1.Transplantation of bone marrow mesenchymal stem cells overexpressing Shootin1 for treatment of spinal cord injury
Wei ZHANG ; Xiaoqi ZHU ; Decai ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(50):7507-7517
BACKGROUND:Genetic modification by Shootin1 aims to effectively improve neural differentiation of bone marrow mesechymal stem cel s (BMSCs) in the injured spinal cord, thereby promoting functional recovery from spinal cord injury after cel transplantation. OBJECTIVE:To explore the nerve regeneration ability of transplanted BMSCs overexpressing Shootin1 in rats with spinal cord injury. METHODS:BMSCs were transfected using adenovirus-Shootin1 for 48 hours. Then, immunofluorescence staining was used to detect Nestin and NeuN expression levels in the transfected cel s under in vitro neuronal induction and differentiation. Animal models of spinal cord injury were made in rats using modified Al en’s method. Thirty minutes later, Shootin1-transfected BMSCs and non-transfected BMSCs were respectively injected into the subarachnoid space of the rats in the transfection and non-transfection groups, respectively. Rats in the model group were given no treatment. Five weeks after modeling, spinal cord samples were taken from each rat to make frozen sections for detection of nerve related markers RESULTS AND CONCLUSION:After 48-hour adenoviral transfection, Shootin1 expression was successful y detected in BMSCs. After 7-day in vitro induction, the cel morphology in the three groups varied, and there was no significant difference in the expression of Nestin and NeuN between the transfection and non-transfection groups. Basso, Beattie and Bresnahan scores were higher in the two cel transplantation groups than the model group. Increased expression levels of Nestin, NeuN, GFAP, MAP-2, ChAT and SYN were observed in both two cel transplantation groups, indicating a strengthened ability of nerve regeneration. Our experimental findings further confirm that BMSCs transplantation for spinal cord injury has achieved good outcomes, and Shootin1 protein plays a certain role in nerve regeneration and functional recovery after spinal cord injury. However, Shootin1 overexpression shows no obvious additional effects in combination with BMSCs transplantation, and further studies on the optimization of BMSCs transplantation for spinal cord injury are necessary.
2.Study of the association between difference in cellular immunity and liver function of hepatitis B virus genotype B,C and interleukin-7 inducing follicular helper T lymphocytes
Dong WANG ; Zhonghua LU ; Qin TANG ; Zheng WANG ; Hao PEI ; Yinfang ZHU ; Decai FU ; Xibing GU
Chinese Journal of Infectious Diseases 2015;(9):522-526
Objective To investigate the association between the difference of specific cytotoxic lymphocyte (CTL) and liver function of patients with hepatitis B virus (HBV) genotype B and C infections and interleukin (IL)‐7 induced follicular helper T lymphocytes (Tfh) .Methods Sixty‐seven patients with chronic hepatitis B (CHB) hospitalized at Wuxi No .5 People′s Hospital from August 2013 to January 2015 were collected and 30 healthy blood donors were set as healthy control group .The peripheral blood IL‐7 , Tfh ,IL‐21 ,HBV specific‐CTL ,nonspecific CTL ,levels of HBV DNA ,alanine aminotransferase (ALT) and total bilirubin (TBil) were compared between patients with genotype B and C infection ,hepatitis B e antigen (HBeAg)‐positive and HBeAg‐negative CHB ,high ALT level and low ALT level .Multivariate regression analysis was performed to determine the factors associated with IL‐7 .The t test was used for quantitative data and chi‐square test was used for categorical data .Results Of the 67 CHB patients with average age of (35 .1 ± 11 .4) ,48 were male and 19 were female;32 were infected with genotype C and 35 were infected with genotype B ;40 were HBeAg‐positive CHB and 27 were HBeAg‐negative CHB ;17 were with high ALT levels and 50 were with low ALT levels .IL‐7 ,Tfh ,IL‐21 and HBV specific‐CTL levels in the peripheral blood of genotype C‐infected patients were (20 .79 ± 4 .82 ) ng/L , (3 .93 ± 0 .82)% ,(24 .77 ± 7 .52) ng/L and (0 .20 ± 0 .04)% ,respectively ,while in genotype B‐infected