Objective To explore the mechanism of Huanglian Ganjiang Decoction in regulating the mitochondrial apoptotic pathway through the NF-κB/CXCL1/CXCR2 pathway in ulcerative colitis(UC).Methods Sixty C57BL/6J mice were divided into normal group,model group,Huanglian Ganzhang Decoction groups(low,medium and high doses)and mesalazine group.Except for the normal group,UC models were established in the other groups.The general conditions of the mice of all groups were recorded.HE staining was used to observe the pathological changes of the colon and rectum;electron microscopy was used to observe the mitochondrial condition;ELISA was used to determine the contents of IL-6,IL-10,D-lactic acid and DAO;Real-time PCR was used to detect the mRNA expression of NF-κB p65,CXCL1,CXCR2,Bcl-2 and Bax;Western blot was used to determine the protein expression levels of NF-κB p65,CXCL1,CXCR2,p-JAK2,p-STAT3,Bcl-2 and Bax.Results Compared with the normal group,the model group of mice showed a decline in condition,shortened length of the colon and rectum,and mitochondrial structural damage,furthermore,the levels of IL-6,DAO and D-lactic acid were increased,IL-10 level was decreased,the mRNA expression of NF-κB p65,CXCL1,CXCR2 and Bax were enhanced,Bcl-2 mRNA expression was weakened,the protein expression of NF-κB p65,CXCL1,CXCR2,p-JAK2,p-STAT3 and Bax were enhanced,Bcl-2 protein expression was weakened.Compared with the model group,the indexes mentioned above were reversed in the drug groups,especially in the high-dose group.Conclusion Huanglian Ganjiang Decoction may exert its therapeutic effect on UC in mice by regulating the NF-κB/CXCL1/CXCR2 pathway and mitochondrial apoptotic pathway.

Objective:To evaluate the therapeutic value of radiotherapy in combined immunotherapy and chemotherapy as first-line treatment for patients with oligometastatic non-small cell lung cancer (NSCLC).Methods:A retrospective analysis was conducted on data from 195 NSCLC patients who lacked epidermal growth factor receptor (EGFR) or anaplastic lymphoma kinase (ALK) mutations and were treated at the Anyang Tumor Hospital and the Fourth Hospital of Hebei Medical University from January 2019 to December 2021. These patients consisted of 166 male and 29 female cases, aged from 28 to 85 years, with an average age of (61.4 ± 9.3) years. These patients were divided into two groups, with each group receiving the radiotherapy and combined immunotherapy and chemotherapy (the radiotherapy and combination group, n = 60) and combined immunotherapy and chemotherapy only (the combination group, n = 135). Then, propensity score matching (PSM) was performed to analyze the differences in prognosis between both groups before and after PSM, as well as the short-term efficacy and adverse reactions after PSM. Results:For the 195 NSCLC patients, the median follow-up time was 31.8 months, with median overall survival (OS) and median progression-free survival (PFS) recorded at 23.8 months and 9.2 months, respectively. The radiotherapy and combination group exhibited enhanced 1-, 2-, and 3-year survival rates of 78.5%, 55.9%, and 45.1%, respectively, significantly higher than the combination group (48.3%, 35.6%, and 26.6%, respectively, χ2 = 14.65, P < 0.001). Similarly, the radiotherapy and combination group displayed 1-, 2-, and 3-year PFS rates of 51.9%, 29.5%, and 22.7%, respectively, exceeding those of the combination group (30.0%, 24.5%, and 16.9%, respectively, χ2=6.09, P=0.014). After PSM, the radiotherapy and combination group manifested an objective response rate (ORR) of 60.0% (33/55) and a disease control rate (DCR) of 89.1% (49/55), which were 16.4% (9/55) and 56.4% (31/55), respectively for the combination group. These results suggested that the radiotherapy and combination group demonstrated significantly higher ORR and DCR ( χ2 = 22.18, 14.85, P<0.001). After PSM, the radiotherapy and combination group yielded 1-, 2-, and 3-year survival rates of 70.9%, 52.3%, and 41.9%, respectively, significantly than the combination group (43.6%, 29.8%, and 27.1%, respectively, χ2=8.95, P=0.003). The radiotherapy and combination group exhibited 1-, 2-, and 3-year PFS rates of 47.3%, 27.3%, and 18.7%, respectively, significantly higher than the combination group (23.6%, 17.6%, and 15.4%, respectively, χ2 = 6.71, P = 0.010). Multivariate Cox regression analysis revealed that independent factors affecting OS included clinical stage, treatment regimen, number of immunotherapy cycles, and treatment efficacy ( HR = 1.88, 2.11, 0.23, 1.79, P < 0.05). Similarly, independent factors affecting PFS consisted of treatment regimen, number of immunotherapy cycles, and treatment efficacy ( HR = 1.62, 0.37, 3.42, P <0.05). There were no statistical differences in the incidence of grade ≥ 2 bone marrow suppression (18.2% vs. 12.7%) and grade ≥ 2 pneumonia (21.8% vs. 14.5%) between both groups ( P>0.05). Conclusions:Introducing radiotherapy into combined immunotherapy and chemotherapy as first-line treatment for oligometastatic NSCLC can optimize both local and systemic disease control and significantly improve patient prognosis without increasing treatment-related adverse reactions.

