OBJECTIVE To investigate the targeting effect of folic acid-modified crebanine nanoparticles （FA-Cre@PEG- PLGA NPs， hereinafter referred to as “NPs”） combined with ultrasound irradiation on M109 cells in vitro and in vivo after administration， and explore the anti-tumor mechanism. METHODS CCK-8 assay was used to detect the inhibitory effect of NPs combined with ultrasound irradiation on the proliferation of M109 cells， and the best ultrasound time was selected. Using human lung cancer A549 cells as a control， the targeting of NPs combined with ultrasound irradiation to M109 cells was evaluated by free folic acid blocking assay and cell uptake assay. The effects of NPs combined with ultrasound irradiation on the migration， invasion， apoptosis， cell cycle and reactive oxygen species （ROS） levels of M109 cells were detected by cell scratch test， Transwell chamber test and flow cytometry at 1 h after 958401536@qq.com administration； the changes of mitochondrial membrane potential （MMP） were observed by fluorescence inverted microscope. A mouse subcutaneous tumor model of M109 cells was constructed， and the in vivo tumor targeting of NPs combined with ultrasound irradiation was investigated by small animal in vivo imaging technology. RESULTS NPs combined with ultrasound irradiation could significantly inhibit the proliferation of M109 cells， and the optimal ultrasound time was 1 h after administration. The free folic acid could antagonize the inhibitory effect of NPs on the proliferation of M109 cells， and combined with ultrasound irradiation could partially reverse this antagonism. Compared with A549 cells， the uptake rate of NPs in M109 cells was significantly higher （P＜0.01）， and ultrasound irradiation could promote cellular uptake. NPs combined with ultrasound irradiation could inhibit the migration and invasion of M109 cells and block the cell cycle in the G0/G1 and G2/M phases. Compared with control group， the apoptosis rate of M109 cells and ROS level were increased significantly （P＜0.01）， while the MMP decreased significantly （P＜0.01） in the different concentration （100， 200， 300 μg/mL） groups of M109 cells. Compared with the mice in non-ultrasound group， the fluorescence intensity and tumor-targeting index of the tumor site in the 0 h ultrasound group were significantly enhanced （P＜0.05 or P＜0.01）. CONCLUSIONS NPs combined with ultrasound irradiation have a strong targeting effect on M109 cells in vitro and in vivo， the anti-tumor mechanism includes inhibiting cell migration and invasion， blocking cell cycle， and inducing apoptosis.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective To investigate the effect of the anti-oxidized low density lipoprotein(ox-LDL)IgM subclass antibody 3A6 on the atherosclerotic plaque formation in ApoE-/-mice.Methods The ApoE-/-mice were randomly divided into three groups:control group(intraperitoneal injection of normal saline),5G8group(intraperitoneal injection of nature anti-LDL IgM subclass antibody)and 3A6group(intraperitoneal injection of natural anti-ox-LDL IgM subclass antibody),with 6mice in each group.The body weight of mice and the level of serum lipid in each group were measured after 12 weeks of high-fat and high-cholesterol diet.hematoxylin-eosin staining was used to analyze the pathological changes and plaque area of the aorta in mice.Results There was no significant difference in body weight,serum lipid level and ox-LDL level among the three groups(P=0.087).Compared with the control group,the area of atherosclerotic plaque was significantly decreased in the 3A6group(P=0.023).Conclusion The natural IgM subclass antibody 3A6 can effectively inhibit the formation of atherosclerotic plaque in ApoE-/-mice.

Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
Animals ; Haplorhini ; Axons ; Motor Neurons ; Interneurons ; Macaca ; Dependovirus/genetics* ; Genetic Vectors

