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ObjectiveMolecular docking and animal experiments were employed to explore the protective effect and mechanism of Da Chengqitang （DCQD） on intestinal barrier in septic mice. MethodText mining method was used to screen the active ingredients in DCQD. AutoDock Tools and Discovery Studio were used to study the interactions of active components with the core target proteins ［claudin-1， tumor necrosis factor （TNF）-α， interleukin （IL）-6， endogenous antimicrobial peptide mCRAMP， Toll-like receptor 4 （TLR4）， and myeloid differentiation primary response gene 88 （MyD88）］ in sepsis. Fifty C57BL/6 mice were randomized into sham， model， low- and high-dose （4 g∙kg-1 and 8 g∙kg-1） DCQD， and ulinastatin groups （n=10）. Before， during， and after the day of modeling surgery， each group was administrated with corresponding drugs. The mice in other groups except the model group were subjected to modeling by cecal ligation and puncture. Enzyme-linked immunosorbent assay （ELISA） was used measure the serum level of D-lactic acid to assess intestinal mucosa permeability. Hematoxylin-eosin staining was employed to observe the histopathological changes in the ileum and assess the intestinal mucosal damage and inflammatory infiltration. Western blotting was employed to determine the expression levels of tight junction proteins claudin-1 and occludin in the ileal tissue， which were indicative of the bowel barrier function. The TNF-α and IL-6 levels were measured by ELISA to assess the intestinal inflammation. The expression of mCRAMP in the ileal tissue was observed by immunohistochemistry. The mRNA levels of mCRAMP， TLR4， and MyD88 in mouse ileal tissue were determined by Real-time polymerase chain reaction， on the basis of which the mechanism of DCQD in protecting the intestinal barrier of septic mice was explored. ResultMolecular docking results showed that most of the 10 active ingredients of DCQD that were screened out by text mining could bind to sepsis targets by van der Waals force， hydrogen bonding， and other conjugated systems. The results of animal experiments showed that compared with the model group， low- or high-dose DCQD lowered the D-lactic acid level in the serum （P<0.01）， alleviated damage to the ileal tissue and mucosal edema， protected the small intestine villus integrity， reduced inflammatory cell infiltration， promoted the expression of claudin-1 （P<0.01）， lowered the IL-6 level （P<0.01）， up-regulated the mRNA and protein levels of mCRAMP （P<0.01）， and down-regulated the mRNA and protein levels of TLR4 and MyD88 （P<0.01） in the ileal tissue. In addition， high-dose DCQD lowered the TNF-α level and promoted the expression of occludin in the ileum tissue （P<0.01）， and low-dose DCQD up-regulated the protein level of occludin in the ileum tissue （P<0.05）. ConclusionDCQD has a protective effect on intestinal barrier in septic mice. It can reduce intestinal inflammation， repair intestinal mucosal damage， improve the tight junction protein level， and reduce intestinal mucosal permeability by up-regulating the mRNA and protein levels of mCRAMP and the down-regulating the expression of genes in the TLR4/MyD88 pathway.
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@#Objective To compare the effects of different signal peptides on the secretion and expression of SARS-CoV-2S1,receptor binding domain(RBD) and RBD dimer proteins in Expisf9 insect cells.Methods The gene sequences of three proteins,SARS-CoV-2 S1(M1-E661),RBD(R319-P545) and RBD dimer(R319-K537 tandem),were selected and divided into 25 groups according to the different N-terminal signal peptide sequences(Endo,honeybee melittin(HBM),GP64,GP67,chitinase(Chi) and HIV-ENV) and C-terminal label sequences.25 recombinant baculoviruses were constructed by Bac-to-Bac system,and 25 groups of tertiary strain banks were prepared.B2 and C4 viruses were inoculated to logarithmic prestage cells(2.8 × 10~6 cells/mL) and logarithmic metaphase cells(1.2 × 10~7 cells/mL),respectively.The viruses of each group were cultured to 100 mL(500 mL shaker) for protein expression,and samples were taken for SDSPAGE electrophoresis,Western-blot and ELISA detection.Two groups with higher expression levels of S1,RBD and RBD dimer proteins were selected for repeated verification.Results When B2 and C4 were inoculated to high cell density,the secretion expression level showed no increase,while there were significant difference between 4 and 5 d after inoculation.The expression level of A7(Endo-S1-tag) was significantly lower than that of A9(HIV-ENV-S1-tag),the expression level of A4(Gp67-S1-tag) was the highest,and the secreted expression level of A1(Endo-Endo-Sl-tag) was significantly lower than that of A7(Endo-S1-tag).The secretion and expression of B6(HIV-ENV-RBD-tag) was signifi-cantly higher than that of B4(Gp67-RBD-tag) and other signal peptide groups,and C4(Gp67-RBD-dimer-tag) expression was significantly higher than that of C3(Gp64-RBD-dimer-tag).Two groups with high expression of each protein were selected separately for repeated verification(A4,A9;B4,B6;C3,C4) and the results showed that A4,B6 and C4 had the highest secretion expression levels.Conclusion The signal peptide for the highest secretion expression of S1 and RBD dimer proteins is the same,which is GP67 signal peptide,while the most suitable signal peptide for RBD protein is HIV-ENV,indicating that the N-terminal sequence can affect protein secretion,signal peptide sequence is universal to a certain extent,but is also related to the target protein sequence to be expressed.
