Continued Evolution of H10N3 Influenza Virus with Adaptive Mutations Poses an Increased Threat to Mammals.

The H10 subtype of avian influenza virus (AIV) poses an ongoing threat to both birds and humans. Notably, fatal human cases of H10N3 and H10N8 infections have drawn public attention. In 2022, we isolated two H10N3 strains (A/chicken/Shandong/0101/2022 and A/chicken/Shandong/0603/2022) from diseased chickens in China. Genome analysis revealed that these strains were genetically associated with human-origin H10N3 virus, with internal genes originating from local H9N2 viruses. Compared to the H10N8 strain (A/chicken/Jiangxi/102/2013), the H10N3 strains exhibited enhanced thermostability, increased viral release from erythrocytes, and accumulation of hemagglutinin (HA) protein. Additionally, we evaluated the pathogenicity of both H10N3 and H10N8 viruses in mice. We found that viral titers could be detected in the lungs and nasal turbinates of mice infected with the two H10N3 viruses, whereas H10N8 virus titers were detectable in the lungs and brains of mice. Notably, the proportion of double HA Q222R and G228S mutations in H10N3 viruses has increased since 2019. However, the functional roles of the Q222R and G228S double mutations in the HA gene of H10N3 viruses remain unknown and warrant further investigation. Our study highlights the potential public health risk posed by the H10N3 virus. A spillover event of AIV to humans could be a foretaste of a looming pandemic. Therefore, it is imperative to continuously monitor the evolution of the H10N3 influenza virus to ensure targeted prevention and control measures against influenza outbreaks.

Species-specific design of artificial promoters by transfer-learning based generative deep-learning model.

Native prokaryotic promoters share common sequence patterns, but are species dependent. For understudied species with limited data, it is challenging to predict the strength of existing promoters and generate novel promoters. Here, we developed PromoGen, a collection of nucleotide language models to generate species-specific functional promoters, across dozens of species in a data and parameter efficient way. Twenty-seven species-specific models in this collection were finetuned from the pretrained model which was trained on multi-species promoters. When systematically compared with native promoters, the Escherichia coli- and Bacillus subtilis-specific artificial PromoGen-generated promoters (PGPs) were demonstrated to hold all distribution patterns of native promoters. A regression model was developed to score generated either by PromoGen or by another competitive neural network, and the overall score of PGPs is higher. Encouraged by in silico analysis, we further experimentally characterized twenty-two B. subtilis PGPs, results showed that four of tested PGPs reached the strong promoter level while all were active. Furthermore, we developed a user-friendly website to generate species-specific promoters for 27 different species by PromoGen. This work presented an efficient deep-learning strategy for de novo species-specific promoter generation even with limited datasets, providing valuable promoter toolboxes especially for the metabolic engineering of understudied microorganisms.

Decision-making ability limitations and brain neural activity changes in healthcare workers after mild COVID-19.

Studies have demonstrated that the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) extensively affects brain function. Although cognitive dysfunction is considered a common manifestation in COVID-19 patients during the recovery period, the potential changes in decision-making ability, are not yet clear. Decision-making functions are essential to the work of healthcare workers. However, there is a lack of a multidimensional assessment of its functioning in COVID-19 cases. Here, we used tests combined with the resting-state functional magnetic resonance imaging (rs-fMRI) stabilization feature amplitude of low-frequency fluctuations (ALFF) to explore decision-making behavior and brain neural activity changes in healthcare workers after mild COVID-19. Participants were divided into the SARS-CoV-2 infected group (SI, n = 41) and healthy controls (HC, n = 42). All participants underwent a series of neuropsychological tests. They performed the Iowa Gambling Task (IGT) and the Game of Dice Task (GDT), followed by fMRI (n = 20) to assess their decision-making ability under ambiguous and risky conditions and changes in brain neural activity. The SI group performed worse in verbal memory than the HC group. Furthermore, the SI group performed worse in the IGT, whereas no significant difference was observed in the GDT. In addition, rs-fMRI showed enhanced spontaneous neural activity in the postcentral gyrus and inferior parietal lobe in the SI group compared to the HC group.

Anterior cervical plating combined with zero-profile anchored spacer for the treatment of cervical facet dislocation in elderly patients.

