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 evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

Hemorrhagic shock is a common clinical emergency that can aggravate cell injury after resuscitation. Astrocytes are crucial for the survival of neurons because they regulate the surrounding ionic microenvironment of neurons. Although hemorrhagic shock and resuscitation (HSR) injury can impair cognition, it remains unclear how this insult directly affects astrocytes. In this study, we established an HSR model by bleeding and re-transfusion in mice. The social interaction test and new object recognition test were applied to evaluate post-operative cognitive changes, and the results suggest that mice experience cognitive impairment following exposure to HSR. In the HSR group, the power spectral density of β and γ oscillations decreased, and the coupling of the θ oscillation phase and γ oscillation amplitude was abnormal, which indicated abnormal neuronal oscillation and cognitive impairment after HSR exposure. In brief, cognitive impairment in mice is strongly correlated with Ca²⁺ signal strength in lateral septum astrocytes following HSR.
Animals ; Astrocytes/metabolism* ; Shock, Hemorrhagic/metabolism* ; Resuscitation/adverse effects* ; Male ; Mice ; Calcium Signaling/physiology* ; Mice, Inbred C57BL ; Septal Nuclei/metabolism* ; Cognitive Dysfunction/etiology* ; Disease Models, Animal ; Cognition Disorders/etiology*

Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.

Objective To explore the characteristics of coagulation and immune function and their clinical significance in children with severe adenovirus pneumonia(SAP).Methods A total of 122 children with SAP and 120 children with non-severe adenovirus pneumonia(NSAP)were enrolled and assigned to severe group and non-severe group.The severe group was further subdivided into good prognosis subgroup(n=105)and poor prognosis subgroup(n=17)according to prognosis.Addition-ally,120 healthy children undergoing physical examination during the same period were included as control group.Coagulation function indicators,including prothrombin time(PT),activated partial thromboplastin time(APTT),fibrinogen(FIB),and thrombin time(TT)were measured.Levels of immunoglobulin A(IgA),immunoglobulin G(IgG),and immunoglobulin M(IgM)were determined using the nephelometric immunoassay.Multivariate Cox regression analysis was conducted to explore the influencing factors of poor prognosis.Results Compared with the control group,children in both the non-severe and severe groups exhibited increased levels of white blood cells(WBC),neutrophils(N),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),interleukin-6(IL-6),procalci-tonin(PCT),FIB,and IgG,as well as decreased levels of B lymphocytes,natural killer(NK)cells,PT,APTT,IgA,and IgM(P＜0.05).Compared with the non-severe group,children in the severe group showed increased levels of WBC,N,CRP,ESR,IL-6,PCT,FIB,and IgG,as well as decreased levels of B lymphocytes,NK cells,PT,APTT,IgA,and IgM(P＜0.05).Compared with the good prognosis subgroup,the poor prognosis subgroup exhibited increased levels of CRP,IL-6,PCT,and IgG,as well as decreased levels of APTT,IgM,and IgA(P＜0.05).The results of multivariate Cox regression analysis showed that PCT,APTT,and IgM were influencing factors of poor prognosis in children with SAP(P＜0.05).Conclusion Children with SAP have decreased levels of PT,APTT,IgA,and IgM,as well as increased levels of FIB and IgG.Coagulation and im-mune function indicators are correlated with the prognosis of SAP.

Objective This study was to investigate the effects of Ginseng Astragalus Tumor Suppressor Formula combined with cisplatin on relevant immune genes and immune functions in mice modeled with hepatocellular carcinoma based on bioinformatics research.Methods Gene expression profiles and clinical data of hepatocellular carcinoma patients were downloaded from TCGA and GEO databases and screened for immune-expressed genes by taking intersections with immune-related genes downloaded from ImmPort database.Immune-related gene pair coefficients(IRGPI)were calculated to construct IRGP prognostic models.The optimal cut-off value of IRGPI for 1-year overall survival of hepatocellular carcinoma patients was determined based on ROC curve analysis,and hepatocellular carcinoma patients were divided into high and low immune risk groups,and the survival status of patients in the two groups was analyzed using the Kaplan-Meier method and the Log-Rank test.Then,we screened the active ingredients and gene targets of Ginseng Astragali Tumor Suppressor Formula for the treatment of hepatocellular carcinoma by network pharmacology,and obtained the intersection genes between Ginseng Astragali Tumor Suppressor Formula and disease,and then extracted the intersection of hepatocellular carcinoma-related immune genes and the intersection genes between Ginseng Astragali Tumor Suppressor Formula and disease through the"VennDiagram"software,and verified it through the animal model.The effects of Astragalus tumor-suppressing formula combined with cisplatin on the immunity genes and immune functions in the tumor tissues of mice with hepatocellular carcinoma were verified in animal models.Results Among 2483 relevant immune genes,84 pairs of immune-related genes were significantly associated with OS in the experimental group(P<0.001),and among the patients categorized into high and low immune risk groups by cut-off value-0.258,the overall survival rate of the high immune risk group was significantly lower than that of the low immune risk group(P<0.001).Univariate Cox analysis showed that IRGP model risk values and clinical characteristics of tumor T-staging had an impact on prognosis.Multifactorial Cox analysis showed that IRGP model risk value and tumor T-stage could be used as independent prognostic factors.266 genes intersecting with hepatocellular carcinoma were screened by network pharmacology technique for Ginseng Astragalus Tumor Suppressor Formula,and further 8 genes were obtained by taking the intersection with 84 pairs of immune-related genes.The experimental results showed that compared with the model group,the tumor mass of mice in each treatment group decreased(P<0.05);the spleen index and thymus index of mice increased(P<0.05);the CD4+/CD8+ratio in serum and spleen tissues of mice decreased;ICAM1,FABP5,IGF2,CDK4,NR1,ADRB2,AR,NR3C2 in tumor tissue of mice mRNA expression were all decreased(P<0.05),and the therapeutic effect of the combined group was significant(P<0.01).Conclusion This study predicted the key immune genes related to hepatocellular carcinoma as well as the prognostic analysis of immune-related genes,and the experimental study verified that Ginseng and Astragalus Tumor Suppressor Formula could effectively reduce the expression of related immune genes in tumor tissues,and improve the proportion of related immune cells in the splenic tissues of mice with hepatocellular carcinoma model as well as improve the immune function of mice.