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@#Abstract: S-palmitoylation, a reversible and dynamic post-translational modification in cells, is involved in regulating the transcription and expression of downstream target genes as well as signal transduction, thereby affecting cell life activities. Studies have shown that thousands of human proteins undergo S-palmitoylation modification, suggesting that S-palmitoylation is closely related to the progression and treatment of diseases. T cells play central roles in anti-tumor immune responses. A variety of T cell immune-related proteins are regulated by S-palmitoylation. In the present study, we focus on the impact of S-palmitoylation on T cell signal transduction and its application in T cell immunotherapy, aiming to provide new ideas for the development of new targets and peptide inhibitors for T cell immunotherapy.
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Objective To study the therapeutic effect of Xuefu Zhuyu Capsule combined with sacubitril valsartan on dilated cardiomyopathy.Methods A total of 70 patients with dilated cardiomyopathy who were hospitalized in the Hospital from January to December 2020 were selected as the research objects and random-ly divided into control group and experimental group,with 35 cases in each group.The control group was only treated with sacubitril valsartan,and the experimental group was treated with Xuefu Zhuyu Capsule combined with sacubitril valsartan.According to the patient's blood pressure and renal function,sacubitril valsartan was titrated from a small dose to the maximum dose.Xuefu Zhuyu Capsule was uesd 2.4 g each time,twice a day,and the treatment time was three months.The symptoms of heart failure,glycosylated hemoglobin(HbA1c),low density lipoprotein(LDL),total cholesterol(TC),triglyceride(TG),ALT,AST,N-terminal pro-brain natriuretic peptide(NT-proBNP),left ventricular end-systolic diameter(LVEDd),left ventricular end-dias-tolic diameter(LVEDs),ejection fraction(EF)and the incidence of arrhythmia were observed in the two groups after treatment.Results After treatment,the level of NT-proBNP in the experimental group was sig-nificantly lower than that in the control group(P＜0.05).There was no significant difference in LDL,TC,TG,LVEDd,LVEDs and EF between the two groups(P＞0.05).The incidence of atrial tachycardia and ven-tricular premature beat in the experimental group was higher than that in the control group(P＜0.05).Con-clusion Xuefu Zhuyu Capsule combined with sacubitril valsartan can significantly improve the symptoms of patients with heart failure and reduce the level of NT-proBNP,but it may increase the proportion of patients with atrial tachycardia and ventricular premature beats.
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Objective To study the clinical effect of levosimendan combined with recombinant human brain natriuretic peptide(rhBNP)on patients with acute heart failure.Methods A total of 100 patients with acute heart failure in the hospital from December 2019 to December 2021 were selected as the research sub-jects.According to different treatment options,the subjects were divided into the control group,levosimendan group,rhBNP group and combined treatment group,with 25 cases in each group.The control group received traditional conventional diuretic,tube expansion and other treatment;the levosimendan group was treated with levosimendan on the basis of the control group;the rhBNP group was treated with rhBNP on the basis of the control group;the combined treatment group was treated with levosimendan and rhBNP on the basis of the control group.The improvement of New York Heart Association(NYHA)classification,death,rehospitaliza-tion rate,6-minute walking distance,improvement of serological indicators and adverse reactions were recor-ded in each group.Results Before treatment,there was no significant difference in baseline data between the groups(P>0.05).On the 1 st and 3 rd day after treatment,the improvement of NYHA classification in the combined treatment group was better than that in the other groups(P<0.05),and the improvement of NY-HA classification in the levosimendan group and rhBNP group was better than that in the control group(P<0.05).The readmission rate within 6 months after treatment in the combined treatment group was lower than that in the other groups(P<0.05).At 5 and 9 days after treatment,the 6-minute walking distance in the combined treatment group was longer than that in the other groups(P<0.05).At 9 days after treatment,the left ventricular ejection fraction(LVEF)in the combined treatment group was higher than that in the other groups(P<0.05),and the level of N-terminal B-type natriuretic peptide(NT-proBNP)in the combined treatment group was lower than that in the other groups(P<0.05).No significant difference was found in the comparison of the occurrence of adverse reactions among the four groups(P>0.05).Conclusion The combina-tion of levosimendan and rhBNP in the treatment of patients with acute heart failure is superior to traditional treatment and monotherapy in early clinical improvement,and dose not increase the incidence of adverse reactions.