patients , those were (29 .13 ± 8 .17) ng/L ,(5 .92 ± 1 .92)% ,(39 .94 ± 24 .00) ng/L and (0 .40 ± 0 .06)% , respectively .Levels of IL‐7 , Tfh ,IL‐21 and HBV specific‐CTL in genotype C‐infected patients were significantly lower than those in genotype B‐infected patients (t= 5 .027 ,5 .595 ,3 .553 and 15 .133 , respectively ;all P<0 .01) .Nonspecific CTL ,HBV DNA ,ALT and TBil levels in the peripheral blood of genotype C‐infected patients were all significantly higher than those in genotype B infected‐patients (t=4 .899 ,6 .815 ,2 .763 and 4 .899 ,respectively ;all P< 0 .01) .IL‐7 ,Tfh ,IL‐21 ,HBV specific‐CTL levels in the peripheral blood of HBeAg‐positive patients were significantly lower than those in HBeAg‐negative patients (all P<0 .01) .Nonspecific CTL ,HBV DNA ,ALT and TBil levels in the peripheral blood of HBeAg‐positive patients were all significantly higher than those in HBeAg‐negative patients (all P<0 .05) .IL‐7 ,Tfh ,IL‐21 ,HBV specific‐CTL levels in the peripheral blood of patients with high ALT levels were all significantly lower than those in patients with low ALT levels (all P<0 .01) .Nonspecific CTL and HBV DNA levels in the peripheral blood of patients with high ALT levels were both significantly higher than those in patients with low ALT levels (both P<0 .01) .HBV DNA ,IL‐21 and nonspecific CTL were all correlated with IL‐7 (all P<0 .01) .Conclusion The differences of HBV specific‐CTL and liver function in CHB patients infected with genotype B and C may be correlated with interleukin‐7 induced Tfhcells.
3.Magnetic resonance tracking of endothelial progenitor cells labeled with superparamagnetic iron ox-ide homing to the site of hepatoma
Xiaoli MAI ; Haijian FAN ; Dan MU ; Decai YU ; Jun YANG ; Bin ZHU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2016;(1):19-24
Objective To track the migration and incorporation of intravenously injected, magneti?cally labeled endothelial progenitor cells ( EPCs) from mouse bone marrow into the blood vessels in a rapid?ly growing HCC model by microMR (7.0 T). Methods This study was approved by the Institutional Com?mittee on Animal Research. H22 hepatic ascitic cancer cells was directly injected into the left liver lobe of BALB/c nude mice ( n=15) . EPCs derived from bone marrow of C57BL/6 mice were isolated and cultured. The third passage EPCs were collected and labeled with 25 μg/ml superparamagnetic iron oxide ( SPIO) and poly?l?lysine (PLL) complex (SPIO?PLL). MTT assay and flow cytometry were used to evaluate the difference of growth curve and apoptosis between labeled and unlabeled EPCs. EPCs labeled with SPIO?PLL were injected into mice via tail vein in experiment group (on the 3rd day after establishing HCC model) (n=15) and control group (n=6). The signal changes of tumor (the 1st, 3rd and 7th day after transplantation) were observed by microMR. Prussian blue staining and immunohistochemistry staining of CD31 were per?formed. MRI findings were confirmed by histomorphology. Two?sample t test was used to analyze the data. Results Single tumor was showed in the liver of all mice 3 d after establishing models. Labeling with SPIO?PLL at a concentration of 25μg/ml did not alter cell growth curve ( measured by MTT assay;t=0.281, P>0.05) and cell apoptosis (analyzed by flow cytometry). The apoptosis rates of SPIO?PLL labeled and un?labled EPCs were (12.31±1.43)% and (11.57±1.24)% in early stage, and (0.55±0.07)% and (0.49± 0?05)% in late stage. No significant differences were observed between them (t=0.967, 1.060; both P>0?05) . Migration and incorporation of transplanted and labeled cells into tumor were documented with in vivo microMR as low signal intensity at the tumor periphery as early as the 3rd day after EPCs administration in preformed tumors (4/5). Prussian blue staining showed iron?positive cells at the sites corresponding to low signal intensity on MRI. The positive cells expressing CD31 existed in intratumoral and peritumoral vessels. There was no signal change in control group at all time points. Conclusions MRI can demonstrate the in?corporation of magnetic labeled mouse EPCs into the implanted hepatoma. It may be helpful for early diagno?sis and therapy of liver tumor.
