Objective To investigate the gene expression and regulatory mechanisms of mouse embryonic fibroblasts (MEFs) under inflammatory conditions, aiming to elucidate the role of MEFs in inflammatory responses and provide a foundation for discovering anti-inflammatory drugs that act by modulating MEF function. Methods MEFs cultured in vitro were divided into the following groups: lipopolysaccharides (LPS)-treated group, inflammatory conditioned medium (CM)-treated group, and control group, which were treated with LPS, CM, and equal volume solvent, respectively. Transcriptome sequencing was used to analyze the effects of two stimuli on gene expression profile of MEFs. Real time fluorescence quantitative PCR (RT-qPCR) was employed to verify the transcription levels of highly expressed genes of MEFs induced by CM. ELISA was performed to determine the concentrations of cytokines in cell supernatants. Finally, the regulatory effects of CM on the activation of signaling pathways in MEFs were analyzed by immunoblotting. Results Transcriptome analysis showed that both LPS and CM induced the transcription of a large number of genes in MEFs. Compared with LPS, CM potentiated the mRNA transcription of some acute phase proteins, inflammatory cytokines, chemokines, matrix metalloproteinases (MMP), prostaglandin synthetases, and colony-stimulating factors. The transcriptome analysis was verified by RT-qPCR. The results of ELISA showed that CM treatment significantly increased the secretion of interleukin 6 (IL-6), C-C motif chemokine ligand (CCL2), and C-X-C motif chemokine ligand (CXCL1) by MEFs compared with LPS. Mechanism study showed that both LPS and CM induced the phosphorylation of nuclear factor-κB p65 (NF-κB p65), p38 mitogen-activated protein kinase (p38 MAPK), extracellular regulated protein kinases 1/2 (ERK1/2), and TANK-binding kinase (TBK) in MEFs, and CM strongly stimulated the phosphorylation of signal transducer and activator of transcription 3 (STAT3) in MEFs. Conclusion Both LPS and CM can induce transcription and protein secretion of various inflammation-related genes in MEFs. CM can partly enhance LPS-induced activation of MEFs, and the mechanism may be related to the enhancement effect of CM on the activation STAT3 signaling pathway.
Animals ; Fibroblasts/immunology* ; Mice ; Lipopolysaccharides/pharmacology* ; Inflammation/metabolism* ; Signal Transduction/drug effects* ; Gene Expression Regulation/drug effects* ; Cytokines/genetics* ; Culture Media, Conditioned/pharmacology* ; Cells, Cultured

Objective To enhance the cyclic peptide compound's membrane permeability,structural stability,and neuroprotective activity,based on the amino acid sequence of peptides of Tat-GluA2-3Y,by designing and synthesizing a serial of cyclic peptides through strategies including polypeptide cyclization,replacement of the cell-penetrating peptide,substitution with D-amino acids,and incorporation of mini polyethylene glycol fragments.Methods The target peptides were synthesized based on standard Fmoc solid-phase method,followed by analysis and purification via reverse phase high-performance liguid chromatography(RP-HPLC).The cytoprotective activity of the peptides was evaluated by using the HT-22 cell model.The transmembrane transport efficiency of the peptides was determined based on the Caco-2 monolayer intestinal epithelial cell model.Plasmatic plasma and metabolic stability of the peptides were measured by in vitro co-incubation experiments with rat plasma and human liver microsomes.Finally,the in vivo neuroprotective activity of the peptides was validated by using a mouse middle cerebral artery occlusion model.Results Seven cyclic peptides were successfully designed and synthesized by using the standard Fmoc solid-phase method,with purities exceeding 90%as confirmed by RP-HPLC.Cytoprotective activity assay demonstrated that both Tat-GluA2-3Y and CMT-C3Y exhibited activity at concentrations above 125 nmol/L,with CMT-C3Y showing superior activity as compared to Tat-GluA2-3Y.The results of the transmembrane assay demonstrated that,compared to Tat-GluA2-3Y,CMT-C3Y exhibited significant transmembrane capabilities at all tested concentrations(P<0.001).CMT-C3Y was classified as a highly permeable compound,whereas Tat-GluA2-3Y was categorized as a moderately permeable compound.Plasma stability studies indicated that over 50%of Tat-GluA2-3Y was metabolized after 4 h of co-incubation with rat plasma.After 8 h of coincubation with CMT-C3Y,the remaining amount was 88.1%,and no obvious degradation phenomenon occurred.In human liver microsomal stability tests,the half-life of Tat-GluA2-3Y was 26.1 min,as compared to 103.8 min for CMT-C3Y,highlighting the enhanced stability of CMT-C3Y.Tat-GluA2-3Y and CMT-C3Y were classified as a fast-metabolizing drug and a moderate-metabolizing drug,respectively.Animal experiments further demonstrated that at a dose of 8 mg/kg the neuroprotective activity of CMT-C3Y was significantly superior to that of Tat-GluA2-3Y(P<0.001).Conclusion The designed bicyclic peptide CMT-C3Y demonstrates significantly higher cell-penetrating efficiency and superior plasma stability as compared to Tat-GluA2-3Y,along with enhanced neuroprotective activity at both cellular and animal levels.

