ObjectiveTo investigate whether Huanglian Jiedutang （HLJD） and its major active constituents （geniposide， baicalin， and berberine） can inhibit the inflammatory response of BV2 cells under lipopolysaccharide （LPS） stimulation via the high-mobility group protein B1 （HMGB1）/Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway， and to explore differences in therapeutic efficacy among the three monomers， their combined formula， and HLJD under equal content ratios. MethodsBV2 microglial cells were used as the primary experimental model. Cell viability was assessed using the cell counting kit-8 （CCK-8） method to examine the effects of different concentrations of dimethyl sulfoxide （DMSO， 0.8%， 0.4%， 0.2%， 0.1%， and 0.05%） on cell viability. IncuCyte was employed to monitor the growth of cells under different concentrations of HLJD （200， 100， 50， 25， 12.5， 6.25 mg·L^-1）. Nitric oxide （NO） assay was used to screen the optimal HLJD concentration. High-performance liquid chromatography （HPLC） determined the content of geniposide， baicalin， and berberine in HLJD， and experimental groups were subsequently established according to the relative proportions of these constituents. CCK-8 assay evaluated cell viability under different treatments. Enzyme-linked immunosorbent assay （ELISA） measured levels of inflammatory factors （TNF-α， IL-1β， IL-6， IL-10） in the supernatant. Flow cytometry assessed the effects of treatments on M1-type polarization of BV2 cells. Western blot determined the expression levels of HMGB1， TLR4， and NF-κB-related proteins. ResultsCompared with the blank group， DMSO at concentrations ≤0.2% did not affect cell viability within 48 h. BV2 cell growth plateaued at 24 h after treatment with 200 mg·L^-1 HLJD. Under stimulation with 2 mg·L^-1 LPS， this concentration of HLJD effectively reduced NO release， and 6 h pre-treatment had a stronger inhibitory effect on NO than direct administration. HPLC results showed that 1 mg of HLJD freeze-dried powder contained approximately 24 μg of geniposide， 15 μg of baicalin， and 30 μg of berberine. Based on these ratios， experimental groups were blank， LPS （2 mg·L^-1）， HLJD （200 mg·L^-1）， monomer combination， geniposide （4.8 mg·L^-1）， baicalin （3 mg·L^-1）， and berberine （6 mg·L^-1）. The monomer combination group consisted of all three active constituents dissolved together. LPS and HLJD or its active constituents did not affect cell viability compared with the blank group. LPS significantly increased TNF-α， IL-1β， IL-6， and IL-10 in the supernatant （P<0.01）. HLJD and its active constituents significantly reduced pro-inflammatory factors TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01） while upregulating anti-inflammatory IL-10 （P<0.01）， with the monomer combination showing the strongest effect （P<0.05， P<0.01）. Compared with the blank group， LPS significantly increased the proportion of CD80⁺CD86⁺ （M1-type） BV2 cells （P<0.01）. HLJD and its constituents partially inhibited M1 polarization （P<0.05， P<0.01）， with the monomer combination exhibiting the most pronounced effect （P<0.05， P<0.01）. Compared with the blank group， LPS upregulated HMGB1， TLR4， and NF-κB-related proteins （P<0.01）， whereas HLJD and its active constituents significantly reduced their expression （P<0.05， P<0.01）， with the monomer combination having the strongest regulatory effect （P<0.05， P<0.01）. ConclusionHLJD and its major active constituents （geniposide， baicalin， berberine） can inhibit LPS-induced inflammatory responses in BV2 cells. The combination of the three active constituents demonstrates the most potent anti-inflammatory effect， significantly attenuating M1-type polarization of BV2 cells via the HMGB1/TLR4/NF-κB signaling pathway.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Objective:To establish an early prediction model based on triglyceride glucose index (TyG) and procalcitonin (PCT) for patients of acute pancreatitis (AP) complicated with acute kidney injury (AKI), and evaluate the diagnostic value of prediction model.Methods:This study was a single center prospective study. AP patients were recruited from the Emergency Department at Peking University People’s Hospital from January to December 2022. The observation endpoint was 14 days after the diagnosis of acute pancreatitis, patients were divided into AKI and control (no AKI) groups according to the observation endpoint. The general characteristics, clinical laboratory examinations, complications, and clinical scores were compared. The risk for AKI development was determined using logistic analyses to establish a risk prediction model. The receiver operating characteristic curve was drawn and the area under the curve (AUC) was calculated. The diagnostic sensitivity and specificity of the model were calculated, and the diagnostic value of the model was compared with that of Ranson score, APACHEⅡ score and BISAP score.Results:A total of 258 patients were selected for this study, including 79 in the AKI group and 179 in the control group. There was no significant difference in serum creatinine and blood urea nitrogen levels between the two groups. Compared with the control group, the AKI group had a higher proportion of males, older age, and had a higher proportion of hypertension. The ratio of neutrophil/lymphocyte ratio, PCT, and TyG were significantly increased. The Ranson score, APACHE Ⅱ score, and BISAP score were higher, and more patients had ARDS and serous fluid accumulation in the later period. Multivariate logistic regression showed that age ( OR=1.071, 95% CI: 1.020-1.125, P=0.006), increased TyG index ( OR=2.632, 95% CI: 1.423-4.866, P=0.002), and elevated PCT ( OR=1.275, 95% CI: 1.067-1.524, P=0.008) were risk factors for AKI in AP patients. According to the risk factors, forecast the AP patients complicated with AKI risk assessment model is established: Logistic (AKI/AP) = -16.697+0.069×age+ 0.968×TyG+0.243×PCT. The sensitivity and specificity of the model for predicting AKI in AP were 79.75% and 96.65%, respectively, and the AUC was 0.856 (95% CI: 0.790-0.922). The predictive ability was better than that of Ranson score, BISAP score and APACHE Ⅱ score (AUC: 0.856 vs. 0.691 vs. 0.745 vs. 0.705, P=0.041). Conclusion:The prediction model based on age, TyG and PCT was valuable for the prediction of AP concurrent AKI in early stage.

Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.

Humans ; Adolescent ; Acne Vulgaris/therapy* ; Skin Care ; Surveys and Questionnaires ; Cognition ; China ; Skin

ObjectiveBased on network pharmacology and transcriptomics， the mechanism of Zishen Qinggan prescription （ZSQGF） in improving glucose and lipid metabolism in type 2 diabetes （T2DM） model rats was explored. MethodBased on network pharmacology analysis of the differential genes between ZSQGF and T2DM， gene ontology（GO）analysis and Kyoto encyclopedia of genes and genomes（KEGG） analysis were conducted， and molecular docking analysis was used to verify the binding between components and targets. A T2DM rat model was established by high-fat feeding and injection of streptozotocin （STZ）. The rats were randomly divided into the control group， model group， metformin （Met， 72 mg·kg^-1） group， and ZSQGF high-， medium-， and low-dose groups （ZSQGF-H， ZSQGF-M， and ZSQGF-L， with 4.8， 2.4， and 1.2 g·kg^-1 raw drug in the solution）. The living status of rats was monitored and the levels of total cholesterol （TC）， total triglycerides （TG）， high density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） in rat serum were detected. The liver tissues were subjected to Hematoxylin eosin（HE） staining and oil red O staining. The differential genes were analyzed through transcriptomics， GO and KEGG analysis， and the protein-protein interaction（PPI） network was obtained to screen key targets. With network pharmacology and transcriptomics analysis results， the protein pathways were identified. The expression levels of nuclear factor-κB （NF-κB）， matrix metalloproteinase（MMP）-1 and MMP-9 proteins in liver tissues were detected by Western blot. The mRNA expression of B-cell lymphoma-2（Bcl-2） modifying factor（BMF）， nicotinamide adenine dinucleotide phosphate （NADPH） oxidase 4 （NOX4）， and fatty acid synthase（FASN） was detected by real-time polymerase chain reaction（Real-time PCR）. The expression of MMP-1 and MMP-9 in the liver was detected by immunofluorescence staining. ResultTranscriptomics and network pharmacology analysis suggested that ZSQGF may protect the liver through the glucose and lipid metabolism pathway and the inflammation pathway. Experiments showed that after 8 weeks of administration， the body weight， blood sugar， serum indicators， and pathological staining results of rats were improved. Western blot results indicated a decrease in the relative expression levels of NF-κB， MMP-1 and MMP-9 proteins in the liver. Real-time PCR results showed a decrease in the transcriptional expression of BMF， NOX4， and FASN in the ZSQGF-H group， while immunofluorescence staining results present decreased expression of MMP-1 and MMP-9 in the ZSQGF groups. ConclusionZSQGF can improve the glucose and lipid metabolism by inhibiting the expression of FASN， reducing lipid synthesis， and regulating the NF-κB signaling pathway.

