), and one long-eared desert hedgehog (Hemiechinus auritus) in the Gurbantunggut Desert. Ticks were identified according to standard morphological keys. Then, they were further analyzed by molecular and phylogenic methods based on two mitochondrial markers, 16S rDNA and cytochrome c oxidase subunit I (COI) genes. Results A total of 889 ticks were collected, representing five species. These included Hyalomma asiaticum (n = 425: 24 larvae, 79 nymphs and 322 adults), Rhipicephalus turanicus (n = 153: 2 nymphs and 151 adults), Haemaphysalis erinacei (n = 298: 4 larvae, 7 nymphs and 287 adults), Ixodes acuminatus (n = 7: 4 nymphs and 3 adults) and Ornithodoros tartakovskyi (6 adults). Based on COI sequences, molecular and phylogenetic analyses showed that (i) I. acuminatus from great gerbils and marbled polecats clustered with I. acuminatus reported from Europe; (ii) O. tartakovskyi found in northwestern China belonged to an independent clade; (iii) Hy. asiaticum, R. turanicus and Ha. erinacei had 100% sequence identities to conspecific ticks sampled previously in China. Conclusions The great gerbil is an important host for the developmental stages of I. acuminatus, O. tartakovskyi, Ha. erinacei, Hy. asiaticum and R. turanicus, thus supporting the life cycle of several tick species which, as adults, parasitize predators (red fox and marble polecat) as well as pastured sheep and hedgehogs in the Gurbantunggut Desert. Ixodes acuminatus and O. tartakovskyi were found for the first time on great gerbil and marbled polecat, respectively. Graphical Abstract

Abstract Background Scutellaria baicalensis Georgi has been extensively used as a medicinal herb in China for over 2000 years. They may be intentionally or inadvertently substituted or blended with comparable species in the local market, threatening clinical medication safety. Molecular markers are effective tools to prevent misidentification and eliminate doping and falsification among Scutellaria plants. This study screened four highly variable regions to identify Scutellaria and its adulterants. In addition, a phylogenetic analysis was performed using the complete cp genome combined with published Scutellaria species samples. Moreover, a comparative analysis of the cp genomes was conducted to investigate the cp genome evolution of S. baicalensis. Results The complete cp genome of five species of Scutellaria was sequenced for the first time, and four previously published Scutellaria species were re-sequenced. They all exhibited a conserved quadripartite structure in their cp genomes, including two distinct regions, namely a small and large single copy region, respectively, and two inverted repeats encompassing the majority of ribosomal RNA genes. Furthermore, the nine species exhibited high conservation from aspects of the genome structure, codon usage, repeat sequences, and gene content. Four highly variable regions (matK-rps16, ndhC-trnV-UAC, psbE-petL, and rps16-trnQ-UUG) may function as potential molecular markers for differentiating S. baicalensis from its adulterants. Additionally, the monophyly of Scutellaria was ascertained and could be reclassified into two subgenera, subgenus Anaspis and subgenus Scutellaria, as evidenced by the phylogenetic analyses on sequences of cp genome and shared protein-coding sequences. According to the molecular clock analysis, it has been inferred that the divergence of Scutellaria occurred at approximately 4.0 Mya during the Pliocene Epoch. Conclusion Our study provides an invaluable theoretical basis for further Scutellaria species identification, phylogenetics, and evolution analysis.

Abstract Background The Aconitum genus is a crucial member of the Ranunculaceae family. There are 350 Aconitum species worldwide, with about 170 species found in China. These species are known for their various pharmacological effects and are commonly used to treat joint pain, cold abdominal pain, and other ailments. Codon usage bias (CUB) analysis contributes to evolutionary relationships and phylogeny. Based on protein-coding sequences (PCGs), we selected 48 species of Aconitum for CUB analysis. Results The results revealed that Aconitum species had less than 50% GC content. Furthermore, the distribution of GC content was irregular and followed a trend of GC1 > GC2 > GC3, indicating a bias towards A/T bases. The relative synonymous codon usage (RSCU) heat map revealed the presence of conservative codons with slight variations within the genus. The effective number of codons (ENC)-Plot and the parity rule 2 (PR2)-bias plot analysis indicate that natural selection is the primary factor influencing the variation in codon usage. As a result, we screened various optimal codons and found that A/T bases were preferred as the last codon. Furthermore, our Maximum Likelihood (ML) analysis based on PCGs among 48 Aconitum species yielded results consistent with those obtained from complete chloroplast (cp.) genome data. This suggests that analyzing mutation in PCGs is an efficient method for demonstrating the phylogeny of species at the genus level. Conclusions The CUB analysis of 48 species of Aconitum was mainly influenced by natural selection. This study reveals the CUB pattern of Aconitum and lays the foundation for future genetic modification and phylogenetic analyses.

Introduction: Zearalenone-14-glucoside (Z14G) is a modified mycotoxin that widely contaminates food across the world. Our preliminary experiment showed that Z14G degrades to zearalenone (ZEN) in the intestine exerting toxicity. Notably, oral administration of Z14G in rats induces intestinal nodular lymphatic hyperplasia. Objectives: To investigate the mechanism of Z14G intestinal toxicity and how it differs from ZEN toxicity. We conducted a precise toxicology study on the intestine of rats exposed to Z14G and ZEN using multi-omics technology. Methods: Rats were exposed to ZEN (5 mg/kg), Z14G-L (5 mg/kg), Z14G-H (10 mg/kg), and pseudo germ free (PGF)-Z14G-H (10 mg/kg) for 14 days. Histopathological studies were performed on intestines from each group and compared. Metagenomic, metabolomic, and proteomic analyses were performed on rat feces, serum, and intestines, respectively. Results: Histopathological studies showed that Z14G exposure resulted in dysplasia of gut-associated lymphoid tissue (GALT) compared to ZEN exposure. The elimination of gut microbes in the PGF-Z14G-H group alleviated or eliminated Z14G-induced intestinal toxicity and GALT dysplasia. Metagenomic analysis revealed that Z14G exposure significantly promoted the proliferation of Bifidobacterium and Bacteroides compared to ZEN. Metabolomic analysis showed that Z14G exposure significantly reduced bile acid, while proteomic analysis found that Z14G exposure significantly reduced the expression of C-type lectins compared to ZEN. Conclusions: Our experimental results and previous research suggest that Z14G is hydrolyzed to ZEN by Bifidobacterium and Bacteroides promoting their co-trophic proliferation. This leads to inactivation of lectins by hyperproliferative Bacteroides when ZEN caused intestinal involvement, resulting in abnormal lymphocyte homing and ultimately GALT dysplasia. It is noteworthy that Z14G is a promising model drug to establish rat models of intestinal nodular lymphatic hyperplasia (INLH), which is of great significance for studying the pathogenesis, drug screening and clinical application of INLH.

