Objective This study investigated the regulatory effect of high mobility group protein B1 (HMGB1) in the peripheral blood of patients with primary immune thrombocytopenia (ITP) on myeloid dendritic cells (mDC) and Th17/regulatory T cells (Treg) balance. Methods The study enrolled 30 newly diagnosed ITP patients and 30 healthy controls.Flow cytometry was used to measure the proportion of mDC, Th17, and Treg cells in the peripheral blood of ITP patients and healthy controls. ELISA was conducted to quantify the serum levels of HMGB1, interleukin 6 (IL-6), IL-23, IL-17, and transforming growth factor β(TGF-β). The mRNA levels of retinoic acid-related orphan receptor γt(RORγt) and forehead box P3(FOXP3) were detected by real-time PCR. The correlation between the abovementioned cells, cytokines, and platelet count was assessed using Pearson linear correlation analysis. Results The proportion of Th17 cells and the expression levels of HMGB1, IL-6, IL-23, IL-17 and the level of RORγt mRNA in the peripheral blood of ITP patients were higher than those in healthy controls. However, the Treg cell proportion and TGF-β level were lower in ITP patients than those in healthy controls. In patients with ITP, the proportion of mDC and the level of FOXP3 mRNA did not show significant changes. The proportion of mDC cells was significantly correlated with the expression of IL-6 and IL-23. Moreover, the expression of HMGB1 showed a significant correlation with the expression of mDC, IL-6, IL-23, RORγt mRNA, and IL-17. Notably, both the proportion of mDC cells and the expression of HMGB1 were negatively correlated with platelet count. Conclusion The high expression of HMGB1 in peripheral blood of ITP patients may induce Th17/Treg imbalance by promoting the maturation of mDC and affecting the secretion of cytokines, thereby potentially playing a role in the immunological mechanism of ITP.
Humans ; Th17 Cells/cytology* ; HMGB1 Protein/genetics* ; T-Lymphocytes, Regulatory/cytology* ; Female ; Male ; Dendritic Cells/metabolism* ; Adult ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic/genetics* ; Nuclear Receptor Subfamily 1, Group F, Member 3/genetics* ; Young Adult ; Interleukin-23/blood* ; Interleukin-17/blood* ; Interleukin-6/blood* ; Forkhead Transcription Factors/genetics* ; Myeloid Cells/cytology* ; Aged

Objective To investigate the synergistic therapeutic effects of HER2 antibody-drug conjugate(HER2-ADC)combined with anti-PD-1 antibody(anti-PD-1)on HER2-expressing bladder cancer and elucidate its regulatory mechanisms on the tumor immune microenvironment.Methods Orthotopic tumor models were established in 40 female C57BL/6 mice(6～8 weeks old,body mass 18～22 g)using MB49 bladder cancer cells overexpressing human HER2.When tumors reached 100 mm3,the mice were randomized into(n=10)control(intraperitoneal injection of 1.0 mL PBS),anti-PD-1(200 μg per mouse every 3 d),HER2-ADC(2.5 mg/kg once weekly),and combination groups(same regimens as above monotherapy).Tumor volume and body mass were measured every 3 d during 28-day treatment.Tumor growth kinetics and survival rates were analyzed.Post-treatment survival was monitored until natural death to determine median survival time(n=5).At day 28,blood and tumor samples(n=5)were collected to detect myeloid-derived suppressor cells(MDSCs;CD11b?Gr1?)and regulatory T cells(Tregs;CD4?CD25?FOXP3?)with flow cytometry,tumor-infiltrating CD3?T,CD8?T,and FOXP3?T cells with immunohistochemical assasy,and liver/kidney functions[alanine aminotransferase(ALT),aspartate aminotransferase(AST),blood urea nitrogen(BUN),creatinine(CRE)]and tissue damage indicators[lactate dehydrogenase isoenzyme(LDH-L)].Results In 28 d after treatment,the combination group obtained significantly smallest tumor volume than the control group and the 2 monotherapy groups(all P<0.01).The longest median survival was observed in the combination group(65 d,P<0.01),followed by the HER2-ADC group(55 d),anti-PD-1 group(53 d)and control group(41 d).After 28 d of treatment,the combination group exhibited obviously the smallest peripheral proportions of MDSCs/Tregs,most tumor-infiltrating CD3?T/CD8?T cells,and less FOXP3?T cells when compared with the 2 monotherapy groups and control group(all P<0.05).While,the 2 monotherapy groups had smaller MDSCs/Tregs proportions than the control group(P<0.05).No significant differences were observed among the 4 groups in serum ALT,AST,BUN,CRE,or LDH-L levels,and all of them were within normal ranges.Conclusion HER2-ADC combined with anti-PD-1 suppresses the growth of orthotopic bladder tumor,probably through their synergic effects on down-regulating MDSCs/Treg and enhancing CD8?T cell infiltration.

