This study aims to explore the mechanism of lung and gut protection by Tanreqing Capsules on the mice infected with influenza virus based on "the lung-gut axis". A total of 110 C57BL/6J mice were randomized into control group, model group, oseltamivir group, and low-and high-dose Tanreqing Capsules groups. Ten mice in each group underwent body weight protection experiments, and the remaining 12 mice underwent experiments for mechanism exploration. Mice were infected with influenza virus A/Puerto Rico/08/1934(PR8) via nasal inhalation for the modeling. The lung tissue was collected on day 3 after gavage, and the lung tissue, colon tissue, and feces were collected on day 7 after gavage for subsequent testing. The results showed that Tanreqing Capsules alleviated the body weight reduction and increased the survival rate caused by PR8 infection. Compared with model group, Tanreqing Capsules can alleviate the lung injury by reducing the lung index, alleviating inflammation and edema in the lung tissue, down-regulating viral gene expression at the late stage of infection, reducing the percentage of neutrophils, and increasing the percentage of T cells. Tanreqing Capsules relieved the gut injury by restoring the colon length, increasing intestinal lumen mucin secretion, alleviating intestinal inflammation, and reducing goblet cell destruction. The gut microbiota analysis showed that Tanreqing Capsules increased species diversity compared with model group. At the phylum level, Tanreqing Capsules significantly increased the abundance of Firmicutes and Actinobacteria, while reducing the abundance of Bacteroidota and Proteobacteria to maintain gut microbiota balance. At the genus level, Tanreqing Capsules significantly increased the abundance of unclassified＿f＿Lachnospiraceae while reducing the abundance of Bacteroides, Eubacterium, and Phocaeicola to maintain gut microbiota balance. In conclusion, Tanreqing Capsules can alleviate mouse lung and gut injury caused by influenza virus infection and restore the balance of gut microbiota. Treating influenza from the lung and gut can provide new ideas for clinical practice.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Lung/metabolism* ; Mice, Inbred C57BL ; Capsules ; Orthomyxoviridae Infections/virology* ; Gastrointestinal Microbiome/drug effects* ; Male ; Humans ; Female ; Influenza A virus/physiology* ; Influenza, Human/virology*

The 2-(2-phenylethyl)chromones were separated from agarwood of Aquilaria agallocha Roxb. and their anti-KRAS mutant non-small cell lung cancer (NSCLC) activities were evaluated. 2-(2-Phenyethyl)chromones in agarwood were separated and purified by silica gel, RP-18 reverse phase silica gel, MCI gel CH20P, Diol, and semi preparative HPLC chromatography techniques, while the structures of the compounds were identified by extensive spectroscopic analysis, such as 1D and 2D NMR and ESI-MS. The cell counting kit 8 (CCK-8) assay was used to screen the anti-tumor activity of the isolated monomeric compounds on three KRAS-mutant NSCLC cells. The cell proliferation, cloning formation, adhesion ability, and cell cycle arrest activity of compound 8 were analyzed. Molecular docking and Western blot experiments were used to study the mechanism of compound 8. The in vivo anti-tumor activity of the compound 8 was evaluated by zebrafish cell derived xenograft (CDX) model. Nineteen known 2-(2-phenylethyl)chromones were isolated from the ethyl acetate extract of agarwood. On A549 (KRAS G12S) cells, compounds 8, 10, and 19 showed good inhibitory activity, on H23 (KRAS G12C) cells, compounds 7, 8, and 19 showed good inhibitory activity, and on H358 (KRAS G12C) cells, compounds 8, 10, and 16 showed good inhibitory activity. Compound 8 had the best inhibitory activity in all three cell lines. It effectively inhibited cell proliferation, clone formation, adhesion ability, and arrested the H23 cell cycle at G2/M phase. Compound 8 could also inhibit the expression of c-Met and its downstream signaling pathways, effectively inhibiting tumor growth in zebrafish CDX model. In conclusion, among the nineteen known 2-(2-phenylethyl)chromones, compound 8 had the best activity and significantly inhibited the proliferation of KRAS-mutant NSCLC cells by arresting the cells in the G2/M phase.