Generative artificial intelligence (GAI) refers to AI technology capable of generating new content such as text, images, or audio from training data. GAI tools not only demonstrate rapid and efficient potential in literature screening, data extraction, and literature appraisal in systematic reviews, but can also be used for guideline evaluation and dissemination, enhancing the readability and promotion efficiency of guidelines. However, the accuracy of content generated by GAI tools, the rationality of cited evidence, the level of evidence, and the reliability of data still need verification. Additionally, data privacy protection and ethical issues are challenges that need to be addressed. This article aims to overview the current status of GAI tools in the formulation, evaluation, dissemination, and implementation of guidelines, explore the feasibility and new models of GAI tools in the field of guidelines, and improve the efficiency and quality of guideline formulation to better serve guideline developers and users.

Objective To investigate the effect of elastic pants on coagulation function and deep vein thrombosis by examining blood coagulation function and deep vein thrombosis in patients with liposuction in the thigh before and after operation.Methods 80 patients with liposuction were randomly divided into Ankle-length elastic pants(Ankle group,A group,n=40)and Knee-length elastic pants(Knee group,K group,n=40)from October 2021 to October 2022.After liposuction surgery,the two groups of patients used elastic bandage to initially compress and bind the thigh.According to the length of the patients'thigh,the patients in A group wore appropriate ankle-length elastic pants and the patients in K group wore appropriate knee-length elastic pants.The index were recorded including the popliteal vein flow rate,the common femoral vein flow rate,the instep temperature,the incidence of deep vein thrombosis(DVT),the incidence of intramuscular vein thrombosis of leg and the coagulation indexes(APTT,PT,Fib,D-dimer).Results Compared with pre-operation,the blood flow rate of popliteal vein and common femoral vein in A group was significantly faster,while the blood flow rate of popliteal vein in K group was significantly slower(P＜0.05).There was no significant change in APTT and PT in the two groups after operation(P＞0.05),but the Fib and D-Dimer increased significantly on the first day after operation,and then decreased gradually(P＜0.05).Compared with K group,the blood flow of popliteal vein and common femoral vein in A group was significantly faster at each time point after operation(P＜0.05).The Fib and D-Dimer of patients in A group were significantly lower than those in K group on the 1st,3rd and 7th day after operation(P＜0.05),and the blood coagulation index between the two groups returned to normal around the 14th day.There was no statistically significant difference in the instep temperature between groups and within groups(P＞0.05).No deep venous thrombosis was found in A group after surgery,while there were 3 cases of deep venous thrombosis without clinical symptoms in K group on the 3rd and 7th day,with no statistical difference(P＞0.05).After surgery,patients in A group and K group had intramuscular venous thrombosis of the leg on the 1st,3rd and 7th day after surgery,and there was statistically significant difference on the 3rd and 7th day between the two groups(P＜0.05).Conclusion Thigh liposuction can lead to deep vein thrombosis without obvious clinical symptoms and intramuscular vein thrombosis of leg.Ankle-length elastic pants are more conducive to blood circulation of patients'legs,improve blood hypercoagulability,and reduce the risk of deep vein thrombosis after operation.

