Objective To construct an in vitro co-culture model of gastric cancer organoids and cancer-associated fibroblasts(CAFs),and investigate the role of cancer-associated fibroblasts in the proliferation and chemotherapy resistance of gastric cancer organoids.Methods Tumor tissues from 12 gastric cancer patients undergoing surgical treatment in Department of General Surgery of Second Affiliated Hospital of Army Medical University from February 2023 to March 2024 were collected to construct gastric cancer organoids using 3D culture.HE staining was used to observe the morphology,and immunohistochemical assay was employed to determine the expression of cytokeratin CK7,carcinoembryonic antigen(CEA),and proliferation marker Ki-67.After CAFs derived from the same patient were cultured,observed for their morphology under a light microscope,and detected for the phenotype by flow cytometry,the cells were co-cultured with gastric cancer organoids in a 1:1 ratio.Phase-contrast microscopy was applied to observe the growth of the organoids and analyze the number,average diameter,and total area.Then,organoids cultured alone served as the control group.After the control and co-culture groups were treated with chemotherapy drugs,5-fluorouracil and oxaliplatin,for 48 h,the viability and apoptosis of organoids were assessed with CellTiter-Glo??3D assay and CellEvent? Caspase 3/7 activity,respectively.Results Gastric cancer organoids and CAFs were successfully established from 10 gastric cancer patient-derived samples.The gastric cancer organoids exhibited morphological characteristics consistent with the corresponding primary tumors,and showed positive expression of CK7,CEA,and Ki-67.CAFs displayed typical spindle-shaped morphology and exhibited the phenotypic markers CD326-,CD45-,CD31-,α-SMA+,CD73+,CD90+,and CD105+.Compared to the organoids cultured alone,the organoids co-cultured with CAFs showed more formation of organoids,in larger average diameter,and taking larger total area(P<0.05).After the treatment of 5-fluorouracil and oxaliplatin,the half-maximal inhibitory concentration(IC50)was 10.66 and 3.26 μmol/L,respectively in the control group,while was 46.23 and 91.11 μmol/L in the co-culture group.Additionally,the number of CellEvent? Caspase 3/7 positive apoptotic cells was significantly less in the co-culture group than the control group.Conclusion Compared with individually cultured gastric cancer organoids,the co-culture model of gastric cancer organoids and CAFs better simulates the pro-tumor proliferation and drug resistance effects of in vivo tumor microenvironment.

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.

OBJECTIVETo investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells.

METHODSThe multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR.

RESULTSThe drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells.

CONCLUSIONERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Phosphorylation ; Up-Regulation

BACKGROUNDThe occurrence of contrast induced acute kidney injury (CIAKI) has a pronounced impact on morbidity and mortality. The aim of the present study was to appraise the diagnostic efficacy of age, estimated glomerular filtration rate (eGFR) and ejection fraction (AGEF) score (age/EF(%)+1 (if eGFR was <60 ml × min(-1)× 1.73 m(-2))) as an predictor of CIAKI in patients with diabetes mellitus (DM) and concomitant chronic kidney disease (CKD).

METHODSThe AGEF score was calculated for 2 998 patients with type 2 DM and concomitant CKD who had undergone coronary/peripheral arterial angiography. CIAKI was defined as an increase in sCr concentration of 0.5 mg/dl (44.2 mmol/L) or 25% above baseline at 72 hours after exposure to the contrast medium. Post hoc analysis was performed by stratifying the rate of CIAKI according to AGEF score tertiles. The diagnostic efficacy of the AGEF score for predicting CIAKI was evaluated with receiver operating characteristic (ROC) analysis.

RESULTSThe AGEF score ranged from 0.49 to 3.09. The AGEF score tertiles were defined as follows: AGEFlow ≤ 0.92 (n = 1 006); 0.92 1.16 (n = 992). The incidence of CIAKI was significantly different in patients with low, middle and high AGEF scores (AGEFlow = 1.1%, AGEFmid = 2.3% and AGEFhigh = 5.8%, P < 0.001). By multivariate analysis, AGEF score was an independent predictor of CIAKI (odds ratio = 4.96, 95% CI: 2.32-10.58, P < 0.01). ROC analysis showed that the area under the curve was 0.70 (95% CI: 0.648-0.753, P < 0.001).