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Objective To analyze the in-hospital and long term prognosis of acute myocardial infarction(AMI)patients with N-terminal B-type brain natriuretic peptide(NT-proBNP)peak value exceeding the up-per limit.Methods A total of 669 patients with AMI diagnosed in a hospital from 2013 to 2018 were selected as research objects.According to the peak value level of NT-proBNP,they were divided into the NT-proBNP peak value exceeding the upper limit group(50 cases)and the NT-proBNP peak value detectable group(619 cases).Propensity score was used for matching(1:2),and the patients were divided into NT-proBNP peak value exceeding the upper limit group(50 cases)and NT-proBNP peak value detectable group(107 cases),and the in-hospital prognosis and long-term prognosis of the two groups were compared,as well as the echo-cardiographic indexes of each group in the acute stage and recovery stage of AMI.Multiple linear regression a-nalysis was used to predict the factors affecting left ventricular ejection fraction in the recovery stage of AMI.Results After matching the propensity score,compared with the NT-proBNP peak value detectable group,the neutrophil to lymphocyte ratio,hypersensitive C reactive protein level and fibrinogen level in the NT-proBNP peak value exceeding the upper limit group were higher at admission,and the differences were statistically sig-nificant(P＜0.05).The proportion of Killip≥ Grade Ⅱ,left ventricular ejection fraction,hospitalization time and major adverse cardiovascular and cerebrovascular events in the NT-proBNP peak value exceeding the up-per limit group were compared with those in the NT-proBNP peak value detectable group,and the differences were statistically significant(P＜0.05).The left ventricular ejection fraction and left ventricular shortening fraction of the NT-proBNP peak value detection group in AMI acute stage were higher than those in the NT-proBNP peak value exceeding the upper limit group in AMI acute stage,and the difference was statistically significant(P＜0.05).Multiple linear regression analysis showed the relationship between the NT-proBNP peak value during hospitalization and the left ventricular ejection fraction of cardiac function during AMI re-covery.The results suggested that the NT-proBNP peak value was not a risk factor affecting the left ventricu-lar ejection fraction of cardiac function during AMI recovery.Conclusion AMI patients with NT-proBNP peak value exceeding the upper limit should be treated with a series of comprehensive treatment strategies to pro-mote their rehabilitation and improve their long-term prognosis.
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Objective To explore the clinical value of chitinase 3-like protein 1(CHI3L1)in the diagnosis of lung cancer.Methods A total of 106 patients with lung cancer admitted to the North District of the First Affiliated Hospital of Anhui Medical University from January to December 2022 were selected as the lung cancer group,76 patients with benign lung disease admitted during the same period were selected as the benign lung disease group and 20 healthy subjects were selected as the control group.Enzyme-linked immunosorbent assay was used to detect CHI3L1 levels.The levels of carcinoembryonic antigen(CEA),neuron-specific eno-lase(NSE),cytokeratin-19 fragment(CYFRA21-1),squamous cell carcinoma antigen(SCC-Ag)and gastrin-releasing peptide precursor(ProGRP)were determined by chemiluminescence assay.Results The levels of CEA,ProGRP,NSE,CYFRA21-1 and CHI3L1 in lung cancer group were significantly higher than those in control group,and the differences were statistically significant(P＜0.05).Serum CEA in lung cancer group was significantly higher than that in benign lung disease group,while serum CHI3L1 was significantly lower than that in benign lung disease group,with statistical significance(P＜0.05).Serum levels of NSE and Pro-GRP were higher in patients with small cell lung cancer than those with lung adenocarcinoma and lung squa-mous cell carcinoma(P＜0.05).Compared with patients with lung adenocarcinoma and small cell lung canc-er,the serum CYFRA21-1 level in patients with lung squamous cell carcinoma was higher,and the difference was statistically significant(P＜0.05).Compared with the control group,the serum levels of NSE,CY-FRA21-1 and CHI3L1 in patients with stage Ⅰ to Ⅱ lung cancer group were significantly increased,and the difference was statistically significant(P＜0.05).Multivariate Logistic stepwise regression analysis was per-formed for CEA,ProGRP,NSE,CYFRA21-1 and CHI3L1,and it was found that NSE and CHI3L1 had an effect on the occurrence of lung cancer.The sensitivity,specificity and area under the curve of CHI3L1 and NSE were 96.2％,90.0％and 0.965 respectively.Conclusion Serum CHI3L1 can assist in the diagnosis and differential diagnosis of lung cancer.The combined detection of CHI3L1 and NSE is helpful for the early de-tection of lung cancer and has good clinical application value.
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Objective To investigate the predictive efficacy of serum aminoterminal brain natriuretic pep-tide precursor(NT-proBNP),hypersensitive C-reactive protein(hs-CRP),D-dimer(D-D)and procalcitonin(PCT)in heart failure after acute myocardial infarction(AMI),Methods A total of 100 AMI patients admit-ted to the hospital from July 2021 to July 2023 were enrolled in the study as the observation group,In addi-tion,100 healthy people who underwent physical examination in the hospital during the same period were en-rolled as the control group,The serum levels of NT-proBNP,hs-CRP,D-D and PCT were detected and com-pared between the observation group and the control group,The AMI patients enrolled in the study were fur-ther divided into the heart failure group(31 cases)and the non-heart failure group(69 cases)according to the presence or absence of heart failure.The serum levels of NT-proBNP,hs-CRP,D-D,and PCT were compared between the two groups,Univariate analysis and multivariate Logistic regression analysis were used to analyze the risk factors of heart failure after AMI,Receiver operating characteristic(ROC)curve and decision curve a-nalysis(DCA)were used to analyze the predictive efficacy of serum NT-proBNP,hs-CRP,D-D and PCT for heart failure after AMI.Results The levels of serum NT-proBNP,hs-CRP,D-D and PCT in the observation group were higher than those in the control group(P＜0.05).The serum levels of NT-proBNP,hs-CRP,D-D and PCT in the complicated heart failure group were higher than those in the non-heart failure group(P＜0.05),Body mass index(BMI),smoking history,hypertension,number of diseased vessels,serum uric acid(SUA),low-density lipoprotein cholesterol(LDL-C),NT-proBNP,hs-CRP,D-D and PCT were risk factors for heart failure after AMI(P＜0.05).ROC curve analysis showed that the area under the curve(AUC)of combined detection of serum NT-proBNP,hs-CRP,D-D and PCT for predicting heart failure after AMI was 0.857(95％CI:0.811-0.948),the sensitivity was 96.12％,and the specificity was 91.28％,which were higher than the corresponding efficacy indexes of single detection(P＜0.05).DCA analysis showed that when the high-risk threshold was 0-0.99,the net benefit rate was greater than 0,which had clinical significance,When the threshold was 0-0.76,the net benefit rate of combined detection of serum NT-proBNP,hs-CRP,D-D and PCT was better than that of serum NT-proBNP,hs-CRP,D-D and PCT alone.Conclusion Combined detection of serum NT-proBNP,hs-CRP,D-D and PCT can improve the predictive efficiency of AMI compli-cated with heart failure,BMI,smoking history,hypertension,number of diseased vessels,SUA,LDL-C,NT-proBNP,hs-CRP,D-D and PCT are risk factors for AMI complicated with heart failure.
