Aim To clarify the mechanism by which Qishen Yixin Granules improved microcirculation vas-cular endothelial dysfunction(VED)in mice,through activating the Nrf2/HO-1 signaling pathway to regulate oxidative stress.Methods C57 mice were randomly divided into six groups:blank group,model group,pos-itive drug group,and low-,medium-,and high-dose groups of Qishen Yixin Granules.The VED model was established by long-term infusion of Ang Ⅱ combined with a high-fat diet.Each treatment group received the corresponding drug intervention.After four weeks of drug intervention,cardiac function was assessed by echocardiography.Carstairs staining was used to ob-serve the formation of microthrombi in myocardial tis-sue.The micro vascular ischemia was evaluated by Hei-denhain staining.The ultrastructure of endothelial cells was observed by electron microscopy.The levels of EMPs,ROS,NO,ET-1,TF,TM,VWF,and TXA2 in serum were measured by ELISA.The expression levels of MDA,SOD,and GSH-Px in mouse heart tissue were determined by chemical methods.Cardiac microvascu-lar density and the expression of Nrf2,Keap1,and HO-1 proteins were detected by Immunohistochemical stai-ning.The protein expressions of Keap1,cytoplasmic Nrf2,nuclear Nrf2,and HO-1 in myocardial tissue were detected by Western blot.Results Qishen Yixin Granules could effectively improve the cardiac function of mice,alleviate the damage of endothelial cells and endothelial function.They could up-regulate serum NO levels and the activities of antioxidant enzymes SOD and GSH-Px,while down-regulating the expression of ROS and vascular inflammatory injury factors such as ET-1,VWF,TXA2,TF,TM,and EMPs.Qishen Yixin Granules also increased the positive counts of CD34,Nrf2,and HO-1,as well as microvessel density.Fur-thermore,they inhibited the expression of MDA,Keap1,and cytoplasmic Nrf2 protein in myocardial tis-sue,while increasing the expression of nuclear proteins HO-1 and Nrf2.Conclusions Qishen Yixin Granules may inhibit oxidative stress and inflammatory response by regulating the Nrf2/HO-1 signaling pathway,thereby improving vascular endothelial damage and cardiac function in VED mice.

Aim To reveal the mechanism of CIP4 homologs protein 1(RICH1)are involved in the regu-lation of myocardial fibrosis.Methods Mouse cardiac fibroblasts(MCFs)cells were treated with transforming growth factor-β(TGF-β1)to induce the formation of a myocardial fibrosis cell model;the level of the target protein was detected by Western blotting;and the RICH1 gene was detected by transfection of the cells with plasmid.The RICH1 gene was overexpressed(RICH 1 OE)using plasmid transfection;the RICH1 gene was silenced using siRNA fragment(siRICH1);and the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3 a1,and Acta2,were de-tected using RT-qPCR.Results RICH1 was signifi-cantly down-regulated in TGF-β1-treated MCFs;the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3a1,and Acta2,were down-regu-lated in the RICH1 OE+TGF-β1 group;and in the siRICH1+TGF-β1 group,myocardial fibrosis marker genes,such as Col1 a1,Col3a1 and Acta2 were up-regulated at the expression level;phosphorylated SMAD2(p-SMAD2)and phosphorylated SMAD3(p-SMAD3)levels were down-regulated in the siRICH1 OE+TGF-β1 group.p-SMAD2 and P-SMAD3 levels were upregulated in the siRICH1+TGF-β1 group.Conclusion RICH1 inhibits TGF-β1-induced myo-cardial fibrosis;RICH1 inhibits TGF-β1-induced myo-cardial fibrosis by negatively regulating the SMAD2/3 signaling pathway.