Objective To explore the mechanism of Huanglian Ganjiang Decoction in regulating the mitochondrial apoptotic pathway through the NF-κB/CXCL1/CXCR2 pathway in ulcerative colitis(UC).Methods Sixty C57BL/6J mice were divided into normal group,model group,Huanglian Ganzhang Decoction groups(low,medium and high doses)and mesalazine group.Except for the normal group,UC models were established in the other groups.The general conditions of the mice of all groups were recorded.HE staining was used to observe the pathological changes of the colon and rectum;electron microscopy was used to observe the mitochondrial condition;ELISA was used to determine the contents of IL-6,IL-10,D-lactic acid and DAO;Real-time PCR was used to detect the mRNA expression of NF-κB p65,CXCL1,CXCR2,Bcl-2 and Bax;Western blot was used to determine the protein expression levels of NF-κB p65,CXCL1,CXCR2,p-JAK2,p-STAT3,Bcl-2 and Bax.Results Compared with the normal group,the model group of mice showed a decline in condition,shortened length of the colon and rectum,and mitochondrial structural damage,furthermore,the levels of IL-6,DAO and D-lactic acid were increased,IL-10 level was decreased,the mRNA expression of NF-κB p65,CXCL1,CXCR2 and Bax were enhanced,Bcl-2 mRNA expression was weakened,the protein expression of NF-κB p65,CXCL1,CXCR2,p-JAK2,p-STAT3 and Bax were enhanced,Bcl-2 protein expression was weakened.Compared with the model group,the indexes mentioned above were reversed in the drug groups,especially in the high-dose group.Conclusion Huanglian Ganjiang Decoction may exert its therapeutic effect on UC in mice by regulating the NF-κB/CXCL1/CXCR2 pathway and mitochondrial apoptotic pathway.

ObjectiveThis study aims to explore the potential mechanism of the Xiezhuo Jiedu formula in regulating pyroptosis for the treatment of ulcerative colitis （UC） using bioinformatics and in vivo animal experiments. MethodsDifferentially expressed genes （DEGs） in colon tissues of UC patients were retrieved from the Gene Expression Omnibus （GEO） database. Pyroptosis-related genes were obtained from the GEO and GeneCards databases. The intersection of these datasets yielded pyroptosis-related DEGs （Pyro-DEGs）. Pyro-DEGs were subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using the Metascape database. A protein-protein interaction （PPI） network was constructed using the STRING database. Least absolute shrinkage and selection operator （LASSO） prediction model and receiver operating characteristic （ROC） analysis were conducted to identify core Pyro-DEGs with diagnostic and therapeutic potential. Immune infiltration analysis of the UC datasets was performed using the deconvolution method （CIBERSORT）， along with correlation analysis with core Pyro-DEGs. Sixty male Sprague-Dawley （SD） rats were randomly divided into a control group， a model group， high-， medium-， and low-dose groups of Xiezhuo Jiedu formula （26.64， 13.32， 6.66 g·kg^-1）， and a mesalazine group （0.27 g·kg^-1）， with 10 rats in each group. UC was established by intrarectal administration of 3，5-trinitrobenzenesulfonic acid （TNBS） dissolved in ethanol. The control and model groups were given distilled water by gavage， while the treatment groups were administered the corresponding drugs for 7 consecutive days. Hematoxylin-eosin （HE） staining was used to observe the colon histopathology. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors such as interleukin-1β （IL-1β）， IL-10， IL-18， and transforming growth factor-β （TGF-β）. Immunohistochemistry （IHC） and Western blot were applied to detect the expression of Caspase-1， gap junction alpha-1 protein （GJA1）， peroxisome proliferator-activated receptor gamma （PPARG）， and S100 calcium-binding protein A8 （S100A8）. Real-time quantitative polymerase chain reaction （Real-time PCR） was utilized to measure mRNA expression of Caspase-1， GJA1， PPARG， and S100A8. Western blot was performed to assess protein expression levels of Caspase-1， GJA1， PPARG， and S100A8. ResultsGEO datasets GSE87466 and GSE87473 yielded 64 Pyro-DEGs. KEGG analysis indicated that these genes were enriched in the NOD-like receptor signaling pathway， tumor necrosis factor （TNF） signaling pathway， and hypoxia-inducible factor 1 （HIF-1） signaling