The Ili River Basin (IRB) is located in the northwest of China. With its large-scale zone and abundant resources, it is believed to be a “wet island” and a biotic resource storehouse in the dry regions of Eurasia. The IRB has stable ecological conditions and abundant water resources, providing natural conditions for agricultural production and human settlements. With the population increasing and economic development advancing, the competition for land resources is becoming fierce, leading to some ecological problems in this region. Therefore, understanding the spatiotemporal changes and driving mechanisms of the “three-zone space” (TZS) in the IRB is of significant practical importance for promoting sustainable development and optimizing the territorial spatial pattern. This study first analyzes the characteristics and intensity of the TZS changes from 2000 to 2020. Then, it utilizes the optimized parameter Geodetector (OPGD) to analyze the driving mechanisms behind these changes. The results show the following. Firstly, the agricultural space (AS) increased by a total of 837.5 km2, the urban space (US) increased by 519.64 km2, and other ecological space (OES) increased by 1518.83 km2. Green ecological space (GES) decreased by 2875.97 km2. Secondly, intensity analysis indicated that the total TZS change in IRB was 11.07%. At the spatial-type level, the increased intensities of OES, US, and AS were active. In spatial transformation intensity, US and OES tended to transform into AS; AS tended to transform into US; and OES and GES had a mutual transformation tendency. Thirdly, AS converted into US around emerging cities like Khorgas and Cocodala. The conversion towards GES was scattered. The mutual conversion between OES and GES showed spatial distribution consistency, mainly occurring in the Borohoro ranges and the Halik ranges. Lastly, regarding the driving mechanisms, the evolution of US in the IRB was driven by social and economic factors. Location and climate factors accelerated agricultural development, facilitating the transformation of GES and OES into AS. Climate and economic factors played a crucial role in the scale of conversions between OES and GES. The findings can provide a basis for the governance and protection of the IRB, help to form a rational territorial spatial pattern, and offer scientific guidance for sustainable land management.

Abstract Background In the new International League Against Epilepsy (ILAE) classification of seizure types, generalized seizures such as absence seizures (ASs) may originate from a focal point and rapidly spread to the bilaterally distributed brain network. Increasing evidence from animal and clinical studies has indicated that focal changes may occur prior to ASs; however, the relationship of ASs with epileptogenic lesions remains unclear. Methods We retrospectively collected and analyzed the clinical, imaging, and electrophysiological data of 16 patients who had ASs and structural lesions with seizure-free outcomes after lesion resection. Results In semiology analysis, nine patients displayed focal onset; only two patients showed simple ASs, and seizure types other than ASs were observed in the remaining patients. On ictal electroencephalography (EEG), four patients showed bilateral synchronous symmetric 3 Hz generalized spike-wave discharges (GSWDs), and the remaining patients showed bilateral 1.5–2.5 Hz GSWDs. Moreover, most patients (13/16, 81.3%) exhibited focal features in addition to ASs, while interictal EEG was the same in 12 patients. Furthermore, on stereoelectroencephalogram (SEEG), 2/5 patients showed focal discharges before bilateral burst GSWDs. Additionally, all patients had structural lesions on imaging. In four typical AS patients, the lesions were located in deep brain regions. Notably, in 9 patients (9/16, 56%), the lesions were located in the posterior cortex. All patients underwent lesion resection and had seizure-free outcomes during follow-up, and intelligence quotient (IQ) also improved by 10.71 ± 3.90 one year after surgery. Conclusions Patients with lesion-related epilepsy may present with ASs that have a focal onset and are associated with good surgical outcomes.

Abstract Urothelial bladder cancer (UBC) is one of the most prevalent malignancies worldwide, with striking tumor heterogeneity. Elucidating the molecular mechanisms that can be exploited for the treatment of aggressive UBC is a particularly relevant goal. Protein ubiquitination is a critical post-translational modification (PTM) that mediates the degradation of target protein via the proteasome. However, the roles of aberrant protein ubiquitination in UBC development and the underlying mechanisms by which it drives tumor progression remain unclear. In this study, taking advantage of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein (Cas) 9 technology, we identified the ubiquitin E3 ligase ANAPC11, a critical subunit of the anaphase-promoting complex/cyclosome (APC/C), as a potential oncogenic molecule in UBC cells. Our clinical analysis showed that elevated expression of ANAPC11 was significantly correlated with high T stage, positive lymph node (LN) metastasis, and poor outcomes in UBC patients. By employing a series of in vitro experiments, we demonstrated that ANAPC11 enhanced the proliferation and invasiveness of UBC cells, while knockout of ANAPC11 inhibited the growth and LN metastasis of UBC cells in vivo. By conducting immunoprecipitation coupled with mass spectrometry, we confirmed that ANAPC11 increased the ubiquitination level of the Forkhead transcription factor FOXO3. The resulting decrease in FOXO3 protein stability led to the downregulation of the cell cycle regulator p21 and decreased expression of GULP1, a downstream effector of androgen receptor signaling. Taken together, these findings indicated that ANAPC11 plays an oncogenic role in UBC by modulating FOXO3 protein degradation. The ANAPC11–FOXO3 regulatory axis might serve as a novel therapeutic target for UBC.

Abstract The Santos Basin, located in the southeastern waters of Brazil, is a passive continental margin basin with the most abundant deepwater petroleum resources in the world discovered to date. However, few studies have been conducted on the present geothermal fields of the Santos Basin, which severely restricts the oil and gas resource evaluation of the basin. This study first utilizes 35 temperature data from 16 post-salt drilling wells and 370 temperature data from 31 pre-salt drilling wells to calculate the post-salt and pre-salt geothermal gradients and terrestrial heat flows in the Santos Basin. Then, the basin simulation software BasinMod 1D is used to quantitatively evaluate the impacts of salt rock sedimentation on the present geothermal fields and the maturity of pre-salt hydrocarbon source rocks. The results demonstrate that the present post-salt geothermal gradient in the Santos Basin is 2.20–3.97 °C/100 m, with an average value of 2.99 °C /100 m, and the post-salt terrestrial heat flow is 54.00–97.32 mW/m2, with an average value of 73.36 mW/m2, while the present pre-salt geothermal gradient is 2.21–2.95 °C/100 m, with an average value of 2.53 °C/100 m, and the pre-salt terrestrial heat flow is 61.85–82.59 mW/m2, with an average value of 70.69 mW/m2. These values are characteristic of a low-temperature geothermal field in a zone with a stable structure. The sedimentation of the salt rock causes a decrease in the temperature of the pre-salt strata, which inhibits pre-salt hydrocarbon source rock maturity, with an inhibition rate of up to 1.32%. The inhibition degree decreases with increasing salt rock thickness. At the same time, the salt rock thickness is positively correlated with the present surface heat flow. The unique distribution of the salt rock and related salt structures lead to present terrestrial heat flow differences among different structural units in the basin. This study is of great significance for evaluating and exploring the pre-salt oil and gas resources in the Santos Basin.

