Acute lung injury (ALI) is a severe disease caused by viral infection that triggers an uncontrolled inflammatory response. This study investigated the capacity of jasurolignoside (JO), a natural compound, to bind to Toll-like receptor 4 (TLR4) and treat ALI. The anti-inflammatory properties of JO were evaluated in vitro through Western blotting, enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining, and co-immunoprecipitation. The investigation utilized a lipopolysaccharide (LPS)-induced ALI animal model to examine the therapeutic efficacy and mechanism of JO in vivo. JO attenuated inflammatory symptoms in infected cells and tissues by modulating the NOD-like receptor family pyrin domain containing protein 3 (NLRP3) inflammasome and the nuclear factor κB (NF-κB)/mitogen-activated protein kinase (MAPK) pathway. Molecular docking simulations revealed JO binding to TLR4 active sites, confirmed by cellular thermal shift assay. Surface plasmon resonance (SPR) demonstrated direct interaction between JO and TLR4 with a Kd value of 35.1 μmol·L^-1. Moreover, JO inhibited tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and IL-6 secretion and reduced leukocyte, neutrophil, lymphocyte, and macrophage infiltration in ALI-affected mice. JO also enhanced lung function and reduced ALI-related mortality. Immunohistochemical staining demonstrated JO's ability to suppress TLR4 expression in ALI-affected mouse lung tissue. This study establishes that JO can bind to TLR4 and effectively treat ALI, indicating its potential as a therapeutic agent for clinical applications.
Toll-Like Receptor 4/chemistry* ; Animals ; Acute Lung Injury/chemically induced* ; Mice ; Humans ; Ilex/chemistry* ; Molecular Docking Simulation ; Male ; NF-kappa B/immunology* ; Mice, Inbred C57BL ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Tumor Necrosis Factor-alpha/genetics* ; Interleukin-1beta/genetics* ; RAW 264.7 Cells ; Disease Models, Animal

Objective:To investigate impacts of usnic acid(UA)on malignant behavior of gastric cancer cells by regulating the chemokine(C-C motif)ligand 2(CCL2)-CCL2 receptor(CCR2)signal axis.Methods:SGC-7901 cells,a well growing human gastric cancer cell line,were treated with different concentrations of UA,which were grouped into low concentration(UA-L)group(62.5 μmol/L UA),medium concentration(UA-M)group(125 μmol/L UA)and high concentration(UA-H)group(250 μmol/L UA);meantime,the cells were transfected with CCL2 overexpression vector(pc DNA3.1 CCL2),empty vector(pc DNA3.1),silenced CCL2(si CCL2)and negative control(si control),and SGC-7901 cells were treated with 250 μmol/L UA,labeled as UA-H+pc DNA3.1 CCL2 group,UA-H+pc DNA3.1 group,UA-H+si control group and UA-H+si CCL2 group,another untreated SGC-7901 cells were taken as the control group.Flow cytometry,MTT and qRT-PCR were applied to detect cell apoptosis,proliferation,and expres-sion levels of CCL2 and CCR2 mRNA;Western blot was applied to detect expression levels of PD-L1,apoptotic protein(Bax),proli-ferative protein(CyclinD1,CCL2,CCR2)and immune escape related protein(B7H1);after co-culturing with CD8+T cells isolated and cultured in vitro,ELISA was applied to detect levels of IL-4,IFN-γ and IL-10 in the supernatant.Gastric cancer cells in each group were co-cultured with activated peripheral blood mononuclear cell(PBMC)1∶1 for 72 hours,and the sensitivity of gastric can-cer cells in each group to T-cell-mediated killing was compared.Results:Compared with control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-L group,UA-M group and UA-H group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels,Bax protein expression were obviously increased(P<0.05);compared with UA-H+pc DNA3.1 group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+pc DNA3.1 CCL2 group were obviously increased,while apoptosis rate,IL-4 and IFN-γ levels,and Bax protein ex-pression were obviously reduced(P<0.05);compared with UA-H+si control group,cell proliferation rate,IL-10 level,CyclinD1,PD-L1,CCL2,CCR2 and B7H1 protein and mRNA expressions,cell counts after co-culturing with activated PBMC 1∶1 for 72 hours in UA-H+si CCL2 group were obviously reduced,while apoptosis rate,IL-4 and IFN-γ levels and Bax protein expression were obviously increased(P<0.05).Conclusion:UA can inhibit gastric cancer cells proliferation,immune escape,and induce apoptosis,which may be related to the inhibition of the CCL2-CCR2 signaling axis.