Objective:To clarify the application effect of information-guided enteral nutrition-associated diarrhea (ENAD) management process in patients with chronic obstructive pulmonary disease (COPD) undergoing non-invasive assisted ventilation.Methods:A mixed cohort study of pre- and post-control was conducted. Thirty-nine patients with COPD who were admitted to the emergency intensive care unit (ICU) of Huzhou First People's Hospital from July 1, 2021 to July 31, 2022 were enrolled. Taking the completion of the software development of ENAD management software for critically ill patients on January 28, 2022 as the time node, 20 patients admitted from July 1, 2021 to January 28, 2022 were set as the control group, and 19 patients admitted from January 29 to July 31, 2022 were set as the observation group. The two groups of patients received the same enteral nutrition support treatment, and the control group implemented the conventional ENAD treatment process with enteral nutrition intolerance disposal process as the core. On the basis of the control group, the observation group implemented the information-guided ENAD treatment process, and the system software actively captured the information of ENAD patients and reminded the medical team to improve the patient's diarrhea-related examination and provide alternative treatment plans. The duration of antidiarrhea, feeding interruption rate, and energy and protein intake, blood biochemical indexes, incidence of abnormal blood electrolyte metabolism, daily continuous non-invasive assisted ventilation and endotracheal intubation after 7 days of targeted diarrhea intervention were compared between the two groups.Results:Except for the basal pulse rate, there were no significant differences in gender distribution, age, and vital signs, basic nutritional status, arterial blood gas analysis and blood biochemistry at admission between the two groups, indicating comparability between the two groups. When ENAD occurred, the patients in the observation group obtained earlier cessation of diarrhea than those in the control group [days: 3.00 (2.00, 3.25) vs. 4.00 (3.00, 5.00), P < 0.01], and the feeding interruption rate was significantly lower than that in the control group [10.53% (2/19) vs. 65.00% (13/20), P < 0.01]. After 7 days of diarrhea intervention, the energy intake of the observation group was significantly higher than that of the control group [kJ·kg -1·d -1: 66.28 (43.34, 70.36) vs. 47.88 (34.60, 52.32), P < 0.01], the levels of hemoglobin (Hb), albumin (Alb) and serum prealbumin (PAB) were significantly higher than those in the control group [Hb (g/L): 119.79±10.04 vs. 110.20±7.75, Alb (g/L): 36.00 (33.75, 37.25) vs. 31.00 (30.00, 33.00), PAB (mg/L): 155.79±25.78 vs. 140.95±14.97, all P < 0.05], the daily continuous non-invasive assisted ventilation duration was significantly shorter than that of the control group [hours: 14 (12, 16) vs. 16 (14, 18), P < 0.01], and the arterial partial pressure of carbon dioxide (PaCO 2) was significantly lower than that of the control group [mmHg (1 mmHg ≈ 0.133 kPa): 66.00 (62.00, 70.00) vs. 68.00 (67.50, 70.05), P < 0.05]. However, there were no significant differences in protein intake, incidence of abnormal electrolyte metabolism, and incidence of endotracheal intubation due to acute respiratory failure between the two groups. Conclusion:The information-guided ENAD treatment process can enable the COPD patients undergoing continuous non-invasive assisted ventilation who experience ENAD to receive earlier cessation of diarrhea, and improve the protein energy metabolism and respiratory function of the patients.

Objective To investigate the clinical efficacy of tiotropium bromide in the treatment of patients with acute exacerbation of chronic obstructive pulmonary disease(AECOPD)complicated with acute respiratory failure.Methods A total of 106 patients with AECOPD complicated with acute respiratory failure admitted to the First People's Hospital of Huzhou from March 2019 to March 2022 were selected and divided into observation group and control group according to random number table method,with 53 cases in each group.Both groups were given routine treatment.The control group was treated with noninvasive biphasic positive airway pressure ventilation.The observation group was treated with tiotropium bromide on the basis of control group.Both groups were treated for 14 days.The clinical efficacy,lung function,blood gas index,inflammatory factors and adverse reactions were compared between two groups.Results The total effective rate of observation group was significantly higher than that of control group(94.34%vs.79.25%,χ2=5.267,P=0.023).After treatment,forced expiratory volume in one second(FEV1),FEV1/forced vital capacity,peak expiratory flow,pH,percutaneous arterial oxygen saturation and arterial partial pressure of oxygen in observation group were significantly higher than those in control group(P<0.05),while arterial partial pressure of carbon dioxide,interleukin-6,interleukin-8,tumor necrosis factor-α and hypersensitive C-reaction protein were lower than those in control group(P<0.05).There were no significant differences in the incidence of adverse drug reactions and ventilator adverse reactions between two groups(P>0.05).Conclusion Titropium bromide in the treatment of AECOPD complicated with acute respiratory failure can significantly improve the curative effect,improve pulmonary function and arterial blood gas,but attention should be paid to the timely treatment of adverse reactions of dry mouth.