This study was aimed at exploring the interaction between the nucleoprotein(NP)of influenza A virus(IAV)and TRIM25.The physicochemical properties and protein structure of IAV NP protein were analyzed through bioinformatics methods.The interaction between IAV NP and TRIM25 proteins was simulated with molecular docking techniques,and the in-teraction sites were predicted.With the cDNA of the A/Puerto Rico/8/1934(H1N1)PR8 strain as the template,the NP pro-tein was cloned into the eukaryotic expression vector pCMV-C-Flag through PCR amplification,the eukaryotic expression re-combinant plasmid pCMV-Flag-NP was constructed,and the expression was further verified.The protein expression levels of pCMV-Flag-NP and pCMV-HA-TRIM25 were detected at various time periods.The interaction between NP protein and TRIM25 protein was verified by co-immunoprecipitation.The co-localization of NP protein and TRIM25 protein in cells was ob-served with laser confocal microscopy.Bioinformatics analysis revealed that the NP protein consists of 498 amino acids and 20 amino acids,and is an unstable hydrophilic protein.The NP protein has multiple phosphorylation sites,as well as N-glycosyla-tion and O-glycosylation sites,but no transmembrane domain or signal peptide domain.Additionally,the NP protein's second-ary structure consists of a high proportion of alpha-helices and random coils.The molecular docking prediction results indicated that IAV NP interacts with TRIM25 protein and has multiple potential interaction sites,including the 233rd alanine,234th ala-nine,236th lysine,and 440th alanine of the NP protein.After successfully constructing and expressing the IAV NP protein,we verified the interaction between IAV NP and TRIM25 protein by immunoprecipitation and laser confocal microscopy obser-vations.Our results together suggested that the structure of the IAV NP protein is closely related to its function,and its im-portance to the virus is clear.In addition,the interaction between IAV NP and TRIM25 protein may be associated with TRIM25's anti-influenza virus mechanism.Further in-depth research may provide new ideas for anti-influenza virus strategies.

This study was aimed at exploring the interaction between the nucleoprotein(NP)of influenza A virus(IAV)and TRIM25.The physicochemical properties and protein structure of IAV NP protein were analyzed through bioinformatics methods.The interaction between IAV NP and TRIM25 proteins was simulated with molecular docking techniques,and the in-teraction sites were predicted.With the cDNA of the A/Puerto Rico/8/1934(H1N1)PR8 strain as the template,the NP pro-tein was cloned into the eukaryotic expression vector pCMV-C-Flag through PCR amplification,the eukaryotic expression re-combinant plasmid pCMV-Flag-NP was constructed,and the expression was further verified.The protein expression levels of pCMV-Flag-NP and pCMV-HA-TRIM25 were detected at various time periods.The interaction between NP protein and TRIM25 protein was verified by co-immunoprecipitation.The co-localization of NP protein and TRIM25 protein in cells was ob-served with laser confocal microscopy.Bioinformatics analysis revealed that the NP protein consists of 498 amino acids and 20 amino acids,and is an unstable hydrophilic protein.The NP protein has multiple phosphorylation sites,as well as N-glycosyla-tion and O-glycosylation sites,but no transmembrane domain or signal peptide domain.Additionally,the NP protein's second-ary structure consists of a high proportion of alpha-helices and random coils.The molecular docking prediction results indicated that IAV NP interacts with TRIM25 protein and has multiple potential interaction sites,including the 233rd alanine,234th ala-nine,236th lysine,and 440th alanine of the NP protein.After successfully constructing and expressing the IAV NP protein,we verified the interaction between IAV NP and TRIM25 protein by immunoprecipitation and laser confocal microscopy obser-vations.Our results together suggested that the structure of the IAV NP protein is closely related to its function,and its im-portance to the virus is clear.In addition,the interaction between IAV NP and TRIM25 protein may be associated with TRIM25's anti-influenza virus mechanism.Further in-depth research may provide new ideas for anti-influenza virus strategies.