Seven compounds were isolated from Onychium japonicum by macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography and semi-preparative HPLC. Their structures were identified by NMR, MS and other spectroscopic methods as onychone A (1), quercetin (2), quercetin-3-O-α-L-rhamnoside (3), kaempferol-7-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnopyranoside (5), (-)-prunin (6), and norathyriol (7). Compound 1 is a novel macrocyclic flavonoid, and all the others are reported from this plant for the first time. In vitro cytotoxic activities of compounds 1-7 were evaluated by MTS testing with five cancer cell lines. Compound 7 exhibited weak cytotoxicity against tumor cell lines A549, SMMC-7721, and SW480.

Objective:To learn about the genotyping of human Brucella isolated from Sichuan Province. Methods:BCSP31-PCR and AMOS-PCR were used to identify the genus and biotype of the 66 strains isolated from confirmed human brucellosis cases in Sichuan Province from 2014 to 2020, respectively. The isolated strains were genotyped by multi-locus sequence typing (MLST)-9. The sequence type (ST) was compared trough the online MLST database. A minimum spanning tree (MST) was constructed to cluster the newly discovered and known ST using the BioNumerics software version 7.6.Results:The 66 strains isolated from human cases of brucellosis in Sichuan Province from 2014 to 2020 were Brucella, and 65 of them were Brucella melitensis while one strain was Brucella abortus. The MLST method identified three known STs (ST-8, ST-39 and ST-2) and one newly type (ST-101). Among them, ST-8 was the main ST in Sichuan Province (90.91%, 60/66), another 4 strains of Brucella melitensis were ST-39, and 1 strain of Brucella abortus was ST-2. The newly type ST-101 was isolated from Leshan City in 2019, belonging to the Brucella melitensis and closely related to the evolution of ST-8. Conclusion:Brucella melitensis is the main epidemic Brucella strain in Sichuan Province, ST-8 is predominant genotype, with a small amount of ST-39, ST-101 and ST-2.

Objective: To examine the clinical value of fluorescence-guided indocyanine green (ICG) laparoscopic anatomical hepatectomy in the treatment of primary hepatocellular carcinoma. Methods: Data from patients diagnosed with hepatocellular carcinoma and who underwent laparoscopic hepatectomy with ICG fluorescence navigation in the Department of Liver Surgery and Liver Transplantation Center of West China Hospital between September 2020 and May 2022 were retrospectively collected. There were 53 males and 19 females, with an age of (55.5±12.9)years(range:42.6 to 68.4 years). Among them, 13 of the cases underwent laparoscopic anatomical liver resection(LALR) guided by tans-arterial ICG,43 of the cases received LAIR guided by portal vein negative ICG, and 16 of the cases received LALR positive by portal vein. Comparison among the three groups was performed by one-way ANOVA; and the rank sum test was used for comparison between groups. The counting data was expressed as percentage,and the χ² test or Fisher's exact probability method was used for comparison between groups. Results: (1) Postoperative pathology: Resection R0 was achieved in all operations. The maximum tumor diameter of the patients in the arterial staining group, the reverse staining group, and the positive staining group(M (IQR)) was 2.5 (2.4) cm, 3.0 (2.5) cm and 3.0(2.4) cm,respectively. There were no statistically significant differences in the maximum tumor diameter between the three groups (P=0.364). The minimum tumor margin was 1.1 (1.1) cm, 1.0 (1.0) cm, 1.1 (1.6) cm in the the arterial staining group, reverse staining group and the positive staining group, respectively. There was no significant difference in the margin among the three groups (P=0.878). (2) Operation conditions: the operation time of the arterial staining group, the negative staining group, and the positive portal staining group was (348±93)minutes,(277±112)minutes,and (295±116)minutes,respectively. There were no significant differences in operation time among the three groups (P=0.134). The intraoperative blood loss of the three groups was 80(150)ml,200(350)ml,and 100(150)ml,respectively. There was no statistically significant difference in intraoperative bleeding volume between the three groups(P=0.743). All cases were not transfused during the operation and were not converted to laparotomy. ALT in the arterial staining group was higher than in the negative staining group in the first two days after the operation ((559±398)IU/L307(257) IU/L, q=235.5,P=0.004;(611±389)IU/L(331±242) IU/L, q=265.2, P=0.002). There was only one case of a grade III complication (Clavien-Dindo grading system) postoperative complication in the negative and positive staining group of the portal vein, respectively. Tumor markers in all patients decreased to the normal range after 2 months of operation. Conclusion: Laparoscopic anatomical hepatectomy guided by ICG fluorescence through arterial staining and portal vein staining is safe and feasible for primary hepatocellular carcinoma treatment.