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Objective:To investigate associations between cardiac biomarkers with stroke severity and short-term outcome in patients with acute ischemic stroke (AIS).Methods:Patients with AIS admitted to the Affiliated Hospital of Qingdao University from June 2018 to February 2024 whose etiological classification was large artery atherosclerosis (LAA), small vessel occlusion (SVO) or cardioembolism (CE) were included retrospectively. According to the National Institutes of Health Stroke Scale score at admission, patients were divided into mild stroke group (≤8) and moderate to severe stroke group (>8). According to the modified Rankin Scale score at discharge, patients were divided into good outcome group (≤2) and poor outcome group (>2). Multivariate logistic regression analysis was used to determine the independent correlation between cardiac biomarkers and short-term outcome. The predictive value of cardiac biomarkers for poor outcome in patients with AIS and different stroke etiology subtypes were evaluated using receiver operating characteristic (ROC) curves. Results:A total of 2 151 patients with AIS were enrolled, including 1 256 males (58.4%), aged 67.40±11.34 years. 1 079 patents were LAA type (50.2%), 679 were SVO type (31.6%), and 393 were CE type (18.3%); 1 223 were mild stroke (56.86%) and 928 (43.14%) were moderate to severe stroke; 1 357 patients (63.09%) had good short-term outcome, and 794 (36.91%) had poor short-term outcome. Multivariate logistic regression analysis showed that N-terminal pro-B type natriuretic peptide (NT-proBNP), NT-proBNP/creatine kinase (CK) isoenzyme MB (CK-MB) ratio, and CK-MB/CK ratio were independent risk factors for poor short-term outcome. ROC curve analysis shows that the CK-MB/CK ratio had a higher predictive value for short-term poor outcome in patients with AIS (the area under the curve, 0.859, 95% confidence interval 0.839-0.879). Various cardiac biomarkers had a higher predictive value for short-term outcome of CE type and LAA type, but the predictive value for short-term outcome of SVO type was lower. Conclusions:Cardiac biomarkers are associated with the severity and poor outcome of AIS. NT-proBNP/CK-MB and CK-MB/CK ratios have higher predictive value for short-term poor outcome of AIS, especially in patients with CE type.
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Objective:To prepare the anti-programmed death-ligand 1 (PD-L1) nanoantibody P3C8-C3Fab by ligating with C3Fab and to investigate its role in plasma half-life.Methods:The C3Fab peptide derived from protein G was molecularly fused with the nanobody P3C8 by DNA recombination technology. The nanoantibody P3C8-C3Fab was inducibly expressed and purified in the E. coli BL21 strain, and the binding of it to PD-L1 protein, mouse IgG, and PD-L1-expressing tumor cells was detected by enzyme-linked immunosorbent assay (ELISA). The residual P3C8-C3Fab was detected in mouse serum at different times using double-antibody sandwich ELISA to assess the prolongation of the plasma half-life of PD-L1 nanobodies by C3Fab. Results:The nanoantibody P3C8-C3Fab was successfully constructed, and it could efficiently express itself in soluble form in BL21. The purified NbP3C8-C3Fab protein was obtained with a mass fraction of about 90% at a yield of 7.18 mg/L. The affinity of P3C8-C3Fab for PD-L1 protein and mouse IgG gradually increased with increasing mass concentration and showed a concentration correlation. The binding of P3C8-C3Fab to lung cancer A549 cells showed a concentration correlation. The concentration standard curve of P3C8-C3Fab in mouse serum showed a typical S-shape with a concentration correlation. The plasma half-life of P3C8 was only 0.44 h, while the plasma half-life of P3C8-C3Fab was 21.27-fold higher, up to 9.36 h.Conclusions:The linkage of C3Fab to the nanobodies of P3C8 can significantly prolong the plasma half-life of P3C8, which is valuable for the improvement of in vivo nanobody effects.
