Objective:To investigate the existence of side population cells with the potency of stem cells in human gallbladder carcinoma cell line GBC-SD and the differences in ABCG2,Oct-4 and CD34 expression among SP cells,non-SP cells and GBC-SD cells.Methods:SP and non-SP cells were sorted from GBC-SD cells by fluorescence-activated cell sorting(FACS).The expression of ABCG2,Oct-4 and CD34 in SP cells,non-SP cells,and GBC-SD cells was detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blot,flow cytometry(FCM)and immunofluorescence chemistry.Results:SP cells with stem cell potency were isolated from GBC-SD cells with a proportion of(0.64±0.08)%.The metastatic ability of SP cells was obviously higher than that of non-SP cells and GBC-SD cells(P<0.05).The expression of ABCG2 was significantly higher in SP cells than in non-SP cells and GBC-SD cells[(89.56±3.86)%vs.(1.32±0.49)%and(12.37±1.61)%,P=0.001].The expression of Oct-4 in these cells was(94.87±1.40)%,(88.16±2.34)%and(90.17±1.61)%,respectively(P>0.05).CD34 was neady absent in these cells on protein level[(1.78±0.51)%vs.(0.63±0.21)%and(0.96±0.381)%,P>0.05)],but it was highly expressed in non-SP cells and GBC-SD cells and absent in SP cells off mRNA leve;.Conclusion:SP calls which hava the potency of stem cells,exist in human gallbladder carcinoma GBC-SD cell line and have the phenotype of ABCG2+Oct-4+CD34-.

Objective To apply Windows presentation foundation (WPF) to rapid access and report printing of medical data.Methods Data access was realized by abstracting database to form data structure through entity framework (EF) and mapping datasheet as attribute.Multipage printing was executed by using Flow Document Scroll Viewer as the container and putting contents into different paragraphs with the block element.Results EF of WPF decreased the frequency of database access by the program greatly,and the reaction speed of the program was increased significantly.Flow document could be used for data printing.Conclusion WPF enhances the efficiency of system development and the beauty of man-machine interface,and thus is worthy promoting for medical system and software development.

Objective To evaluate the safety and efficacy of multi-band mucosectomy (MBM) for early esophageal cancer and precancerous lesions.Methods Data of 28 patients with early esophageal cancer or precancerous lesions undergoing MBM were reviewed in regarding of procedure complications and follow-up results.Results A total of 32 lesions were resected successfully by MBM in one session,with mean procedure time of 28.3 minutes.The mean diameter of specimens was 12mm.No residual neoplasm was found at the base of any resected specimens.The post-MBM pathological findings consisted of 2 cases of intramucosal cancer,1 case of submucosal cancer,and 25 cases of moderate-severe dysplasia.No perforation,delayed hemorrhage or subcutaneous emphysema occurred.Intraoperative bleeding occurred in 23 cases,including 3 cases of pulsatile bleeding,which were controlled with metal clip,and 20 cases of minor bleeding which were managed with APC or halted automatically at the end of procedure.Chest pain after the procedure occurred in 5 cases and were relieved soon.The patient with submucosal cancer underwent subsequent surgical resection,with no residual cancer in surgical specimen or lymph node metastasis.Twenty seven other cases were followed up endoscopically for 2-12 months.Esophageal stricture occurred in 2 cases,and were successfully relieved by dilatation with stent or bougienage.No recurrent lesion or metastasis were revealed.Conclusion MBM is a relatively safe and effective endoscopic technique for treatment of early esophageal intramucosal cancer and precancerous lesions,but further studies are needed to evaluate the long-term results.

Objective:To study performed of develop a mice model of allergic diseases induced by crude extractings from populus pollen.Methods: A total of 60 BALB/c mice were divided randomly into there groups:normal control group,Albumin Egg(OVA) group and populus pollen model group with 20 in each.Mices were repeatedly sensitized by intraperitoneal injections of OVA or crude populus pollen extract every 5 d for four doses.Five days after the last sensitization,mices were repeatedly challenged by once daily antigen from 21-25 d.The changes of inflammatory cells in bronchoalveolar lavage fluid(BALF) were stained with hematoxylin and eosin(HE) to evaluate the degree of allergic inflammation.PAS staining was used to observe the secretion of airway mucus;The changes of the nasal mucosas and lungs of mice were stained with HE to evaluate the degree of allergic inflammation.And the average optical density of IL-4 and IFN-γ positive cells in lung tissue was measured by immunohistochemistry.The total IgE in the serum was also measured by enzyme-linked immunoassay(ELISA).Results: Compared with the mice in normal control group,those in OVA group and model group developed obvious allergic inflammation in the nasal mucosas and lungs,and increased airway mucus secretion.The number of inflammatory cells including eosinophil and neutrophils markedly increased in BALF smear.The average optical density of IL-4 positive cells in lung tissue was all increased in OVA group and model group compared with those in normal control group,and the average optical density of IFN-γ in lung tissue was on the contrary.The total IgE in the serum were all increased in OVA group and model group compared with those in normal control group,and the IFN-γ in the serum was significantly reduced in OVA group and model group compared with those in normal control group.Conclusion: Taken together,crude populus pollen extract can successfully induce a mice model of allergic diseases.This model is a useful tool in studying the mechanisms of allergic disease.

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.

