BACKGROUND:Traditional Chinese medicine has rich experience and unique advantages in the empirical treatment of phlegm-heat and Fu-organs excess syndrome of ischemic stroke.In order to further explore the therapeutic targets and mechanisms of traditional Chinese medicine for this disease,it is crucial to establish a stable and reliable animal model of phlegm-heat and Fu-organs excess syndrome combined with empirical symptoms of ischemic stroke. OBJECTIVE:To explore the establishment method and evaluation system of the rat model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome. METHODS:Sixty male Sprague-Dawley rats were randomly divided into four groups:blank control group(n=12),ischemic stroke group(n=18),disease+syndrome group(n=18),phlegm-heat and Fu-organ excess syndrome group(n=12),all of which were given high-fat diet for 25 days.On the 26th day,the rats in the blank control group and ischemic stroke group were intragastrically given normal saline and high fat diet,while those in the other two groups were intragastrically given autologous feces suspension and high fat diet for 3 continuous days.After gavage,ischemic stroke models were established using the suture method in the ischemic stroke group and disease+syndrome group.The changes in diet,water intake,body mass,body temperature,fecal traits,nasal secretions,sputum in the throat,and tongue image were recorded.Neurological deficits,tongue image,blood lipid levels,morphological changes of brain tissue and carotid artery,and the serum levels of motilin and somatostatin were detected. RESULTS AND CONCLUSION:Compared with the control group,the rats in the disease+syndrome group had shortness of breath,listlessness,irritability,bradykinesia,a large number of secretions around the nose,audible and heavy sputum in the throat,decreased diet and water intake,increased body mass,body temperature,and slingual vein score,decreased fecal pellet count,Bristol score and fecal moisture content,increased serum total cholesterol,triglyceride,low-density lipoprotein and somatostatin levels,decreased motilin level,increased neurological deficit score,significant pathological changes of the carotid artery,and significant morphological changes of the brain tissue.The ischemic stroke group only showed pathological changes of ischemic brain tissue,without the characteristics of phlegm-heat and Fu-organ excess syndrome.The phlegm-heat and Fu-organ excess syndrome group could present with the typical characteristics of traditional Chinese medicine syndromes,without the pathological changes of brain tissue with ischemic stroke.To conclude,the compound modeling method of high-fat induction combined with suture method and autologous feces gavage can establish an animal model of ischemic stroke with phlegm-heat and Fu-organ excess syndrome.

Objective To investigate the regulatory role and mechanism of lobetyolin(LBT,a poly-acetylene glycoside isolated from the roots of Codonopsis pilosula)in the proliferation and apop-tosis of brain glioma cells based on the Akt/GSK-3β/Snail signaling pathway.Methods Human brain glioma cell line U-373MG was randomly divided into normal,SC79(Akt activator),LBT,and LBT+SC79 groups.After corresponding interventions,CCK-8 assay,colony formation assay,and flow cytometry were used to detect the proliferation and apoptosis of the cells.Western blot-ting was employed to measure the protein expression levels of the molecules related to prolifera-tion,apoptosis,and Akt/GSK-3β/Snail signaling pathway.After tumor xenograft nude mouse model of U-373MG cells was established,followed by grouping and interventions as above cell experiments,the tumor weight and volume were measured.Immunohistochemical assay and TUNEL assay were performed to detect the proliferation and apoptosis of tumor cells.Western blotting was applied to detect Akt/GSK-3β/Snail signaling pathway related proteins in the nude mouse groups.Results In the LBT+SC79 group,cell viability,number of formed colonies,pro-tein levels of cyclin D1,Bcl-2 and Snail,p-Akt/Akt and p-GSK-3β/GSK-3β,tumor weight and vol-ume,and positive ratios of Ki67,cyclin D1 and Bcl-2 in transplanted tumors were increased(P＜0.05),and cell apoptotic rate[(3.20±1.14)％vs(46.15±1.52)％,P＜0.05],Bax protein level(0.51±0.07 vs 0.89±0.06,P＜0.05),and positive ratios of TUNEL[(51.56±7.13)％vs(74.95±8.61)％,P＜0.05]and Bax[(32.71±5.43)％vs(41.86±4.90),P＜0.05]in transplanted tumors were declined when compared with the LBT group.Conclusion LBT can induce apoptosis and inhibit proliferation of brain glioma cells in vitro and in vivo by blocking activation of the Akt/GSK-3β/Snail signaling pathway.