pathway. Four core Pyro-DEGs （Caspase-1， GJA1， PPARG， and S100A8） were identified. Immune infiltration analysis showed that expression of these genes was positively correlated with mast cells， neutrophils， M0 macrophages， M1 macrophages， and dendritic cells. Animal experimental results indicated that compared with the control group， the model group had significantly increased levels of IL-1β and IL-18， significantly decreased levels of IL-10 and TGF-β. The model group showed enhanced Caspase-1， GJA1， and S100A8 staining， and significantly increased mRNA and protein expression of Caspase-1， GJA1， and S100A8 （P<0.01）. In contrast， the expression of PPARG was reduced in the model group （P<0.01）. After treatment， all dosage groups showed varying degrees of improvement （P<0.05， P<0.01）， with the high-dose group showing the most significant improvement （P<0.01）. ConclusionCaspase-1， GJA1， PPARG， and S100A8 are core Pyro-DEGs closely associated with the pathogenesis of UC. These genes may collaborate with immune cells such as mast cells， neutrophils， and M0 macrophages to mediate disease development. The Xiezhuo Jiedu formula may regulate the expression of core Pyro-DEGs through the NOD-like receptor， TNF， and HIF-1 core signaling pathways， thereby modulating immune homeostasis in UC rats and effectively alleviating UC.

AIM:To construct and breed infertility model mice with ATP/GTP binding carboxypeptidase 1(Agtpbp1)gene knockout homozygote(Agtpbp1-/-)by CRISPR/Cas9 technology,so as to provide an animal model for the subsequent exploration of the pathogenesis of Agtpbp1 gene in male sterility.METHODS:The CRISPR/Cas9 technology was used to obtain Agtpbp1 gene knockout heterozygote(Agtpbp1+/-)mice according to the data of the main protein functional region of Agtpbp1 gene and combined with Cas9 nuclease.The obtained Agtpbp1+/-mice were mated,and their offspring were genotyped by PCR and agarose gel electrophoresis.The expression of Agtpbp1 at different levels was detected by RT-PCR,Western blot and immunohistochemical staining to support the identification results.The HE staining was used to observe the mouse cerebellum and eyeball structure to analyze the effect of Agtpbp1 gene knockout on Purkinje cells and photoreceptor cells.The symptoms of ataxia in mice were observed in combination with behavioral tests.The growth of mice was observed,and the changes of testicular tissue volume and weight of male mice were analyzed.The HE staining was used to observe the changes of testicular structure,and PAS staining was used to observe the changes of testicular germ cell cycle.Finally,sperm analyzer was used to analyze the sperm motility,so as to analyze the growth and develop-ment of the mice.RESULTS:The male and female Agtpbp1+/-mice could continue to mate,and three genotypes,Agtpbp1 wild-type(Agtpbp1+/+),Agtpbp1+/-and Agtpbp1-/-,were obtained.The genotypes of the offspring mice were successfully identified by PCR.The results of RT-PCR,Western blot and immunohistochemical staining verified the successful con-struction of Agtpbp1-/-mouse model at different levels(P<0.05).The results of HE staining showed that Purkinje cells were lost in the cerebellum of Agtpbp1-/-mice and the number of photoreceptor cells in the eyeball was reduced.Behavioral tests confirmed that Agtpbp1-/-mice had ataxia symptoms such as motor dysfunction and uncoordinated movements.Com-pared with control group,the testicular volume and weight of Agtpbp1-/-mice were significantly reduced.The results of HE staining showed a very small amount of sperm in the testis of Agtpbp1-/-mice.Combined with the sperm analyzer,it was ob-served that the sperm motility,vitality and movement rate of Agtpbp1-/-mice were significantly lower than those of the con-trol mice.Testicular sections with PAS staining showed cell cycle arrest of the sperm from Agtpbp1-/-mice.CONCLU-SION:In this study,Agtpbp1 knockout mice were successfully bred.The deletion of Agtpbp1 caused the arrest of sper-matogenic cell differentiation and the decrease in sperm motility in adult male mice,resulting in infertility.At the same time,it provides a new experimental tool for further exploring the molecular mechanism of Agtpbp1-induced male sterility.