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Objective To observe the influence of Qishen Yiqi Guttate Pills(mainly composed of Astragali Radix,Salviae Miltiorrhizae Radix et Rhizoma,Notoginseng Radix et Rhizoma,and Dalbergiae Odoriferae Lignum)on the clinical efficacy of patients with acute myocardial infarction after percutaneous coronary intervention(PCI).Methods Sixty post-PCI patients with acute myocardial infarction of qi deficiency and blood stasis type who met the inclusion criteria were randomly divided into a treatment group and a control group,with 30 patients in each group.The control group was treated with conventional western medicine,and the treatment group was treated with Qishen Yiqi Guttate Pills on the basis of treatment for the control group.The course of treatment for the two groups lasted for 3 months.The changes of cardiac function indicators and serum levels of hypersensitive C-reactive protein(hs-CRP)and N-terminal B-type natriuretic peptide precursor(NT-pro BNP)were observed before and after the treatment in the two groups,and the incidence of cardiovascular adverse events during the treatment in the two groups were also compared.Results(1)After treatment,the serum hs-CRP and NT-pro BNP levels of patients in the two groups were significantly decreased(P＜0.05)and the left ventricular ejection fraction(LVEF)was significantly increased(P＜0.05)compared with those before treatment.And the effects on lowering the levels of serum hs-CRP and NT-pro BNP and on increasing LVEF of the treatment group were significantly superior to those of the control group,the differences being statistically significant(P＜0.05).(2)During the treatment period,the incidence of cardiovascular adverse events in the treatment group was 6.67%(2/30),which was significantly lower than 26.67%(8/30)of the control group,and the difference was statistically significant when comparing the two groups(P＜0.05).Conclusion Qishen Yiqi Guttate Pills can effectively improve cardiac function,decrease serum hs-CRP and NT-pro BNP levels,and reduce the occurrence of adverse cardiovascular events in post-PCI patients with acute myocardial infarction of qi deficiency and blood stasis type.
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Objective To investigate the effect of glucagon-like peptide 1(GLP-1)receptor agonists exendin-4 on the secretion of cyclophilin A(CyPA)to inhibit atherosclerosis(AS)and vascular calcification in mice role of the process.Methods Twenty ApoE-/-mice were randomly divided into model group and exendin-4 group,10 mice in each group,and were fed with high-fat diet to establish AS model,another 10 wild-type C57BL/6J mice were taken as the control group,and the exendin-4 group was intraperitoneally injected with the GLP-1R agonist exendin-4,1/d,for 8 weeks.After 8 weeks,the ELISA method was used to determine the level of triglyceride(TG),total cholesterol(TC),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)and CyPA,serum calcium level was detected by methylthymol blue colorimetric method,oil red O staining to detect the development of atherosclerotic plaques in the aorta,HE staining was used to observe the pathological changes of the aorta,Von Kossa staining was used to observe the calcium deposition in the aorta,immunohistochemical staining,Real-time PCR and Western blotting were used to detect the expression levels of aortic RUNX2 and bone morphogenetic protein 2(BMP-2),immunofluorescent staining was used to detect the positive expression of CyPA in aortic tissue.Results Compared with the control group,the serum levels of TG,TC,LDL-C,Ca and CyPA in the model group increased(P<0.05),the atherosclerotic plaque areas of the aorta increased(P<0.05),the aortic wall was thickened significantly and a large number of inflammatory cells were infiltrated,a large amount of calcium deposits were deposited in the aortic parietal membrane,the positive expression area ratio of RUNX2 and BMP-2,the relative mRNA expression of RUNX2 and BMP-2,the relative protein expression of RUNX2 and BMP-2 in aortic tissue all increased(P<0.05),and the red fluorescence of CyPA expression in aortic tissue was enhanced significantly.Compared with the model group,the serum levels of TG,TC,LDL-C,Ca and CyPA in the exendin-4 group decreased(P<0.05),the atherosclerotic plaque areas of the aorta decreased(P<0.05),the thickening of the aortic wall and the infiltration of inflammatory cells were alleviated significantly,the calcium deposition in the aortic wall was reduced,the positive expression area ratio of RUNX2 and BMP-2,the relative mRNA expression of RUNX2 and BMP-2,the relative protein expression of RUNX2 and BMP-2 in aortic tissue all decreased(P<0.05),and at the same time,the red fluorescence of CyPA expression in aortic tissue was weakened significantly.Conclusion GLP-1 receptor agonists exendin-4 can inhibit atherosclerosis and vascular calcification in mice,and the mechanism may be related to the reduction of CyPA secretion.