5.Short-term prognostic value of soluble Axl receptor tyrosine kinase for elderly patients with acute decompensated heart failure
Tongliang MA ; Ruodong HAN ; Yu HUANG ; Decai ZHU
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(3):262-266
Objective To investigate the clinical value of serum soluble Axl receptor tyrosine ki-nase(sAxlTK)in evaluating short-term prognosis in patients with acute decompensated heart failure(ADHF).Methods A total of 238 elderly ADHF patients admitted to Bozhou People's Hospital from January 1,2018 to October 1,2021 were recruited and divided into poor prognosis group(45 patients)and good prognosis group(193 patients)according to the occurrence of com-plex events within 90 d of follow-up.Based on the optimal cut-off value of serum sAxlTK level,they were also assigned into high level group(80 cases)and low level group(158 cases).Serum levels of sAxlTK,troponin Ⅰ(cTnⅠ)and N-terminal B-type natriuretic peptide precursor(NT-proBNP)were detected.Results Serum sAxlTK level was significantly higher in the poor prognosis group than the good prognosis group[43.89(33.95,51.44)μg/L vs 23.89(18.73,33.92)μg/L,P<0.01].Multivariate logistic regression analysis showed that serum cTnⅠ and sAxlTK levels were independent risk factors for short-term poor prognosis in ADHF patients(OR=1.922,95%CI:1.035-3.568,P=0.039;OR=1.021,95%CI:1.008-1.034,P=0.001).ROC curve analysis indicated that the AUC value of combined serum sAxlTK,cTnⅠ and NT-proBNP levels to predict short-term poor prognosis was 0.836(95%CI:0.778-0.895).The incidence of complex events within 90 d was significantly higher in the high level group than the low level group(45.0%vs 5.7%,P<0.05).Kaplan-Meier curve analysis revealed that the cumulative inci-dence of complex events was also higher in the high level group than the low level group(X2=66.991,Plog rank<0.01).The high level group had significantly lower overall survival rate and worse survival prognosis than the low level group(X2=16.899,Plog rank<0.01).Conclusion High serum sAxlTK level in elderly ADHF patients at admission is associated with a higher risk of 90-day short-term poor prognosis.Serum sAxlTK has the potential to become a useful tool for early prediction of short-term poor prognosis,and its combination with cTnⅠ and NT-proBNP can fur-ther improve the accuracy of prognosis prediction.
6. The clinical application value of ultrasound-guided percutaneous lung biopsy in the diagnosis of peripheral lung lesions of silicosis
Decai ZENG ; Ji WU ; Linping ZHU ; Hui CHEN ; Ting ZHANG ; Ying TAN ; Xueyu CHE
Chinese Journal of Ultrasonography 2018;27(6):524-528
Objective:
To determine the clinical application value of percutaneous lung biopsy guided by ultrasound in the diagnosis of peripheral lung lesions of silicosis.