Objective To explore the association between cardiorespiratory fitness and total atherosclerotic burden of brain-and heart-arteries in patients with ischemic stroke.Methods This study retrospectively enrolled consecutive patients with ischemic stroke of the large-artery atherosclerosis or small-artery occlusion subtypes accompanied by cervicocephalic atherosclerosis,who were admitted to the Department of Neurology,Xuanwu Hospital,Capital Medical University,between May 2023 and December 2024.General and clinical data,including age(＜60years,≥60years),sex,admission blood pressure,body mass index,history of hypertension,diabetes mellitus,hyperlipidemia,ischemic stroke,symptomatic coronary artery disease,smoking,alcohol consumption,and medication use(including antiplatelet drugs,anticoagulants,lipid-lowering drugs,antidiabetic drugs,antihypertensive drugs,and β-blockers),characteristics of ischemic stroke(including the duration of onset,ischemic side[left,right,bilateral],and ischemic location[anterior circulation,posterior circulation,bilateral circulation])were collected from all patients enrolled.On the morning after admission,fasting venous blood samples were collected to measure the levels of triglycerides,total cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,fasting blood glucose,glycated hemoglobin,homocysteine,fibrinogen,D-dimer,C-reactive protein,high-sensitivity C-reactive protein,N-terminal pro-B-type natriuretic peptide,creatine kinase-MB,cardiac troponin T,and myoglobin.Transthoracic echocardiography was performed within 7 days after admission to evaluate cardiac function.Left ventricular ejection fraction and cardiac output were recorded,and cardiac index were calculated.Upon admission,neurological impairment,disability,motor function,and balance ability were assessed using the National Institutes of Health stroke scale(NIHSS),modified Rankin scale(mRS),Fugl-Meyer assessment(FMA),and Berg balance scale,respectively.The Saltin-Grimby physical activity level scale was used to evaluate the intensity of habitual physical activity prior to stroke onset.Cardiopulmonary exercise testing was performed within 7 days after admission to assess cardiopulmonary fitness.During the test,peak oxygen uptake(VO2peak),percentage of predicted VO2peak(VO2peak%pred),and metabolic equivalent of task(MET)were recorded.Based on the minimum oxygen uptake required for independent living(15 ml/[kg·min]),VO2peak was classified into low VO2peak(＜15 ml/[kg·min])and high VO2peak(≥15 ml/[kg·min]).With VO2peak%pred＜0.60 as the cutoff,VO2peak%pred was divided into low VO2peak%pred(＜0.60)and high VO2peak%pred(≥0.60).MET were categorized into low MET(＜mean MET)and high MET(≥mean MET)according to the mean value of MET in this study population.Imaging data of cardio-cerebral arteries were obtained by simultaneous CT arteriography within 7 days after admission.Total atherosclerotic burden of brain-and heart-supplying arteries(TAB-BHAs)was used to quantitatively evaluate the overall degree of cardio-cerebral atherosclerosis.And patients were stratified into high TAB-BHAs(≥4 points)and low TAB-BHAs(＜4 points)groups.The differences in general and clinical characteristics,as well as cardiopulmonary fitness indices,were assessed among different groups of TAB-BHAs.Spearman' s correlation was used in the preliminary analysis of the relationship between cardiorespiratory fitness indicators and TAB-BHAs.Multivariate Logistic regression was conducted with VO2peak,low VO2peak%pred,and low METs as independent variables,and high TAB-BHAs as the dependent variable to assess the association between cardiopulmonary fitness and TAB-BHAs.Covariates were selected through regressions stratified by age≥60 years,male,the least absolute shrinkage and selection operator(LASSO),and the variables exhibit statistically significant differences between the low and high TAB-BHAs groups,respectively.Results A total of 104 patients with ischemic stroke were enrolled.Among all patients enrolled,60patients(with ages ranging from 30 to 72years and a mean age of[54±11]years)met the inclusion and exclusion criteria,and were ultimately included.Amid the 60 patients,52 were male and 8 were female.There were 20 patients(33.3%)with low VO2peak,28 patients(46.7%)with low VO2peak%pred,and 31 patients(51.7%)with low MET(MET＜5.0).45 patients were classified into the low TAB-BHAs group and 15 into the high TAB-BHAs group.