OBJECTIVE: The purpose of this study was to evaluate the efficacy of anterior cervical plating combined with zero-profile (Z-P) anchored spacer for the treatment of cervical facet dislocation in elderly patients. METHODS: This is a retrospective study. Twelve elderly patients (from 57 to 77 years old, averaged 65 years) with unilateral or bilateral facet dislocation of sub-axial cervical spine from September 2015 to September 2019 surgically treated at the authors' hospital were enrolled in this study. The patients with osteoporosis or osteopenia were all surgically treated by anterior-only procedure using cervical plating combined with zero-profile anchored spacer after closed manual reduction under general anesthesia and spinal cord monitoring. The operation times (OT), estimated blood loss (EBL), perioperative complications, were recorded. The clinical evaluation included visual analogue scales (VAS) and the American Spinal Injury Association (ASIA) scale. The radiographic evaluation included kyphotic angle (KA) and disc height (DH) and the fusion rate. RESULTS: Anterior discectomy, interbody fusion and fixation were performed in all patients after the disloctions were reduced by manual maneuver. The average OT was 66 minutes, with a range from 45 to 110 minutes. The EBL averaged 42 ml per surgical procedure, with a range from 20 to 60 ml. The VAS, ASIA, KA were improved significantly after surgery (P<0.05). The average follow-up time was 24.2 months, with a range from 12 to 38 months. There were no statistical differences between the immediately post-op KA and KA at the last follow-up (P>0.05). No disc space subsidence was observed statistically (P<0.05) Interbody fusion was obtained in all patients. Two patient experienced slight difficulty in swallowing, which were improved 6 weeks later. There were no hardware failure, no segmental instability, no wound infection or other complications. CONCLUSIONS: Manual reduction with spinal cord monitoring under general anesthesia is a safe and efficient option and the anterior cervical plating combined with Z-P spacer could achieve reliable fixation for the patients with cervical facet dislocation in the elderly patients with osteoporosis or osteopenia.

PELO facilitates PLK1-induced the ubiquitination and degradation of Smad4 and promotes the progression of prostate cancer.

PLK1 and Smad4 are two important factors in prostate cancer initiation and progression. They have been reported to play the opposite role in Pten-deleted mice, one is an oncogene, the other is a tumor suppressor. Moreover, they could reversely regulate the PI3K/AKT/mTOR pathway and the activation of MYC. However, the connections between PLK1 and Smad4 have never been studied. Here, we showed that PLK1 could interact with Smad4 and promote the ubiquitination and degradation of Smad4 in PCa cells. PLK1 and PELO could bind to different domains of Smad4 and formed a protein complex. PELO facilitated the degradation of Smad4 through cooperating with PLK1, thereby resulting in proliferation and metastasis of prostate cancer cell. Changes in protein levels of Smad4 led to the alteration of biological function that caused by PLK1 in prostate cancer cells. Further studies showed that PELO upregulation was positively associated with high grade PCa and knockdown of PELO expression significantly decreased PCa cell proliferation and metastasis in vitro and vivo. PELO knockdown in PCa cells could enhance the tumor suppressive role of PLK1 inhibitor. In addition, blocking the interaction between PELO and Smad4 by using specific peptide could effectively inhibit PCa cell metastasis ability in vitro and vivo. Overall, these findings identified a novel regulatory relationship among PLK1, Smad4 and PELO, and provided a potential therapeutic strategy for advanced PCa therapy by co-targeting PLK1 and PELO.

Actinoplanes aureus sp. nov., a novel protease-producing actinobacterium isolated from soil.

A novel protease-producing actinobacterium, designated strain NEAU-A11T, was isolated from soil collected from Aohan banner, Chifeng, Inner Mongolia Autonomous Region, China, and characterised using a polyphasic approach. The hydrolytic enzymes, such as proteases, played critical roles in destruction of fungi by degrading the protein linkages to disrupt integrity in the cell wall. This suggested that the isolate could be a good biocontrol candidate against pathogens to control fungal diseases. On the basis of 16S rRNA gene sequence analysis, strain NEAU-A11T was indicated to belong to the genus Actinoplanes and was most closely related to Actinoplanes rectilineatus JCM 3194 T (98.9%). Cell walls contained meso-diaminopimelic acid as the diagnostic diamino acid and the whole-cell sugars were arabinose, xylose and glucose. The phospholipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and two phosphatidylinositol mannosides. The predominant menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8). The major fatty acids were C18:0, C16:0, C18:1 ω9c, C17:0 and C15:0. Genome sequencing revealed a genome size of 10,742,096 bp, a G + C content of 70.5% and 9,514 protein-coding genes (CDS), including 102 genes coding for protease. Moreover, Genome analysis showed that strain NEAU-A11T contained 255 glycoside hydrolases (GHs), 152 glycosyl transferases (GTs), 40 carbohydrate esterases (CEs), 26 polysaccharide lyases (PLs), and 12 auxiliary activities (AAs) genes. Genome mining analysis using antiSMASH 5.0 led to the identification of 20 putative gene clusters responsible for the production of diverse secondary metabolites. Phylogenetic analysis using the 16S rRNA gene sequences showed that the strain formed a stable clade with A. rectilineatus JCM 3194 T in the genus Actinoplanes. Whole-genome phylogeny showed strain NEAU-A11T formed a stable phyletic line with Actinoplanes lutulentus DSM 45883 T (97.6%). However, whole-genome average nucleotide identity value between strain NEAU-A11T and its reference strains A. rectilineatus JCM 3194 T and A. lutulentus DSM 45883 T were found to be 81.1% and 81.6%, respectively. The levels of digital DNA-DNA hybridization between them were 24.6% (22.2-27.0%) and 24.8% (22.5-27.3%), respectively. The values were well below the criteria for species delineation of 70% for dDDH and 95-96% for ANI, suggesting that the isolate differed genetically from its closely related type strain. The content of G + C in genomic DNA was 70.5%, within the range of 67-76%. In addition, evidences from phenotypic, chemotaxonomic and genotypic studies indicated that strain NEAU-A11T represents a novel species of the genus Actinoplanes, for which the name Actinoplanes aureus sp. nov. is proposed, with NEAU-A11T (= CCTCC AA 2019063 T = JCM 33971 T) as the type strain.

