[Objective] To investigate the infection status and characteristics of HEV among voluntary blood donors in Ningbo, and to provide a basis for improving the blood screening strategy. [Methods] A total of 12 227 blood samples from voluntary blood donors in Ningbo from June 2022 to May 2023 were tested for HEV serology, enzymology, and nucleic acid testing. Furthermore, HEV gene sequencing was performed for genotyping analysis, and donors with reactive nucleic acid testing results were followed up to confirm their infection status. [Results] The reactivity rate of HEV Ag, anti-HEV IgM and anti-HEV IgG was 0.098%, 0.899% and 29.198%, respectively. There was no difference in the reactivity of anti-HEV IgM and anti-HEV IgG between genders, donation frequencies and donation types (P>0.05). The reactivity rate increased significantly with age (P<0.05). The rate of ALT disqualification (ALT>50U/L) was significantly higher than that in non-reactive samples (P<0.05). The HEV Ag reactivity rate (0.098%) was not correlated with gender, donation frequency, donation type or age. One HEV RNA positive case was found, with a positive rate of 0.008%(1/12 227). It was confirmed to be hepatitis E virus genotype 3 by sequencing analysis. Apart from HEV Ag reactivity, all other blood safety screening items were non-reactive, suggesting this case might be in the acute infection phase. The follow-up results showed that all indicators of the donor's previous blood donation were non-reactive. [Conclusion] Pre-donation ALT detection can reduce the risk of transfusion-transmitted HEV (TT-HEV) to a certain extent, and the effective way to prevent TT-HEV is to detect HEV RNA and serology of donor blood.

OBJECTIVE To evaluate the quality of diagnosis and treatment guidelines and expert consensuses on childhood immune thrombocytopenic purpura （ITP） published domestically and internationally， in order to provide reference for clinical practice and future guideline/expert consensus development and improvement. METHODS A systematic search was conducted across multiple databases， including PubMed， Cochrane Library， Embase， CNKI， Wanfang data， VIP， CBM； additionally， supplementary searches were carried out on websites such as Medlive， the Chinese Medical Association’s official website， and National Institute for Health and Clinical Excellence in the UK. The retrieval time ranged from the inception to September 2， 2024. Researchers who had undergone systematic training independently evaluated the methodology and report quality included in the guideline/consensus using the Appraisal of Guidelines Research and Evaluation Ⅱ （AGREE Ⅱ） and the Reporting Items for Practice Guidelines in Healthcare （RIGHT）. RESULTS A total of 11 guidelines/consensuses were included. The average scores for the six domains of AGREE Ⅱ tool respectively were “range and purpose” （[ 66.67±17.98）% ]， “participants” [58.33% （13.89%，73.61%）]， “rigor” （[ 41.81±23.85）% ]， “clarity”（[ 69.57±19.35）%]， “applicability” （[ 35.98±17.83）%]， and “independence” [27.08% （0，75.00%）]； out of 11 articles， 9 had a recommendation level of B， 2 had a recommendation level of C， and there were no A-level articles. The average reporting rates of the 7 areas in the RIGHT tool were “basic information” （[ 72.35±12.95）% ]， “background” （[ 54.55±15.40）%]，“ evidence” （[ 36.36±24.81）%]，“ recommended opinions” （[ 53.25±19.20）%]，“ review and quality assurance” [0 （0， 25.00%）]， “funding and conflict of interest statement and management” [12.50%（0，25.00%）]， and other aspects [8.33%（0， 50.00%）]. In addition， there was no statistically significant difference in the AGREE Ⅱ and RIGHT scores between the guidelines and consensuses （P＞0.05）. CONCLUSIONS The overall quality of the guidelines and consensuses included in this study is not high， with a recommended level of B or C. It is recommended that clinical decision-making prioritize referring to the relatively high-quality guideline/consensus among them. The quality of evidence in the existing traditional Chinese medicine guidelines for children with ITP needs to be improved， and there is no integrated guideline/consensus for traditional Chinese and Western medicine. It is recommended to revise or write relevant guideline/consensus according to the requirements of AGREE Ⅱ and RIGHT in various fields to guide clinical practice.