Objective To investigate the effect and mechanism of shikonin on the proliferation,migration,invasion and apoptosis of human gastric cancer MGC803 cells.Methods The MGC803 cells in the logarithmic growth phase were randomly divided into the blank control group,shikonin group,shikonin+insulin-like growth factor-1(IGF-1)group,and shikonin+LY294002 group.Cells in the blank control group were cultured in drug-free medium,cells in the shikonin group were cultured in the medium containing shikonin with a final concentration of 10 μmol·L-1,cells in the shikonin+IGF-1 group were cultured in the medium containing shikonin with a final concentration of 10 μmol·L-1 and IGF-1 with a final concentration of 10 μmol·L-1,and cells in the shikonin+LY294002 group were cultured in the medium containing shikonin with a final concentration of 10 μmol·L-1 and LY294002 with a final concentration of 30 μmol·L-1.After 24 h of culture,the cell proliferation was detected by cell counting kit-8,the cell apoptosis was detected by flow cytometry,the cell migration was detected by scratch assay,and the cell invasion was detected by Transwell assay.The expression levels of B cell lymphoma-2(Bcl-2),Bcl-2 related X protein(Bax),cytochrome C(Cyt C),cleaved caspase-3,cleaved caspase-9,phosphoinositide 3 kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(PKB),and phosphorylated PKB(p-PKB)proteins were measured by using Western blot.Results The MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin group were significantly higher than those in the blank control group(P＜0.05);the MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin+IGF-1 group were significantly lower than those in the shikonin group(P＜0.05);and the MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin+LY294002 group were significantly higher than those in the shikonin group(P＜0.05).The MGC803 cell scratch healing rate and the number of invasive cells in the shikonin group were significantly lower than those in the blank control group(P＜0.05);the MGC803 cell scratch healing rate and the number of invasive cells in the shikonin+IGF-1 group were significantly higher than those in the shikonin group(P＜0.05);and the MGC803 cell scratch healing rate and the number of invasive cells in the shikonin+LY294002 group were significantly lower than those in the shikonin group(P＜0.05).The relative expression level of Bcl-2 protein in MGC803 cells in the shikonin group was significantly lower than that in the blank control group(P＜0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly higher than those in the blank control group(P＜0.05);the relative expression level of Bcl-2 protein in MGC803 cells in the shikonin+IGF-1 group was significantly higher than that in the shikonin group(P＜0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly lower than those in the shikonin group(P＜0.05);and the relative expression level of Bcl-2 protein in MGC803 cells in the shikonin+LY294002 group was significantly lower than that in the shikonin group(P＜0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly higher than those in the shikonin group(P＜0.05).The relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin group were significantly lower than those in the blank control group(P＜0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin group and the blank control group(P＞0.05);the relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin+IGF-1 group were significantly higher than those in the shikonin group(P＜0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin+IGF-1 group and the shikonin group(P＞0.05);and the relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin+LY294002 group were significantly lower than those in the shikonin group(P＜0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin+LY294002 group and the shikonin group(P＞0.05).Conclusion Shikonin can inhibit the proliferation,migration and invasion and promote the apoptosis of human gastric cancer MGC803 cells,which may be related to its inhibition of the PI3K/PKB signaling pathway.