Methods:
Experimental silicosis was produced in rabbits by the intratracheal administration of silica with non-exposure method. Imaging changes were observed in 36 rabbits on 60 days after intratracheal instillation of silica. To contrast with CT results, percutaneous lung biopsy of peripheral lesions was guided by ultrasound. The success rate of sufficient material, the diagnosis rate of coincidence between biopsy and pathology, and the incidence of complications were calculated. The biopsy with sufficient material, biopsy findings coincided with pathological results and no complications were defined as strictly success of the puncture. The baseline data and monitoring index were compared between successful biopsy group and unsuccessful biopsy group. Each rabbit was intravenously administrated by 10 000 U of heparin for the antiocoagulation and sacrificed by fast injection of 10% KCl through jugular vein catheterization. Specimens from lung tissue were collected and stained with hematoxylin-eosin. Pathological changes of lung tissue were observed through an optical microscope.
Results:
Of 36 silicosis rabbits, peripheral lung lesions of silicosis were observed in 30 rabbits. Biopsy procedures were performed with ultrasound guidance in 30 rabbits. The total success rate of biopsy was 70% (21/30). The success rate of sufficient material was 93% (28/30), the diagnosis rate of coincidence between biopsy and pathology 86%(24/28), and the incidence of complications was 10% (3/30) respectively. Compared with failure group, peripheral lesions in successful biopsy group were bigger in size, closer to the chest wall, and lower respiratory rate, the difference was statistically significant (
7.Clinical value of split domino donor auxiliary liver transplantation
Wenjie ZHANG ; Qingxiang XU ; Guoqiang LI ; Decai YU ; Yang YUE ; Xinhua ZHU ; Qiaoyu LIU ; Heng CUI ; Beicheng SUN
Chinese Journal of Digestive Surgery 2022;21(2):287-294
Objective:To investigate the clinical value of split domino donor auxiliary liver transplantation.Methods:The retrospective and descriptive study was conducted. The clinco-pathological data of 3 liver transplantation recipients who were admitted to Nanjing Drum Tower Hospital affiliated to Nanjing University Medical School and 1 liver transplantation recipient who was admitted to external hospital in September 2018 were collected. The first case was male, aged 22 years, who was diagnosed as type II citrullinemia (CTLN2). The second case undergoing liver transplantation in external hospital was male, aged 59 years, who was diagnosed as decompensated alcoholic cirrhosis. The third case was female, aged 52 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The fourth case was female, aged 51 years, who was diagnosed as hepatocellular carcinoma of right lobe of liver. The donor liver from a brain and cardiac death donor was split in vitro into the left liver and the right liver, in which the right liver without middle hepatic vein, and the modified piggyback liver transplantation using the left liver and the classical orthotropic liver transplantation using the right liver was conducted on the first and the second case, respectively. The original liver of the first case was split in vivo into the left liver and the right liver, and the piggyback auxiliary liver transplantation using the left liver and the piggyback auxiliary liver transplantation using the right liver was conducted on the third and the fourth case who underwent extended right hemihepatectomy, respectively. Observation indicators: (1) intraoperative situations; (2) follow-up. Follow-up was conducted using outpatient examination and telephone interview to detect liver function, liver imaging, complication and survival of recipients up to October 2021.Results:(1) Intraoperative