Objective This study was to investigate the effects of Ginseng Astragalus Tumor Suppressor Formula combined with cisplatin on relevant immune genes and immune functions in mice modeled with hepatocellular carcinoma based on bioinformatics research.Methods Gene expression profiles and clinical data of hepatocellular carcinoma patients were downloaded from TCGA and GEO databases and screened for immune-expressed genes by taking intersections with immune-related genes downloaded from ImmPort database.Immune-related gene pair coefficients(IRGPI)were calculated to construct IRGP prognostic models.The optimal cut-off value of IRGPI for 1-year overall survival of hepatocellular carcinoma patients was determined based on ROC curve analysis,and hepatocellular carcinoma patients were divided into high and low immune risk groups,and the survival status of patients in the two groups was analyzed using the Kaplan-Meier method and the Log-Rank test.Then,we screened the active ingredients and gene targets of Ginseng Astragali Tumor Suppressor Formula for the treatment of hepatocellular carcinoma by network pharmacology,and obtained the intersection genes between Ginseng Astragali Tumor Suppressor Formula and disease,and then extracted the intersection of hepatocellular carcinoma-related immune genes and the intersection genes between Ginseng Astragali Tumor Suppressor Formula and disease through the"VennDiagram"software,and verified it through the animal model.The effects of Astragalus tumor-suppressing formula combined with cisplatin on the immunity genes and immune functions in the tumor tissues of mice with hepatocellular carcinoma were verified in animal models.Results Among 2483 relevant immune genes,84 pairs of immune-related genes were significantly associated with OS in the experimental group(P<0.001),and among the patients categorized into high and low immune risk groups by cut-off value-0.258,the overall survival rate of the high immune risk group was significantly lower than that of the low immune risk group(P<0.001).Univariate Cox analysis showed that IRGP model risk values and clinical characteristics of tumor T-staging had an impact on prognosis.Multifactorial Cox analysis showed that IRGP model risk value and tumor T-stage could be used as independent prognostic factors.266 genes intersecting with hepatocellular carcinoma were screened by network pharmacology technique for Ginseng Astragalus Tumor Suppressor Formula,and further 8 genes were obtained by taking the intersection with 84 pairs of immune-related genes.The experimental results showed that compared with the model group,the tumor mass of mice in each treatment group decreased(P<0.05);the spleen index and thymus index of mice increased(P<0.05);the CD4+/CD8+ratio in serum and spleen tissues of mice decreased;ICAM1,FABP5,IGF2,CDK4,NR1,ADRB2,AR,NR3C2 in tumor tissue of mice mRNA expression were all decreased(P<0.05),and the therapeutic effect of the combined group was significant(P<0.01).Conclusion This study predicted the key immune genes related to hepatocellular carcinoma as well as the prognostic analysis of immune-related genes,and the experimental study verified that Ginseng and Astragalus Tumor Suppressor Formula could effectively reduce the expression of related immune genes in tumor tissues,and improve the proportion of related immune cells in the splenic tissues of mice with hepatocellular carcinoma model as well as improve the immune function of mice.

Paraplegia caused by spinal cord injury is a serious neurological complication, for which surgery is currently the main treatment method. Due to different surgical approaches, patients are usually expected to maintain a passive prone position for a long time or switch between the supine and prone positions. Affected by multiple factors such as neurogenic sensory disorders, pathological changes in muscle tone and operative duration, the risk of intraoperative acquired pressure injury (IAPI) is significantly increased. Current clinical prevention strategies for IAPI in these patients predominantly focus on localized pressure relief during positioning, lacking systematic, standardized comprehensive prevention protocols or evidence-based guidelines. To address it, Department of Nursing, Orthopedics Branch, China International Exchange and Promotive Association for Medical and Health Care, Spinal Trauma Professional Committee, Orthopedics Branch, Chinese Medical Doctor Association, Nursing Group of Spine and Spinal Cord Professional Committee of Chinese Association of Rehabilitation Medicine organized experts in relevant fields to formulate Guideline for the prevention of intraoperative acquired pressure injury in paraplegic patients with spinal cord injury ( version 2025), based on evidence-based medical evidence and latest research results and clinical practice at home and abroad. Eleven recommendations were put forward from the aspects of preoperative risk assessment, intraoperative prevention strategies, postoperative handover and monitoring, and supportive mechanisms for IAPI prevention, aiming to standardize the prevention measures and management strategies of IAPI in paraplegic patients with spinal cord injury and accelerate the recovery of patients and improve the therapeutic effect.