The liver has diverse functions such as metabolism， detoxification， and immune defense， and the maintenance of hepatic microenvironment homeostasis is crucial for overall bodily health. The hepatic microenvironment consists of the components such as parenchymal cells， non-parenchymal cells， and non-cellular components. Chronic inflammatory responses induced by various etiological factors may promote the formation and progression of liver fibrosis. During the dynamic progression of liver fibrosis， from the early to advanced stages， various components within the hepatic microenvironment undergo a series of changes， which can promote the malignant progression of liver fibrosis. An in-depth exploration of the mechanisms underlying such changes in each component of the liver fibrosis microenvironment is of great significance for understanding the pathogenesis of liver fibrosis and discovering potential treatment strategies.

ObjectiveTo investigate the changes in coagulation system in acute decompensated cirrhosis （ADC） patients with or without sepsis and the association of these changes with short-term prognosis. MethodsA prospective study was conducted among 116 ADC patients who were hospitalized in Nanfang Hospital from January 2021 to July 2023， among whom there were 86 patients with sepsis and 30 patients without sepsis， and 54 patients with sepsis alone who had no chronic liver disease were enrolled as control group. Thromboelastography （TEG） and other conventional coagulation parameters were used to comprehensively evaluate the coagulation function of patients. The data including TEG results and short-term prognosis were collected， and a correlation analysis was performed. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups； the chi-square test was used for comparison of categorical data between two groups. The Spearman correlation coefficient was calculated to investigate the correlation between different variables. The Logistic regression model was used to perform the univariate and multivariate analyses. ResultsFor the ADC patients with sepsis， the lungs and bloodstream were the main infection sites， and bacteria were the main pathogenic microorganism. TEG results showed that compared with the patients with sepsis alone， the patients with ADC and sepsis had a significant reduction in median maximum amplitude （MA）， a significant increase in coagulation time （K time）， and a significant reduction in α angle （all P<0.05）； the patients with ADC and sepsis had a significantly longer reaction time （R time） than those with ADC alone （P=0.02）， and the patients with sepsis alone had a significantly longer R time than those with ADC and sepsis （P=0.04）. There was no correlation between MA and platelet count in the patients with ADC and sepsis （r=-0.133， P=0.057）， while there was a significant correlation between MA and platelet count in the patients with sepsis alone （r=0.595， P=0.001）. SOFA score was negatively correlated with MA in sepsis patients with or without ADC （r=-0.503 and -0.561， both P<0.001）， and for the patients with ADC and sepsis， R time， K time， and α angle were weakly correlated with SOFA score and had a relatively strong correlation with APTT （all P<0.05）. The patients with ADC alone all survived within 90 days， and compared with the death group， the patients with sepsis alone who survived had significantly higher values of MA and α angle （all P<0.05）； there was a significant difference in α angle on day 90 between the survival group and the death group， no matter whether the patients were comorbid with ADC or not （both P<0.01）， while for the patients with ADC and sepsis， there was no significant difference in MA value on day 90 between the survival group and the death group （P>0.05）. ConclusionFor ADC patients comorbid with sepsis， coagulation function assessment and monitoring should be taken seriously in clinical practice， and TEG parameters and SOFA score should be monitored when necessary to develop individualized treatment regimens.

Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells (DCs) and their exosomes (DEXs). Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) to differentiate into DCs and were subsequently matured with tumor necrosis factor α(TNF-α). Exosomes were isolated by ultracentrifugation, and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry, which were also used to quantify the expression of immune membrane proteins on DCs and DEXs. Results On the 10th day of culture, DCs displayed high surface expression of CD11c, CD80, CD86, major histocompatibility complex class I (MHC-I), and MHC-II. Expression peaked at day 18(CD11c: 78.66%±20.33%, CD80: 76.41%±10.02%, CD86: 96.43%±0.43%, MHC-I: 84.71%±2.96%, MHC-II: 80.01%±7.03%). After day 24, the overall expression showed a declining trend, with statistically significant differences observed for all markers except CD80 and MHC-II. By day 30, 80% of the DCs still expressed CD80, CD86, and MHC-II. The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18, followed by an overall decline with prolonged culture time, with statistically significant differences observed for all markers except CD80. Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces (CD11c: r=0.98; CD80: r=0.65; CD86: r=0.82; MHC-I: r=0.86; MHC-II: r=0.93). Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period. The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins, with temporal trends aligned with those of the parent DCs.
Humans ; Dendritic Cells/immunology* ; Exosomes/immunology* ; Monocytes/metabolism* ; Cells, Cultured ; Time Factors ; B7-1 Antigen/metabolism* ; Membrane Proteins/immunology* ; Cell Culture Techniques/methods* ; B7-2 Antigen/metabolism* ; Cell Differentiation ; CD11c Antigen/metabolism* ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

Objective:Progressive bone resorption and destruction is one of the most critical clinical features of middle ear cholesteatoma,potentially leading to various intracranial and extracranial complications.However,the mechanisms underlying bone destruction in middle ear cholesteatoma remain unclear.This study aims to explore the role of parathyroid hormone-related protein(PTHrP)in bone destruction associated with middle ear cholesteatoma. Methods:A total of 25 cholesteatoma specimens and 13 normal external auditory canal skin specimens were collected from patients with acquired middle ear cholesteatoma.Immunohistochemical staining was used to detect the expressions of PTHrP,receptor activator for nuclear factor-kappa B ligand(RANKL),and osteoprotegerin(OPG)in cholesteatoma and normal tissues.Tartrate-resistant acid phosphatase(TRAP)staining was used to detect the presence of TRAP positive multi-nucleated macrophages in cholesteatoma and normal tissues.Mono-nuclear macrophage RAW264.7 cells were subjected to interventions,divided into a RANKL intervention group and a PTHrP+RANKL co-intervention group.TRAP staining was used to detect osteoclast formation in the 2 groups.The mRNA expression levels of osteoclast-related genes,including TRAP,cathepsin K(CTSK),and nuclear factor of activated T cell cytoplasmic 1(NFATc1),were measured using real-time polymerase chain reaction(real-time PCR)after the interventions.Bone resorption function of osteoclasts was assessed using a bone resorption pit analysis. Results:Immunohistochemical staining showed significantly increased expression of PTHrP and RANKL and decreased expression of OPG in cholesteatoma tissues(all P<0.05).PTHrP expression was significantly positively correlated with RANKL,the RANKL/OPG ratio,and negatively correlated with OPG expression(r=0.385,r=0.417,r=-0.316,all P<0.05).Additionally,the expression levels of PTHrP and RANKL were significantly positively correlated with the degree of bone destruction in cholesteatoma(r=0.413,r=0.505,both P<0.05).TRAP staining revealed a large number of TRAP-positive cells,including multi-nucleated osteoclasts with three or more nuclei,in the stroma surrounding the cholesteatoma epithelium.After 5 days of RANKL or PTHrP+RANKL co-intervention,the number of osteoclasts was significantly greater in the PTHrP+RANKL co-intervention group than that in the RANKL group(P<0.05),with increased mRNA expression levels of TRAP,CTSK,and NFATc1(all P<0.05).Scanning electron microscopy of bone resorption pits showed that the number(P<0.05)and size of bone resorption pits on bone slices were significantly greater in the PTHrP+RANKL co-intervention group compared with the RANKL group. Conclusion:PTHrP may promote the differentiation of macrophages in the surrounding stroma of cholesteatoma into osteoclasts through RANKL induction,contributing to bone destruction in middle ear cholesteatoma.