Taxifolin (TAX) is a natural compound known for its liver protection effect, but the mechanism remains unknown. Phosphorylated proteomics analyses discovered that the phosphorylation level of NDRG1 at T328 was a key event of TAX-improved liver fibrosis. We established models with NDRG1 knockout (KO) in vivo and in vitro, demonstrating that NDRG1 KO attenuated the development of hepatocyte injury, and combining NDRG1 KO and TAX administration did not result in a reduction in protection against liver injury. Cellular thermal shift assay and surface plasma resonance analysis showed that TAX directly binds to NDRG1 rather than its upstream kinase, subsequently demonstrating that TAX regulated phosphorylation of NDRG1 at T328 through binding to its C289 site. NDRG1 T328A (phosphorylated mutation) and T328E (mimic phosphorylation) in vivo and in vitro confirmed that pNDRG1_T328 exacerbates hepatocyte injury along with DNA damage, inflammatory response, and apoptosis, thereby contributing to hepatic stellate cells (HSCs) activation. In contrast, TAX can inhibit the above pathological abnormalities and block hepatocyte injury-triggered HSCs activation and fibrosis. Overall, TAX is a potent liver protection drug primarily targeting NDRG1 and inhibiting pNDRG1_T328 in hepatocytes.

Gene therapy, harnessing the power of CRISPR-Cas9 and/or DNAzyme systems, stands as a pivotal approach in cancer therapy, enabling the meticulous manipulation of genes pivotal to tumorigenesis and immunity. However, the pursuit of precise gene therapy encounters formidable hurdles. Herein, a near-infrared upconversion theranostic nanomachine is devised and tailors for CRISPR-Cas9/DNAzyme systems mediate precise gene therapy. An ingenious logic DNAzyme system consists of Chain 1 (C1)/Chain 2 (C2) and endogenous lncRNA is designed. We employ manganese modified upconversion nanoparticles for carrying ultraviolet-responsive C1-PC linker-C2 (C₂P) chain and Cas9 ribonucleoprotein (RNP), with outermost coats with hyaluronic acid. Upon reaching tumor microenvironment (TME), the released Mn²⁺ ions orchestrate a trifecta: facilitating endosomal escape, activating cGAS-STING signaling, and enabling T1-magnetic resonance imaging. Under near-infrared irradiation, Cas9 RNP/C₂P complex dissociates, releasing Cas9 RNP into the nucleus to perform gene editing of Ptpn2, while C1/C2 chains self-assemble with endogenous lncRNA to form a functional DNAzyme system, targeting PD-L1 mRNA for gene silencing. This strategy remodels the TME by activating cGAS-STING signaling and dual immune checkpoints blockade, thus realizing tumor elimination. Our theranostic nanomachine armed with the CRISPR-Cas9/DNAzyme logic systems, represents a resourceful and promising strategy for advancing cancer systemic immunotherapy and precise gene therapy.

Given the increasing concern regarding antibacterial resistance, the antimicrobial properties of naphthoquinones have recently attracted significant attention. While 1,4-naphthoquinone and its derivatives have been extensively studied, the antibacterial properties of 5,8-dihydroxy-1,4-naphthoquinone derivatives remain relatively unexplored. This study presents a comprehensive in vitro and in vivo analysis of the antibacterial activity of 35 naturally sourced and chemically synthesized derivatives of 5,8-dihydroxy-1,4-naphthoquinone. Kirby-Bauer antibiotic testing identified three compounds with activity against methicillin-resistant Staphylococcus aureus (MRSA), with one compound (PNP-02) demonstrating activity comparable to vancomycin in minimum inhibitory concentration, minimum bactericidal concentration (MBC), and time-kill assays. Microscopic and biochemical analyses revealed that PNP-02 adversely affects the cell wall and cell membrane of MRSA. Mechanistic investigations, including proteomic sequencing analyses, Western blotting, and RT-qPCR assays, indicated that PNP-02 compromises cell membrane integrity by inhibiting arginine biosynthesis and pyrimidine metabolism pathways, thereby increasing membrane permeability and inducing bacterial death. In an in vivo mouse model of skin wound healing, PNP-02 exhibited antibacterial efficacy similar to vancomycin. The compound demonstrated low toxicity to cultured human cells and in hemolysis assays and remained stable during serum incubation. These findings suggest that PNP-02 possesses promising bioactivity against MRSA and represents a potential novel antibacterial agent.
Methicillin-Resistant Staphylococcus aureus/genetics* ; Anti-Bacterial Agents/chemistry* ; Naphthoquinones/administration & dosage* ; Animals ; Microbial Sensitivity Tests ; Mice ; Humans ; Staphylococcal Infections/microbiology* ; Molecular Structure