situations. Liver transplantation was conducted successfully on the first, third and fourth case, with the operation time, the volume of intraoperative blood loss, the donor liver cold ischemia time, the graft-to-recipient weight ratio were 400 minutes, 370 minutes, 390 minutes, 600 mL, 1 300 mL, 1 600 mL, 230 minutes, 152 minutes, 135 minutes, 1.2%, 0.8%, 1.1%. (2) Follow-up. B-ultrasound examination of the first, third and fourth case after liver transplantation showed that the blood flow was normal, and all the 3 cases discharged and were followed up at postoperative 1, 6 and 12 month. The liver function, the level of blood ammonia and citrulline were normal of the first, third and fourth case at postoperative 1 week. Imaging examina-tion showed normal liver morphology of the first and third case, and a transplanted liver atrophy caused by portal vein steal of the fourth case. ① The level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), direct bilirubin (DBil) of the first case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 22.8 U/L, 404.1 U/L, 355.5 U/L, 289.6 U/L, 31.0 U/L, 23.1 U/L, 42.1 U/L and 25.8 U/L, 31.5 U/L, 517.7 U/L, 327.6 U/L, 172.9 U/L, 15.9 U/L, 21.4 U/L, 47.5 U/L and 29.7 U/L, 3.8 μmol/L, 92.1 μmol/L, 87.4 μmol/L, 79.7 μmol/L, 90.1 μmol/L, 130.6 μmol/L, 33.8 μmol/L and 25.4 μmol/L, 2.3 μmol/L, 47.0 μmol/L, 44.1 μmol/L, 47.1 μmol/L, 57.4 μmol/L, 70.9 μmol/L, 24.7 μmol/L and 9.7 μmol/L, respectively. The level of citrulline and blood ammonia of the first case before and after liver transplantation were 999.0 μmol/L, 196.0 μmol/L and 14.6 μmol/L, 9.0 μmol/L, respectively. The first case was followed up for 3 years and survived without any liver transplantation related complication. ② The level of ALT, AST, TBil, DBil of the third case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 21.3 U/L, 143.9 U/L, 182.0 U/L, 132.0 U/L, 17.2 U/L, 10.1 U/L, 17.6 U/L and 16.8 U/L,20.0 U/L, 291.0 U/L, 227.5 U/L, 106.4 U/L, 15.8 U/L, 10.8 U/L, 17.1 U/L and 19.4 U/L, 6.8 μmol/L, 50.9 μmol/L, 45.0 μmol/L, 34.0 μmol/L, 32.4 μmol/L, 22.3 μmol/L, 12.8 μmol/L and 14.9 μmol/L, 2.5 μmol/L, 18.4 μmol/L, 17.2 μmol/L, 14.9 μmol/L, 14.8 μmol/L, 12.1 μmol/L, 3.6 μmol/L and 4.4 μmol/L. The level of citrulline and blood ammonia of the third case after liver transplantation were 24.9 μmol/L and 16.0 μmol/L. The third case was followed up for 3 years and survived without any liver transplantation related complication. ③ The level of ALT, AST, TBil, DBil of the fourth case before liver transplantation, at postoperative 1 day, 2 day, 3 day, 7 day, 10 day, 6 month and 1 year were 35.0 U/L, 268.7 U/L, 682.0 U/L, 425.8 U/L, 57.5 U/L, 34.0 U/L, 29.4 U/L and 18.1 U/L, 37.0 U/L, 419.1 U/L, 436.2 U/L, 139.5 U/L, 35.2 U/L, 32.4 U/L, 54.7 U/L and 32.8 U/L, 7.1 μmol/L, 64.2 μmol/L, 41.4 μmol/L, 17.6 μmol/L, 34.2 μmol/L, 48.7 μmol/L, 14.1 μmol/L and 21.8 μmol/L, 2.8 μmol/L, 18.9 μmol/L, 16.1 μmol/L, 6.0 μmol/L, 14.6 μmol/L, 26.7 μmol/L, 3.9 μmol/L, 11.8 μmol/L. The level of citrulline and blood ammonia of the fourth case after liver transplantation were 8.4 μmol/L and 47.0 μmol/L. One week after surgery, the transplanted right liver of the fourth case occurred atrophy due to blood stealing from the right branch of the portal vein. B-ultrasound examination showed that the reflux of the hepatic artery and hepatic vein was unobstructed. Immunosuppressants were discontinued 3 months after operation on the fourth case and there was no complication such as rejection, bile leakage, biliary stricture, thrombosis and vascular stricture during follow-up. The fourth case died of lung metastasis 19 months after operation.Conclusion:Split domino donor auxiliary liver transplantation can be used for the treatment of metabolic liver disease and advanced hepatocellular carcinoma.