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Objective To investigate the expression level and clinical significance of cyclic citrullinated peptide antigen-specific T cells(CCP/AST)in synovial fluid and synovial tissue of rheumatoid arthritis(RA)patients.Methods A total of 128 RA patients in Shijiazhuang Hospital of Traditional Chinese Medicine from January to December 2021 were selected as the RA group,and 50 patients who needed arthroscopy for joint pain in the hospital during the same period were selected as the control group.Among the RA group,there were 46 cases in the mild group,52 cases in the moderate group,and 30 cases in the severe group.The protein expression levels of rheumatoid factors(RF)and anticitrullinated protein antibodies(ACPA)in synovial tissues of the subjects in each group were analyzed by Western blot.The frequency of CCP/AST in the synovial fluid of the subjects was analyzed by flow cytometry.The intensity of the staining of CCP/AST in synovial tissues was observed by double immunofluorescence staining/laser confocal scanning.Pearson correlation analysis was used to assess the correlation between the CCP/AST expression of synovial fluid and synovial tissue and RF and ACPA.Logistic regression was used to analyze the risk factors for the development of rheumatoid arthritis.Results In the order of control,mild,moderate and severe groups,RF(1.01±0.01,1.53±0.03,2.01±0.08,2.66±0.12 kDa)and ACPA proteins(1.03±0.01,1.61±0.03,2.04±0.10,2.59±0.13 kDa)in synovial tissues of patients were sequentially elevated,and the differences were all statistically significant(F=14.207,12.446,all P＜0.05).The expression of CCP/AST in synovial fluid of patients in the control,mild,moderate and severe groups was increased sequentially(8.26%±1.68%,22.46%±3.28%,33.58%±4.37%,46.15%±5.44%),and the difference was statistically significant(F=25.306,P＜0.05).Meanwhile,the intensity of CCP/AST staining in synovial tissues of patients in the control,mild,moderate and severe groups was also increased sequentially(1.05±0.26,1.35±0.89,2.04±0.56,2.78±0.15 score),and the difference was statistically significant(F=70.67,P＜0.05).The expression of CCP/AST in the synovial fluid and synovial tissues of patients with RA was positively correlated with RF(r=0.861,0.934,all P＜0.05)and ACPA in synovial fluid and synovial tissue(r=0.854,0.913,all P＜0.05).Logistic regression analysis showed that hypertension(OR=3.241,95%CI:1.491～6.752),diabetes mellitus(OR=2.565,95%CI:1.126～5.813),synovial fluid(OR=4.450,95%CI:1.652～11.622),and CCP/AST expression in synovial tissues(OR=5.629,95%CI:2.474～12.390)were independent risk factors for the development of RA(P＜0.05).Conclusion CCP/AST showed high expression in synovial fluid and synovial tissue of RA patients and related to disease activity and joint destruction,which can be used to assess the clinical joint mobility and bone destruction degree in such patients.
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BACKGROUND:In the process of exploring the mechanism of Alzheimer's disease,the important role of bioinformatics for common target screening has been revealed,enabling the use of its screening results as a basis for exploring the therapeutic effects of drugs on the disease. OBJECTIVE:To predict the targets of liraglutide,a glucagon-like peptide-1 receptor agonist,in the treatment of Alzheimer's disease by bioinformatics and molecular biology. METHODS:DisGeNET database and SEA database were used to obtain the common genes of Alzheimer's disease and liraglutide.GO/KEGG enrichment analysis of common targets was conducted using DAVID online database.Protein-protein interaction networks were constructed using STRING database.The optimal dosage of liraglutide was determined using cell counting kit-8 assay.Expression of key proteins was analyzed using immunofluorescence and immunoblotting techniques.The mouse hippocampal neuron HT22 cell line was used for ex vivo experiments,and the cells were randomly divided into three groups:HT22 group,HT22+Aβ group,and HT22+Aβ+Lir group.No special treatment was done in the HT22 group,while Aβ1-42 was used to intervene in the HT22 cell line for 24 hours to construct an Aβ injury cell model in the HT22+Aβ group.In additional to modeling,liraglutide was added to the HT22+Aβ+Lir group for 12 hours. RESULTS AND CONCLUSION:A total of 3 333 genes associated with Alzheimer's disease were screened from DisGeNET database.Then 147 potential targets of liraglutide were obtained from SEA database.Finally,64 common targets of Alzheimer's disease and Liraglutide were determined using R packets.GO/KEGG analysis of common targets using DAVID online database suggested that common targets were mainly enriched in the following biological processes:neuroactive ligand-receptor interaction,renin-angiotensin system,bladder cancer,endopeptidase activity,peptide receptor activity,G protein-coupled peptide receptor activity,and transport vesicles.The obtained 64 common target proteins were imported into SRTING online database for protein-protein interaction network construction,and the top three genes,matrix metalloproteinases 2,9 and interleukin 1β,were obtained.The activity of cultured cells was detected by the cell counting kit-8 kit.Liraglutide at 100 nmol/L was the optimal concentration for antagonizing Aβ1-42.In the western blot and immunofluorescence assays,the expression of matrix metalloproteinases 2,9 and interleukin 1β was significantly increased in the HT22+Aβ group compared with the HT22 group(P＜0.05)but significantly decreased in the HT22+Aβ+Lir group compared with the HT22+Aβ group(P＜0.05).To conclude,the above bioinformatics data and secondary validation of differential genes in the GEO database suggest that both matrix metalloproteinases 2,9 and interleukin 1β could be potential targets of liraglutide in the treatment of Alzheimer's disease.