Objective:To analyze the current situation of medical consumables management policy in China,and to provide a reference for the refined management of medical consumables.Methods:Through the policy triangle model and policy tool theory,it comprehensively analyzes the reimbursement medical consumables catalogue and payment management policy of medical insurance in China,covering the policy background,content,process,and participant dimensions.Results:The use frequency of medical consumables policy tools is not balanced,the payment management rules need to be refined,and the participation of multi-stakeholders such as patients is lacking.Conclusion:It is necessary to further strengthen the foundational management of reimbursement medical consumables catalogue,improve the access mechanism of medical consumables for medical insurance,and explore the formulation of categorized payment standards and innovative payment mechanisms.

Objective To investigate the effect of ventilation with different tidal volumes on respiratory mechanics and haemodynamics in children with severe pneumonia.Methods A total of 104 children with severe pneumonia were divided into the control group and the observation group using a random number table method,with 52 cases in each group.Both groups underwent mechanical ventilation treatment.The control group was treated with conventional tidal volume of 10-12 mL/kg,while the observation group was given a low tidal volume of 6-8 mL/kg.The respiratory mechanics indicators,hemodynamic indicators and inflammatory factor indicators before and after treatment were compared between the two groups.Results After treatment,the airway resistance(AR)and airway closing pressure(AOP)were lower in the observation group than those of the control group(P＜0.05),and pulmonary dynamic compliance(Cdyn)was higher in the observation group than that of the control group(P＜0.05).There were no significant differences in heart rate(HR),mean arterial pressure(MAP)and central venous pressure(CVP)after treatment between the two groups(P＜0.05).After treatment,the levels of interleukin(IL)-6,IL-8 and tumor necrosis factor-α(TNF-α)were all lower in the observation group than those in the control group(P＜0.05).Conclusion Small tidal volume mechanical ventilation is effective in improving the respiratory mechanics of children with severe pneumonia without causing large fluctuations in their hemodynamics.At the same time,it can reduce the level of inflammatory factors in the organism.

Objective To explore the expression of leukemia inhibitory factor receptor(LIFR)in patients with non-ST-segment elevation myocardial infarction(NSTEMI)and its possible associa-tion with myocardial remodeling.Methods A total of 188 patients with acute myocardial infarc-tion who underwent coronary angiography in our hospital from January 2023 to November 2024 were prospectively enrolled and divided into a control group(94 cases)and an NSTEMI group(94 cases)according to being diagnosed with NSTEMI or not.General clinical data of the patients were collected,and the correlation between serum LIFR level and other indicators was analyzed using linear regression analysis.Results Compared with the control group,the NSTEMI group had significantly higher ratios of smoking history,elevated levels of LIFR,NT-proBNP,cTnⅠ,Cr and UA,increased WBC count,but lower LVEF value[48.94％vs 13.83％,P＜0.01;5.82(4.23,8.11)mmol/L vs 0.97(0.60,1.41)mmol/L,P＜0.01;2.53(1.24,9.71)pg/L vs 0.03(0.02,0.04),P＜0.01;18.57(4.11,250.00)ng/L vs 0.00(0.00,0.00)ng/L,P＜0.01;82.50(68.00,121.25)μmol/L vs 68.50(53.00,88.25)μmol/L,P＜0.01;411.00(349.00,521.25)μmol/L vs 337.00(286.75,406.00)μmol/L,P＜0.01;10.21(8.71,13.09)× 109/L vs 6.22(4.67,7.46)× 109/L,P＜0.01;47.00(38.00,54.00)％vs 59.00(56.00,60.00)％,P＜0.01].Serum LIFR level in the patients was posi-tively correlated with NT-proBNP,cTnⅠ,Cr and WBC count(β=1.403,95％CI:10 597.867-17 327.087,P=0.000;β=0.232,95％CI:114.558-1769.808,P=0.026;β=0.336,95％CI:0.164-0.617,P=0.001).Conclusion LIFR may be involved in the development of myocardial remode-ling and heart failure after myocardial infarction through its role in inflammation.