8.Prognostic value of procalcitonin and C-reactive protein combined with sequential organ failure assessment score in elderly patients with sepsis induced by pulmonary infection
Xiaolan QIN ; Quanlai GUO ; Yuntao LIU ; Decai ZHU ; Jun LI ; Danwen ZHENG ; Junsheng TONG
Chinese Critical Care Medicine 2019;31(5):562-565
Objective To investigate the prognostic value of procalcitonin (PCT) and C-reactive protein (CRP) combined with sequential organ failure assessment (SOFA) score in elderly patients with sepsis induced by pulmonary infection. Methods A retrospective study was conducted. The elderly patients aged over 60 years old with sepsis induced by pulmonary infection admitted to Guangdong Provincial Hospital of Chinese Medicine from October 2015 to December 2018 were enrolled. The PCT, CRP and SOFA scores of patients within 24 hours after admission and 28-day prognosis were recorded. The patients were divided into groups according to the severity of the disease and 28-day prognosis. The differences in above parameters among all the groups were compared. Receiver operator characteristic (ROC) curve was drawn to analyze the prognostic value of the above indicators in elderly patients with sepsis induced by pulmonary infection alone or in combination. Results A total of 265 patients were enrolled in the study. According to the severity of the disease, the patients were divided into sepsis group (n = 194) and septic shock group (n = 71). According to the 28-day prognosis, the patients were divided into survival group (n = 186) and non-survival group (n = 79). Compared with the sepsis group, the PCT, CRP and SOFA scores of patients in the septic shock group were significantly increased [PCT (μg/L): 6.16 (1.94, 19.60) vs. 1.56 (0.34, 7.32), CRP (mg/L): 128.90 (54.93, 198.70) vs. 91.45 (30.15, 175.30), SOFA score: 9.0 (7.0, 12.0) vs. 4.0 (3.0, 5.0)] with significant differences (all P < 0.05). Compared with the survival group, the PCT, CRP and SOFA scores of sepsis patients in the non-survival group were significantly increased [PCT (μg/L): 4.80 (1.06, 19.60) vs. 1.82 (0.34, 7.24), CRP (mg/L): 135.20 (58.10, 225.50) vs. 91.45 (31.50, 172.53), SOFA score: 7.0 (4.0, 11.0) vs. 4.0 (3.0, 6.0)] with significant differences (all P < 0.01). ROC curve analysis showed that the area under the ROC curve (AUC) of PCT, CRP, SOFA score and CRP+PCT+SOFA score was 0.641, 0.607, 0.697, and 0.712, indicating that above parameters had certain predictive value for 28-day prognosis of elderly patients with sepsis induced by pulmonary infection, and the combined predictive value of them was the greatest with the sensitivity of 55.1% and the specificity of 80.1%. Conclusion PCT, CRP and SOFA score are commonly used to evaluate the prognosis of the elderly patients with sepsis induced by pulmonary infection, and the combination of them has higher evaluation value.