BACKGROUND:Semaphone 3A(Sema3A)is an important neurovascular growth inhibitor.It is not clear how Sema3A is involved in the pathogenesis of discogenic low back pain.Exploring the potential mechanism of Sema3A in intervertebral disc degeneration can provide a new target and theoretical basis for the prevention and treatment of discogenic low back pain. OBJECTIVE:To explore the mechanism of interleukin-1β inhibiting the expression of Sema3A by activating the nuclear factor-κB signaling pathway to induce intervertebral disc degeneration in rats. METHODS:RT-qPCR was used to detect the expression of Sema3A mRNA in normal and degenerative human nucleus pulposus tissues.Nucleus pulposus cells of Sprague-Dawley rats were isolated,cultured,and passaged to the 3rd generation.Then,passage 3 cells were divided into three groups:the blank control group was routinely cultured for 48 hours,the degeneration group was intervened with 10 ng/mL interleukin 1β for 48 hours,and the degeneration+inhibitor group was treated by 5 μmol/L nuclear factor-κB signaling pathway-specific inhibitor BAY11-7082 for 1 hour,followed by interleukin-1β for 48 hours.At the end of the intervention,cell viability was detected by cell counting kit-8,cell apoptosis was detected by Annexin V/FITC staining,mRNA expression of cellular matrix,vascular and neural markers and Sema3A was detected by RT-qPCR,and protein expression of marker proteins,p65 and p-p65 was detected by western blot. RESULTS AND CONCLUSION:RT-qPCR assay showed that the expression of Sema3A mRNA was lower in degenerative human nucleus pulposus tissue than in normal human nucleus pulposus tissue(P＜0.05).Compared with the blank control group,the nucleus pulposus cell viability decreased and the apoptotic rate increased in the degeneration group(P＜0.05);compared with the degeneration group,the nucleus pulposus cell viability increased and the apoptotic rate decreased in the degeneration + inhibitor group(P＜0.05).Compared with the blank control group,mRNA expression of type Ⅱ collagen,polyproteoglycan,and Sema3A was decreased in the degeneration group(P＜0.05),while mRNA expression of CD31 and neurofilament 200 was increased(P＜0.05).Compared with the degeneration group,mRNA expression of type Ⅱ collagen,polyproteoglycan,and Sema3A was elevated in the degeneration+inhibitor group(P＜0.05)and mRNA expression of CD31 and neurofilament 200 decreased(P＜0.05).Compared with the blank control group,the protein expression of type Ⅱ collagen,polyproteoglycan,and Sema3A was decreased in the degeneration group(P＜0.05),and the protein expression of CD31,neurofilament protein 200,p65,and p-p65 was elevated(P＜0.05);compared with the degeneration group,the protein expression of type Ⅱ collagen,polyproteoglycan,and Sema3A was elevated in the degeneration+inhibitor group(P＜0.05),and protein expression of CD31,neurofilament 200,p65,and p-p65 was decreased(P＜0.05).To conclude,interleukin-1β does inhibit the expression of Sema3A by activating the nuclear factor-κB signaling pathway,which can also increase the degradation of extracellular matrix,promote the innervation and angiogenesis in degenerative intervertebral disc,and may be one of potential factors that contribute to intervertebral disc degeneration and discogenic low back pain.

In order to study the compounds from Glechoma longituba (Nakai) Kupr. and explore the substance bases of its dissipating blood stasis, MCI, silica gel, Sephadex LH-20, ODS column chromatography and preparative thin layer chromatography were used to isolate and purify the compounds. The structures were identified by MS, 1D NMR, and 2D NMR spectra analysis, etc. Eight triterpenoids were isolated from dichloromethane fraction of ethanol extract from G. longituba and identified as 2α,3α,16β-trihydroxy-13α,27-cyclours-11-en-28-oic acid (1), 28-norurs-12-ene-3β,17β-diol (2), 28-norolean-12-ene-3β,17β-diol (3), oleanonic acid (4), 3β,13β-dihydroxyurs-11-en-28-oic acid (5), uvaol (6), oleanolic acid (7), ursolic acid (8). Compound 1 is a new hexacyclic triterpene with 13α,27-cyclopropane structure, named euscaphic acid H; compounds 2 and 3 were isolated from Lamiaceae for the first time while compounds 4 and 5 were isolated from this genus. The in vitro anticoagulant activity of triterpenoids was evaluated by four coagulation indexes (activated partial thromboplastin time, APTT; thrombin time, TT; prothrombin time, PT; fibrinogen, FIB). Among them, compounds 1 and 6 showed obvious anticoagulant effects.