Objective To investigate the effects of zalcitabine(ddC),a mitochondrial DNA polymerase γ(POLG)inhibitor,on the migration,invasion,and to preliminarily explore mitochondrial biogenesis of human tri-ple-negative breast cancer MDA-MB-231 cells.Methods The effect of ddC on cell viability was detected using the MTT assay.The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell in-vasion assays.Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit.The protein expression of POLG,NADH dehydrogenase subunit Ⅰ(NADH1),NADH dehydrogenase subunit Ⅱ(NADH2),ATP synthase subunit 6(ATPase6),cytochrome c oxidase subunit Ⅰ(COX-1)and cytochrome c ox-idase subunit Ⅲ(COX-3)were determined using Western blot.The POLG mRNA level and mtDNA copy number were determined using qPCR.The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit.MDA-MB-231 cells were transfected with pcDNA3.1-EG-FP-POLG plasmids to overexpress POLG.The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells.Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P＜0.01).ddC inhibited cell viability in a dose-dependent man-ner.ddC inhibited the migration(P＜0.01)and invasion(P＜0.01)of MDA-MB-231 cells;however,it dis-played no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A)at the same concentration.ddC downregulated the protein(P＜0.01)and mRNA(P＜0.01)levels of POLG,reduced mtD-NA copy number(P＜0.01)and downregulated mtDNA-coded NADH1,NADH2,ATPase6,COX-1 and COX-3 protein expression(P＜0.01)in MDA-MB-231 cells.Furthermore ddC inhibited mitochondrial content(P＜0.01)and ATP(P＜0.01)levels in MDA-MB-231 cells.POLG overexpression increased the migration(P＜0.05)and invasion(P＜0.05)abilities of MDA-MB-231 cells,while ddC did not significantly inhibit the migra-tion and invasion abilities of MDA-MB-231 cells overexpressing POLG.Conclusion ddC downregulates POLG ex-pression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels,thereby inhibiting the migra-tion and invasion of MDA-MB-231 cells.

Objective To compare the incidence of superior facet joint violation in patients with different preoperative diagnoses in posterior lumbar interbody fusion.Methods The clinical data of 320 patients who underwent posterior lumbar interbody fusion from January 2018 to December 2021 in our hospital were retrospectively analyzed,and the preoperative diagnoses were including the lumbar disc herniation/lumbar spinal canal stenosis,degenerative lumbar spondylolisthesis,and isthmic lumbar spondylolisthesis.The superior facet joint violation was graded based on the postoperative lumbar CT,the incidences of superior facet joint intrusion of patients with different preoperative diagnosis were compared,and the correlations between relevant parameters(including the apical vertebrae of fixation segments,facet joint angle,depth of lamina,introversion angle of pedicle screw,axial diameter of facet joint,and slippage length of vertebrae)with superior facet joint violation and preoperative diagnoses were investigated.Results The incidence of superior facet joint violation in the patients with isthmic lumbar spondylolisthesis(57.4%)was higher than that in the patients with lumbar disc herniation/lumbar spinal canal stenosis(41.2%)and degenerative lumbar spondylolisthesis(35.9%),with statistically significant differences(P<0.05).The apical vertebrae of fixation segments,facet joint angle,axial diameter of facet joint and introversion angle of pedicle screw were related with the occurrence of superior facet joint violation(P<0.05).There were statistically significant differences in the facet joint angle,axial diameter of facet joint and apical vertebrae of fixation segments among patients with different preoperative diagnoses(P<0.05).Conclusion Patients with isthmiclumbar spondylolisthesis are more likely to occur superior facet joint violation than patients with lumbar disc herniation/lumbar spinal canal stenosis and degenerative lumbar spondylolisthesis.