OBJECTIVE To optimize hematopoietic stem cell transplantation therapy and provide support for drug research by investigating the dynamic process of hematopoietic and immune system reconstitution after bone marrow transplantation(BMT)in mice.METHODS CD45.2+C57BL/6 mice were used as recipient mice and randomly divided into the normal control group and transplantation group,with 30 mice in each.The transplantation group was irradiated by a lethal dose of cobalt-60 rays.Bone marrow cells were prepared from CD45.1+C57BL/6 mice and transfused into recipient mice through the tail vein.Peripheral blood,spleens,lymph nodes,thymuses and bone marrow were collected at 1,2,4,8 and 16 weeks after transplantation.Blood routine examination was performed with peripheral blood and total cell numbers in suspensions of other organs were counted by an automated cell counter.Cell classification analysis of white blood cells in peripheral blood,cell suspensions of other organs was performed by flow cytometry.RESULTS Four weeks after BMT,the numbers of white blood cells and red blood cells in peripheral blood of recipient mice returned to the same level of or higher level than normal control(P<0.05).Although the number of platelets recovered significantly,it was still mark-edly lower than that of normal control until 16 weeks post BMT(P<0.05).In addition,the percentages of myeloid leukocytes and B cells in peripheral blood,spleens,lymph nodes,and bone marrow,as well as megakaryocytes and erythrocyte progenitor cells in bone marrow also returned to normal,and the majority of myeloid leukocytes and B cells were CD45.1+cells from the donors.Eight weeks after BMT,T cells in peripheral blood,spleens,lymph nodes,thymuses,and bone marrow of recipient mice returned to normal,and CD45.1+T cells were dominating.CONCLUSION The hematopoietic and immune reconstitution of recipient mice is nearly completed eight weeks after BMT.However,the reconstruction speed of different kinds of cells and the reconstruction status of same kind of cell in different organs vary widely.

Objective To investigate the diagnostic value of computer tomography (CT) combined with serum secreted frizzled-related protein 4 (SFRP4) and soluble suppression of tumorigenicity 2 (sST2) for pancreatitis after biliary duct stent implantation in patients with malignant biliary obstruction (MBO). Methods A total of 243 MBO patients with biliary duct stent implantation were selected as research objects, and they were divided into pancreatitis group with 105 cases and no pancreatitis group with 138 cases according to condition of postoperative complicated pancreatitis. Enzyme-linked immunosorbent assay was used to detect serum levels of SFRP4 and sST2, and the receiver operating characteristic (ROC) curve and the four table method were used to analyze the diagnostic values of serum SFRP4 or sST2 alone and their combination with CT for postoperative complicated pancreatitis. Results The serum levels of SFRP4 and sST2 in the pancreatitis group were significantly higher than those in the no pancreatitis group (P < 0.05); the values of area under the curve (AUC) of serum SFRP4 and sST2 levels for diagnosis of postoperative complicated pancreatitis were 0.694 and 0.667, respectively. The sensitivity and specificity of CT for diagnosis of postoperative complicated pancreatitis were 76.19% and 60.87%, respectively; the sensitivity and accuracy of CT combined with serum SFRP4 and sST2 levels in diagnosing postoperative complicated pancreatitis were 98.10% and 76.54%, respectively; the sensitivity and accuracy of the combination of the three indexes for diagnosis were significantly higher than those of CT, SFRP4 or sST2 alone (P < 0.05). Conclusion MBO patients with pancreatitis after biliary duct stent implantation have higher serum levels of SFRP4 and sST2, and CT combined with serum levels of SFRP4 and sST2 has high diagnostic value for postoperative complicated pancreatitis.

OBJECTIVE: To study the chemical constituents of the roots of Angelica dahurica, a well-known Chinese herbal medicine named Baizhi in Chinese. METHODS: Compounds were separated by various chromatographies, and the structures of new compounds were elucidated based on the analysis of their spectroscopic and spectrometric data (1D, 2D NMR, HRESI MS, IR, and UV). The absolute configurations of new compounds were determined by the calculated electronic circular dichroism and chemical derivatization. The inhibitory activities of all isolates against nitric oxide (NO) production were evaluated using lipopolysaccharide-activated RAW 264.7 macrophage cells. RESULTS: Seven new 3,4-dihydro-furanocoumarin derivatives ( 1a/ 1b, 2a/ 2b, 3a/ 3b, 4) together with a known furanocoumarin ( 5) were isolated from the roots of A. dahurica. The new compounds included three pairs of enantiomers, (4S, 2''R)-angelicadin A ( 1a)/(4R, 2''S)-angelicadin A ( 1b), (4S, 2''S)-angelicadin A ( 2a)/(4R, 2''R)-angelicadin A ( 2b), and (4S, 2''S)-secoangelicadin A ( 3a)/(4R, 2''R)-secoangelicadin A ( 3b), together with (4R, 2''R)-secoangelicadin A methyl ester ( 4). The known xanthotoxol ( 5) inhibited the NO production with the half-maximal inhibitory concentration (IC₅₀) value of (32.8 ± 0.8) µmol/L, but all the new compounds showed no inhibitory activities at the concentration of 100 µmol/L. CONCLUSION This is the first report of the discovery of 3,4-dihydro-furanocoumarins from A. dahurica. The results are not only meaningful for the understanding of the chemical constituents of A. dahurica, but also enrich the reservoir of natural products.