Abstract Background Programmed cell death‐1/ligand 1 inhibitors are a new treatment strategy for advanced urothelial carcinoma. Therefore, a comparative evaluation of their efficacy and toxicity compared with chemotherapy is necessary. Methods We comprehensively searched PubMed, Web of Science, Embase, and Cochrane Library databases and performed a meta‐analysis of randomized controlled trials up to July 2021. We considered overall survival as the primary outcome, and progression‐free survival, objective response rate, and treatment‐related adverse events as secondary outcomes. Results Overall, 3584 patients from five studies were evaluated. Compared with first‐line chemotherapy, programmed cell death‐1/ligand 1 inhibitors were significantly associated with worse progression‐free survival (p

Infections with spotted fever group rickettsiae represent a worldwide health problem, characterized by persistent high fever, headache, and rash in humans, domestic animals, and wildlife. To date, the occurrence of Rickettsia species in hard ticks has not been thoroughly studied, especially in eastern and southern Kazakhstan. A total of 1,245 adult ticks, comprising 734 Dermacentor marginatus, 219 Hyalomma scupense, 144 Hyalomma asiaticum, 84 Hyalomma marginatum, 48 Rhipicephalus turanicus, and 16 Haemaphysalis erinacei, collected from East Kazakhstan, Abay, Jetsu, Almaty, Jambyl, South Kazakhstan and Qyzylorda oblasts of Kazakhstan, were used to screen rickettsial agents using molecular methods. Rickettsia raoultii, Rickettsia slovaca, Rickettsia aeschlimannii and Rickettsia heilongjiangensis were identified using sequencing, and 31.5% (392/1245) of ticks carried rickettsial agents. The difference in the natural landscapes explains the variety of the collected ticks and expands our knowledge of Rickettsia species and their geographical distribution in Kazakhstan. To the best of our knowledge, this study reports the first finding of R. heilongjiangensis in Kazakhstan.

Abstract The development of geothermal resources in the Dongpu Depression can improve not only the economic benefits of the oilfield but also the ecological environment. Therefore, it is necessary to evaluate the geothermal resources in the region. Based on the heat flow, geothermal gradient and thermal properties, the temperatures and their distribution in different strata are calculated using geothermal methods, and the geothermal resource types of the Dongpu Depression are identified. The results show that the geothermal resources include low-temperature, medium-temperature and high-temperature geothermal resources in the Dongpu Depression. The Minghuazhen and Guantao Formations mainly include low-temperature and medium-temperature geothermal resources; the Dongying and Shahejie Formations include low-temperature, medium-temperature and high-temperature geothermal resources; the Ordovician rocks mainly include medium-temperature and high-temperature geothermal resources. The Minghuazhen, Guantao and Dongying Formations can form good geothermal reservoirs and are favorable layers for exploring low-temperature and medium-temperature geothermal resources. The geothermal reservoir of the Shahejie Formation is relatively poor, and the thermal reservoirs may be developed in the western slope zone and the central uplift. The Ordovician carbonate strata can provide thermal reservoirs for geothermal resources, and the Cenozoic bottom temperature is more than 150 °C except for most of the western gentle slope zone. In addition, for the same stratum, the geothermal temperatures in the southern Dongpu Depression are higher than those in the northern depression.

Abstract Background Tropical water lily is an aquatic plant with high ornamental value, but it cannot overwinter naturally at high latitudes. The temperature drop has become a key factor restricting the development and promotion of the industry. Results The responses of Nymphaea lotus and Nymphaea rubra to cold stress were analyzed from the perspective of physiology and transcriptomics. Under the cold stress, Nymphaea rubra had obvious leaf edge curling and chlorosis. The degree of peroxidation of its membrane was higher than that of Nymphaea lotus, and the content of photosynthetic pigments also decreased more than that of Nymphaea lotus. The soluble sugar content, SOD enzyme activity and CAT enzyme activity of Nymphaea lotus were higher than those of Nymphaea rubra. This indicated that there were significant differences in the cold sensitivity of the two varieties. GO enrichment and KEGG pathway analysis showed that many stress response genes and pathways were affected and enriched to varying degrees under the cold stress, especially plant hormone signal transduction, metabolic pathways and some transcription factor genes were from ZAT gene family or WKRY gene family. The key transcription factor ZAT12 protein in the cold stress response process has a C2H2 conserved domain, and the protein is localized in the nucleus. Under the cold stress, overexpression of the NlZAT12 gene in Arabidopsis thaliana increased the expression of some cold-responsive protein genes. The content of reactive oxygen species and MDA in transgenic Arabidopsis thaliana was lower, and the content of soluble sugar was higher, indicating that overexpression of NlZAT12 can improve the cold tolerance of Arabidopsis thaliana. Conclusion We demonstrate that ethylene signalling and reactive oxygen species signalling play critical roles in the response of the two cultivars to cold stress. The key gene NlZAT12 for improving cold tolerance was identified. Our study provides a theoretical basis for revealing the molecular mechanism of tropical water lily in response to cold stress.

Abstract This study investigated the degradation characteristics of different biodegradable film and its effects on soil nutrients in tillage layer, growth and development of taro and yield formation. Field experiment with biodegradable films, including poly-(butylene adipate-co-butylene terephthalate) PBAT, (poly-carbon dioxide) PCO2, (poly propylene carbonate) PPC, as well as common mulch film (CK1) and uncovered mulch film (CK2) were conducted on Longxiang taro in 2020 and 2021 respectively. The degradation rate of the three biodegradable films was PBAT > PPC > PCO2. Compared with CK1, the alkali-hydrolyzed N of PBAT at the growth stage and fruiting stage significantly increased in 2020 and 2021, respectively (both, P

In this work, a green analytical method was established for the simultaneous extraction and detection of 20 analytes–10 neonicotinoid insecticides and their 10 major toxic metabolites in edible herbs. QuEChERS and LC-MS/MS were used to analyze the 20 analytes in five edible herbs. The residues of the 20 neonicotinoid insecticides and their metabolites in 109 herbal samples were detected, of which 90 samples were positive, and the residue of total neonicotinoid insecticides ranged from 0.26 to 139.28 μg/kg. Acetamiprid (77.06 %, ≤85.95 μg/kg), imidacloprid (67.89 %, ≤32.49 μg/kg) and their metabolites (N-desmethyl-acetamiprid (44.04 %, ≤18.42 μg/kg) and desnitro imidacloprid (48.62 %, ≤16.55 μg/kg) were most frequently detected in herbs. Significant positive correlations were found between imidacloprid/acetamiprid and their metabolites in Lycii fructus and Citri reticulatae pericarpium. Therefore, more attention may be given to the neonicotinoid insecticide residues in edible herbs in the future.

