The amino acid sequences of the OmpA protein isolated from Escherichia coli QML2206-1(E.coli QML2206-1)in our laboratory were analyzed for homology with different strains of OmpA proteins using bioinformatics software,and the OmpA protein was analyzed for its physicochemical properties,transmembrane structure and signal peptide prediction,B-cell anti-genic epitope prediction,secondary and tertiary structure prediction.The OmpA gene fragment was ligated with pET-32a vector to construct a prokaryotic expression vector,which was purified by a nickel column affinity purification system after prokaryotic expression and optimization of ex-pression conditions in BL21(DE3).The purified recombinant protein was fully mixed with Freund's adjuvant to immunize mice,and the levels of mouse-specific IgG antibody and the expression levels of cytokines CD4,CD8 and IL-4 in mouse serum were detected by ELISA,and the immuno-protective effect was evaluated by mouse attack protection test.OmpA protein is a hydrophilic protein with no transmembrane structural domains and a secondary structure consisting mainly of irregular coils(47.98％)and α-helices(29.77％),with 12 antigenic epitopes that can bind to anti-bodies produced by B cells.The recombinant protein OmpA with a relative molecular mass of a-bout 55 kDa was successfully obtained by prokaryotic expression,and the highest expression was induced by IPTG concentration of 0.000 4 mmol/L for 6 h at 37 ℃.The serum-specific IgG anti-body potency of recombinant protein immunized mice was up to 1∶32 000;the expression levels of CD4,CD8 and IL-4 in the serum of immunized mice were elevated compared with those of the con-trol group.The survival rate of mice was 80％and 40％after attack with minimum lethal dose(MLD)and 2 times minimum lethal dose(2MLD),respectively.OmpA recombinant protein has good antigenicity and certain immunoprotective effects,and this study provides a technical basis for the next step in the development of a genetically engineered subunit vaccine against yak-appli-cable E.coli based on OmpA protein.

ObjectiveThis study aims to investigate the effects of Linggui Zhugantang （LGZGT） on ventricular remodeling （VR） in mice with myocardial infarction （MI） and its impact on the Ras homologgene A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway. MethodsThe MI model of mice was established by ligating the left anterior descending coronary artery （LAD）. They were divided into the sham-operated group， the model group， the low-dose， medium-dose， and high-dose groups of LGZGT （2.34， 4.68， 9.36 g·kg^-1）， and the captopril group （3.25 mg·kg^-1）， with 10 mice in each group. After four weeks of continuous drug administration by gavage， the level of cardiac function in each group of mice was examined using small animal Doppler ultrasound. Hematoxylin-eosin （HE） staining and Masson staining was used to assess the morphological changes of myocardial tissue and calculate the rate of collagen fiber deposition in mouse myocardial tissue. Wheat germ agglutinin （WGA） staining was employed to compare the cross-sectional area of cardiomyocytes in each group of mice. The expression levels of α-smooth muscle actin （α-SMA）， matrix metalloproteinase-2 （MMP-2）， type Ⅰcollagen （Col Ⅰ）， Col Ⅲ， tissue inhibitor of metalloproteinase 1（TIMP1）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Bcl-2， Caspase-3， and cleaved Caspase-3 were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to evaluate the mRNA levels of the pathway-related genes RhoA， ROCK1， and ROCK2. The protein expression levels of RhoA， ROCK1， and ROCK2 were tested by Western blot. ResultsThe level of cardiac function was markedly declined in the model group compared to the sham-operated group（P<0.01）. Myocardial tissue morphology changed significantly. The cross-sectional area of cardiomyocytes was significantly enlarged. The expression of α-SMA， MMP-2， Col Ⅰ， and Col Ⅲ was significantly upregulated（P<0.01）， and TIMP1 protein expression was significantly reduced（P<0.01）. The expressions of apoptosis-related proteins Bax were significantly up-regulated（P<0.01）， while the expression of Bcl-2 protein was significantly decreased（P<0.01）. The mRNA expression of RhoA， ROCK1， and ROCK2 were significantly upregulated （P<0.01）. Compared to the model group， the low-dose， medium-dose， and high-dose groups of LGZGT and the captopril group significantly reversed the experimental results of the model group in a dose-dependent manner （P<0.05， P<0.01）. ConclusionLGZGT significantly attenuated myocardial fibrosis， myocardial hypertrophy， and cardiomyocyte apoptosis after MI in mice and effectively reversed VR， the mechanism of which may be related to the modulation of the RhoA/ROCK signaling pathway.