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BACKGROUND:Frog active peptides have rich activities,such as antibacterial and anti-tumor,and are expected to solve the problem of antibiotic resistance. OBJECTIVE:The active peptide QUB2984 was discovered in the skin secretions of Agalychnis callidryas.Its structure and properties were simulated by bioinformatics.The peptide was synthesized,purified,and identified and its biological functions were investigated. METHODS:Agalychnis callidryas skin secretions were collected by electrostimulation.The sequence of QUB2984 was obtained through constructing a cDNA library with isolated mRNA.BLAST was used for peptide sequence alignment.Besides that,Iterative Threading ASSEmbly Refinement(I-TASSER)and HeliQuest tools were used for protein secondary structure simulation.It was synthesized by solid-phase peptide synthesis,purified by reverse-phase high-performance liquid chromatography,and structurally confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.The purified peptide was used to evaluate its biological activity.Its antibacterial effect was evaluated by the minimum inhibitory concentration method.Its cytotoxic effect was detected by MTT assay.Its safety was investigated by a hemolysis test. RESULTS AND CONCLUSION:(1)Peptide QUB2984 had basically α-spiral structure,with a relatively intact hydrophobic surface,and a certain destructive ability to biofilm.The third amino acid position of QUB2984 was composed of W and had a G-X-G structure.(2)The minimum inhibitory concentration of QUB2984 against gram-positive Staphylococcus aureus was 2 μmol/L,the minimum inhibitory concentration against gram-negative Escherichia coli was 2 μmol/L,and the minimum inhibitory concentration against the fungus Candida albicans was 8 μmol/L.(3)The active peptide QUB2984 had obvious inhibitory effect on human non-small cell lung cancer cells NCI-H838 at 10-5 mol/L concentration,and the hemolytic effect on horse red cells at 64 μmol/L concentration was 50%.(4)The results showed that QUB2984 had anti-bacterial and anti-cancer activity,and it had a positive charge of +3,which was conducive to contact with bacteria or cells.
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OBJECTIVE:At present,there are many reports on the related factors associated with the incidence of cervical spine instability in patients with rheumatoid arthritis,but there are problems such as small sample size and many confounding factors,and the research results of various studies on the same related factors are also different.This article analyzed the factors related to cervical spine instability in patients with rheumatoid arthritis by means of a systematic review. METHODS:Articles related to cervical spine instability in patients with rheumatoid arthritis were collected by searching both Chinese and English databases until March 2023.The outcome of cervical spine instability in patients with rheumatoid arthritis was used as the grouping criterion to abstract basic information,baseline patient characteristics,laboratory-related tests,medication use,and other relevant risk factors.Meta-analysis was done using Stata 14.0 software. RESULTS:(1)Sixteen relevant studies,all of moderate or above quality,were included,including seven studies with case-control studies and nine with cross-sectional studies.The overall incidence of cervical spine instability in patients with rheumatoid arthritis was 43.08%.(2)Meta-analysis showed:Related risk factors included female(OR=0.60,95%CI:0.44-0.82,P=0.002);age at disease onset(SMD=-0.52,95%CI:-0.86 to-0.18,P=0.003);duration of disease(SMD=0.58,95%CI:0.14-1.02,P=0.01);body mass index(OR=0.74,95%CI:0.63-0.88,P=0.001);rheumatoid factors positive univariate analysis subgroup(OR=1.33,95%CI:1.02 to 1.72,P=0.04),C-reactive protein(SMD=0.26,95%CI:0.16-0.35,P=0.00),erythrocyte sedimentation rate(SMD=0.15,95%CI:0.002-0.29,P=0.047),anti-cyclic-citrullinated peptide antibodies(OR=1.73,95%CI:1.19-2.51,P=0.004),28-joint Disease Activity Score(SMD=0.20,95%CI:0.04-0.37,P=0.02),destruction of peripheral joints(OR=2.48,95%CI:1.60-3.85,P=0.00),and corticosteroids(OR=1.91,95%CI:1.54-2.37,P=0.00)were strongly associated with the development of rheumatoid arthritis-cervical spine instability.Female and corticosteroid use were independently associated with the occurrence of rheumatoid arthritis-cervical spine instability. CONCLUSION:Based on clinical evidence from 16 observational studies,the overall incidence of rheumatoid arthritis-cervical spine instability was 43.08%.However,the incidence of cervical spine instability in rheumatoid arthritis patients varied greatly among different studies.Gender(female)and the use of corticosteroids were confirmed as independent correlation factors for the onset of cervical spine instability in patients with rheumatoid arthritis.The results of this study still provide some guidance for early clinical recognition,diagnosis,and prevention of rheumatoid arthritis-cervical spine instability.
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Bronchopulmonary dysplasia（BPD）is a significant complication that greatly impacts the prognosis of preterm infants.The incidence of pulmonary hypertension（PH）in preterms with moderate to severe BPD is relatively high，which further increases the mortality among infants with BPD.Diagnosis of BPD could be made until 36 weeks corrected gestational age，and the clinical manifestations of BPD-PH are also not specific enough for accurate identification.Cardiac catheterization is considered the gold standard for diagnosis，which is an invasive procedure，while echocardiography heavily relies on the experience of operator，limiting early detection of both BPD and BPD-PH.N-terminal pro-brain natriuretic peptide（NT-proBNP），secreted by ventricular myocytes in response to volume or pressure overload，has been widely utilized in assessing cardiac load in cases of congenital heart disease.Recent studies have shown that the blood NT-proBNP levels significantly elevate in infants with BPD within one week after birth and during the development stage of BPD.Additionally，there is a substantial increase in NT-proBNP levels from 28 to 36 weeks corrected gestational age，which has both high sensitivity and high specificity in predicting the occurrence of BPD-PH.Therefore，NT-proBNP is expected to serve as a biomarker for the early prediction of BPD and BPD-PH.This review summarized the biological function，metabolic characteristics，and current research advancements regarding NT-proBNP in BPD and BPD-PH.The aim is to utilize NT-proBNP as an early predictor for BPD and BPD-PH，enabling timely identification of premature infants with high risk factors of these diseases，and facilitating prompt intervention to ultimately enhance prognosis.