Abstract Background Laryngeal masks airway (LMA) has been increasingly used in surgical patients. However, the use of LMA in laparoscopic surgeries remains controversial. The major concerns include the potential risk of esophageal regurgitation, aspiration, and difficulties to achieve effective ventilation. The aim of this study was to evaluate the safety and effectiveness of the LMA® Protector™ in patients undergoing laparoscopic surgery. Methods Patients aged 18 to 70 years, scheduled for laparoscopic surgeries were included. The insertion time, successful insertion rate, and oropharyngeal leak pressure were measured. Airway complications and airway manipulations during the procedure were documented. Effective ventilation rate was calculated. Visible bloodstains and reflux content in the drainage channel were documented after the removal of LMA® Protector™. Results Three hundred patients were enrolled. The insertion of LMA® Protector™ failed in seven patients resulting with a successful insertion rate of 97.7%. During the maintenance of anesthesia, airway manipulation was required in 19 patients (19/293, 6.48%), in three of whom the LMA was replaced with endotracheal intubation resulting with an effective ventilation rate of 96.7% (290/300). The oropharyngeal leak pressure was 30.18 ± 5.88 cmH2O. Seventy-five patients (25.86%) reported mild sore throat on the first day after surgery. Bloodstains on study devices were noticed in 58 patients (20%). Seventy-five patients (25.86%) reported mild sore throat on the first day after surgery. Gastric reflux was noticed in the drainage tube in 5 patients (1.72%) with no signs of aspiration in any of those patients. Conclusions The LMA® Protector™ was shown to be safe and effective in patients undergoing laparoscopic surgeries. Although minor complications that require no further treatment, no clinically diagnosed aspiration was noticed in our study. Gastric reflux was noticed in the drainage tube in five patients undergoing laparoscopic gynecology surgery. Further research is needed to verify whether LMA® Protector™ is suitable for procedures in Trendelenburg position or other situations that a high risk of gastroesophageal reflux exists. Trial registration The trial was registered at the Chinese Clinical Trial Registry ( ChiCTR1800018300 , date of registration: September 2018).

We aim to detect the presence of Tacheng tick virus 2 (TcTV-2) in ticks of southeastern Kazakhstan. A total of 205 ticks were collected and separated into four species, namely Hyalomma scupense, Dermacentor marginatus, Hyalomma asiaticum and Hyalomma anatolicum. The partial S segment of TcTV-2 was detected in individual RNA of separated ticks by reverse transcriptase polymerase chain reaction. 11.22% (23/205) of ticks were positive to the viral S segment. This is first report of presence of the TcTV-2 in Hy. scupense and Hy. anatolicum from Kazakhstan. This finding extends tick species and the geographic distribution of TcTV-2.

In recent years, the residues of neonicotinoid insecticide in food and environmental samples have attracted extensive attention. Neonicotinoids have many adverse effects on human health, such as cancer, chronic disease, birth defects, and infertility. They have substantial toxicity to some non-target organisms (especially bees). Hence, monitoring the residues of neonicotinoid insecticides in foodstuffs is necessary to guarantee public health and ecological stability. This review aims to summarize and assess the metabolic features, residue status, sample pretreatment methods (solid-phase extraction (SPE), Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS), and some novel pretreatment methods), and detection methods (instrument detection, immunoassay, and some innovative detection methods) for neonicotinoid insecticide residues in food and environmental samples. This review provides detailed references and discussion for the analysis of neonicotinoid insecticide residues, which can effectively promote the establishment of innovative detection methods for neonicotinoid insecticide residues.

Zearalenone (ZEN), a prevalent mycotoxin contaminating food and known for its intestinal toxicity, has been suggested as a potential risk factor for inflammatory bowel disease (IBD), although the exact relationship between ZEN exposure and IBD remains unclear. In this study, we established a rat model of colon toxicity induced by ZEN exposure to investigate the key targets of ZEN-induced colon toxicity and explore the underlying connection between ZEN exposure and IBD. Histological staining of the rat colon revealed significant pathological changes resulting from ZEN exposure (p < 0.01). Furthermore, the proteomic analysis demonstrated a notable upregulation of protein expression levels, specifically STAT2 (0.12 ± 0.0186), STAT6 (0.36 ± 0.0475) and ISG15 (0.43 ± 0.0226) in the rat colon (p < 0.05). Utilizing bioinformatics analysis, we combined ZEN exposure and IBD clinical sample databases to reveal that ZEN exposure may increase the risk of IBD through activation of the STAT-ISG15 pathway. This study identified novel targets for ZEN-induced intestinal toxicity, providing the basis for further study of ZEN exposure to IBD.

As a precious soil resource, black soils in Northeast China are currently facing severe land degradation. Visible and near-infrared spectroscopy (vis-NIR, 350–2500 nm) and mid-infrared spectroscopy (MIR, 2500–25,000 nm) have shown great potential to predict soil properties. However, there is still limited research on using MIR in situ. The aim of this study was to explore the feasibility of in situ MIR for the prediction of soil total nitrogen (TN) and total phosphorus (TP) and to compare its performance with the use of laboratory MIR, as well as the use of in situ and laboratory vis-NIR. A total of 450 samples from 90 soil profiles, along with their in situ and laboratory spectra of MIR and vis-NIR, were collected in a field with ten different tillage and management practices in a typical black soil area of Northeast China. Partial least square regression (PLSR), random forest (RF) and multivariate adaptive regression splines (MARS) were used to generate the calibrations between the spectra and the two properties. The results showed that both MIR and vis-NIR were able to predict the TN whether in laboratory or in situ conditions, but neither of them could predict the TP quantitatively since there was no sensitive band on both spectra regarding the TP. The prediction accuracy of the TN with laboratory spectra was higher than that with in situ spectra, for both vis-NIR and MIR. The optimal prediction accuracy of the TN with laboratory MIR (RMSE = 0.11 g/kg, RPD = 3.12) was higher than that of laboratory vis-NIR (RMSE = 0.14 g/kg, RPD = 2.45). The optimal prediction accuracy of in situ MIR (RMSE = 0.20 g/kg, RPD = 1.80) was lower than that of in situ vis-NIR (RMSE = 0.16 g/kg, RPD = 2.14). The prediction performance of the spectra followed laboratory MIR > laboratory vis-NIR > in situ vis-NIR > in situ MIR. The performance of in situ MIR was relatively poor, mainly due to the fact that MIR was more influenced by soil moisture. This study verified the feasibility of in situ MIR for soil property prediction and provided an approach for obtaining rapid soil information and a reference for soil research and management in black soil areas.