ObjectiveThis study aims to investigate the effects of Linggui Zhugantang （LGZGT） on ventricular remodeling （VR） in mice with myocardial infarction （MI） and its impact on the Ras homologgene A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway. MethodsThe MI model of mice was established by ligating the left anterior descending coronary artery （LAD）. They were divided into the sham-operated group， the model group， the low-dose， medium-dose， and high-dose groups of LGZGT （2.34， 4.68， 9.36 g·kg^-1）， and the captopril group （3.25 mg·kg^-1）， with 10 mice in each group. After four weeks of continuous drug administration by gavage， the level of cardiac function in each group of mice was examined using small animal Doppler ultrasound. Hematoxylin-eosin （HE） staining and Masson staining was used to assess the morphological changes of myocardial tissue and calculate the rate of collagen fiber deposition in mouse myocardial tissue. Wheat germ agglutinin （WGA） staining was employed to compare the cross-sectional area of cardiomyocytes in each group of mice. The expression levels of α-smooth muscle actin （α-SMA）， matrix metalloproteinase-2 （MMP-2）， type Ⅰcollagen （Col Ⅰ）， Col Ⅲ， tissue inhibitor of metalloproteinase 1（TIMP1）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Bcl-2， Caspase-3， and cleaved Caspase-3 were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to evaluate the mRNA levels of the pathway-related genes RhoA， ROCK1， and ROCK2. The protein expression levels of RhoA， ROCK1， and ROCK2 were tested by Western blot. ResultsThe level of cardiac function was markedly declined in the model group compared to the sham-operated group（P<0.01）. Myocardial tissue morphology changed significantly. The cross-sectional area of cardiomyocytes was significantly enlarged. The expression of α-SMA， MMP-2， Col Ⅰ， and Col Ⅲ was significantly upregulated（P<0.01）， and TIMP1 protein expression was significantly reduced（P<0.01）. The expressions of apoptosis-related proteins Bax were significantly up-regulated（P<0.01）， while the expression of Bcl-2 protein was significantly decreased（P<0.01）. The mRNA expression of RhoA， ROCK1， and ROCK2 were significantly upregulated （P<0.01）. Compared to the model group， the low-dose， medium-dose， and high-dose groups of LGZGT and the captopril group significantly reversed the experimental results of the model group in a dose-dependent manner （P<0.05， P<0.01）. ConclusionLGZGT significantly attenuated myocardial fibrosis， myocardial hypertrophy， and cardiomyocyte apoptosis after MI in mice and effectively reversed VR， the mechanism of which may be related to the modulation of the RhoA/ROCK signaling pathway.

Fracture healing is a complex process that necessitates the synergistic action of various cells and molecules. Macrophages play an indispensable and crucial regulatory role in the process of fracture repair, influencing stages such as inflammatory modulation, angiogenesis, and tissue remodeling. This article delves into the functional characteristics of macrophages and their roles at different stages of fracture healing. Additionally, it explores the impact of aging macrophages on the healing process. Furthermore, the potential of emerging therapies, such as hydrogel-based treatments and exosomes, in modulating macrophage responses is analyzed. This study provides a theoretical foundation for the development of innovative therapies aimed at enhancing the efficacy of fracture healing.