(1)Compared with the low TAB-BHAs group,patients in the high TAB-BHAs group were significantly older and had a higher proportion of patients aged≥60 years,they also showed significantly higher fibrinogen levels and a history of antihypertensive drug use(all P＜0.05).No significant differences were observed in other general or clinical characteristics between the two groups(all P＞0.05).Regarding cardiopulmonary fitness parameters,patients in the high TAB-BHAs group had lower VO2peak and MET levels,and higher proportions of patients with low VO2peak,low VO2peak%pred,and low MET compared with those in the low TAB-BHAs group(all P＜0.05).(2)Based on the Spearman's correlation analysis,VO2peak(r=-0.298,P=0.021),VO2peak%pred(r=-0.305,P=0.018),MET(r=-0.303,P=0.018)all exhibited negative correlations between and TAB-BHAs.(3)LASSO regression was used to identify the most pertinent variables for high TAB-BHAs,including low VO2peak,low VO2peak%pred,and fibrinogen.Multivariate Logistic regression analysis was then performed with low VO2peak,low VO2peak%pred,and low MET as independent variables and high TAB-BHAs as the dependent variable.The results showed that after adjusting for age≥60 years,male,and fibrinogen,both low VO2peak(adjusted OR,4.420,95%CI 1.052-18.573,P=0.042)and low VO2peak%pred(adjusted OR,7.423,95%CI 1.411-39.046,P=0.018)were independently associated with high TAB-BHAs in ischemic stroke patients.After further adjusting for variables that differed among high and low TAB-BHAs groups(including age≥60 years,male,fibrinogen,and history of using antihypertensive drugs),multivariate Logistic regression showed that low VO2peak%pred remained independently associated with high TAB-BHAs in ischemic stroke patients(adjusted OR,6.347,95%CI 1.170-34.418,P=0.032).Conclusion Poor cardiopulmonary fitness might be associated with high degree of overall cardio-cerebral atherosclerosis,suggesting cardiorespiratory fitness as a potential correlated indicator that could provide clues for improving early screening of patients with heavy cardio-cerebral atherosclerotic burden and for exploring the underlying mechanisms.

AIM:To construct and breed infertility model mice with ATP/GTP binding carboxypeptidase 1(Agtpbp1)gene knockout homozygote(Agtpbp1-/-)by CRISPR/Cas9 technology,so as to provide an animal model for the subsequent exploration of the pathogenesis of Agtpbp1 gene in male sterility.METHODS:The CRISPR/Cas9 technology was used to obtain Agtpbp1 gene knockout heterozygote(Agtpbp1+/-)mice according to the data of the main protein functional region of Agtpbp1 gene and combined with Cas9 nuclease.The obtained Agtpbp1+/-mice were mated,and their offspring were genotyped by PCR and agarose gel electrophoresis.The expression of Agtpbp1 at different levels was detected by RT-PCR,Western blot and immunohistochemical staining to support the identification results.The HE staining was used to observe the mouse cerebellum and eyeball structure to analyze the effect of Agtpbp1 gene knockout on Purkinje cells and photoreceptor cells.The symptoms of ataxia in mice were observed in combination with behavioral tests.The growth of mice was observed,and the changes of testicular tissue volume and weight of male mice were analyzed.The HE staining was used to observe the changes of testicular structure,and PAS staining was used to observe the changes of testicular germ cell cycle.Finally,sperm analyzer was used to analyze the sperm motility,so as to analyze the growth and develop-ment of the mice.RESULTS:The male and female Agtpbp1+/-mice could continue to mate,and three genotypes,Agtpbp1 wild-type(Agtpbp1+/+),Agtpbp1+/-and Agtpbp1-/-,were obtained.The genotypes of the offspring mice were successfully identified by PCR.The results of RT-PCR,Western blot and immunohistochemical staining verified the successful con-struction of Agtpbp1-/-mouse model at different levels(P<0.05).The results of HE staining showed that Purkinje cells were lost in the cerebellum of Agtpbp1-/-mice and the number of photoreceptor cells in the eyeball was reduced.Behavioral tests confirmed that Agtpbp1-/-mice had ataxia symptoms such as motor dysfunction and uncoordinated movements.Com-pared with control group,the testicular volume and weight of Agtpbp1-/-mice were significantly reduced.The results of HE staining showed a very small amount of sperm in the testis of Agtpbp1-/-mice.Combined with the sperm analyzer,it was ob-served that the sperm motility,vitality and movement rate of Agtpbp1-/-mice were significantly lower than those of the con-trol mice.Testicular sections with PAS staining showed cell cycle arrest of the sperm from Agtpbp1-/-mice.CONCLU-SION:In this study,Agtpbp1 knockout mice were successfully bred.The deletion of Agtpbp1 caused the arrest of sper-matogenic cell differentiation and the decrease in sperm motility in adult male mice,resulting in infertility.At the same time,it provides a new experimental tool for further exploring the molecular mechanism of Agtpbp1-induced male sterility.