We used metagenomic analysis to identify Tacheng tick virus 2 infection in a patient with a history of tick bite in northwestern China. We confirmed the virus with reverse transcription-PCR, virus isolation, and genomic analysis. We detected viral RNA in 9.6% of ticks collected from the same region.

Environmental pollution and medicine safety have aroused increasing public concerns due to human health. Amongst various contaminants, mercury is of special attention owing to their environmental persistence and biogeochemical recycling and ecological risks. Herein, a simple and highly parallel electrochemical biosensor for Hg determination was designed and investigated. The proposed biosensor was prepared and compared between (1) DTT/MB-DNA/Au with configuration occupation approach and (2) MCH/MB-DNA/Au with passivation approach. According to the combined results of scanning electrochemical microscope (SECM) and Randles-Sevcik equation, the DTT modified electrode exhibited high uniformity on DNA distribution and superb stability on electron transfer in Hg2+ detection. Evidentially, the response value of proposed DTT/MB-DNA/Au was increased from 57.518% to 97.607%, while RSD% between duplicate runs had dropped from 22.658% to 0.223% (n = 3). Moreover, the increased proportion of effective working area was 467.380% compared with general sensors. Besides, DTT concentration, DNA concentration as well as assembly time were optimized, utilizing electrochemical impedance spectroscopy (EIS), Cyclic Voltammetry (CV) and Square Wave Anode Stripping Voltammetry (SWASV). This optimized biosensor exhibited an excellent selectivity toward Hg2+ over Cu2+, As2+, Cd2+, Pb2+, Cr3+, Ni2+ and Zn2+ etc., and the stability of DTT/MB-DNA/Au were at least two times better even after 3 days under room temperature. Also, a linear relation was observed between the peak current and Hg2+concentrations in a range from 0.25 nM to 2.00 μM with a detection limit of 53.00 pM under optimal conditions. Finally, DTT/MB-DNA/Au was applied for plants and medical products analysis. In all, this optimized DTT/MB-DNA/Au with advantages of high repeatability and sensitivity would provide a new insight into the design and application of biosensor for reliable sensing in safeguarding plant protection and medicinal safety.

Brucella utilizes diverse virulence factors to modulate the infectious cycle and lifestyle associated with eukaryotic hosts. Lipopolysaccharides (LPS) play an important role in the establishment of persistent infections of Brucella, but its mechanism in host cell to interfere with the host"s specific signaling pathway related to the elimination of Brucella is still not clear. LpsA, a glycosyl transferase, involves in the biosynthesis of LPS, and may aff ect the intracellular survival of Brucella. This study aimed to investigate the eff ects of lpsA on pyroptosis in Brucella-infected macrophage cells. We constructed the lpsA mutant strain (M5-90ΔlpsA) and the complementary strain (M5-90ΔlpsA-C) of Brucella melitensis strain M5-90 infected mouse macrophages to detect pyroptosis. We found that the inactivation of the lpsA gene weakened the ability of LPS gains access to cytosol during Brucella mutant infections, and reduced the survival of Brucella. Western blot and quantitative real-time PCR assays showed that the mRNA and protein levels of caspase-11 and NOD-like receptor family pyrin domain-containing 3 in M5-90ΔlpsA group were higher than those observed in M5-90 group. These results indicated M5-90ΔlpsA promoted the pyroptosis of RAW264.7 cells (a mouse macrophage cell line) after Brucella infection. The finding reveals that Brucella lpsA plays a partial role in innate immunity and infl ammatory response by inhibiting the LPS-induced atypical pyroptosis pathway.

Abstract The high prevalence and spread of arthropod-borne viruses (arboviruses) make them an important cause of viral encephalitis in humans. Most epidemic viral encephalitides have an etiology associated with arboviruses. Among various arboviruses, the Japanese encephalitis virus, West Nile virus, Zika virus, Dengue virus and Chikungunya virus can induce seizures. Arboviruses of the genus Flavivirus are usually transmitted by mosquitoes and other host animals. These vector-borne pathogens can cause epidemic viral encephalitis. Seizures may not be the major manifestation in these viral encephalitides, but may predict a poor prognosis. In this article, we discuss the relationships between these viruses and seizures from perspectives of clinical characteristics, pathogenesis, prognosis and treatments of each.

Abstract We consider the following coupled fractional Schrödinger system: { ( − Δ ) s u + λ 1 u = μ 1 | u | 2 p − 2 u + β | v | p | u | p − 2 u , ( − Δ ) s v + λ 2 v = μ 2 | v | 2 p − 2 v + β | u | p | v | p − 2 v in R N , $$ \textstyle\begin{cases} (-\Delta )^{s}u+\lambda _{1}u=\mu _{1} \vert u \vert ^{2p-2}u+ \beta \vert v \vert ^{p} \vert u \vert ^{p-2}u, \\ (-\Delta )^{s}v+\lambda _{2}v=\mu _{2} \vert v \vert ^{2p-2}v+\beta \vert u \vert ^{p} \vert v \vert ^{p-2}v \end{cases}\displaystyle \quad \text{in } {\mathbb{R}^{N}}, $$ with 0 < s < 1 $0< s 0 $\beta >0$ .

Twenty-five dog fleas (Ctenocephalides canis) and five ticks identified as Hyalomma asiaticum were collected from a Eurasian lynx (Lynx lynx) pup in northwestern China. Molecular analyses of four genetic markers showed the presence of Rickettsia aeschlimannii DNA in 5 out of 20 fleas. Only rickettsial 17-kDa gene was detected in the blood sample of the lynx. In addition, 2 out of 20 fleas were positive to Wolbachia endosymbiont by targeting 16S rDNA while there was no Bartonella DNA found both in 5 ticks and 20 fleas by using gltA and 16S-23S ITS. Our findings suggest that i) C. canis parasitizing wild Eurasian lynx harbors R. aeschlimannii and Wolbachia endosymbiont in the China-Kazakhstan border, and ii) Wolbachia endosymbiont in present study is closer to that in C. canis infesting sheltered dogs in Turkey.