Objective:To investigate the changes of nitric oxide(NO),endothelin-1(ET-1),thromboxane B2(TXB2)levels in patients with chronic heart failure(CHF)and their correlation with plasma brain natriuretic peptide(BNP).Methods:We enrolled 110 CHF patients admitted in the Second Hospital of Qinhuangdao between January and December 2022.According to the New York Heart Association(NYHA)cardiac function classification,the patients were divided into cardiac function class Ⅱ group(n=22),cardiac function class Ⅲ group(n=50),and cardiac function class Ⅳ group(n=38).Baseline data,levels of NO,ET-1,TXB2,BNP and other related laboratory indexes were compared among three groups.Pearson correlation analysis was used to analyze the association of NO,ET-1,TXB2 with plasma BNP in CHF patients.Multivariate linear regression was employed to analyze related factors of elevated BNP in CHF patients.Results:Compared with patients in class Ⅱ group and class Ⅲ group,those in class Ⅳ group had significant higher uric acid[(467.39±32.60)μmol/L vs.(367.25±22.39)μmol/L vs.(421.42±28.34)μmol/L],total bilirubin[(17.36±3.10)μmol/L vs.(10.65±1.39)μmol/L vs.(11.12±2.01)μmol/L],BNP[(897.60±50.11)ng/L vs.(381.37±31.25)ng/L vs.(527.60±47.84)ng/L],NO[(50.12±5.95)μmol/L vs.(25.36±2.14)μmol/L vs.(37.92±4.84)μmol/L],ET-1[(114.10±10.53)pg/L vs.(80.25±7.38)pg/L vs.(97.03±8.40)pg/L],TXB2[(417.98±29.35)pg/ml vs.(302.63±19.63)pg/ml vs.(381.29±26.44)pg/ml](P＜0.001 all).Compared with those in class Ⅱ group,those in class Ⅲ group had significant higher above-mentioned indexes(except total bilirubin)(P＜0.001 all).Pearson correlation analysis indicated that plasma NO,ET-1,TXB2 were positively associated with BNP(r=0.828,0.750,0.720,P＜0.001 all).Multivariate linear regression analysis indicated that uric acid,total bilirubin,plasma NO,ET-1 and TXB2 levels were independent risk factors for elevated BNP level in CHF patients(B=0.555～20.550,P＜0.05 or＜0.01),while the use of angiotensin converting enzyme inhibitors(ACEI)/angiotensin Ⅱ receptor blockers(ARB)was an independent protective factor(B=-46.222,P=0.027).Conclusion:The levels of NO,ET-1,and TXB2 are closely related to the occurrence and development of CHF,and they show an increasing trend with the progression of CHF,and is closely related to BNP.

Aim To reveal the mechanism of CIP4 homologs protein 1(RICH1)are involved in the regu-lation of myocardial fibrosis.Methods Mouse cardiac fibroblasts(MCFs)cells were treated with transforming growth factor-β(TGF-β1)to induce the formation of a myocardial fibrosis cell model;the level of the target protein was detected by Western blotting;and the RICH1 gene was detected by transfection of the cells with plasmid.The RICH1 gene was overexpressed(RICH 1 OE)using plasmid transfection;the RICH1 gene was silenced using siRNA fragment(siRICH1);and the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3 a1,and Acta2,were de-tected using RT-qPCR.Results RICH1 was signifi-cantly down-regulated in TGF-β1-treated MCFs;the expression levels of myocardial fibrosis marker genes,such as Col1 a1,Col3a1,and Acta2,were down-regu-lated in the RICH1 OE+TGF-β1 group;and in the siRICH1+TGF-β1 group,myocardial fibrosis marker genes,such as Col1 a1,Col3a1 and Acta2 were up-regulated at the expression level;phosphorylated SMAD2(p-SMAD2)and phosphorylated SMAD3(p-SMAD3)levels were down-regulated in the siRICH1 OE+TGF-β1 group.p-SMAD2 and P-SMAD3 levels were upregulated in the siRICH1+TGF-β1 group.Conclusion RICH1 inhibits TGF-β1-induced myo-cardial fibrosis;RICH1 inhibits TGF-β1-induced myo-cardial fibrosis by negatively regulating the SMAD2/3 signaling pathway.

Idiopathic pulmonary fibrosis(IPF)and lung cancer(LC)pose a serious threat to human health,the prognosis of IPF is poor,the 5-year survival rate is low,and the incidence rate and mortality of LC are high.At present,IPF combined with LC(IPF-LC)has attracted much attention,but the pathogenesis is unclear.Epidemiology shows that,the risk of LC in IPF patients is 7 times than that of the general population,the incidence rate varies in different regions,which is related to smoking rate and diagnostic criteria.In terms of pathogenesis,chronic inflammation,angiogenesis,signaling pathways,aging,gene mutations,epithelial mesenchymal transition(EMT),and immune dysfunction are common pathogenic factors of both,and pathological changes such as smoking and pulmonary fibrosis increase the risk of LC.Thoroughly studying these mechanisms is of great significance for the early diagnosis,treatment strategy formulation,and improvement of patient prognosis of IPF-LC.