Fingolimod inhibits proliferation and epithelial-mesenchymal transition in sacral chordoma by inactivating IL-6/STAT3 signalling.

PURPOSE: To explore the sensitivity of the immunosuppressive agent fingolimod (FTY720) in chordoma and determine whether it can serve as an appropriate alternate treatment for unresectable tumours in patients after incomplete surgery. METHODS: Cell viability assays, colony formation assays and EdU assays were performed to evaluate the sensitivity of chordoma cell lines to FTY720. Transwell invasion assays, wound healing assays, flow cytometry, cell cycle analysis, immunofluorescence analysis, Western blotting analysis and enzyme-linked immunosorbent assays (ELISAs) were performed to evaluate cell invasion, epithelial-mesenchymal transition (EMT) and activation of related pathways after treatment with FTY720. The effect of FTY720 was also evaluated in vivo in a xenograft model. RESULTS: We found that FTY720 inhibited the proliferation, invasion and metastasis of sacral chordoma cells (P < 0.01). FTY720 also inhibited the proliferation of tumour cells in a xenograft model using sacral chordoma cell lines (P < 0.01). The mechanism was related to the EMT and apoptosis of chordoma cells and inactivation of IL-6/STAT3 signalling in vitro and in vivo. CONCLUSIONS: Our findings indicate that FTY720 may be an effective therapeutic agent against chordoma. These findings suggest that FTY720 is a novel agent that can treat locally advanced and metastatic chordoma.

miR­329­3p regulates neural stem cell proliferation by targeting E2F1.

Neural stem cells (NSCs) are a class of self­renewing and undifferentiated progenitor cells that retain the ability to differentiate to neurons, astrocytes and oligodendrocytes. MicroRNAs (miRNAs) are small noncoding RNAs that serve crucial roles in regulating a number of cellular processes, including cell proliferation, differentiation and apoptosis. Our previous GeneChip data indicated that the expression of miR­329­3p was increased in neurons compared with NSCs. However, whether miRNA­329­3p participates in regulating NSC function remains to be elucidated. In the present study, it was identified that the expression of miR­329­3p was upregulated in NSCs during neuronal differentiation, whereas expression of transcription factor E2F1 (E2F1), a putative target gene of miR­329­3p, was downregulated. Using luciferase reporter assays, it was confirmed that miR­329­3p regulated E2F1 expression. As differentiation has been demonstrated to limit the proliferative capacity of NSCs, the effects of miR­329­3p and E2F1 modulation on NSC proliferation were examined. Forced overexpression of miR­329­3p or RNA­mediated silencing of E2F1 inhibited NSC proliferation, and overexpression of miR­329­3p also inhibited E2F1 expression. Notably, ectopic expression of E2F1 reversed the inhibition of NSC proliferation induced by miR­329­3p overexpression. These results indicated that miR­329­3p may serve crucial roles in regulating the proliferation of NSCs, at least in part via inhibition of E2F1 expression. These data improve the understanding of the microRNA­mRNA regulatory network that controls NSC proliferation.

Gas Sensors Based on Nano/Microstructured Organic Field-Effect Transistors.