The amino acid sequences of the OmpA protein isolated from Escherichia coli QML2206-1(E.coli QML2206-1)in our laboratory were analyzed for homology with different strains of OmpA proteins using bioinformatics software,and the OmpA protein was analyzed for its physicochemical properties,transmembrane structure and signal peptide prediction,B-cell anti-genic epitope prediction,secondary and tertiary structure prediction.The OmpA gene fragment was ligated with pET-32a vector to construct a prokaryotic expression vector,which was purified by a nickel column affinity purification system after prokaryotic expression and optimization of ex-pression conditions in BL21(DE3).The purified recombinant protein was fully mixed with Freund's adjuvant to immunize mice,and the levels of mouse-specific IgG antibody and the expression levels of cytokines CD4,CD8 and IL-4 in mouse serum were detected by ELISA,and the immuno-protective effect was evaluated by mouse attack protection test.OmpA protein is a hydrophilic protein with no transmembrane structural domains and a secondary structure consisting mainly of irregular coils(47.98％)and α-helices(29.77％),with 12 antigenic epitopes that can bind to anti-bodies produced by B cells.The recombinant protein OmpA with a relative molecular mass of a-bout 55 kDa was successfully obtained by prokaryotic expression,and the highest expression was induced by IPTG concentration of 0.000 4 mmol/L for 6 h at 37 ℃.The serum-specific IgG anti-body potency of recombinant protein immunized mice was up to 1∶32 000;the expression levels of CD4,CD8 and IL-4 in the serum of immunized mice were elevated compared with those of the con-trol group.The survival rate of mice was 80％and 40％after attack with minimum lethal dose(MLD)and 2 times minimum lethal dose(2MLD),respectively.OmpA recombinant protein has good antigenicity and certain immunoprotective effects,and this study provides a technical basis for the next step in the development of a genetically engineered subunit vaccine against yak-appli-cable E.coli based on OmpA protein.

Objective:To examine the prevention effecacy and safety of preprotein convertase subtilisin-kexin 9 (PCSK9) inhibitors in stroke in patients with atherosclerotic cardiovascular disease (ASCVD) at primary and secondary prevention.Methods:PubMed, Embase, Web of Science, Cochrane Library, and Wanfang and CNKI databases were searched for randomized controlled trials comparing evolocumab, alirocumab, tafolecimab or inclisiran (experimental group) with placebo or conventional therapy (control group) in hyperlipidemia and ASCVD from inception to March 2024. Valid data were extracted after screening and applying Cochrane Literature quality assessment tool to assess the literature quality. Efficacy outcome (incidences of stroke and ischemic stroke) and safety outcome (cardiovascular mortality, and incidences of aminotransferase increased by more than 3 times, creatine kinase increased by more than 3 times, allergic reaction and hemorrhagic stroke) were recorded. Meta analysis of the extracted data was conducted using Stata software to calculate the risk difference ( RD). Results:Twenty articles (21 randomized controlled trials) were included with 62 799 patients. For primary prevention, no significant difference was found between PCSK9 inhibitors and control groups in stroke incidence ( RD=0.000, 95% CI: -0.002-0.003, P=0.905) or ischemic stroke incidence ( RD=0.001, 95% CI: -0.005-0.006, P=0.824); incidence of creatine kinase increased by more than 3 times in the PCSK9 inhibitors group was significantly decreased compared with that in the control group ( RD=-0.005, 95% CI: -0.010-0.000, P=0.039). For secondary prevention, PCSK9 inhibitors group had significantly reduced stroke incidence ( RD=-0.004, 95% CI: -0.006--0.002, P<0.001) and ischemic stroke incidence ( RD=-0.003, 95% CI: -0.005--0.002, P<0.001) compared with control group; no significant differences in cardiovascular mortality, or incidences of aminotransferase increased by more than 3 times, creatine kinase increased by more than 3 times, allergic reaction and hemorrhagic stroke were noted between the PCSK9 inhibitors group and control group ( P>0.05). Conclusion:PCSK9 inhibitors in primary prevention have no significant effect on stroke or ischemic stroke incidences, but can decrease the incidence of creatine kinase increased by more than 3 times; PCSK9 inhibitors in secondary prevention can reduce stroke and ischemic stroke incidences without increasing complications and thus enjoying certain safety.