CONCLUSIONThe AGEF score is effective for stratifying risk of CIAKI in patients with DM and CKD undergoing coronary/peripheral arterial angiography. (Clinical Trial identifier: NCT00786136).

Acute Kidney Injury ; physiopathology ; Contrast Media ; Female ; Glomerular Filtration Rate ; physiology ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Renal Insufficiency, Chronic ; physiopathology

Objective To investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells. Methods The multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR. Results The drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells. Conclusion ERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

Objective To investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells. Methods The multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR. Results The drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells. Conclusion ERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

ObjectiveTo explore the imaging of the thrombosis after pharmacological triggering of plaque rupture in atherosclerotic rabbit model by using 3.0 T high-resolution magnetic resonance imaging.MethodsTwenty male New Zealand white rabbits were divided into an experimental group (n = 16) and a control group (n = 4).The aortic wall injuries were induced by an intravascular balloon in experimental group rabbits after high cholesterol diet.The pharmacological triggering with Russell's viper venom and histamine was performed after 3 months of establishment of model.All of the animals underwent pre-trigger and post-trigger MR examinations including 3D time of fight (3D TOF),T1 WI,T2WI and post contrast T1 WI.Euthanasia was performed in all rabbits and gross anatomy and histological specimen of aorta were obtained.Comparing the location and length of the thrombus between MRI images and histopathology was used Pearson test.Comparing the calculated indexes of abdominal aorta between rabbits with and without thrombosis was used AVONA test and LSD-t test.Results After triggering,8 in 14 survived rabbits developed thrombosis in experimental group,meanwhile,no thrombus was found in control group.The accuracy of multi-sequences MRI for detecting of thrombus was 87.1％ (27/31).MRI data correlated with the histopathology regarding thrombus length ( r = 0.85,P ＜ 0.01 ) and thrombus location ( r = 0.94,P＜0.01 ).Compared with rabbits without thrombosis,the rabbits with thrombosis had narrower lumen of abdominal aorta in the pre-triggered MR images [ ( 5.71 ± 2.38 )mm2 vs.( 8.93 ± 5.36) mm2,P ＜ 0.01 ].ConclusionMRI is useful tool to determine the thrombosis and plaque rupture in atherosclerotic rabbit model.

OBJECTIVE: To explore and summarize the experience of surgical treatment for primary retroperitoneal tumor (PRT)in children. METHODS: Clinical data of 17 patients with PRT treated from January 2001 to January 2008 were retrospectively analyzed, including image examination, pathologic examination and surgical procedure. RESULTS: Seventeen patients underwent complete resection; 8 benign PRT, and 9 malignant PRT were diagnosed by operation and postoperative pathologic examination. Vascular surgery was done on 11 patients, 6 cases of multi-visceral resection, 1 vascular transplant, and 1 multi-visceral resection. Two patients had recurrent malignant PRT. CONCLUSION For pediatric complex retroperitoneal tumors, complete resection can reduce the recurrence and improve the long-term survival.
Child ; Child, Preschool ; Female ; Germinoma ; surgery ; Humans ; Lymphoma ; surgery ; Male ; Retroperitoneal Neoplasms ; surgery ; Retrospective Studies ; Teratoma ; surgery

Objective To investigate immunoprophylactic potential of genetic engineering vaccines of enterohaemorrhagie Escherichia coli O157:H7 in BALB/c mice after immunization with these vaccines. Methods Sixty BALB/c mice (3 weeks old) were randomized averagely into 5 groups. Group 1-3 were im-munized respectively with IntiminC300, Stx2B and HIyAN436, group 4 with a combination of these three vaccines, and group 5 with PBS. Each mouse was immunized with vaccine(100 μg)and Al(OH)3 adjuvant (100 μg) for 3 times. After 7 d of the second and third immunization, serum of each mouse was collected and the different antibodies were detected. After 10 d of the last immunization, all mice were given drinking water containing streptomycin for 3 d before and following oral challenge with O157:H7 (109 CFU), and treated with clinical, microbiological and pathological examination. Results The three vaccines elicited high titer antiserum, and some mice were died after infection with O157. The livability of group 1-4 was re-spectively 73%, 64%, 36% and 91%. And these vaccines depressed fecal and colon shedding with O157. Condusion IntiminC300, Stx2B and HIyAN436 have certain protective efficacy for infection of O157, and combined immunization was more effective than single vaccine.