Objective:Middle ear cholesteatoma is a non-tumorous condition that typically leads to hearing loss,bone destruction,and other severe complications.Despite surgery being the primary treatment,the recurrence rate remains high.Therefore,exploring the molecular mechanisms underlying cholesteatoma is crucial for discovering new therapeutic approaches.This study aims to explore the involvement of N6-methyladenosine(m6A)methylation in long non-coding RNAs(lncRNAs)in the biological functions and related pathways of middle ear cholesteatoma. Methods:The m6A modification patterns of lncRNA in middle ear cholesteatoma tissues(n=5)and normal post-auricular skin tissues(n=5)were analyzed using an lncRNA m6A transcriptome microarray.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were conducted to identify potential biological functions and signaling pathways involved in the pathogenesis of middle ear cholesteatoma.Methylated RNA immunoprecipitation(MeRIP)-PCR was used to validate the m6A modifications in cholesteatoma and normal skin tissues. Results:Compared with normal skin tissues,1 525 lncRNAs were differentially methylated in middle ear cholesteatoma tissues,with 1 048 showing hypermethylation and 477 showing hypomethylation[fold change(FC)≥3 or<1/3,P<0.05].GO enrichment analysis indicated that hypermethylated lncRNAs were involved in protein phosphatase inhibitor activity,neuron-neuron synapse,and regulation of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid(AMPA)receptor activity.Hypomethylated lncRNAs were associated with mRNA methyltransferase activity,secretory granule membrane,and mRNA methylation.KEGG analysis revealed that hypermethylated lncRNAs were mainly associated with 5 pathways:the Hedgehog signaling pathway,viral protein interaction with cytokines and cytokine receptors,mitogen-activated protein kinase(MAPK)signaling pathway,cytokine-cytokine receptor interaction,and adrenergic signaling in cardiomyocytes.Hypomethylated lncRNAs were mainly involved in 4 pathways:Renal cell carcinoma,tumor necrosis factor signaling pathway,transcriptional misregulation in cancer,and cytokine-cytokine receptor interaction.Additionally,MeRIP-PCR confirmed the changes in m6A methylation levels in NR_033339,NR_122111,NR_130744,and NR_026800,consistent with microarray analysis.Real-time PCR also confirmed the significant upregulation of MAPK1 and NF-κB,key genes in the MAPK signaling pathway. Conclusion:This study reveals the m6A modification patterns of lncRNAs in middle ear cholesteatoma,suggests a direction for further research into the role of lncRNA m6A modification in the etiology of cholesteatoma.The findings provide potential therapeutic targets for the treatment of middle ear cholesteatoma.

Objective To investigate the protective effect and mechanism of J147 on the injury of human neuroblastoma cells(SH-SY5Y)induced by high glucose(HG).Methods We established HG-induced SH-SY5Y cell injury model.Then SH-SY5Y cells were divided into blank control(Con)group HG group,HG+J147 0.5 μmol/L(HG+J147 0.5)group,HG+J147 1 μmol/L(HG+J147 1)group,HG+J147 2 μmol/L(HG+J147 2)group,HG+PI3K/AKT inhibitor LY294002(LY)10 μmol/L(HG+LY)group,HG+ERK1/2 inhibitor U0126(U0)5 μmol/L(HG+U0)group,HG+J147 2 μmol/L+LY 10 μmol/L(HG+J147 2+LY)group,HG+J147 2 μmol/L+U0 5 μmol/L(HG+J147 2+U0)group.Cell viability was detected by MTS cell proliferation and toxicity detection kit;LDH activity was tested by lactate dehydrogenase kit;morphological changes of SH-SY5Y cells were evaluated by microscope;cell apoptosis was detected by flow cytometry;and apoptosis-related proteins(Bcl-2,Bax)and signaling pathway-related proteins(p-AKT,AKT,p-ERK1/2,ERK1/2,p-CREB,CREB,BDNF)were detected by Western blot.Results Compared with Con group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT,p-ERK/ERK,p-CREB/CREB and BDNF protein expressions decreased(P<0.01),while LDH activity and apoptosis rate increased in HG group(P<0.01).Compared with HG group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT,p-ERK/ERK,p-CREB/CREB and BDNF protein expressions increased(P<0.01),while LDH activity and apoptosis rate decreased in HG+J147 2 group(P<0.01).Compared with HG+J147 2 group,the cell viability,Bcl-2/Bax ratio,p-AKT/AKT and BDNF protein expression decreased(P<0.05 or P<0.01),while LDH activity and apoptosis rate increased(P<0.05 or P<0.01),the expression of p-ERK/ERK protein in HG+J147 2+LY group decreased(P<0.05),and the expression of p-CREB/CREB protein in HG+J147 2+U0 group decreased in HG+J147 2+LY and HG+J147 2+U0 groups(P<0.05).Conclusions J147 can alleviate HG-induced SH-SY5Y cell damage,and the mechanism may be related to the activation of PI3K/AKT and ERK1/2 signaling and the reduction of apoptosis.