Objective:To evaluate the therapeutic value of radiotherapy in combined immunotherapy and chemotherapy as first-line treatment for patients with oligometastatic non-small cell lung cancer (NSCLC).Methods:A retrospective analysis was conducted on data from 195 NSCLC patients who lacked epidermal growth factor receptor (EGFR) or anaplastic lymphoma kinase (ALK) mutations and were treated at the Anyang Tumor Hospital and the Fourth Hospital of Hebei Medical University from January 2019 to December 2021. These patients consisted of 166 male and 29 female cases, aged from 28 to 85 years, with an average age of (61.4 ± 9.3) years. These patients were divided into two groups, with each group receiving the radiotherapy and combined immunotherapy and chemotherapy (the radiotherapy and combination group, n = 60) and combined immunotherapy and chemotherapy only (the combination group, n = 135). Then, propensity score matching (PSM) was performed to analyze the differences in prognosis between both groups before and after PSM, as well as the short-term efficacy and adverse reactions after PSM. Results:For the 195 NSCLC patients, the median follow-up time was 31.8 months, with median overall survival (OS) and median progression-free survival (PFS) recorded at 23.8 months and 9.2 months, respectively. The radiotherapy and combination group exhibited enhanced 1-, 2-, and 3-year survival rates of 78.5%, 55.9%, and 45.1%, respectively, significantly higher than the combination group (48.3%, 35.6%, and 26.6%, respectively, χ2 = 14.65, P < 0.001). Similarly, the radiotherapy and combination group displayed 1-, 2-, and 3-year PFS rates of 51.9%, 29.5%, and 22.7%, respectively, exceeding those of the combination group (30.0%, 24.5%, and 16.9%, respectively, χ2=6.09, P=0.014). After PSM, the radiotherapy and combination group manifested an objective response rate (ORR) of 60.0% (33/55) and a disease control rate (DCR) of 89.1% (49/55), which were 16.4% (9/55) and 56.4% (31/55), respectively for the combination group. These results suggested that the radiotherapy and combination group demonstrated significantly higher ORR and DCR ( χ2 = 22.18, 14.85, P<0.001). After PSM, the radiotherapy and combination group yielded 1-, 2-, and 3-year survival rates of 70.9%, 52.3%, and 41.9%, respectively, significantly than the combination group (43.6%, 29.8%, and 27.1%, respectively, χ2=8.95, P=0.003). The radiotherapy and combination group exhibited 1-, 2-, and 3-year PFS rates of 47.3%, 27.3%, and 18.7%, respectively, significantly higher than the combination group (23.6%, 17.6%, and 15.4%, respectively, χ2 = 6.71, P = 0.010). Multivariate Cox regression analysis revealed that independent factors affecting OS included clinical stage, treatment regimen, number of immunotherapy cycles, and treatment efficacy ( HR = 1.88, 2.11, 0.23, 1.79, P < 0.05). Similarly, independent factors affecting PFS consisted of treatment regimen, number of immunotherapy cycles, and treatment efficacy ( HR = 1.62, 0.37, 3.42, P <0.05). There were no statistical differences in the incidence of grade ≥ 2 bone marrow suppression (18.2% vs. 12.7%) and grade ≥ 2 pneumonia (21.8% vs. 14.5%) between both groups ( P>0.05). Conclusions:Introducing radiotherapy into combined immunotherapy and chemotherapy as first-line treatment for oligometastatic NSCLC can optimize both local and systemic disease control and significantly improve patient prognosis without increasing treatment-related adverse reactions.