Objective:To analyze the epidemiological and clinical features of infectious diseases of the central nervous system (CNS).Methods:A cross-sectional study and analysis were conducted to summarize the epidemiological and clinical characteristics of 9 918 patients with CNS infectious diseases, who were diagnosed and treated at 29 hospitals across China from January 1, 2001 to December 31, 2020. Data collected included demographic data, clinical manifestations, health economic indicators, and prognostic outcomes.Results:Among the 9 918 collected cases of CNS infectious diseases, 5 559 were male (56.0%) and 4 359 were female (44.0%), with an onset age of 38 (25, 53) years. Education level: slightly more junior high school education (2 651 cases, 26.7%), and less elementary school education and below (2 181 cases, 22.0%) were found. Occupational distribution: farmers were found predominant (3 215 cases, 32.4%), followed by workers (1 826 cases, 18.4%) and students (1 633 cases, 16.5%). Clinical manifestations: headache (6 074 cases, 61.2%), fever (5 869 cases, 59.2%) and positive meningeal irritation signs (2 273 cases, 22.9%) were the 3 most common clinical manifestations, followed by nausea and (or) vomiting (2 095 cases, 21.1%), impaired consciousness (2 077 cases, 20.9%), psychiatric symptom (1 866 cases, 18.8%) and epilepsy (1 627 cases, 16.4%), etc., and cranial nerve involvement was found in 669 cases (6.7%). Major pathogens included viruses in 6 814 cases (68.7%), Mycobacterium tuberculosis in 1 677 cases (16.9%), common bacteria in 864 cases (8.7%), fungi in 254 cases (2.6%), spirochetes of syphilis in 183 cases (1.8%), parasites in 121 cases (1.2%), and rickettsiae in 5 cases (0.1%). Urban-rural distribution: slightly more cases were found in the countryside (5 418 cases, 54.6%) than in the towns (4 500 cases, 45.4%). Distribution of onset by season: 2 412 cases (24.3%) fell ill in spring, 2 835 cases (28.6%) in summer, 2 187 cases (22.1%) in fall, and 2 484 cases (25.0%) in winter. Health economics: the duration of hospitalization was 15 (8, 27) days, and the cost of hospitalization was 1.53 (0.91, 3.02)×10 000 yuan. Prognosis: 9 531 cases (96.1%) were cured or improved, and 92 cases (0.9%) died. Conclusions:The pathogens responsible for CNS infectious diseases are predominantly viruses. Although the incidence is slightly higher during the summer months, the overall seasonal pattern is not particularly pronounced. These infections are more commonly observed in young and middle-aged males and present with a diverse range of clinical manifestations, contributing to a significant disease burden.

Objective:To explore the potential risk factors for cancer in patients with adenomatous polyps based on tumor markers and polyp lesion characteristics.Methods:A retrospective analysis was conducted to collect clinical data of 115 patients with adenomatous intestinal polyps who visited Nanjing Jiangbei Hospital from November 2022 to November 2024. They were divided into a cancerous group (17 cases) and a non cancerous group (98 cases) based on whether they were cancerous or not. Clinical data such as tissue type and polyp site and tumor marker levels such as carcinoembryonic antigen (CEA) and cancer antigen 72-4 (CA72-4) were collected at the first visit of all patients. The potential risk factors of adenomatous intestinal polyp canceration were investigated by Logistic regression analysis.Results:Univariate analysis revealed that the proportion of villous tubular adenomas, central depression of polyps, and lobulated polyps in the cancerous group were higher than those in the non cancerous group. Serum levels of CEA and CA72-4 were also higher in the cancerous group than in the non cancerous group : 13/17 vs.47.96% (47/98), 7/17 vs. 15.31% (15/98), 6/17 vs. 8.16% (8/98), (6.41 ± 1.81) μg/L vs. (4.23 ± 1.48) μg/L, (6.98 ± 1.83) kU/L vs. (5.66 ± 1.78) kU/L, respectively. The difference was statistically significant ( P<0.05). The results of Logistic regression analysis showed that the histological subtype of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 were independent risk factors for cancer in patients with adenomatous intestinal polyps ( P<0.05). A nomogram risk model was constructed based on the influencing factors of canceration in patients with adenomatous intestinal polyps. The calibration curve was drawn, and the calibration curve was similar to the Y-X straight line, suggesting that the evaluation results of the nomogram risk model were highly consistent with the actual observation results. The receiver operating characteristic (ROC) curve was drawn. The results showed that the area under the curve (AUC) of the nomogram risk model for evaluating the canceration of patients with adenomatous intestinal polyps was 0.956, and the evaluation value was high. The decision curve was drawn, with the threshold of high risk as the horizontal coordinate and the net rate of return as the vertical coordinate. The results showed that when the threshold was in the range of 0 - 0.85, 0.96 - 0.99, the net benefit rate of predicting the cancer risk of patients with adenomatous intestinal polyps was greater than 0 and the maximum net benefit rate was 0.148. Conclusions:The histological classification of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 are independent risk factors for cancer in patients with adenomatous intestinal polyps; The evaluation efficiency of the column chart risk model constructed based on the above factors is good.