Idiopathic pulmonary fibrosis(IPF)and lung cancer(LC)pose a serious threat to human health,the prognosis of IPF is poor,the 5-year survival rate is low,and the incidence rate and mortality of LC are high.At present,IPF combined with LC(IPF-LC)has attracted much attention,but the pathogenesis is unclear.Epidemiology shows that,the risk of LC in IPF patients is 7 times than that of the general population,the incidence rate varies in different regions,which is related to smoking rate and diagnostic criteria.In terms of pathogenesis,chronic inflammation,angiogenesis,signaling pathways,aging,gene mutations,epithelial mesenchymal transition(EMT),and immune dysfunction are common pathogenic factors of both,and pathological changes such as smoking and pulmonary fibrosis increase the risk of LC.Thoroughly studying these mechanisms is of great significance for the early diagnosis,treatment strategy formulation,and improvement of patient prognosis of IPF-LC.

Intraoperative cell salvage (IOCS) has been widely applied as an important blood conservation measure in surgical operations. However, there is currently a lack of clinical practice guidelines for the implementation of IOCS in patients with malignant tumors. This report aims to provide clinicians with recommendations on the use of IOCS in patients with malignant tumors based on the review and assessment of the existed evidence. Data were derived from databases such as PubMed, Embase, the Cochrane Library and Wanfang. The guideline development team formulated recommendations based on the quality of evidence, balance of benefits and harms, patient preferences, and health economic assessments. This study constructed seven major clinical questions. The main conclusions of this guideline are as follows: 1) Compared with no perioperative allogeneic blood transfusion (NPABT), perioperative allogeneic blood transfusion (PABT) leads to a more unfavorable prognosis in cancer patients (Recommended); 2) Compared with the transfusion of allogeneic blood or no transfusion, IOCS does not lead to a more unfavorable prognosis in cancer patients (Recommended); 3) The implementation of IOCS in cancer patients is economically feasible (Recommended); 4) Leukocyte depletion filters (LDF) should be used when implementing IOCS in cancer patients (Strongly Recommended); 5) Irradiation treatment of autologous blood to be reinfused can be used when implementing IOCS in cancer patients (Recommended); 6) A careful assessment of the condition of cancer patients (meeting indications and excluding contraindications) should be conducted before implementing IOCS (Strongly Recommended); 7) Informed consent from cancer patients should be obtained when implementing IOCS, with a thorough pre-assessment of the patient's condition and the likelihood of blood loss, adherence to standardized internally audited management procedures, meeting corresponding conditions, and obtaining corresponding qualifications (Recommended). In brief, current evidence indicates that IOCS can be implemented for some malignant tumor patients who need allogeneic blood transfusion after physician full evaluation, and LDF or irradiation should be used during the implementation process.

Aim To clarify the mechanism by which Qishen Yixin Granules improved microcirculation vas-cular endothelial dysfunction(VED)in mice,through activating the Nrf2/HO-1 signaling pathway to regulate oxidative stress.Methods C57 mice were randomly divided into six groups:blank group,model group,pos-itive drug group,and low-,medium-,and high-dose groups of Qishen Yixin Granules.The VED model was established by long-term infusion of Ang Ⅱ combined with a high-fat diet.Each treatment group received the corresponding drug intervention.After four weeks of drug intervention,cardiac function was assessed by echocardiography.Carstairs staining was used to ob-serve the formation of microthrombi in myocardial tis-sue.The micro vascular ischemia was evaluated by Hei-denhain staining.The ultrastructure of endothelial cells was observed by electron microscopy.The levels of EMPs,ROS,NO,ET-1,TF,TM,VWF,and TXA2 in serum were measured by ELISA.The expression levels of MDA,SOD,and GSH-Px in mouse heart tissue were determined by chemical methods.Cardiac microvascu-lar density and the expression of Nrf2,Keap1,and HO-1 proteins were detected by Immunohistochemical stai-ning.The protein expressions of Keap1,cytoplasmic Nrf2,nuclear Nrf2,and HO-1 in myocardial tissue were detected by Western blot.Results Qishen Yixin Granules could effectively improve the cardiac function of mice,alleviate the damage of endothelial cells and endothelial function.They could up-regulate serum NO levels and the activities of antioxidant enzymes SOD and GSH-Px,while down-regulating the expression of ROS and vascular inflammatory injury factors such as ET-1,VWF,TXA2,TF,TM,and EMPs.Qishen Yixin Granules also increased the positive counts of CD34,Nrf2,and HO-1,as well as microvessel density.Fur-thermore,they inhibited the expression of MDA,Keap1,and cytoplasmic Nrf2 protein in myocardial tis-sue,while increasing the expression of nuclear proteins HO-1 and Nrf2.Conclusions Qishen Yixin Granules may inhibit oxidative stress and inflammatory response by regulating the Nrf2/HO-1 signaling pathway,thereby improving vascular endothelial damage and cardiac function in VED mice.