Objective To explore the association between cardiorespiratory fitness and total atherosclerotic burden of brain-and heart-arteries in patients with ischemic stroke.Methods This study retrospectively enrolled consecutive patients with ischemic stroke of the large-artery atherosclerosis or small-artery occlusion subtypes accompanied by cervicocephalic atherosclerosis,who were admitted to the Department of Neurology,Xuanwu Hospital,Capital Medical University,between May 2023 and December 2024.General and clinical data,including age(＜60years,≥60years),sex,admission blood pressure,body mass index,history of hypertension,diabetes mellitus,hyperlipidemia,ischemic stroke,symptomatic coronary artery disease,smoking,alcohol consumption,and medication use(including antiplatelet drugs,anticoagulants,lipid-lowering drugs,antidiabetic drugs,antihypertensive drugs,and β-blockers),characteristics of ischemic stroke(including the duration of onset,ischemic side[left,right,bilateral],and ischemic location[anterior circulation,posterior circulation,bilateral circulation])were collected from all patients enrolled.On the morning after admission,fasting venous blood samples were collected to measure the levels of triglycerides,total cholesterol,low-density lipoprotein cholesterol,high-density lipoprotein cholesterol,fasting blood glucose,glycated hemoglobin,homocysteine,fibrinogen,D-dimer,C-reactive protein,high-sensitivity C-reactive protein,N-terminal pro-B-type natriuretic peptide,creatine kinase-MB,cardiac troponin T,and myoglobin.Transthoracic echocardiography was performed within 7 days after admission to evaluate cardiac function.Left ventricular ejection fraction and cardiac output were recorded,and cardiac index were calculated.Upon admission,neurological impairment,disability,motor function,and balance ability were assessed using the National Institutes of Health stroke scale(NIHSS),modified Rankin scale(mRS),Fugl-Meyer assessment(FMA),and Berg balance scale,respectively.The Saltin-Grimby physical activity level scale was used to evaluate the intensity of habitual physical activity prior to stroke onset.Cardiopulmonary exercise testing was performed within 7 days after admission to assess cardiopulmonary fitness.During the test,peak oxygen uptake(VO2peak),percentage of predicted VO2peak(VO2peak%pred),and metabolic equivalent of task(MET)were recorded.Based on the minimum oxygen uptake required for independent living(15 ml/[kg·min]),VO2peak was classified into low VO2peak(＜15 ml/[kg·min])and high VO2peak(≥15 ml/[kg·min]).With VO2peak%pred＜0.60 as the cutoff,VO2peak%pred was divided into low VO2peak%pred(＜0.60)and high VO2peak%pred(≥0.60).MET were categorized into low MET(＜mean MET)and high MET(≥mean MET)according to the mean value of MET in this study population.Imaging data of cardio-cerebral arteries were obtained by simultaneous CT arteriography within 7 days after admission.Total atherosclerotic burden of brain-and heart-supplying arteries(TAB-BHAs)was used to quantitatively evaluate the overall degree of cardio-cerebral atherosclerosis.And patients were stratified into high TAB-BHAs(≥4 points)and low TAB-BHAs(＜4 points)groups.The differences in general and clinical characteristics,as well as cardiopulmonary fitness indices,were assessed among different groups of TAB-BHAs.Spearman' s correlation was used in the preliminary analysis of the relationship between cardiorespiratory fitness indicators and TAB-BHAs.Multivariate Logistic regression was conducted with VO2peak,low VO2peak%pred,and low METs as independent variables,and high TAB-BHAs as the dependent variable to assess the association between cardiopulmonary fitness and TAB-BHAs.Covariates were selected through regressions stratified by age≥60 years,male,the least absolute shrinkage and selection operator(LASSO),and the variables exhibit statistically significant differences between the low and high TAB-BHAs groups,respectively.Results A total of 104 patients with ischemic stroke were enrolled.Among all patients enrolled,60patients(with ages ranging from 30 to 72years and a mean age of[54±11]years)met the inclusion and exclusion criteria,and were ultimately included.Amid the 60 patients,52 were male and 8 were female.There were 20 patients(33.3%)with low VO2peak,28 patients(46.7%)with low VO2peak%pred,and 31 patients(51.7%)with low MET(MET＜5.0).45 patients were classified into the low TAB-BHAs group and 15 into the high TAB-BHAs group.(1)Compared with the low TAB-BHAs group,patients in the high TAB-BHAs group were significantly older and had a higher proportion of patients aged≥60 years,they also showed significantly higher fibrinogen levels and a history of antihypertensive drug use(all P＜0.05).No significant differences were observed in other general or clinical characteristics between the two groups(all P＞0.05).Regarding cardiopulmonary fitness parameters,patients in the high TAB-BHAs group had lower VO2peak and MET levels,and higher proportions of patients with low VO2peak,low VO2peak%pred,and low MET compared with those in the low TAB-BHAs group(all P＜0.05).(2)Based on the Spearman's correlation analysis,VO2peak(r=-0.298,P=0.021),VO2peak%pred(r=-0.305,P=0.018),MET(r=-0.303,P=0.018)all exhibited negative correlations between and TAB-BHAs.(3)LASSO regression was used to identify the most pertinent variables for high TAB-BHAs,including low VO2peak,low VO2peak%pred,and fibrinogen.Multivariate Logistic regression analysis was then performed with low VO2peak,low VO2peak%pred,and low MET as independent variables and high TAB-BHAs as the dependent variable.The results showed that after adjusting for age≥60 years,male,and fibrinogen,both low VO2peak(adjusted OR,4.420,95%CI 1.052-18.573,P=0.042)and low VO2peak%pred(adjusted OR,7.423,95%CI 1.411-39.046,P=0.018)were independently associated with high TAB-BHAs in ischemic stroke patients.After further adjusting for variables that differed among high and low TAB-BHAs groups(including age≥60 years,male,fibrinogen,and history of using antihypertensive drugs),multivariate Logistic regression showed that low VO2peak%pred remained independently associated with high TAB-BHAs in ischemic stroke patients(adjusted OR,6.347,95%CI 1.170-34.418,P=0.032).Conclusion Poor cardiopulmonary fitness might be associated with high degree of overall cardio-cerebral atherosclerosis,suggesting cardiorespiratory fitness as a potential correlated indicator that could provide clues for improving early screening of patients with heavy cardio-cerebral atherosclerotic burden and for exploring the underlying mechanisms.