Flea distribution in northern region of Xinjiang Uygur Autonomous Region (XUAR) and fluctuations of the annual fleas index in Alataw Pass were investigated. During a 4-year (2015–2018) study, 5789 fleas were collected directly from 15 mammals at eight counties in northern XUAR of northwestern China. Nineteen flea species, belonging to sixteen genera and seven families, were further confirmed by four genetic markers (18S rDNA, 28S rDNA, COI and COII) after morphological observation. Pulex irritans and Paraceras crispum parasitizing Asian badgers (Meles leucurus) were recorded for the first time. In addition, the fluctuations of the annual fleas index in Alataw Pass were surveyed. Xenopsylla gerbilli minax, Xenopsylla conformis conformis and Nosopsyllus laeviceps laeviceps were highly detected in the warm season while Paradoxopsyllus repandus, Ctenophthalmus dolichus dolichus and Coptopsylla lamellifer ardua were only found in the cold season. These findings extend our knowledge of flea species, distribution and annual fluctuations especially in China-Kazakhstan border. Keywords: Flea distribution, XUAR, Fluctuations, The annual fleas index, Wild mammals

Circular RNAs (circRNAs) are broad and diverse endogenous non-coding RNAs. Emerging evidence has revealed that circRNAs play pivotal roles in cancers, regulating the gene expression by acting as a microRNA (miRNA) sponge. However, the biological functions of circRNAs in bladder cancer (BCa) remain largely unknown. In this study, we identified an altered circRNA, termed circFUT8, by screening RNA sequencing data generated from three BCa tissues and matched adjacent normal bladder tissues. Quantitative real-time PCR analysis demonstrated that circFUT8 was downregulated in BCa tissues and correlated with patients’ prognosis, histological grade, and lymph node (LN) metastasis. Functionally, gain- and loss-of-function assays indicated that circFUT8 inhibited the migration and invasion of BCa cell lines in vitro and LN metastasis in vivo. Mechanistically, circFUT8 directly bound to miR-570-3p and partially abrogated its oncogenic role, and miR-570-3p could suppress the expression of tumor suppressor gene Krüpple-like factor 10 (KLF10) by binding its 3′ untranslated region (3′ UTR). Moreover, we found that circFUT8 promoted the expression of KLF10 by competitively sponging miR-570-3p. In conclusion, circFUT8 functions as a tumor suppressor in BCa cells by targeting the miR-570-3p/KLF10 axis and may serve as a potential biomarker and therapeutic target for the management of BCa patients with LN metastasis. Keywords: circFUT8, bladder cancer, lymph node metastasis, miR-570-3p, KLF10, Slug

Abstract Background Increasing molecular evidence supports that bats and/or their ectoparasites may harbor vector-borne bacteria, such as bartonellae and borreliae. However, the simultaneous occurrence of rickettsiae in bats and bat ticks has been poorly studied. Methods In this study, 54 bat carcasses and their infesting soft ticks (n = 67) were collected in Shihezi City, northwestern China. The heart, liver, spleen, lung, kidney, small intestine and large intestine of bats were dissected, followed by DNA extraction. Soft ticks were identified both morphologically and molecularly. All samples were examined for the presence of rickettsiae by amplifying four genetic markers (17-kDa, gltA, ompA and ompB). Results All bats were identified as Pipistrellus pipistrellus, and their ticks as Argas vespertilionis. Molecular analyses showed that DNA of Rickettsia parkeri, R. lusitaniae, R. slovaca and R. raoultii was present in bat organs/tissues. In addition, nine of the 67 bat soft ticks (13.43%) were positive for R. raoultii (n = 5) and R. rickettsii (n = 4). In the phylogenetic analysis, these bat-associated rickettsiae clustered together with conspecific sequences reported from other host and tick species, confirming the above results. Conclusions To the best of our knowledge, DNA of R. parkeri, R. slovaca and R. raoultii was detected for the first time in bat organs/tissues. This is also the first molecular evidence for the presence of R. raoultii and R. rickettsii in bat ticks. To our knowledge, R. parkeri was not known to occur in Asia. Our results highlight the need to assess rickettsial agents in a broader range of bat species and associated tick species.

Urothelial carcinoma of the bladder (UCB) is a major type of bladder cancer with a distinct tumor microenvironment (TME). Although neutrophils are the main component of myeloid cells in the TME, the clinical significance and function of the neutrophils remain unclear in UCB. Here, we observed CD66b+ neutrophils were predominantly enriched in the stroma of UCB tissues and their levels emerged as an independent prognostic factor for overall survival (P = 0.006, n = 237), and were positively associated with age (P = 0.033), tumor stage (P < 0.0001), nodal metastasis (P = 0.045), and histological grade (P < 0.0001). Furthermore, we found that CD66b+ neutrophils were frequently co-localized with CD4+ T cells (R=0.35, P = 0.0067), CD8+ T cells (R=0.52, P

Alternaria mycotoxins are ubiquitous mycotoxins that contaminate food and animal feed. Here, an UPLC-MS/MS was developed and used for the detection of seven Alternaria mycotoxins in 19 different edible and medicinal herbs. Extensive optimization resulted in a simple and convenient sample preparation procedure with satisfactory extraction and a lower matrix effect. LOQs ranged from 0.01 to 2.0 ng/mL. Recoveries varied between 71.44% and 112.65%, with RSD less than 12%. The method was successfully applied for use in the mycotoxin analysis of 260 samples. A high percentage (28.46%) of samples were contaminated by 1–5 mycotoxins. Alternariol mono methylether was the predominant mycotoxin with high percentage of positive samples (37.5%), followed by alternariol (22.5%), alternariol (17.5%), tentoxin (10.83%), altertoxin Ⅰ (7.5%), and altenusin (4.17%). Collectively, the natural incidence data obtained from this study will help with better, validated risk assessments and efforts towards more comprehensive, future regulation.