Objective:To explore the potential risk factors for cancer in patients with adenomatous polyps based on tumor markers and polyp lesion characteristics.Methods:A retrospective analysis was conducted to collect clinical data of 115 patients with adenomatous intestinal polyps who visited Nanjing Jiangbei Hospital from November 2022 to November 2024. They were divided into a cancerous group (17 cases) and a non cancerous group (98 cases) based on whether they were cancerous or not. Clinical data such as tissue type and polyp site and tumor marker levels such as carcinoembryonic antigen (CEA) and cancer antigen 72-4 (CA72-4) were collected at the first visit of all patients. The potential risk factors of adenomatous intestinal polyp canceration were investigated by Logistic regression analysis.Results:Univariate analysis revealed that the proportion of villous tubular adenomas, central depression of polyps, and lobulated polyps in the cancerous group were higher than those in the non cancerous group. Serum levels of CEA and CA72-4 were also higher in the cancerous group than in the non cancerous group : 13/17 vs.47.96% (47/98), 7/17 vs. 15.31% (15/98), 6/17 vs. 8.16% (8/98), (6.41 ± 1.81) μg/L vs. (4.23 ± 1.48) μg/L, (6.98 ± 1.83) kU/L vs. (5.66 ± 1.78) kU/L, respectively. The difference was statistically significant ( P<0.05). The results of Logistic regression analysis showed that the histological subtype of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 were independent risk factors for cancer in patients with adenomatous intestinal polyps ( P<0.05). A nomogram risk model was constructed based on the influencing factors of canceration in patients with adenomatous intestinal polyps. The calibration curve was drawn, and the calibration curve was similar to the Y-X straight line, suggesting that the evaluation results of the nomogram risk model were highly consistent with the actual observation results. The receiver operating characteristic (ROC) curve was drawn. The results showed that the area under the curve (AUC) of the nomogram risk model for evaluating the canceration of patients with adenomatous intestinal polyps was 0.956, and the evaluation value was high. The decision curve was drawn, with the threshold of high risk as the horizontal coordinate and the net rate of return as the vertical coordinate. The results showed that when the threshold was in the range of 0 - 0.85, 0.96 - 0.99, the net benefit rate of predicting the cancer risk of patients with adenomatous intestinal polyps was greater than 0 and the maximum net benefit rate was 0.148. Conclusions:The histological classification of villous tubular adenoma, central depression of polyps, lobulated polyps, and high levels of CEA and CA72-4 are independent risk factors for cancer in patients with adenomatous intestinal polyps; The evaluation efficiency of the column chart risk model constructed based on the above factors is good.

Objective To investigate the regulatory role and mechanism of lobetyolin(LBT,a poly-acetylene glycoside isolated from the roots of Codonopsis pilosula)in the proliferation and apop-tosis of brain glioma cells based on the Akt/GSK-3β/Snail signaling pathway.Methods Human brain glioma cell line U-373MG was randomly divided into normal,SC79(Akt activator),LBT,and LBT+SC79 groups.After corresponding interventions,CCK-8 assay,colony formation assay,and flow cytometry were used to detect the proliferation and apoptosis of the cells.Western blot-ting was employed to measure the protein expression levels of the molecules related to prolifera-tion,apoptosis,and Akt/GSK-3β/Snail signaling pathway.After tumor xenograft nude mouse model of U-373MG cells was established,followed by grouping and interventions as above cell experiments,the tumor weight and volume were measured.Immunohistochemical assay and TUNEL assay were performed to detect the proliferation and apoptosis of tumor cells.Western blotting was applied to detect Akt/GSK-3β/Snail signaling pathway related proteins in the nude mouse groups.Results In the LBT+SC79 group,cell viability,number of formed colonies,pro-tein levels of cyclin D1,Bcl-2 and Snail,p-Akt/Akt and p-GSK-3β/GSK-3β,tumor weight and vol-ume,and positive ratios of Ki67,cyclin D1 and Bcl-2 in transplanted tumors were increased(P＜0.05),and cell apoptotic rate[(3.20±1.14)％vs(46.15±1.52)％,P＜0.05],Bax protein level(0.51±0.07 vs 0.89±0.06,P＜0.05),and positive ratios of TUNEL[(51.56±7.13)％vs(74.95±8.61)％,P＜0.05]and Bax[(32.71±5.43)％vs(41.86±4.90),P＜0.05]in transplanted tumors were declined when compared with the LBT group.Conclusion LBT can induce apoptosis and inhibit proliferation of brain glioma cells in vitro and in vivo by blocking activation of the Akt/GSK-3β/Snail signaling pathway.