Objective To investigate the mechanism of kaempferol(KPF)in the treatment of macular edema sec-ondary to retinal vein occlusion(RVO-ME).Methods RVO model was established in SD rats using photocoagulation.Twenty SD rats were randomly divided into:Control group(normal rats,saline injection),Model group(RVO model rats,saline injection),Low KPF group[RVO model rats,KPF injection(8 mg·kg-1·d-1)],High KPF group[RVO model rats,KPF injection(16 mg·kg-1·d-1)],with 5 rats per group for 14 days.Fundus photography observed retinal vessels;HE staining evaluated retinal structural changes.HRMECs were randomly divided into:control group(no intervention),CoCl2 group(300 μmol·L-1 CoCl2),Low-dose KPF group(300 μmol·L-1 CoCl2+20 μmol·L-1 KPF),High-dose KPF group(300 μmol·L-1 CoCl2+30 μmol·L-1 KPF)and pathway inhibitor group(DAPT group)(300 μmol·L-1 CoCl2+20 μmol·L-1 DAPT),with 24-hour intervention.CCK-8 assay detected cell viability;Scratch test measured cell migration rate;ELISA quantified inflammatory factors[interleukin(IL)-6,tumor necrosis factor(TNF)-α];FITC-dextran assay evalu-ated endothelial monolayer permeability;Immunofluorescence detected zonula occludens(ZO)-1 expression;Western blot and RT-PCR measured protein and mRNA levels of ZO-1,Occludin,Notch1,and Hes1.Results Fundus photography showed interrupted venous blood flow and retinal edema in Model group,alleviated by KPF.HE staining revealed disorder-ed retinal arrangement and severe edema in the Model group,improved by KPF.Compared with CoCl2 group,Low-and High-dose KPF groups showed reduced cell migration rate,increased cell vitality(both P＜0.05),IL-6 and TNF-α levels de-creased in Low-and High-dose KPF groups and DAPT group(all P＜0.05),permeability coefficient decreased in Low-and High-dose KPF groups and DAPT group(all P＜0.05),ZO-1 expression increased in Low-and High-dose KPF groups and DAPT group(all P＜0.05),ZO-1 and Occludin protein/mRNA levels increased;Notch1 and Hes1 protein/mRNA levels de-creased in Low-and High-dose KPF groups and DAPT group(all P＜0.05).Conclusion KPF treat RVO-ME by inhibiting the Notch1/Hes1 pathway,exerting anti-inflammatory effects and improving intercellular tight junctions.

Objective To develop and validate an efficient and simple liquid chromatography with tandem mass spectrometry(LC-MS/MS)method for determination of polymyxin E in human plasma,and apply the established method in therapeutic drug monitoring(TDM)of polymyxin E.Methods The LC-MS/MS platform was based on AB SCIEX HPLC-4500MD system.Gradient elution was performed with 0.2％formic acid in water and 0.2％formic acid in acetonitrile.Phenomenex Kinetex XB-C18 column(100 mm × 2.1 mm,2.6 μm)were used.The analytes were detected by electrospray ionization(ESI)positive multiple reaction monitoring mode.The ion pairs for analytes(polymyxins E1,E2)and internal standard(polymyxins B1)were m/z 390.7→101.3,m/z 386.0→101.2,and m/z 402.3→101.2,respectively.Plasma samples were processed with protein precipitation method.Results Polymyxin E1 and E2 showed good linearity in the range of 0.031 2-6.24 mg/L and 0.006 15-1.23 mg/L,respectively.The within-run accuracy of polymyxin E1 and E2 in plasma ranged from 89.4％to 99.8％and 91.5％to 108.2％,respectively,while the between-run accuracy ranged from 91.8％to 104.7％and 95.6％to 105.2％,respectively.The within-run precision of polymyxin E1 and E2 in plasma ranged from 4.9％to 8.9％and 2.8％to 8.5％,respectively,while the between-run precision ranged from 4.1％to 7.6％and 4.2％to 9.8％,respectively.The average internal standard normalized matrix effect factors of polymyxins E1 and E2 were 96.9％-111.2％and 106.1％-112.8％in blank plasma samples from 6 different sources,102.5％-106.8％and 98.8％-105.2％in lipemic plasma,respectively,107.8％-108.9％and 106.9％-1 07.4％in hemolyzed plasma,respectively.The precision of matrix effects was less than 15.0％.The average recovery rate was 102.9％-107.5％for polymyxin E1 and E2,and 107.0％for internal standard polymyxin B1.The precision was less than 3.7％.Conclusions In this study,a simple and efficient LC-MS/MS method was established for determination of polymyxin E1 and E2 in human plasma,which is reliable in the therapeutic drug monitoring and pharmacokinetic study of polymyxin E.

Objective To investigate the effect of ventilation with different tidal volumes on respiratory mechanics and haemodynamics in children with severe pneumonia.Methods A total of 104 children with severe pneumonia were divided into the control group and the observation group using a random number table method,with 52 cases in each group.Both groups underwent mechanical ventilation treatment.The control group was treated with conventional tidal volume of 10-12 mL/kg,while the observation group was given a low tidal volume of 6-8 mL/kg.The respiratory mechanics indicators,hemodynamic indicators and inflammatory factor indicators before and after treatment were compared between the two groups.Results After treatment,the airway resistance(AR)and airway closing pressure(AOP)were lower in the observation group than those of the control group(P＜0.05),and pulmonary dynamic compliance(Cdyn)was higher in the observation group than that of the control group(P＜0.05).There were no significant differences in heart rate(HR),mean arterial pressure(MAP)and central venous pressure(CVP)after treatment between the two groups(P＜0.05).After treatment,the levels of interleukin(IL)-6,IL-8 and tumor necrosis factor-α(TNF-α)were all lower in the observation group than those in the control group(P＜0.05).Conclusion Small tidal volume mechanical ventilation is effective in improving the respiratory mechanics of children with severe pneumonia without causing large fluctuations in their hemodynamics.At the same time,it can reduce the level of inflammatory factors in the organism.