Robust soil organic matter (SOM) mapping is required by farms, but their generation requires a large number of samples to be chemically analyzed, which is cost prohibitive. Recently, research has shown that visible and near-infrared (vis-NIR) reflectance spectroscopy is a fast and accurate technique for estimating SOM in a cost-effective manner. However, few studies have focused on using vis-NIR spectroscopy as a covariate to improve the accuracy of spatial modeling. In this study, our objective was to compare the mapping accuracy from a spatial model using kriging methods with and without the covariate of vis-NIR spectroscopy. We split the 261 samples into a calibration set (104) for building the spectral predictive model, a test set for generating the vis-NIR augmented set from the prediction of the fitted spectral predictive model (131), and a validation set (26) for evaluating map accuracy. We used two datasets (235 samples) for Kriging: a laboratory-based dataset (Ld, observations from calibration and test datasets) and a laboratory-based dataset with vis-NIR augmented predictions (Au.p, observations from calibration and predictions from test dataset), a laboratory-based dataset with vis-NIR spectra as the covariance (Ld.co) and augmented dataset with predictions using vis-NIR with vis-NIR spectra for the covariance (Au.p.co). The first one to seven accumulated principal components of vis-NIR spectra were used as the covariates when we used the measurement of Ld.co and Au.p.co. The map accuracy was evaluated by the validation set for the four datasets using Kriging. The results indicated that adding vis-NIR spectra as covariates had great potential in improving the map accuracy using kriging, and much higher accuracies were observed for Ld.p.co (RMSE of 5.51 g kg−1) and Au.p.co (RMSE of 5.66 g kg−1) than without using vis-NIR spectra as covariates for Ld (RMSE of 7.12 g kg−1) and Au.p (RMSE of 7.69 g kg−1). With a similar model performance to Ld.p.co, Au.p.co can reduce the cost of laboratory analysis for 60% of soil samples, demonstrating its advantage in cost-efficiency for spatial modeling of soil information. Therefore, we conclude that vis-NIR spectra can be used as a cost-effective technique to obtain augmented data to improve fine-resolution spatial mapping of soil information.

Mycotoxins can occur naturally in a variety of agriculture products, including cereals, feeds, and Chinese herbal medicines (TCMs), via pre- and post-harvest contamination and are regulated worldwide. However, risk mitigation by monitoring for multiple mycotoxins remains a challenge using existing methods due to their complex matrices. A multi-toxin method for 22 mycotoxins (aflatoxin B1, B2, G1, G2, M1, M2; ochratoxin A, B, C; Fumonisin B1, B2, B3; 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, diace-toxyscirpenol, HT-2, T-2, deepoxy-deoxynivalenol, deoxynivalenol, neosolaniol, zearalenone, and sterigmatocystin) using centrifugation-assisted solid-phase extraction (SPE) clean-up prior to ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis for Arecae Semen and its processed products was developed and validated. Several experimental parameters affecting the extraction and clean-up efficiency were systematically optimized. The results indicated good linearity in the range of 0.1–1000 μg/kg (r2 > 0.99), low limits of detection (ranging from 0.04 μg/kg to 1.5 μg/kg), acceptable precisions, and satisfactory recoveries for the selected mycotoxins. The validated method was then applied to investigate mycotoxin contamination levels in Areca catechu and its processed products. The mycotoxins frequently contaminating Areca catechu were aflatoxins (AFs), and the average contamination level and number of co-occurring mycotoxins in the Arecae Semen slices (Binlangpian) were higher than those in commercially whole Arecae Semen and Arecae Semen Tostum (Jiaobinlang). Sterigmatocystin was detected in 5 out of 30 Arecae Semen slices. None of the investigated mycotoxins were detected in Arecae pericarpium (Dafupi). The results demonstrated that centrifugation-assisted SPE coupled with UHPLC-MS/MS can be a useful tool for the analysis of multiple mycotoxins in Areca catechu and its processed products.

Improvement of understanding of the safety profile and biological significance of antidiabetic agents in breast cancer (BC) progression may shed new light on minimizing the unexpected side effect of antidiabetic reagents in diabetic patients with BC. Our recent finding showed that Saxagliptin (Sax) and Sitagliptin (Sit), two common antidiabetic dipeptidyl peptidase-4 inhibitors (DPP-4i) compounds, promoted murine BC 4T1 metastasis via a ROS–NRF2–HO-1 axis in nonobese diabetic–severe combined immunodeficiency (NOD-SCID) mice. However, the potential role of DPP-4i in BC progression under immune-competent status remains largely unknown. Herein, we extended our investigation and revealed that Sax and Sit also accelerated murine BC 4T1 metastasis in orthotopic, syngeneic, and immune-competent BALB/c mice. Mechanically, we found that DPP-4i not only activated ROS–NRF2–HO-1 axis but also triggered reactive oxygen species (ROS)-dependent nuclear factor kappa B (NF-κB) activation and its downstream metastasis-associated gene levels in vitro and in vivo, while NF-кB inhibition significantly abrogated DPP-4i-driven BC metastasis in vitro. Meanwhile, inhibition of NRF2–HO-1 activation attenuated DPP-4i-driven NF-кB activation, while NRF2 activator ALA enhanced NF-кB activation, indicating an essential role of ROS–NRF2–HO-1 axis in DPP-4i-driven NF-кB activation. Furthermore, we also found that DPP-4i increased tumor-infiltrating CD45, MPO, F4/80, CD4, and Foxp3-positive cells and myeloid-derived suppressor cells (MDSCs), and decreased CD8-positive lymphocytes in metastatic sites, but did not significantly alter cell viability, apoptosis, differentiation, and suppressive activation of 4T1-induced splenic MDSCs. Moreover, we revealed that DPP-4i triggered ROS-NF-κB-dependent NLRP3 inflammasome activation in BC cells, leading to increase in inflammation cytokines such as interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), intercellular cell adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), IL-1β and IL-33, and MDSCs inductors granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF, and M-CSF, which play a crucial role in the remodeling of tumor immune-suppressive microenvironment. Thus, our findings suggest that antidiabetic DPP-4i reprograms tumor microenvironment that facilitates murine BC metastasis by interaction with BC cells via a ROS–NRF2–HO-1–NF-κB–NLRP3 axis. This finding not only provides a mechanistic insight into the oncogenic ROS–NRF2–HO-1 in DPP-4i-driven BC progression but also offers novel insights relevant for the improvement of tumor microenvironment to alleviate DPP-4i-induced BC metastasis.

We isolated Brucella melitensis biovar 3 from the spleen of an Asian badger (Meles leucurus) in Nilka County, northwestern China. Our investigation showed that this isolate had a common multilocus variable-number tandem-repeat analysis 16 genotype, similar to bacterial isolates from local aborted sheep fetuses.