Objective To investigate the value of echocardiography in assessing right ventricular function of primary light-chain amyloidosis(AL)-cardiac amyloidosis(CA)patients.Methods A total of 25 patients biopsy diagnosed as AL-CA,who were admitted to the First Affiliated Hospital of Wenzhou Medical University from January 2018 to December 2021,were enrolled in this study as amyloidosis group.And 25 heathy volunteers were set as control group.The routine right ventricular morphology and functional parameters were acquired by standard echocardiography.Right ventricular global longitudinal strain(RVGLS)and right ventricular free wall longitudinal strain(RVfwLS)were obtained by two-dimensional speckle tracking imagine.Tricuspid annular plane systolic excursion(TAPSE)/pulmonary artery systolic pressure(PASP),RVGLS/PASP,RVfwLS/PASP ratio were calculated as the noninvasively parameters of right ventricular-pulmonary artery coupling.Results ①Compared with control group,patients in amyloidosis group showed significantly larger right heart size,and significantly thicker right ventricular free wall[(7.16±1.84)mm vs.(3.76±0.66)mm],the difference was statistically significant(P<0.001).②Compared with control group,RVGLS[(17.43±2.45)%vs.(23.22±3.73)%]and RVfwLS[(18.44±2.10)%vs.(22.62±3.64)%]were significantly impaired in amyloidosis group,the difference was statistically significant(P<0.001).The ratios of TAPSE/PASP,RVGLS/PASP,RVfwLS/PASP were found to be lower in amyloidosis group,the difference was statistically significant(P<0.001).③TAPSE/PASP showed mild correlations with N-terminal brain natriuretic peptide precursor(NT-proBNP),highly sensitive T cardiac troponin(hs-TNT)(r=0.45 and 0.40)there were statistically significant(P<0.05).Conclusion Echocardiography can effectively evaluate the right heart function impairment in AL-CA patients,providing valuable information for a deeper understanding of the pathological and physiological mechanisms of the disease.

Objective:To evaluate the clinical and echocardiographic characteristics of valvular involvement in SLE patients and its predictive value for valvular outcome.Methods:A retrospective study included 58 adult SLE patients with valvular involvement diagnosed for the first time by echocardiography in the First Affiliated Hospital of Wenzhou Medical University. According to gender and age, 1∶2 SLE patients without valvular involvement were selected as the control group. Clinical and echocardiographic data within 24 hours after initial admission were collected. The predictive factors were analyzed using cox parametric regression analysis.Results:①Among 58 patients,significant valve lesions were detected by echocardiography in 50 and presented as valve thickening with regurgitation. The mitral valve was the most commonly affected, followed by the aortic valves. ②Compared with the non-valvular involvement group, the percent of CRP, SLC-R, IgA、aCL-IgG were higher in the valvular involvement group [CRP:60.3% vs 30.4%, χ2=10.54, P=0.001; SLC-R: 63.8% vs 19.8%, χ2=33.08, P<0.001; IgA:27.6% vs 22.4%, χ2=16.04, P<0.001; aCL-IgG: 27.6% vs 2.6%, χ2=24.84, P<0.001]. ③Cox parametric regression analysis showed that CRP [ HR (95% CI)=2.641(1.499, 4.654), P=0.001,SLC-R HR(95% CI)=3.234(11.776, 5.888), P<0.001], and aCL-IgG[ HR(95% CI)=2.166 (1.148, 4.087) , P=0.017] were independent predictors of valvular involvement. Conclusion:SLE patients with increased levels of CRP、SLC-R、IgA and aCL-IgG have a high risk of valvular involvement. CRP、SLC-R and aCL-IgG are independent predictors of valvular involvement.