To study the prevalence of depression, anxiety, and insomnia among social workers during the prolonged battle against the COVID-19 pandemic and explore the associated risk factors.

Sleep is an important activity of daily life for individuals,and sleep disorders can seriously affect their physical and mental health.This article summarizes the impact of vestibular stimulation on sleep,and reviews feasible and effective intervention methods from swing movement,galvanic vestibular stimulation,and weighted blankets stimulation,in order to provide new ideas and methods for the diagnosis and treatment of sleep disorders.

Objective:To investigate the association between body mass index (BMI) and the risk of chronic obstructive pulmonary disease (COPD).Methods:This study employed a single-center, cross-sectional design combined with online Mendelian randomization (MR) analysis. A total of 16 187 pieces of lung function data for COPD screening from the “Happy Breathing” project in the Jingzhou area between September 2021 and October 2023 were initially collected. Cases with missing questionnaire information, failed quality control, or isolated restrictive or small airway dysfunction were excluded, resulting in a final dataset of 3 999 cases, comprising 2 330 non-COPD and 1 669 COPD cases. Binary COPD status was set as the outcome variable, with BMI as the exposure variable. A smooth curve plot was used to depict the relationship between BMI and COPD prevalence; if non-linearity was observed, a two-piece linear regression model was employed to calculate threshold effects, adjusting for potential confounders such as age, sex, smoking history, biomass fuel exposure, and COPD awareness. Subsequently, online genome-wide association study (GWAS) data were utilized for MR analysis to explore any potential causal link between high BMI and COPD development.Results:Analysis of the “Happy Breathing” COPD screening data from the Jingzhou area revealed a nonlinear relationship between BMI and COPD risk. For BMI values <23.7 kg/m2, each 1 kg/m2 increase in BMI was associated with an 18% decrease in COPD risk ( OR=0.82, 95% CI: 0.79-0.85). However, for BMI >23.7 kg/m2, no significant association was found between increasing BMI and COPD risk ( OR=1.00, 95% CI: 0.96-1.05). MR analysis using online GWAS data suggested no potential causal relationship between high BMI and COPD occurrence, with both heterogeneity and pleiotropy tests yielding P>0.05. Conclusion:There may be a nonlinear relationship between BMI and the risk of COPD, with no apparent association between BMI and COPD risk for values above 23.7 kg/m2.

Objective:To evaluate the effect of patient-controlled intravenous analgesia (PCIA) with esketamine mixed with hydromorphone on the postoperative sleep quality in pediatric patients.Methods:This was a prospective randomised controlled study. One hundred pediatric patients of both sexes, aged 4-12 yr, with a body mass index of 13-26 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, undergoing elective abdominal surgery with general anesthesia, who agreed to use postoperative patient-controlled intravenous analgesia in our hospital from January to November 2023, were divided into esktamine mixed with hydromorphone group (EH group) and hydromorphone group (H group), with 50 patients in each group. PCIA solution contained esketamine 1 mg/kg and hydromorphon 0.1 mg/kg in 100 ml of 0.9% sodium chloride in EH group and hydromorphone 0.2 mg/kg in 100 ml of 0.9% sodium chloride in H group. The Pittsburgh Sleep Quality Index was used to evaluate the preoperative sleep quality by asking pediatric patients themselves or their guardians. On the morning of 1-3 days after surgery, the numeric rating scale (NRS) was used to evaluate the sleep quality of the first 3 nights after surgery. At 24 and 48 h after surgery, the FLACC pain scale was used to evaluate the intensity of pain. The consumption of hydromorphone within 48 h after surgery and the incidence of postoperative adverse reactions such as nausea, vomiting, itching and delirium were recorded. Results:Compared with group H, the NRS score was significantly increased on the 1st postoperative night, the ratio of poor sleep quality was decreased ( P<0.05), and no significant change was found in the NRS score on the 2nd or 3rd postoperative night and ratio of poor sleep quality ( P>0.05), and the FLACC pain scale score at 24 and 48 h after surgery and consumption of hydromorphone within 48 h after surgery were decreased in group EH ( P<0.05). There was no significant difference in the incidence of postoperative adverse reactions between the two groups ( P>0.05). Conclusions:PCIA with esketamine mixed with hydromorphone can improve the postoperative sleep quality in pediatric patients.