Objective To construct a prediction model for post-neoadjuvant therapy recurrent laryngeal nerve lymph node(RLN LN)status via clinical and CT image data in esophageal cancer patients pre-neoadjuvant therapy.Methods A retrospective analysis was conducted on 403 patients with locally advanced esophageal cancer who received neoadjuvant therapy and radical resection for esophageal cancer.All patients were divided into a training cohort(n=270)and a validation cohort(n=133)randomly according to a 2:1 ratio.Clinical and imaging features associated with positive RLN LN pathology were selected by univariate analysis.Multivariate logistic stepwise regression model was used to construct the prediction model.The prediction ability of the model was evaluated by receiver operating characteristic(ROC)curve.Results The basic model included neoadjuvant therapy and RLN LN short diameter,with an area under the curve(AUC)of 0.7(training cohort)and 0.65(validation cohort).The final prediction model included neoadjuvant therapy,human albumin,platelet count,largest lymph node enhancement characteristics,whether the largest lymph node was in the recurrent laryngeal region,and RLN LN short diameter,with AUC of 0.83[95％confidence interval(CI)0.768-0.899]and 0.76(95％CI 0.645-0.887)for the training and validation cohorts,respectively.Conclusion The model based on clinical data and imaging features pre-neoadjuvant therapy for esophageal cancer can assist in clinically predicting the post-neoadjuvant therapy RLN LN status.

Objective:To explore the effects of dexmedetomidine (DEX) on postoperative pain, oxidative stress and adverse reactions in patients undergoing radical mastectomy.Methods:A total of 90 patients with breast cancer who received radical surgical treatment in our hospital from Jun. 2022 to Jun. 2023 were prospectively included as research objects and randomly divided into 3 groups with 30 patients in each group. DEX group was applied before, during and after surgery, respectively. The levels of pain visual analogue scale (VAS), Richmonation sedation score (RASS), superoxide dismu-tase (SOD) and malondialdehyde (MDA) were recorded.Results:The recovery time and extubation time in preoperative and intraoperative DEX group were significantly lower than those in postoperative DEX group, and the awakening time and extubation time in preoperative DEX group were significantly lower than those in intraoperative DEX group ( F value was 48.62 and 53.98, respectively, P<0.001). At 1 h, 6 h and 12 h after surgery, the VAS and RASS scores of patients in the preoperative and intraoperative DEX group were significantly lower than those in the postoperative DEX group, compared with those in the intraoperative DEX group. The VAS and RASS scores in the DEX group were significantly decreased ( F value: 62.34, 55.24, 69.26, 36.82, 24.20, 39.97, P<0.001). At 24h after surgery, there was no significant difference in VAS and RASS scores among the three groups ( F value was 0.45 and 0.81, respectively, P value was 0.613 and 0.418). Immediately after surgery, 24 h after surgery, 72 h after surgery, the SOD level of DEX group was significantly higher than that of DEX group before and during surgery ( F value was 29.37, 33.24, 10.35, P<0.001). MDA levels were significantly lower than those in postoperative DEX group ( F value was 30.52, 41.27, 8.26, P<0.001). There was no significant difference in the incidence of postoperative adverse reactions among all groups ( P>0.05) . Conclusion:Preoperative and intraoperative application of DEX can reduce postoperative pain and oxidative stress in breast cancer patients, help patients recover quickly after surgery, and preoperative application is superior to intraoperative application.