Benefiting from the advantages of organic field-effect transistors (OFETs), including synthetic versatility of organic molecular design and environmental sensitivity, gas sensors based on OFETs have drawn much attention in recent years. Potential applications focus on the detection of specific gas species such as explosive, toxic gases, or volatile organic compounds (VOCs) that play vital roles in environmental monitoring, industrial manufacturing, smart health care, food security, and national defense. To achieve high sensitivity, selectivity, and ambient stability with rapid response and recovery speed, the regulation and adjustment of the nano/microstructure of the organic semiconductor (OSC) layer has proven to be an effective strategy. Here, the progress of OFET gas sensors with nano/microstructure is selectively presented. Devices based on OSC films one dimensional (1D) single crystal nanowires, nanorods, and nanofibers are introduced. Then, devices based on two dimensional (2D) and ultrathin OSC films, fabricated by methods such as thermal evaporation, dip-coating, spin-coating, and solution-shearing methods are presented, followed by an introduction of porous OFET sensors. Additionally, the applications of nanostructured receptors in OFET sensors are given. Finally, an outlook in view of the current research state is presented and eight further challenges for gas sensors based on OFETs are suggested.

Polyketide derivatives from the sponge associated fungus Aspergillus europaeus with antioxidant and NO inhibitory activities.

DPPH assay of the in-house marine-derived fungi uncovered that the EtOAc extract of the cultured fungus Aspergillus europaeus WZXY-SX-4-1, which was isolated from the marine sponge Xestospongia testudinaria, possesses radical scavenging activity. Chromatographic separation of the bioactive extract resulted in the isolation of 20 polyketide derivatives, including six new compounds namely eurobenzophenones A-C (1-3), euroxanthones A-B (4-5), and (+)1-O-demethylvariecolorquinones A (6). The structures of new compounds were determined on the basis of the analyses of spectroscopic data, including the Snatzke method for the configurational assignment. Benzophenones 3, 9 and 10 exhibited potent radical scavenging activity against DPPH. All polyketides were evaluated for the inhibitory effects toward the LPS induced nitric oxide (NO) production in mouse microglia BV2 cells and the NF-κB activation in human colon carcinoma cell line SW480. Compound 9 with the significant DPPH radical scavenging activity is corresponded to the potent inhibition against NF-κB in SW480 cells induced by LPS. Compounds 2, 4, 16-18 exerted remarked down-regulation of NF-κB in LPS-induced SW480 cells with weak inhibitory effects against NO production and the DPPH radical scavenging activity.

Anticancer effect of miR-96 inhibitor in bladder cancer cell lines.

The present study aimed to investigate the role of microRNA-96 (miR-96) in the proliferation, invasion and apoptosis of bladder cancer cell lines, and the associated mechanisms. The expression of miR-96 and human ether-à-go-go-related (HERG1) potassium channel in the normal uroepithelium SV-HUC-1 cell line, and bladder cancer T24 and 5637 cell lines were examined using reverse transcription-polymerase chain reaction or/and western blotting. Transfection with miR-96 inhibitor or scrambled control (SC) was used to study the biological activities of miR-96 in bladder cancer cell lines. MTT, flow cytometric and Transwell assays were applied to detect cell viability, apoptosis and invasion, respectively. A dual-luciferase reporter assay was applied to determine the association between miR-96 and HERG1 expression. As demonstrated, miR-96 was highly expressed in the two bladder cancer cell lines, particularly in T24 cells. Following transfection with miR-96 inhibitor, miR-96 expression was significantly reduced in the T24 cell line, compared with SC. The miR-96 inhibitor suppressed cell proliferation and invasion, promoted apoptosis and arrested the cell cycle at the G1 phase. Consistently, HERG1 was also highly expressed in the two bladder cancer cell lines at the mRNA and protein level, but not in the normal uroepithelium cell line. The miR-96 inhibitor also significantly decreased HERG1 expression compared with SC. The results of the dual-luciferase reporter assay indicated that miR-96 directly targeted wild-type HERG1. In conclusion, miR-96 inhibitor exhibited anticancer effects on bladder cancer cells by inhibiting proliferation and invasion of cells, and promoting their apoptosis. HERG1 was an important target of miR-96. These results provided experimental evidence supporting miR-96 as a therapeutic target for patients with bladder cancer.

Multi-walled carbon nanotube modified dummy-template magnetic molecularly imprinted microspheres as solid-phase extraction material for the determination of polychlorinated biphenyls in fish.