Objective:To investigate the effects of phillyrin(PHN)on the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of glioma U251 cells by adjusting the high mobility group protein B1(HMGB1)/receptor of advanced glycation endproduct(RAGE)signaling pathway.Methods:Human glioma U251cells were assigned into the PHN-0 group(treated with 0 μmol/L PHN),the low,medium,and high-dose PHN groups(PHN-50、PHN-100、PHN-200 groups,treated with 50,100,and 200 μmol/L PHN respectively),the PHN+pcDNA-NC group(treated with 200 μmol/L PHN after transfection of pcDNA-NC plasmid),and the PHN+HMGB1 group(treated with 200 μmol/L PHN after transfection of overexpressed HMGB1 plasmid).The proliferation ability of cells in each group was detected by the CCK-8 method and the clone formation assay.The apoptosis level of cells in each group was detected by flow cytometry.The migration and invasion abilities of cells in each group were detected by the Transwell assay.ELISA was used to detect the IL-8 secretion level of cells in each group.Immunofluorescence was used to detect the positive rates of N-cadherin and E-cadherin in cells of each group.WB assay was performed to detect the expression levels of Toll like receptor 4(TLR4),nuclear factor-kappa B(NF-κ B),HMGB1,RAGE,N-cadherin,E-cadherin,cell cycle protein D1(cyclin D1),cyclin dependent kinase 2(CDK2),B-lymphoblastoma-2(Bcl-2),Bcl-2 associated X protein(BAX)proteins in cells of each group.Results:Compared with those in the PHN-0 group,the proliferation activity,the number of clone formation,the numbers of invasion and migration,IL-8 secretion levels,the positive rate and protein expression of N-cadherin,and the expressions of TLR4,NF-κB,HMGB1,RAGE,cyclin D1 and CDK2 protein in the PHN-50,PHN-100,and PHN-200 groups decreased significantly(all P<0.05);and the apoptosis rate,the positivity rate and protein expression of E-cadherin,and the BAX/Bcl-2 ratio increased significantly(all P<0.05).At the same time,overexpression of HMGB1 could reverse the inhibitory effects of PHN on the proliferation,migration,invasion and EMT of U251 cells,as well as its promoting effect on the apoptosis(all P<0.05).Conclusion:PHN inhibits the proliferation,invasion and EMT progression of glioma U251 cells through the HMGB1/RAGE signaling pathway.

Objective:To analyze the epidemiological and clinical features of infectious diseases of the central nervous system (CNS).Methods:A cross-sectional study and analysis were conducted to summarize the epidemiological and clinical characteristics of 9 918 patients with CNS infectious diseases, who were diagnosed and treated at 29 hospitals across China from January 1, 2001 to December 31, 2020. Data collected included demographic data, clinical manifestations, health economic indicators, and prognostic outcomes.Results:Among the 9 918 collected cases of CNS infectious diseases, 5 559 were male (56.0%) and 4 359 were female (44.0%), with an onset age of 38 (25, 53) years. Education level: slightly more junior high school education (2 651 cases, 26.7%), and less elementary school education and below (2 181 cases, 22.0%) were found. Occupational distribution: farmers were found predominant (3 215 cases, 32.4%), followed by workers (1 826 cases, 18.4%) and students (1 633 cases, 16.5%). Clinical manifestations: headache (6 074 cases, 61.2%), fever (5 869 cases, 59.2%) and positive meningeal irritation signs (2 273 cases, 22.9%) were the 3 most common clinical manifestations, followed by nausea and (or) vomiting (2 095 cases, 21.1%), impaired consciousness (2 077 cases, 20.9%), psychiatric symptom (1 866 cases, 18.8%) and epilepsy (1 627 cases, 16.4%), etc., and cranial nerve involvement was found in 669 cases (6.7%). Major pathogens included viruses in 6 814 cases (68.7%), Mycobacterium tuberculosis in 1 677 cases (16.9%), common bacteria in 864 cases (8.7%), fungi in 254 cases (2.6%), spirochetes of syphilis in 183 cases (1.8%), parasites in 121 cases (1.2%), and rickettsiae in 5 cases (0.1%). Urban-rural distribution: slightly more cases were found in the countryside (5 418 cases, 54.6%) than in the towns (4 500 cases, 45.4%). Distribution of onset by season: 2 412 cases (24.3%) fell ill in spring, 2 835 cases (28.6%) in summer, 2 187 cases (22.1%) in fall, and 2 484 cases (25.0%) in winter. Health economics: the duration of hospitalization was 15 (8, 27) days, and the cost of hospitalization was 1.53 (0.91, 3.02)×10 000 yuan. Prognosis: 9 531 cases (96.1%) were cured or improved, and 92 cases (0.9%) died. Conclusions:The pathogens responsible for CNS infectious diseases are predominantly viruses. Although the incidence is slightly higher during the summer months, the overall seasonal pattern is not particularly pronounced. These infections are more commonly observed in young and middle-aged males and present with a diverse range of clinical manifestations, contributing to a significant disease burden.