Objective To elucidate the influence of two procedures aortic root remodeling using a straight tubular artificial vessel while preserving the aortic valve and the Florida sleeve procedure on the biomechanics of the aortic root.Methods Five finite element models of the aortic root were reconstructed using computed tomography angiography images,including two cases of aortic root remodeling(A1 and A2),two cases of the Florida sleeve procedure(B1 and B2),and one control group without aortic root pathology(C).Numerical simulations were performed to obtain the blood flow and pressure distribution result to assess the differences in the hemodynamics of the aortic root.Results There were no significant differences in the peak systolic velocity between the two procedures and the control.However,the flow velocity after aortic root remodeling was smoother,similar to the model of the control group,with a more stable average aortic pressure and wall shear stress.In the Florida sleeve procedure,high-speed blood flow affected the vessel wall,leading to various degrees of wall shear stress and pressure concentrations along the aortic wall.Conclusions After aortic root replacement with valve preservation,blood flow patterns in the reconstructed aortic root depended on postoperative changes in sinus geometry.Both surgical procedures showed favorable blood flow patterns;however,the flow pattern after aortic root remodeling was more stable than that after the Florida sleeve procedure.

ObjectiveTo evaluate the clinical effectiveness and safety of Lianling Formula （连苓方） in the treatment of adolescent atopic dermatitis （AD） exacerbation stage with heart fire and spleen deficiency syndrome. MethodsNinety-two patients were randomly divided into a treatment group and a control group， with 46 cases in each group. Patients in the control group took oral ebastine tablets， and patients in the treatment group took Lianling Formula orally， and both groups used 0.03% topical tacrolimus for 4 weeks. Before and after treatment， the improvement of scoring atopic dermatitis index by 50% （SCORAD50） and 75% （SCORAD75）， scoring atopic dermatitis index （SCORAD）， visual analog scale （VAS）， traditional Chinese medicine （TCM） symptom score， the children's dermatology life quality index （CDLQI）， Athens insomnia scale （AIS） score of both groups were evaluated， and the adverse reactions in both groups were recorded. ResultsFour cases in the treatment group and five cases in the control group were dropped， and intention-to-treat analysis was used. Finally 46 cases in each group were included in the statistics. The SCORAD scores after treatment were lower than those before treatment， and the scores in treatment group were lower than those of the control group （P＜0.01）； the attainment rates of SCORAD50 and SCORAD75 after treatment in the treatment group were 80.43% and 47.83%， which were significantly higher than those in the control group of 63.04% and 26.09%， respectively （P＜0.05）. VAS scores， TCM symptom scores， CDLQI scores， and AIS scores were lower than those before treatment （P＜0.01）， and VAS scores， TCM symptom scores， and CDLQI scores of the treatment group were lower than those of the control group after treatment （P＜0.05 or P＜0.01）， while AIS scores were higher than those of the control group （P＜0.05）. The erythema， papule/edema， oozing/crusting， epidermal peeling， and dryness in the SCORAD scores of the treatment group after treatment were lower than those before treatment （P＜0.05 or P＜0.01）， and the erythema， papule/edema， oozing/crusting， and dryness of children in the control group were lower than those before treatment （P＜0.05 or P＜0.01）. The scores of erythema， oozing/crusting， and epidermal peeling in the treatment group after treatment were all lower than those in the control group （P＜0.05）. The scores of vexation， itching， dry mouth， poor appetite and irregular bowel movements in terms of TCM symptoms in the treatment group after treatment were lower than those before treatment （P＜0.01）， and the scores of itching， vexation， dark urine in the control group were lower than those before treatment （P＜0.05 or P＜0.01）. The scores of itching， dry mouth， poor appetite， irregular bowel movements， and dark urine in the treatment group after treatment were all lower than that in the control group （P＜0.05 or P＜0.01）. No adverse reactions occurred in both groups of children during treatment. ConclusionLianling Formula combined with topical tacrolimus can effectively improve the clinical symptoms of adolescent AD exacerbation with heart fire and spleen deficiency syndrome， and the formula is effective and safe in improving the children's skin lesions， reducing itching， relieving the accompanied TCM symptoms， and improving the quality of life.