Objective To investigate the effect of preparation of shear-thinning CPC/LP nano-composite hydrogel for wound healing by supramolecular interaction of polydopamine(LP)and phy-cocyanin(CPC)based on nano-clayoxidation.Methods LP aggregates were formed by polymeriza-tion of dopamine monomers in weak alkaline oxygen-containing environment of nano-clay dispersion,and then mixed it with multifunctional CPC to form shear-thinning nanocomposite hydrogels by electro-static action and Schiff base.The microstructure,rheological properties,blood compatibility and wound healing effect of hydrogel were studied systematically.Results The gelation performance of CPC/LP nanocomposite hydrogel was related to LP content.When LP content was 3％and CPC con-tent was 0.5％,stable gel could be formed;rheological test results showed that the nanocomposite hydrogel had the properties of shear thinning and self-healing;the results of blood compatibility test showed that the hemolysis rate of the nanocomposite hydrogel was less than 5％,and it had good blood compatibility.In addition,the nanocomposite hydrogel could be injected into the injury site through a smaller needle to promote wound healing.Conclusion CPC/LP nanocomposite hydrogel exhibits excellent injectable properties and can effectively promote wound healing.

Objective To investigate the effect of preparation of shear-thinning CPC/LP nano-composite hydrogel for wound healing by supramolecular interaction of polydopamine(LP)and phy-cocyanin(CPC)based on nano-clayoxidation.Methods LP aggregates were formed by polymeriza-tion of dopamine monomers in weak alkaline oxygen-containing environment of nano-clay dispersion,and then mixed it with multifunctional CPC to form shear-thinning nanocomposite hydrogels by electro-static action and Schiff base.The microstructure,rheological properties,blood compatibility and wound healing effect of hydrogel were studied systematically.Results The gelation performance of CPC/LP nanocomposite hydrogel was related to LP content.When LP content was 3％and CPC con-tent was 0.5％,stable gel could be formed;rheological test results showed that the nanocomposite hydrogel had the properties of shear thinning and self-healing;the results of blood compatibility test showed that the hemolysis rate of the nanocomposite hydrogel was less than 5％,and it had good blood compatibility.In addition,the nanocomposite hydrogel could be injected into the injury site through a smaller needle to promote wound healing.Conclusion CPC/LP nanocomposite hydrogel exhibits excellent injectable properties and can effectively promote wound healing.

To efficiently remove all recurrent lymph nodes (rLNs) and minimize complications, we developed a combination approach that consisted of ⁶⁸Gallium prostate-specific membrane antigen (PSMA) ligand positron emission tomography (PET)/computed tomography (CT) and integrated indocyanine green (ICG)-guided salvage lymph node dissection (sLND) for rLNs after radical prostatectomy (RP). Nineteen patients were enrolled to receive such treatment. ⁶⁸Ga-PSMA ligand PET/CT was used to identify rLNs, and 5 mg of ICG was injected into the space between the rectum and bladder before surgery. Fluorescent laparoscopy was used to perform sLND. While extensive LN dissection was performed at level I, another 5 mg of ICG was injected via the intravenous route to intensify the fluorescent signal, and laparoscopy was introduced to intensively target stained LNs along levels I and II, specifically around suspicious LNs, with ⁶⁸Ga-PSMA ligand PET/CT. Next, both lateral peritonea were exposed longitudinally to facilitate the removal of fluorescently stained LNs at levels III and IV. In total, pathological analysis confirmed that 42 nodes were rLNs. Among 145 positive LNs stained with ICG, 24 suspicious LNs identified with ⁶⁸Ga-PSMA ligand PET/CT were included. The sensitivity and specificity of ⁶⁸Ga-PSMA ligand PET/CT for detecting rLNs were 42.9% and 96.6%, respectively. For ICG, the sensitivity was 92.8% and the specificity was 39.1%. At a median follow-up of 15 (interquartile range [IQR]: 6-31) months, 15 patients experienced complete biochemical remission (BR, prostate-specific antigen [PSA] <0.2 ng ml^-1), and 4 patients had a decline in the PSA level, but it remained >0.2 ng ml^-1. Therefore, ⁶⁸Ga-PSMA ligand PET/CT integrating ICG-guided sLND provides efficient sLND with few complications for patients with rLNs after RP.
Gallium Isotopes ; Gallium Radioisotopes ; Humans ; Indocyanine Green ; Ligands ; Lymph Node Excision ; Lymphatic Metastasis/diagnostic imaging* ; Male ; Neoplasm Recurrence, Local/surgery* ; Positron Emission Tomography Computed Tomography ; Prostate ; Prostatectomy ; Prostatic Neoplasms/surgery* ; Salvage Therapy