OBJECTIVES: To investigate the inhibitory effect of GSK484, a PAD4 inhibitor, on H3Cit expression following sepsis and its effects for improving sepsis-induced endothelial dysfunction. METHODS: Eighteen C57BL/6 mice were randomized into sham-operated group, sepsis model group and GSK484 treatment group (n=6), and in the latter two groups, models of sepsis were established by cecal ligation and puncture (CLP). The mice in GSK484 treatment group were given an intraperitoneal injection of GSK484 (4 mg/kg) on the second day following the surgery. Twenty-four hours after the injection, the mice were euthanized for measurement of serum levels of VEGF, ESM-1, IL-6 and IL-1β using ELISA. Lung tissue pathology was observed with HE staining, and pulmonary expressions of F-actin, VE-cadherin, ZO-1 and H3Cit proteins were detected using immunofluorescence staining and Western blotting. In primary cultured of mouse lung microvascular endothelial cells, the effect of stimulation with LPS (10 μg/mL) for 24 h on tube formation, proliferation, apoptosis and expressions of VEGF, ESM-1, IL-6 and IL-1β were assessed using CCK-8 assay, flow cytometry and ELISA. RESULTS: Compared to the sham-operated mice, the septic mice exhibited significant lung tissue pathologies characterized by vascular congestion, alveolar rupture, edema, and neutrophil infiltration. Serum levels of IL-6, IL-1β, VEGF, and ESM-1 were elevated, pulmonary expressions of F-actin, VE-cadherin, and ZO-1 were decreased, and H3Cit expression was increased significantly in the septic mice. GSK484 treatment effectively mitigated these changes in the septic mice. The LPS-stimulated endothelial cells showed increased productions of IL-6, IL-1β, VEGF and ESM-1, which were significantly reduced after treatment with 2.5 μmol/L GSK484. CONCLUSIONS GSK484 treatment effectively suppresses H3Cit expression in septic mice to ameliorate sepsis-induced endothelial dysfunction.
Animals ; Sepsis/drug therapy* ; Mice ; Mice, Inbred C57BL ; Lung Injury/drug therapy* ; Endothelial Cells/drug effects* ; Interleukin-6/metabolism* ; Protein-Arginine Deiminase Type 4/metabolism* ; Lung/metabolism* ; Vascular Endothelial Growth Factor A/metabolism* ; Interleukin-1beta/metabolism* ; Disease Models, Animal ; Cadherins/metabolism* ; Apoptosis/drug effects* ; Imidazoles ; Antigens, CD

This study was performed to explore the differential genes and functions of double-negative T cells in peripheral blood of patients with colorectal cancer.Two colorectal cancer patients and two healthy physical examiners were selected,and peripheral blood double-negative T cells were firstly sorted by flow cytometry,and then sequencing data were obtained using single cell full-length transcriptome(SMART-seq2)sequencing technology to screen differentially expressed genes.The screened differentially expressed genes were subjected to Gene Ontology Enrichment(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.The STRING database was used to construct the protein interaction network and identify key genes by Cytoscape software;RT-qPCR was used to verify the differential expression genes in DNT cells.Compared with healthy subjects,there were 1 276 peripheral blood double-negative T-cell differential genes in colorectal cancer patients,including 141 up-regulated genes and 1 135 down-regulated genes.GO analysis showed that the differential genes were mainly involved in biological functions such as methylation,metabolic processes and transferase activity;KEGG pathway analysis showed that the differential genes were mainly involved in signaling pathways such as autophagy,P53 signaling pathway and phosphatidylinositol metabolism.The protein interaction network contains 1 154 nodes and 1 022 edges,in addition,10 hub genes were identified:PIK3C3,WIPI1,ATG101,PIK3R4,DDX10,RBM28,SDAD1,ATG16L1,UVRAG,ATG7.RT-qPCR validated 10 differentially expressed genes,of which 7 differentially expressed genes showed trends consistent with sequencing results,and 3 genes showed expression inconsistent with sequencing results.DNT cells may be involved in the development of colorectal cancer through methylation,P53 signaling pathway and autophagy,and at the same time,DNT cells may inhibit the development of colorectal cancer through the regulation of genes.This study provides a theoretical basis for further investigation of the function of DNT cells in malignant tumors.

Breast cancer has overtaken lung cancer as the most common malignancy in women. Although the early diagnostic rate has continuously improved, recurrence and metastasis remain a problem to be solved. Therefore, scientists should search for effective prognostic markers for breast cancer patients and adopt individualized programs for different patients. Studies have shown that cancer prognosis, to a certain extent, is related to the nutrition inflammation index. Preoperative prognostic nutritional index (PNI) and controlled nutritional status (CONUT) are indicators that comprehensively reflect the nutritional level and inflammatory state of patients, respectively. Different from other cancers, the incidence of breast cancer is related to nutritional status, and an extremely high or low score is not conducive to the prognosis of breast cancer patients. This paper reviews the research progress of PNI and CONUT in breast cancer.