9.Quantitative magnetic susceptibility imaging sequence for intracranial inflammation in patients with optic neuromyelitis
Xinli WANG ; Ning FENG ; Ningning WANG ; Zhizheng ZHUO ; Haoxiao CHANG ; Ai GUO ; Decai TIAN ; Xiaodong ZHU
Chinese Journal of Postgraduates of Medicine 2023;46(8):679-683
Objective:To identify the potential intracranial inflammation in neuromyelitis optica spectrum disorders(NMOSD) patients without supratentorial MRI lesions using quantitative susceptibility mapping (QSM).Methods:Seventy NMOSD patients and 35 age- and gender-matched healthy controls (NC) underwent QSM, 3D-T 1, diffusion MRI from Beijing Tiantan Hospital during June 2019 to June 2021. Susceptibility was compared among NMOSD patients with acute attack (ANMOSD), NMOSD patients in chronic phase (CNMOSD) and NC. The correlation between susceptibility in several brain regions and the cerebrospinal fluid levels of inflammatory makers were analyzed. Results:NMOSD patients showed different susceptibility in several brain regions including bilateral hippocampus, precuneus, right cuneus, putamen, superior parietal and inferior temporal ( P<0.001) and the posr-hoc showed it is higher than normal. Compared to CNMOSD patients, the ANMOSD patients showed increased susceptibility in the cuneus (0.009 ± 0.004 vs. 0.005 ± 0.004, P<0.05). There was significant positive correlations between susceptibility and CSF levels of sTREM2 which reflect the active of microglial cells ( r = 0.494, P<0.05). Conclusions:Despite the absence of supratentorial lesions on MRI, increased susceptibility suggests underlying inflammation in the cerebral cortex in both patients with ANMOSD and CNMOSD, and some of them are obviously related to inflammatory markers in CSF. QSM sequence can be used to explore the potential inflammation in NMOSD patients without obvious supratentorial lesions.
10.Exploring the Mechanism of Action of Qizhu Kang'ai Formula (芪术抗癌方) for the Treatment of Colorectal Cancer based on Network Pharmacology and Experimental Validation
Ruolan SUN ; Yan LIANG ; Fan ZHAO ; Shijiao ZHU ; Linlu WAN ; Xu WANG ; Qihang YIN ; Gang YIN ; Decai TANG
Journal of Traditional Chinese Medicine 2024;65(4):404-413
ObjectiveTo explore the potential molecular mechanism of Qizhu Kang'ai Formula (芪术抗癌方, QZKAF) for the treatment of colorectal cancer (CRC). MethodsNetwork pharmacology was used to analyze the active ingredients and targets of QZKAF for CRC, and analyze the key targets of QZKAF for the treatment of CRC by gene function annotation (GO) and Kyoto Encyclopedia of Genomes (KEGG) pathway enrichment analysis. Molecular docking was applied to predict the binding activity of the core active ingredients to the key targets. A orthotopic transplantation tumor mice model of CRC was established to validate the key targets of QZKAF for CRC obtained from network pharmacology analysis. Forty-eight mice were randomly divided into the sham operation group, the model group, the 5-fluorouracil (5-Fu) group, and the QZKAF low-, medium-, and high-dose groups, with 8 mice in each group. Except for the sham operation group, the remaining groups underwent colon cancer orthotopic transplantation tumor modeling. The 5-Fu group was given 30 mg/kg of 5-Fu by intraperitoneal injection once every 3 days on the alternate day after modeling, while the QZKAF low-, medium-, and high-dose groups were given 2.925, 5.85, and 11.7 g/(kg·d) of QZKAF by gastric gavage, respectively, and the sham-operation group and the model group were gavaged with 0.1 ml/10 g of normal saline every day, all for 21 days. The in situ tumors mass and the number of liver metastases were compared between the groups. The pathological changes of colon tumor tissues were observed by HE staining, and the protein expression of protein tyrosine phosphatase nonreceptor type 1 (PTPN1), vinculin, integrin subunit αν, integrin subunit β3, and E-cadherin were detected in colon tumor tissues by Western blot. ResultsNetwork pharmacology screening yielded that the top six core active ingredients of QZKAF intervening in CRC were quercetin, kaempferol, apigenin, luteolin, baicalein and ursolic acid. There were 212 targets of action, and the ranked top three were prostaglandin endoperoxide synthase 1 (PTGS1), prostaglandin endoperoxide synthase 2 (PTGS2), and PTPN1, which may be the key targets of QZKAF in the treatment of CRC. These key targets were significantly enriched mainly in phosphatidylinositol 3-kinase/protein kinase B (PI3K-Akt) signaling pathway, focal adhesion and adhesion junction. Molecular docking results: except for PTGS1 with better binding activity to quercetin, kaempferol, and apigenin (binding energy ≥