Objective To construct and appraise a new model for predicting the prognosis of rectal cancer patients using the Lasso-Cox strategy.Methods The clinical pathological data of 599 rectal cancer patients who underwent radical resection were analyzed.Comparison between groups,Lasso and Cox regression were used to select varia-bles and construct a model,and its discrimination,consistency,and clinical benefits were appraised by the receiv-er operating characteristic(ROC),calibration curve,and decision curve analysis.Results Comparison between groups showed that age,body mass index(BMI),preoperational nutrition status,carbohydrate antigen199(CA199),preoperative chemotherapy,intraoperative blood transfusion,vascular or nerve invasion,cancer nod-ules,pathologic T,N,and TNM stages,tumor recurrence or metastasis,radiotherapy and postoperative survival time were associated with grouping of death or survival in rectal cancer patients.Among them,8 variables were se-lected by lasso and contained into the Cox regression model.Age(HR=1.04,P＜0.05),BMI(HR=0.89,P＜0.05),blood transfusion(HR=2.29,P＜0.05),postoperative chemotherapy(HR=0.16,P＜0.01),recur-rence(HR=43.67,P＜0.01),and metastasis(HR=2.75,P＜0.05)were identified as independent prognostic factors,which were used to construct a nomogram model.The area under the curve(AUC)and the 95％confi-dence interval of the receiver operating characteristic(ROC)curve of the predictive model was 0.95(0.91-0.99),P＜0.01.The predicted probability of 1-year and 3-year survival was close to the actual probability.The DCA curve of the model was far away from a decision line parallel to the X-axis and another line with a negative slope.Conclusion The newly established nomogram has good discrimination,consistency and clinical benefits,which help predict the prognosis of rectal cancer after surgery.

Objective: To explore the influential factors on tumor diameters in rectal cancer. Methods : Cross sectional study data were collected from patients undergoing radical resection of rectal cancer.Patients were grouped by median value of tumor diameter, and the clinical pathological characteristics were compared between the two groups.The associations between tumor diameter and continuous variables were analyzed using locally weighted scatterplot smoothing(LOWESS) method.The associating factors of tumor diameter were further analyzed using generalized additive model(GAM). Results : Patients with rectal cancer were divided into two groups with tumor diameter either<4 centimeters or ≥4 centimeters.Proportions of T,TNM stages differed significantly between different groups.Median carcinoembryonic antigen(CEA) in≥4 centimeters group were 4.90 ng/ml(inter-quartile range [IQR] 2.39,9.53 ng/ml),much higher than that in<4 centimeters group(2.93 ng/ml [IQR:1.78,4.90 ng/ml]),P<0.01.LOWESS analysis showed that there were non-linear relationships between tumor diameter and the following variables: age, the distance between tumor and anal verge, CEA,neutrophils, lymphocytes, platelets, neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio.GAM showed that increased blood platelets(P<0.05) and later T stage(P<0.01) were significantly associated with increased tumor diameters, and increased CEA levels were also associated with larger tumors(P<0.01),but there were saturation effects. Conclusion Platelets, T stage and CEA are associated with tumor diameter in rectal cancer, indicating that the above factors should be considered when studying associated factors of tumor diameter in rectal cancer.