Objective:To observe the clinical efficacy and action mechanism of Jujing Decoction(JJD)in the treatment of as-thenoteratozoospermia(ATZ)by comparing JJD with combined administration of the antioxidant stress drug and sperm energy metabo-lism agent.Methods:According to the inclusion criteria,we enrolled 67 male patients with ATZ in this randomized controlled clini-cal study and treated them by oral administration of JJD(the JJD group,n=34)or natural vitamin E combined with L-carnitine solu-tion(the positive control group,n=33),both for 12 weeks.We collected the semen parameters,sperm DNA fragmentation index(DFI),sperm mitochondrial membrane potential(MMP),seminal plasma reactive oxygen species(ROS)and superoxide dismutase(SOD)levels from the patients,observed the ultrastructure of sperm mitochondria under the transmission electron microscope(TEM)before and after treatment,and analyzed the clinical efficacy and action mechanism of JJD by comparing the data obtained between the two groups.Results:Treatment and follow-up were completed in 60 of the cases,30 in the JJD and 30 in the positive control group.The total rate of clinical effectiveness was significantly higher in the JJD than in the positive control group(76.8％vs 43.3％,P＜0.05).Compared with the baseline,the percentages of progressively motile sperm(PMS)and morphologically normal sperm(MNS),DFI and MMP were significantly improved(P＜0.05),the level of seminal plasma ROS decreased(P＞0.05),and that of SOD re-markably increased(P＜0.05)after treatment with JJD;PMS,MNS,DFI and MMP were also improved(P＞0.05),seminal plas-ma ROS decreased(P＞0.05)and SOD increased(P＜0.05)in the positive controls after medication.In comparison with the posi-tive controls,the patients treated with JJD showed even more significant improvement in PMS([29.37±14.56]％vs[42.68±15.86]％,P＜0.05),MNS([1.84±1.32]％vs[3.66±1.72％]％,P＜0.05),DFI([32.66±5.23]％vs[16.61±4.20]％,P＜0.05)and MMP([46.47±9.48]％vs[61.79±8.61]％,P＜0.05),ROS([7.08±0.51]vs[5.06±0.52]μmol/L,P＞0.05),and SOD([100.65±10.59]vs[139.05±14.71]U/ml,P＜0.05).TEM revealed significantly improved ultrastructure of sperm mitochondria after treatment with JJD.No serious adverse reactions were observed in either group dur-ing follow-up.Conclusion:JJD,superior to natural vitamin E and L-carnitine oral solution,can safely and effectively increase the percentages of PMS and MNS,MMP and the level of seminal plasma SOD,reduce sperm DFI and seminal plasma ROS,and improve the ultrastructure of sperm mitochondria in patients with ATZ.The underlying mechanism of action may be related to its ability of im-proving the structure and function of sperm mitochondria via antioxidant stress.

As artificial intelligence technology rapidly advances, its deployment within the medical sector presents substantial ethical challenges. Consequently, it becomes crucial to create a standardized, transparent, and secure framework for processing medical data. This includes setting the ethical boundaries for medical artificial intelligence and safeguarding both patient rights and data integrity. This consensus governs every facet of medical data handling through artificial intelligence, encompassing data gathering, processing, storage, transmission, utilization, and sharing. Its purpose is to ensure the management of medical data adheres to ethical standards and legal requirements, while safeguarding patient privacy and data security. Concurrently, the principles of compliance with the law, patient privacy respect, patient interest protection, and safety and reliability are underscored. Key issues such as informed consent, data usage, intellectual property protection, conflict of interest, and benefit sharing are examined in depth. The enactment of this expert consensus is intended to foster the profound integration and sustainable advancement of artificial intelligence within the medical domain, while simultaneously ensuring that artificial intelligence adheres strictly to the relevant ethical norms and legal frameworks during the processing of medical data.