Antibody-drug conjugates(ADCs)are a new type of targeting antibodies that conjugate with highly toxic anticancer drugs via chemical linkers to exert high specificity and efficient killing of tumor cells,thereby attracting considerable attention in precise oncology therapy.Cetuximab(Cet)is a typical antibody that offers the benefits of good targeting and safety for individuals with advanced and inoperable cutaneous squamous cell carcinoma(cSCC);however,its anti-tumor activity is limited to a single use.Cisplatin(CisPt)shows good curative effects;however,its adverse effects and non-tumor-targeting ability are major drawbacks.In this study,we designed and developed a new ADC based on a new cytotoxic platinum(Ⅳ)prodrug(C8Pt(Ⅳ))and Cet.The so-called antibody-platinum(Ⅳ)prodrugs conjugates,named Cet-C8Pt(Ⅳ),showed excellent tumor targeting in cSCC.Specifically,it accurately delivered C8Pt(Ⅳ)into tumor cells to exert the combined anti-tumor effect of Cet and CisPt.Herein,metabolomic analysis showed that Cet-C8Pt(Ⅳ)promoted cellular apoptosis and increased DNA damage in cSCC cells by affecting the vitamin B6 metabolic pathway in tumor cells,thereby further enhancing the tumor-killing ability and providing a new strategy for clinical cancer treatment using antibody-platinum(Ⅳ)prodrugs conjugates.

Objective: To investigate the potential association among preoperative breast MRI features, axillary nodal burden (ANB), and disease-free survival (DFS) in patients with early-stage breast cancer. Materials and Methods: We retrospectively reviewed 297 patients with early-stage breast cancer (cT1-2N0M0) who underwent preoperative MRI between December 2016 and December 2018. Based on the number of positive axillary lymph nodes (LNs) determined by postoperative pathology, the patients were divided into high nodal burden (HNB; ≥3 positive LNs) and non-HNB (<3 positive LNs) groups. Univariable and multivariable logistic regression analyses were performed to identify independent risk factors associated with ANB. Predictive efficacy was evaluated using the receiver operating characteristic (ROC) curve and area under the curve (AUC). Univariable and multivariable Cox proportional hazards regression analyses were performed to determine preoperative features associated with DFS. Results: We included 47 and 250 patients in the HNB and non-HNB groups, respectively. Multivariable logistic regression analysis revealed that multifocality/multicentricity (adjusted odds ratio [OR] = 3.905, 95% confidence interval [CI]: 1.685– 9.051, P= 0.001) and peritumoral edema (adjusted OR = 3.734, 95% CI: 1.644–8.479, P = 0.002) were independent risk factors for HNB. Combined peritumoral edema and multifocality/multicentricity achieved an AUC of 0.760 (95% CI: 0.707– 0.807) for predicting HNB, with a sensitivity and specificity of 83.0% and 63.2%, respectively. During the median follow-up period of 45 months (range, 5–61 months), 26 cases (8.75%) of breast cancer recurrence were observed. Multivariable Cox proportional hazards regression analysis indicated that younger age (adjusted hazard ratio [HR] = 3.166, 95% CI: 1.200–8.352, P= 0.021), larger tumor size (adjusted HR = 4.370, 95% CI: 1.671–11.428, P= 0.002), and multifocality/multicentricity (adjusted HR = 5.059, 95% CI: 2.166–11.818, P< 0.001) were independently associated with DFS. Conclusion Preoperative breast MRI features may be associated with ANB and DFS in patients with early-stage breast cancer.