Background: Metastasis is a major obstacle in the treatment of bladder cancer (BC). Circular RNAs exert various functions in the aggressive biological behaviour of cancers. In this study, we aimed to elucidate how circPICALM influences BC metastasis. Methods: The expression of circPICALM was analysed by real-time PCR. The tumourigenic properties of BC cells were evaluated using in vitro migration, invasion, and wound healing assays and an in vivo footpad model. The interaction between circPICALM and miR-1265 was confirmed by pull-down and dual-luciferase reporter assays and biotin-labelled miRNA capture. The interaction of STEAP4 and focal adhesion kinase (FAK) was confirmed by co-immunoprecipitation. Findings: CircPICALM was downregulated in BC tissues, and low circPICALM expression was related to advanced T stage, high grade, lymph node positivity and poor overall survival. Overexpression of circPICALM inhibited the metastasis of BC cells, and DHX9 negatively regulated circPICALM levels. CircPICALM colocalized with miR-1265 and acted as a sponge for this miRNA, and the pro-invasion effect of miR-1265 was abolished by circPICALM overexpression. STEAP4, a target of miR-1265, suppressed metastasis; it bound to FAK to prevent autophosphorylation at Y397 and influenced EMT in BC cells. Interpretation: CircPICALM can inhibit BC metastasis and bind to miR-1265 to block its pro-invasion activity. STEAP4 is a target of miR-1265 and is related to FAK phosphorylation and EMT. Fund: This research was supported by National Natural Science Foundation of China, No.81772728, National Natural Science Foundation of China, No.81772719, National Natural Science Foundation of China No.81572514. Keywords: circPICALM, miR-1265, STEAP4, EMT, Metastasis, Bladder cancer

Abstract Background Circular RNAs (circRNAs) have been considered to mediate occurrence and development of human cancers, generally acting as microRNA (miRNA) sponges to regulate downstream genes expression. However, the aberrant expression profile and dysfunction of circRNAs in human bladder cancer remain to be investigated. The present study aims to elucidate the potential role and molecular mechanism of circACVR2A in regulating the proliferation and metastasis of bladder cancer. Methods circACVR2A (hsa_circ_0001073) was identified by RNA-sequencing and validated by quantitative real-time polymerase chain reaction and agarose gel electrophoresis. The role of circACVR2A in bladder cancer was assessed both in vitro and in vivo. Biotin-coupled probe pull down assay, biotin-coupled microRNA capture, dual-luciferase reporter assay, and fluorescence in situ hybridization were conducted to evaluate the interaction between circACVR2A and microRNAs. Results The expression of circACVR2A was lower in bladder cancer tissues and cell lines. The down-regulation of circACVR2A was positively correlated with aggressive clinicopathological characteristics, and circACVR2A served as an independent risk factor for overall survival in bladder cancer patients after cystectomy. Our in vivo and in vitro data indicated that circACVR2A suppressed the proliferation, migration and invasion of bladder cancer cells. Mechanistically, we found that circACVR2A could directly interact with miR-626 and act as a miRNA sponge to regulate EYA4 expression. Conclusions circACVR2A functions as a tumor suppressor to inhibit bladder cancer cell proliferation and metastasis through miR-626/EYA4 axis, suggesting that circACVR2A is a potential prognostic biomarker and therapeutic target for bladder cancer.

Abstract Background Dysregulated miR-7 and aberrant NF-κB activation were reported in various human cancers. However, the expression profile, clinical relevance and dysregulated mechanism of miR-7 and NF-κB RelA/p65 in human gastric cancers (GC) metastasis remain largely unknown. This study is to investigate the expression profile, clinical relevance and dysregulated mechanism of miR-7 and NF-κB RelA/p65 in GC and to explore the potential therapeutic effect of miR-7 to GC distant metastasis. Methods TCGA STAD and NCBI GEO database were used to investigate the expression profile of miR-7 and NF-κB RelA/p65 and clinical relevance. Lentivirus-mediated gene delivery was applied to explore the therapeutic effect of miR-7 in GC. Real-time PCR, FACS, IHC, IF, reporter gene assay, IP, pre-miRNA-7 processing and binding assays were performed. Results Low miR-7 correlated with high RelA/p65 in GC with a clinical relevance that low miR-7 and high RelA/p65 as prognostic indicators of poor survival outcome of GC patients. Moreover, an impaired pre-miR-7 processing caused by dysregulated Dicer1 expression is associated with downregulated miR-7 in GC cells. Functionally, delivery of miR-7 displays therapeutic effects to GC lung and liver metastasis by alleviating hemangiogenesis, lymphangiogenesis as well as inflammation cells infiltration. Mechanistically, miR-7 suppresses NF-κB transcriptional activity and its downstream metastasis-related molecules Vimentin, ICAM-1, VCAM-1, MMP-2, MMP-9 and VEGF by reducing p65 and p-p65-ser536 expression. Pharmacologic prevention of NF-κB activator LPS obviously restored miR-7-suppressed NF-κB transcriptional activation and significantly reverted miR-7-inhibited cell migration and invasion. Conclusions Our data suggest loss of miR-7 in GC promotes p65-mediated aberrant NF-κB activation, facilitating GC metastasis and ultimately resulting in the worse clinical outcome. Thus, miR-7 may act as novel prognostic biomarker and potential therapeutic target for aberrant NF-κB-driven GC distant metastasis.

Fritillariae Cirrhosae Bulbus (FCB) is a well-known and precious traditional Chinese medicine with a medicinal history spanning thousands of years. In recent years, it has been reported that fungal and mycotoxin contamination influenced the safety and quality of FCB. It is essential to systematically study the fungal community for the early warning of fungal and mycotoxin contamination in this herb. A total of 15 FCB samples were collected from five provinces in China, and the fungal communities in the FCB samples were analyzed via amplifying the internal transcribed spacer 2 region through the Illumina Miseq PE300 platform. Furthermore, we compared the differences in fungal community in five groups based on collection areas. Results showed that Ascomycota (41.58–99.66%) and Mucoromycota (0–57.42%) were dominant at the phylum level. Eurotiomycetes (8.49–63.93%), Eurotiales (8.49–63.53%), and Aspergillaceae (8.49–63.51%) were the most abundant at the class, order, and family levels. Aspergillus (8.49–63.41%), Rhizopus (0–57.42%), Fusarium (0–22.81%), Cladosporium (0.16–9.14%), and Alternaria (0.06–17.95%) were the main genera in FCB samples. A total of 34 fungal taxa were identified at the species level, including five potentially toxigenic fungi namely Penicillium brevicompactum, P. citrinum, P. oxalicum, Trichothecium roseum, and Aspergillus restrictus. The differences in fungal community between the five groups were observed. Our findings provide references for the safe utilization and quality improvement of FCB.