ObjectiveKey microRNAs （miRNAs） of colorectal adenoma （CRA） were identified and analyzed by bioinformatics methods， and differentially expressed genes （DEGs） were screened to construct regulatory relationships. The mechanism of Xiezhuo Jiedu recipe in preventing CRA was speculated and verified by animal experiments. MethodThe miRNAs dataset GSE50194 was obtained from the Gene Expression Omnibus （GEO） database of intestinal mucosal tissue of CRA patients， and the differentially expressed miRNAs were screened by GEO2R and Excel. TargetScan， miRTarbase， and miRDB databases were used to predict the target genes of the differentially expressed miRNAs， and an intersection was obtained. Key DEGs were screened through the STRING database and Cytoscape software， and the TRRUST database was used to predict downstream binding transcription factors （TFs）. The mRNA intersection was enriched by gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） in the Metascape database. DIANA TOOLS were applied to perform KEGG enrichment analysis of key miRNAs， and the key signaling pathways were selected for animal experiments. In animal experiments， the CRA mice model was established by using sodium glycan sulfate （DSS） drinking combined with intraperitoneal injection of azomethane oxide （AOM）， and Xiezhuo Jiedu recipe and aspirin were given by intragastric administration at the same time. The experiment lasted for nine weeks. The pathological changes in intestinal tissue were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of miR-34a-5p in adenoma tissue. Protein expression levels of phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， phosphoryl-PI3K （p-PI3K）， phosphoryl-Akt （p-Akt）， and B cell lymphoma （Bcl）-2 were detected by Western blot. The expression of Cyclin D1 （CCND1） was detected by immunohistochemistry （IHC）. In situ terminal transferase labeling （TUNEL） was used to detect apoptosis of adenoma tissue cells. ResultThe GEO database screened the GSE50194 dataset， and miR-34a-5p was selected as the research object from CRA and normal tissue. A total of 93 DEGs were selected. Among them， GO and KEGG enrichment analyses were closely related to biological processes such as transcriptional regulatory complex， RNA polymerase Ⅱ transcriptional regulatory complex， enzyme-linked receptor protein signaling pathway， and DNA-binding transcriptional activator activity， cancer pathway， PI3K/Akt pathway， etc. miR-34a-5p is mainly enriched in PI3K/Akt， cell cycle， and colorectal cancer pathways. Five key DEGs were screened out through the Matescape database， among which Bcl-2 and CCND1 were the key DEGs of miR-34a-5p. Further screening of the TFs of key DEGs revealed that E2F transcription factor 1 （E2F1） and tumor protein P53 （TP53） were the main TFs of Bcl-2 and CCND1. Animal experiments showed that Xiezhuo Jiedu recipe could effectively up-regulate mRNA level of miR-34a-5p， down-regulate the expression of PI3K， Akt， Bcl-2， p-PI3K， and p-Akt proteins in the intestinal tissue of CRA mice， down-regulate the positive expression rate of CCND1， and increase the apoptosis rate of intestinal epithelial cells. ConclusionIt is speculated that Xiezhuo Jiedu recipe may inhibit the abnormal proliferation and promote the apoptosis of intestinal epithelial cells in CRA mice by regulating the miR-34a-5p/PI3K/Akt signaling pathway， thus playing a role in the prevention of CRA.