Objective To explore the expression of leukemia inhibitory factor receptor(LIFR)in patients with non-ST-segment elevation myocardial infarction(NSTEMI)and its possible associa-tion with myocardial remodeling.Methods A total of 188 patients with acute myocardial infarc-tion who underwent coronary angiography in our hospital from January 2023 to November 2024 were prospectively enrolled and divided into a control group(94 cases)and an NSTEMI group(94 cases)according to being diagnosed with NSTEMI or not.General clinical data of the patients were collected,and the correlation between serum LIFR level and other indicators was analyzed using linear regression analysis.Results Compared with the control group,the NSTEMI group had significantly higher ratios of smoking history,elevated levels of LIFR,NT-proBNP,cTnⅠ,Cr and UA,increased WBC count,but lower LVEF value[48.94％vs 13.83％,P＜0.01;5.82(4.23,8.11)mmol/L vs 0.97(0.60,1.41)mmol/L,P＜0.01;2.53(1.24,9.71)pg/L vs 0.03(0.02,0.04),P＜0.01;18.57(4.11,250.00)ng/L vs 0.00(0.00,0.00)ng/L,P＜0.01;82.50(68.00,121.25)μmol/L vs 68.50(53.00,88.25)μmol/L,P＜0.01;411.00(349.00,521.25)μmol/L vs 337.00(286.75,406.00)μmol/L,P＜0.01;10.21(8.71,13.09)× 109/L vs 6.22(4.67,7.46)× 109/L,P＜0.01;47.00(38.00,54.00)％vs 59.00(56.00,60.00)％,P＜0.01].Serum LIFR level in the patients was posi-tively correlated with NT-proBNP,cTnⅠ,Cr and WBC count(β=1.403,95％CI:10 597.867-17 327.087,P=0.000;β=0.232,95％CI:114.558-1769.808,P=0.026;β=0.336,95％CI:0.164-0.617,P=0.001).Conclusion LIFR may be involved in the development of myocardial remode-ling and heart failure after myocardial infarction through its role in inflammation.

Objective To develop and validate an efficient and simple liquid chromatography with tandem mass spectrometry(LC-MS/MS)method for determination of polymyxin E in human plasma,and apply the established method in therapeutic drug monitoring(TDM)of polymyxin E.Methods The LC-MS/MS platform was based on AB SCIEX HPLC-4500MD system.Gradient elution was performed with 0.2％formic acid in water and 0.2％formic acid in acetonitrile.Phenomenex Kinetex XB-C18 column(100 mm × 2.1 mm,2.6 μm)were used.The analytes were detected by electrospray ionization(ESI)positive multiple reaction monitoring mode.The ion pairs for analytes(polymyxins E1,E2)and internal standard(polymyxins B1)were m/z 390.7→101.3,m/z 386.0→101.2,and m/z 402.3→101.2,respectively.Plasma samples were processed with protein precipitation method.Results Polymyxin E1 and E2 showed good linearity in the range of 0.031 2-6.24 mg/L and 0.006 15-1.23 mg/L,respectively.The within-run accuracy of polymyxin E1 and E2 in plasma ranged from 89.4％to 99.8％and 91.5％to 108.2％,respectively,while the between-run accuracy ranged from 91.8％to 104.7％and 95.6％to 105.2％,respectively.The within-run precision of polymyxin E1 and E2 in plasma ranged from 4.9％to 8.9％and 2.8％to 8.5％,respectively,while the between-run precision ranged from 4.1％to 7.6％and 4.2％to 9.8％,respectively.The average internal standard normalized matrix effect factors of polymyxins E1 and E2 were 96.9％-111.2％and 106.1％-112.8％in blank plasma samples from 6 different sources,102.5％-106.8％and 98.8％-105.2％in lipemic plasma,respectively,107.8％-108.9％and 106.9％-1 07.4％in hemolyzed plasma,respectively.The precision of matrix effects was less than 15.0％.The average recovery rate was 102.9％-107.5％for polymyxin E1 and E2,and 107.0％for internal standard polymyxin B1.The precision was less than 3.7％.Conclusions In this study,a simple and efficient LC-MS/MS method was established for determination of polymyxin E1 and E2 in human plasma,which is reliable in the therapeutic drug monitoring and pharmacokinetic study of polymyxin E.