This study was aimed at establishing a sensitive and specific sandwich ELISA detection method for Brucella.We screened monoclonal capture antibodies and detection antibodies for Brucella detection,and optimized and determined the opti-mal antibody coating time and concentration,as well as the optimal blocking solution,blocking time,and yin-yang critical val-ue.The specificity of this method was verified by examination of other bacteria prone to cross-reacting with Brucella.The sen-sitivity of the method was verified by detection of a gradient dilution of inactivated Brucella.Moreover,the sandwich ELISA detection results were compared with test tube agglutination and qPCR results.The selected capture antibody was 4A12,and the selected detection antibody was 6C12.Experimental analysis indicated that the optimal coating concentration for the 4A12 capture antibody was 5 μg/mL,and the optimal dilution ratio for the 6C12 detection antibody was 1∶2000.The optimal coating conditions were overnight at 4℃,and blocking with 5％skim milk powder for 2 hours.The established double antibody sand-wich ELISA method reacted with only Brucella but not other bacteria,thus demonstrating the method's good specificity.Inac-tivated Brucella solution was still detectable after dilution to 1 × 105 CFU/mL,thus demonstrating the method's good sensitiv-ity.The intra-and inter batch coefficients of variation were both below 10％,thus indicating the method's good repeatability.Thus,this study successfully established a dual antibody sandwich ELISA method for Brucella detection,which has good spe-cificity and sensitivity,and might provide an effective approach for the precise diagnosis and effective prevention and control of brucellosis.

Objective:To investigate the changes of nitric oxide(NO),endothelin-1(ET-1),thromboxane B2(TXB2)levels in patients with chronic heart failure(CHF)and their correlation with plasma brain natriuretic peptide(BNP).Methods:We enrolled 110 CHF patients admitted in the Second Hospital of Qinhuangdao between January and December 2022.According to the New York Heart Association(NYHA)cardiac function classification,the patients were divided into cardiac function class Ⅱ group(n=22),cardiac function class Ⅲ group(n=50),and cardiac function class Ⅳ group(n=38).Baseline data,levels of NO,ET-1,TXB2,BNP and other related laboratory indexes were compared among three groups.Pearson correlation analysis was used to analyze the association of NO,ET-1,TXB2 with plasma BNP in CHF patients.Multivariate linear regression was employed to analyze related factors of elevated BNP in CHF patients.Results:Compared with patients in class Ⅱ group and class Ⅲ group,those in class Ⅳ group had significant higher uric acid[(467.39±32.60)μmol/L vs.(367.25±22.39)μmol/L vs.(421.42±28.34)μmol/L],total bilirubin[(17.36±3.10)μmol/L vs.(10.65±1.39)μmol/L vs.(11.12±2.01)μmol/L],BNP[(897.60±50.11)ng/L vs.(381.37±31.25)ng/L vs.(527.60±47.84)ng/L],NO[(50.12±5.95)μmol/L vs.(25.36±2.14)μmol/L vs.(37.92±4.84)μmol/L],ET-1[(114.10±10.53)pg/L vs.(80.25±7.38)pg/L vs.(97.03±8.40)pg/L],TXB2[(417.98±29.35)pg/ml vs.(302.63±19.63)pg/ml vs.(381.29±26.44)pg/ml](P＜0.001 all).Compared with those in class Ⅱ group,those in class Ⅲ group had significant higher above-mentioned indexes(except total bilirubin)(P＜0.001 all).Pearson correlation analysis indicated that plasma NO,ET-1,TXB2 were positively associated with BNP(r=0.828,0.750,0.720,P＜0.001 all).Multivariate linear regression analysis indicated that uric acid,total bilirubin,plasma NO,ET-1 and TXB2 levels were independent risk factors for elevated BNP level in CHF patients(B=0.555～20.550,P＜0.05 or＜0.01),while the use of angiotensin converting enzyme inhibitors(ACEI)/angiotensin Ⅱ receptor blockers(ARB)was an independent protective factor(B=-46.222,P=0.027).Conclusion:The levels of NO,ET-1,and TXB2 are closely related to the occurrence and development of CHF,and they show an increasing trend with the progression of CHF,and is closely related to BNP.