Artificial Intelligence/legislation & jurisprudence* ; Humans ; Consensus ; Computer Security/standards* ; Confidentiality/ethics* ; Informed Consent/ethics*

OBJECTIVE: To explore whether the protein Deglycase protein 1 (DJ1) can ameliorate Alzheimer's disease (AD)-like pathology in Amyloid Precursor Protein/Presenilin 1 (APP/PS1) double transgenic mice and its possible mechanism to provide a theoretical basis for exploring the pathogenesis of AD. METHODS: Adeno-associated viral vectors (AAV) of DJ1-overexpression or DJ1-knockdown were injected into the hippocampus of 7-month-old APP/PS1 mice to construct models of overexpression or knockdown. Mice were divided into the AD model control group (MC), AAV vector control group (NC), DJ1-overexpression group (DJ1 ⁺), and DJ1-knockdown group (DJ1 ^-). After 21 days, the Morris water maze test, immunohistochemistry, immunofluorescence, and western blotting were used to evaluate the effects of DJ1 on mice. RESULTS: DJ1 ⁺ overexpression decreased the latency and increased the number of platform traversals in the water maze test. DJ1 ^- cells were cured and atrophied, and the intercellular structure was relaxed; the number of age spots and the expression of AD-related proteins were significantly increased. DJ1 ⁺ increased the protein expression of Nuclear factor erythroid 2-related factor 2 (NRF2), heme oxygenase-1 (HO-1), light chain 3 (LC3), phosphorylated AMPK (p-AMPK), and B cell lymphoma-2 (BCL-2), as well as the antioxidant levels of total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC), and Glutathione peroxidase (GSH-PX), while decreasing the levels of Kelch-like hydrates-associated protein 1 (Keap1), mammalian target of rapamycin (mTOR), p62/sequestosome1 (p62/SQSTM1), Caspase3, and malondialdehyde (MDA). CONCLUSION DJ1-overexpression can ameliorate learning, memory, and AD-like pathology in APP/PS1 mice, which may be related to the activation of the NRF2/HO-1 and AMPK/mTOR pathways by DJ1.
Animals ; Mice ; Alzheimer Disease/therapy* ; AMP-Activated Protein Kinases/metabolism* ; Amyloid beta-Protein Precursor/metabolism* ; Antioxidants/metabolism* ; Disease Models, Animal ; Hippocampus/metabolism* ; Kelch-Like ECH-Associated Protein 1/metabolism* ; Mammals/metabolism* ; Mice, Inbred C57BL ; Mice, Transgenic ; NF-E2-Related Factor 2/metabolism* ; Presenilin-1/metabolism* ; TOR Serine-Threonine Kinases/metabolism*

Objective: To explore the pathogenic mechanism of the miR-340/high mobility group box 1 (HMGB1) axis in the formation of liver fibrosis. Methods: A rat liver fibrosis model was established by injecting CCl(4) intraperitoneally. miRNAs targeting and validating HMGB1 were selected with gene microarrays after screening the differentially expressed miRNAs in rats with normal and hepatic fibrosis. The effect of miRNA expressional changes on HMGB1 levels was detected by qPCR. Dual luciferase gene reporter assays (LUC) was used to verify the targeting relationship between miR-340 and HMGB1. The proliferative activity of the hepatic stellate cell line HSC-T6 was detected by thiazolyl blue tetrazolium bromide (MTT) assay after co-transfection of miRNA mimics and HMGB1 overexpression vector, and the expression of extracellular matrix (ECM) proteins type I collagen and α-smooth muscle actin (SMA) was detected by western blot. Statistical analysis was performed by analysis of variance and the LSD-t test. Results: Hematoxylin-eosin and Masson staining results showed that the rat model of liver fibrosis was successfully established. Gene microarray analysis and bioinformatics prediction had detected eight miRNAs possibly targeting HMGB1, and animal model validation had detected miR-340. qPCR detection results showed that miR-340 had inhibited the expression of HMGB1, and a luciferase complementation assay suggested that miR-340 had targeted HMGB1. Functional experiments results showed that HMGB1 overexpression had enhanced cell proliferation activity and the expression of type I collagen and α-SMA, while miR-340 mimics had not only inhibited cell proliferation activity and the expression of HMGB1, type I collagen, and α-SMA, but also partially reversed the promoting effect of HMGB1 on cell proliferation and ECM synthesis. Conclusion: miR-340 targets HMGB1 to inhibit the proliferation and ECM deposition in hepatic stellate cells and plays a protective role during the process of liver fibrosis.
Animals ; Rats ; Cell Proliferation ; Collagen Type I/metabolism* ; Fibrosis ; Hepatic Stellate Cells ; HMGB1 Protein/genetics* ; Liver Cirrhosis/pathology* ; MicroRNAs/metabolism*