A patient aged 68 years old presented urinary frequency, urgency, and gross hematuria for 1 month, with initial PSA of 72.72 ng/ml and alkaline phosphatase (ALP)of 114 U/L. Prostate biopsy pathology showed small cell neuroendocrine carcinoma of prostate. The patient was immediately administered 6 cycle of chemotherapy including etoposide and cisplatin combined with medical castration. The CDK4 gene was detected 1.99 times amplification by peripheral blood free DNA (cfDNA)gene analysis. The chemotherapy was followed by parbosini therapy. The number and density of bone metastases continued to decrease significantly by bone scan at 3 and 6 months after treatment, with a continuous decline of ALP and PSA. After 1 year of follow-up, pelvic MRI and bone systemic imaging indicated stable lesions, with PSA of 0.05 ng/ml and ALP of 59 U/L.

BACKGROUND: Intervertebral disk (IVD) degeneration, which can cause lower back pain, is a major predisposing factor for disability and can be managed through multiple approaches. However, there is no satisfactory strategy currently available to reconstruct and recover the natural properties of IVDs after degeneration. As tissue engineering develops, scaffolds with embedded cell cultures have proved critical for the successful regeneration of IVDs. METHODS: In this study, an integrated scaffold for IVD replacement was developed. Through scanning electron microscopy and other mechanical measurements, we characterized the physical properties of different hydrogels. In addition, we simulated the physiological structure of natural IVDs. Nucleus pulposus (NP) cells and annulus fibrosusderived stem cells (AFSCs) were seeded in gelatin methacrylate (GelMA) hydrogel at different concentrations to evaluate cell viability and matrix expression. RESULTS: It was found that different concentrations of GelMA hydrogel can provide a suitable environment for cell survival. However, hydrogels with different mechanical properties influence cell adhesion and extracellular matrix component type I collagen, type II collagen, and aggrecan expression. CONCLUSION This tissue-engineered IVD implant had a similar structure and function as the native IVD, with the inner area mimicking the NP tissue and the outer area mimicking the stratified annulus fibrosus tissue. The new integrated scaffold demonstrated a good simulation of disc structure. The preparation of efficient and regeneration-promoting tissueengineered scaffolds is an important issue that needs to be explored in the future. It is hoped that this work will provide new ideas and methods for the further construction of functional tissue replacement discs.

Objective:To investigate the relationship between renal function and the total cerebral small vessel disease(CSVD)score in elderly patients with a mild stroke.Methods:Patients with a mild stroke(National Institutes of Health Stroke Scale score≤3)and aged 60 years and above hospitalized in our hospital from March to December 2019 were consecutively enrolled.Total CSVD scores were evaluated based on enlarged perivascular spaces, cerebral microbleeds, white matter hyperintensities of presumed vascular origin and lacunes of presumed vascular origin.The estimated glomerular filtration rate(eGFR)was derived by using the formula of chronic kidney disease epidemiology collaboration based on serum creatinine(CKD-EPI Scr). The relationship between eGFR and the total CSVD score was analyzed. Results:Of the 152 patients enrolled, 35, 38, 39, 28 and 12 had total CSVD scores of 0, 1, 2, 3 and 4, respectively.The age of patients increased and eGFR decreased with the increase of total CSVD scores(CSVD from 0 to 4, [(67.43±6.59), (68.42±6.83), (72.18±7.94), (76.46±7.75) vs.(79.92±6.17)years old, F=12.018, P<0.001]; [(92.94±12.45)ml·min -1·1.73m -2, (90.52±8.62)ml·min -1·1.73m -2, (89.45±8.48)ml·min -1·1.73m -2 and(83.90±7.19)ml·min -1·1.73m -2vs.(79.16±7.77)ml·min -1·1.73m -2, F=7.210, P<0.001]. Spearman correlation analysis showed that eGFR was negatively correlated with the total CSVD score( r=-0.399, P<0.001). After adjusting for risk factors including age, gender and hypertension, multivariate ordinary regression analysis showed that decreased eGFR was an independent risk factor for the total CSVD score( OR=0.957, 95% CI: 0.924-0.990, P=0.012). Conclusions:In elderly patients with a mild stroke, eGFR is an independent risk factor for the total CSVD score, which shows an increased burden as eGFR decreases.