[Objective] To analyze the changes of platelet extracellular vesicles (PEVs) content in apheresis at different storage periods, track the PEVs and transfusion efficacy of patients, and explore the mechanism of PEVs in relation to platelet storage damage and transfusion efficacy. [Methods] From November 2022 to April 2024, a total of 85 apheresis platelets samples were collected at Jiading District Blood Station. Flow cytometry was used to detect the content of PEVs on the day of collection and before transfusion. The changes in the content of PEVs stored for 1 to 5 days were observed. Patients with acute leukemia were selected as transfusion recipients, and their content of platelet count and PEVs before transfusion and within 24 hours after transfusion, and the efficacy were tracked to explore the mechanism of PEVs in relation to storage lesion and transfusion efficacy. [Results] The content of PEVs before platelet infusion (8.73±4.84) was significantly higher than that on the day of platelet collection (5.11±3.33), and the difference was statistically significant (P<0.01). The increase in the content of PEVs in storage for 2 to 5 days was 2.55±1.38, 3.49±2.63, 3.86±3.55 and 4.50±3.91, with statistically significant difference (P<0.05). The PEVs content in patients after blood transfusion was positively correlated with that before transfusion and that in apheresis platelet bags during transfusion (P<0.001) A total of 85 cases of apheresis platelet transfusion were conducted, with 61 effective transfusions having a CCI value of (13.43±4.70), and 24 transfusion refractoriness cases having a CCI value of (2.27±3.67), showing a statistically significant difference (P<0.001). The transfusion effectiveness rates for patients receiving different storage periods of apheresis platelets of 2 to 5 days were 88.89%, 68.42%, 68.18% and 57.14% respectively, with corresponding CCI values of 11.18±6.10, 10.43±6.77, 9.53±6.75 and 9.48±8.86, and there was no significant difference between CCI groups (P>0.05). There was no significant correlation between the PEVs content before and after transfusion and the efficacy of CCI (P>0.05). There was no significant correlation between the PEVs content before apheresis platelet transfusion and the efficacy of CCI (P>0.05). [Conclusion] The content of PEVs increased with the prolongation of storage time, which could be used as a potential blood quality evaluation and monitoring index during the storage period, but it was not significantly correlated with the efficacy of CCI in transfusion.

Hip fracture in the elderly is characterized by high incidence, high disability rate, and high mortality and has been recognized as a public health issue threatening their health. Surgery is the preferred choice for the treatment of elderly patients with hip fracture. However, lower extremity deep venous thrombosis (DVT) has an extremely high incidence rate during the perioperative period, and may significantly increase the risk of patients′ death once it progresses to pulmonary embolism. In response to this issue, the clinical guidelines and expert consensuses all emphasize active application of comprehensive preventive measures, including basic prevention, physical prevention, and pharmacological prevention. In this prevention system, basic prevention is the basis of physical and pharmacological prevention. However,there is a lack of unified and definite recommendations for basic preventive measures in clinical practice. To this end, the Orthopedic Nursing Professional Committee of the Chinese Nursing Association and Nursing Department of the Orthopedic Branch of the China International Exchange and Promotive Association for Medical and Health Care organized relevant nursing experts to formulate Expert consensus on perioperative basic prevention for lower extremity deep venous thrombosis in elderly patients with hip fracture ( version 2024) . A total of 10 recommendations were proposed, aiming to standardize the basic preventive measures for lower extremity DVT in elderly patients with hip fractures during the perioperative period and promote their subsequent rehabilitation.

Objective To establish a bone knife deformation prediction model for periacetabular osteotomy and quickly and accurately predict bone knife deformation.Methods A finite element numerical model of a pelvic bone knife containing both cortical and cancellous bones was established,and the correlation between nodal strain and deformation was analyzed.The strains of 5 nodes with the strongest integrated correlation were selected as the inputs,and the displacement increments of the nodes on the blade part were established as the outputs.After training the model with the dataset,a deep learning neural network regression model based on the finite element dataset was used to establish a prediction model for the strain deformation of the bone knife.After the model prediction was completed,a binocular visual localization system was used to determine the spatially accurate position of the bone knife during the osteotomy procedure as a means of intraoperative tracking of the bone knife.Results The R2 value of the prediction model was 0.987 81 and the average deformation error after discretizing the bone knife into nodes was 0.07 mm.The prediction model quickly and accurately acquired bone knife deformation and showed great potential for reducing PAO surgical accidents.Conclusions The bone knife deformation prediction model developed in this study rapidly predicted bone knife deformation from strain information.Thus,it can avoid injuring tissues,such as nerves and blood vessels around the tissue,reduce the difficulty and risk of periacetabular osteotomy,and provide theoretical support for clinical application.