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Objective:To analyze the clinical value of dynamic electrocardiogram combined with serum brain natriuretic peptide (BNP) and creatine kinase isoenzyme (CK-MB) levels in evaluating the severity of respiratory tract infection complicated with viral myocarditis.Methods:A total of 125 patients with respiratory tract infection who were admitted to the Affiliated Run Run Shaw Hospital of Medical School, Zhejiang University from January 2018 to December 2022 were selected retrospectively. They were divided into the complication group (71 cases) and the non-complication group (54 cases) according to whether they were complicated with viral myocarditis. Dynamic and conventional electrocardiogram characteristics were compared. Serum BNP and CK-MB levels in the complication group and the non-complication group at admission were analyzed. Dynamic electrocardiograms and serum BNP and CK-MB levels of patients with different severity of myocarditis were comparatively analyzed. The value of combined diagnosis was analyzed.Results:The detection rates of atrial premature beats and sinus tachycardia by dynamic electrocardiogram were higher than those by conventional electrocardiogram: 38.03% (27/71) vs. 22.54% (16/71), 28.17% (20/71) vs. 14.08% (10/71) ( P<0.05). Serum BNP and CK-MB levels in the complication group were higher than those in the non-complication group: (104.26 ± 10.75) ng/L vs. (54.11 ± 5.69) ng/L, (17.89 ± 1.86) U/L vs. (13.46 ± 1.42) U/L ( P<0.05). The detection rates of atrial premature beats, sinus tachycardia, burst ventricular tachycardia, first-degree atrioventricular block, ST segment change and low QRS voltage, serum BNP and CK-MB levels in patients with respiratory tract infection complicated with severe viral myocarditis (37 cases) were higher than those in patients with mild viral myocarditis (34 cases): 51.35% (19/37) vs. 23.53% (8/34), 43.24% (16/37) vs. 11.76% (4/34), 32.43% (12/37) vs. 11.76% (4/34), 29.73% (11/37) vs. 8.82% (3/34), 43.24% (16/37) vs. 14.71% (5/34), 24.32% (9/37) vs. 5.88% (2/34), (107.19 ± 10.56) ng/L vs. (101.08 ± 10.18) ng/L, (18.33 ± 1.85) U/L vs. (17.41 ± 1.76) U/L ( P<0.05). Compared with clinical pathological diagnosis, Kappa values of dynamic electrocardiogram, BNP at admission, CK-MB at admission and combination of the three for diagnosing acute upper respiratory tract infection complicated with viral myocarditis were 0.784, 0.765, 0.733 and 0.879. The sensitivity and accuracy of combined diagnosis were higher. Conclusions:Dynamic electrocardiogram combined with serum BNP and CK-MB can better help to evaluate the severity of respiratory tract infection complicated with myocarditis. Therefore, they are worthy of monitoring.
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Tirzepatide,a new glucagon-like peptide-1/glucose dependent insulin stimulating polypeptide(GLP-1/GIP)double receptor agonist,has been clinically shown to have a strong hypoglycemic effect and a very significant effect on reducing body mass.It can improve insulin sensitivity,and has superior cardiovascular protection and the effect of improving nonalcoholic fatty liver disease/nonalcoholic steatohepatitis(NAFLD/NASH),with few side effects and good compliance.As a dual gut hor-mone agonist,tirzepatide shows strong potential to improve metabolic levels.This article reviews the mechanism of action and clini-cal studies of the GLP-1/GIP dual receptor agonist tirzepatide.
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Objective:To study the correlation between serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) level and echocardiographic parameters in patients with chronic Keshan disease, providing reference for diagnosis and prognosis evaluation of chronic Keshan disease.Methods:Ninety-nine patients with chronic Keshan disease who received standardized treatment at Jingchuan County People's Hospital in Pingliang City, Gansu Province from January to December 2020 were selected. Among them, 16 patients were classified as cardiac function grade Ⅱ according to New York Heart Association (NYHA), 69 as grade Ⅲ and 14 as grade Ⅳ. The patients underwent echocardiography and their serum NT-proBNP level was measured using fluorescence immunochromatography. The differences in serum NT-proBNP levels among patients with different cardiac function grades were compared, and the correlation between cardiac function grades, serum NT-proBNP level and echocardiographic parameters was analyzed.Results:The serum NT-proBNP levels in patients with cardiac function grades Ⅱ, Ⅲ, and Ⅳ were (1 107.26 ± 268.03), (2 125.98 ± 293.02), and (8 268.59 ± 2 659.50) pg/ml, respectively. The differences among the three groups were statistically significant ( F = 13.94, P < 0.001). The serum NT-proBNP level was positively correlated with cardiac function grades ( r = 0.44, P < 0.001), left ventricular end-diastolic diameter, left ventricular end-systolic diameter, and left atrial diameter ( r = 0.45, 0.52, 0.38, P < 0.001), and negatively correlated with fractional shortening and left ventricular ejection fraction ( r = - 0.39, - 0.46, P < 0.001). Conclusions:The serum NT-proBNP level in patients with chronic Keshan disease with different cardiac function grades is different, and is positively correlated with echocardiographic parameters reflecting the degree of cardiac structural and functional impairment. The NT-proBNP level may become an early diagnostic, grading, and prognostic indicator for chronic Keshan disease.