Novel multi-walled carbon nanotube modified dummy-template molecularly imprinted microspheres (MWCNTs@DMMIPs) were successfully synthesized as adsorbents for six kinds of polychlorinated biphenyls (PCBs). MWCNTs@DMMIPs were prepared by a surface molecular imprinting technique. Core-shell Fe3 O4 @SiO2 nanoparticles were employed as magnetic support. 3,4-Dichlorobenzene acetic acid was used as a dummy template instead of PCBs, methacrylic acid was used as functional monomer and ethylene glycol dimethacrylate was used as the cross-linker. The resulting absorbent was characterized by various methods. The adsorbent was employed for extracting PCBs and exhibited good selectivity and high adsorption efficiency. Furthermore, it was reusable and capable of magnetic separation. Adsorption kinetics fit well with a pseudo-second-order kinetic equation and also exhibited a three-stage intra-particle diffusion model. The Freundlich model was used to describe the adsorption isotherms. The materials were successfully applied to the magnetic dispersive solid-phase extraction of six kinds of PCBs followed by gas chromatography with mass spectrometry determination in fish samples, the limit of detection of six kinds of PCBs were 0.0028-0.0068 µg/L and spiked recoveries ranged between 73.41 and 114.21%. The prepared adsorbent was expected to be a new material for the removal and recovery of PCBs from contaminated foods.

Experimental study of therapy of bone marrow mesenchymal stem cells or muscle-like cells/calcium alginate composite gel for the treatment of stress urinary incontinence.

OBJECTIVE: To explore the myoblast formation around the urethra and increase in urethral resistance of bone marrow mesenchymal stem cells or muscle-like cells/calcium alginate composite gel injection therapy and effect on LPP in SUI rat model. METHODS: Isolation, cultivation, and identification of SD rat bone marrow mesenchymal stem cell were performed. 5-Azacytidine was introduced to induce muscle-like cells. SUI was produced in 72 6-week-old female Sprague-Dawley rats, which were divided into four groups: stem cell-gel group, muscle-like cell-gel group, Gel group, and mock control group. One, 4, and 8 weeks after injection, the leak point pressure (LPP) was measured. HE staining of Desmin and α-skeletal muscle actin (α-SMA) were performed. RESULTS: At 4 and 8 weeks after injection in stem cell-gel group and muscle-like cell-gel group, growth of blood vessels gradually increased at gel edge, BMSC, and muscle-like cells gathered around the new blood vessels observed by fluorescence tracer, muscle-like cells grew into elongated spindle-like cells, Desmin, and α-SMA staining were obviously positive expression. LPP determinations of the mock control group compared with the Gel groups were significantly different. CONCLUSIONS: Compound of BMSC, muscle-like cells, and calcium alginate composite gel has the potential to differentiate into muscle cells in the microenvironment of SUI rat model. It is found by LPP measurement that the correlation between the increase in urethral resistance and the volume effect of calcium alginate gel is high.

An Ultrasensitive Electrochemical Immunosensor for HIV p24 Based on Fe3O4@SiO2 Nanomagnetic Probes and Nanogold Colloid-Labeled Enzyme-Antibody Copolymer as Signal Tag.

An ultrasensitive portable electrochemical immunosensor for human immunodeficiency virus p24 (HIV p24) antigen detection has been developed, whereby the detection sensitivity was 1000 times higher than that of the ELISA method. Firstly, a novel HRP enzyme-antibody copolymer (EV-p24 Ab2) was synthesized through an EnVision regent (EV, a dextrin amine skeleton anchoring more than 100 molecules of HRP and 15 molecules of anti IgG), then incubated in the secondary antibody of p24. Secondly, the copolymer was immobilized on the gold nanocolloids (AuNPs) to fabricate a novel signal tag (AuNPs/EV-p24 Ab2). Subsequently, a sandwich-type immunoreaction would take place between the capture probe (silicon dioxide-coated magnetic Fe3O4 nanoparticles (MNPs) labeled with the primary p24 antibody (MNPs-p24 Ab1)), p24 (different concentrations) and the signal tag [AuNPs/EV-p24 Ab2)] to form the immunocomplex. Finally, the immunocomplex was absorbed on the surface of screen printed carbon electrode (SPCE) by a magnet and immersed in the o-hydroxyl phenol (HQ) and H2O2. The large amounts of HRP on the signal tag can catalyze the oxidation of HQ by H2O2, which can induce an amplified reductive current. Moreover, the capture probe could improve the accumulation ability of p24 and facilitate its separation from the substrate through the magnet. Under optimal conditions, the proposed immunoassay exhibited good sensitivity to p24 within a certain concentration range from 0.001 to 10.00 ng/mL, with a detection limit of 0.5 pg/mL (S/N = 3). The proposed method can be used for real-time and early detection of HIV-infected people.