Objective To study the difference of expression of proteins in pancreatic cancer and adjacent fissue.Methods Proteomes of eight pairs of pancreatic ductal adenocarcinoma tissue samples and adjacent tissue samples were obtained by high resolution two-dimensional gel electrophoresis(2-DE).Comprehensive analyses of proteins were focused on total protein spots exhibiting statistical alternations between the two groups.Protein identification was done by peptide mass fingerprinting with tandem mass spectrometry(MS/MS).In addition,Western blotting and immunohistochemistry were performed to verify the expression of certain candidate protein.Results A total of 28 protein spot-features were found to be significantly increased and 17 significantly decreased in tumor tissues.Thirty of these protein spots were identified,which included enzymes,antioxidant proteins,signal transduction proteins,calcium-binding protein,structural proteins,chaperones and others.Western blotting and IHC further validated up-regulated expressions of one candidate protein(annexin II)in tumor tissues.Conclusions The analysis of proteomics with 2-DE on human tissue is a useful method for discovering valuable cancer marker candidates.These differential expressed proteins may serve as biomarkers for early detection and therapeutic targets to pancreatic cancer.

Objective To compare the clinical features and endoscopic findings of Crohn's disease(CD) and intestinal tuberculosis(ITB) in order to differentiate CD from ITB. Methods The clinical and endoscopic data from 168 patients with CD and 156 patients with ITB between June 2003 and February 2009 were retrospectively analyzed. Results The salient features of CD were male patients in predominance (male : female was 108 :60) and high incidence of colectomy (CD 33.3% vs ITB 10.9%, P<0.01). Diarrhea (66.1%), hematochezia (32.1%), perianal disease (16.1%), intestinal obstruction (28.0%) were more frequent in CD patients than in ITB patients (47.0%, 7.7%, 3.4%, 9.4% respectively, all P values<0.05). The salient features of ITB were night sweating, pulmonary tuberculosis, ascites, hyperglobulin, increased erythrocyte sedimentation rate and the positive serum antibody to mycobacterium. The endoscopic examination showed that the fissure-shape ulcer, grid-shape ulcer, cobblestone sign and intestinal stricture were more frequent in CD patients than in ITB patients (all P values <0.05). Whereas the circular ulcer and involved ileocecal valve with fixed bouche shape were more common in ITB patients (P<0.05). Conclusions The clinical characteristics are different in CD and ITB patients. The endoscopic findings including fissure-shape ulcer, grid-shape ulcer, circular ulcer, cobblestone sign and the status of involved ileocecal valve are important in the differentiation of ITB from CD.

Objectives To isolate cancer stem cells (CSCs) in pancreatic cancer cell lines PANC1 and ASPC-1 with serum-free medium( SFM ), and to detect the expression of miR-590-3p in CSCs. Methods PANC1 and ASPC-1 cells was cultured in serum-free medium. The monoclonal formation, differentiation and cell cycle, half inhibitory concentration ( IC50 ), and the expression of the surface markers CD24 + , CD44 + were detected. qRT-PCR was used to detect the expression of miR-590-3p. Results After SFM culture, (0.94 ±0.53 ) ％ of ASPC-1 and (0.57 + 0. 12 ) ％ PANC1 survived, and they formed spheres, and could continuously passage in vitro. Cell spheres differentiation recurred when serum was supplemented in SFM. The G0/G1 stage proportion, CD24+ , CD44 + , CD24+ CD44+ cells proportion, IC50 in ASPC-1 cell were (75.3 ± 5.4)％,0.96％ ～ 2.01％, 27.52％ ～ 34.47％, 0.35％ ～ 0.44％ and (224.37 ± 5.71 ) μg/ml, which were significantly higher than that those in parent cell [ (43.7 ± 3.8 ) ％, 0. 38％ ～ 0.42％, 17.65％ ～ 18.25％,0.05％ ～0.08％, (11.43 ±2.10)μg/ml, P＜0.05]. The G0/G1 stage proportion, CD24+ ,CD44+ ,CD24 +CD44 + cells proportion, IC50 in PANC 1 cell were ( 80. 1 ± 4.7) ％, 5.31％ ～ 9.84％, 72.05％ ～ 93.06％,4.91％ ～5.21％, (296.58±4.27) μg/ml, which were significantly higher than that those in parent cell [ (46.1 ±5.3)％, 4.09％ ～4.97％, 47.71％ ～55.66％, 1.48％ ～2.63％, (26.17 ±3.81)μg/ml, P＜0.05]. The expression of miR-590-3p in ASPC-1, PANC1 spheres was 4.67 and 4.52 times higher than the expression in parent cell lines. Conclusions Pancreatic cancer cell spheres can be isolated from ASPC-1, PANC1 by culture with SFM. miR-590-3p is up-regulated and may play an important role in regulating biological characteristics of pancreatic cancer stem cells.

OBJECTIVE: To explore the expression of vascular endothelial growth factor (VEGF) C and D in gastric cancer and its relationship with tumor angiogenesis and lymph node metastasis. METHODS: Immunohistochemistry(SABC) and real-time PCR were used to detect the expression of VEGF-C, VEGF-D protein and mRNA in 32 gastric cancer tissues and 32 normal gastric tissues. RESULTS: The positive expression rate of VEGF-C and VEGF-D in gastric cancer tissue was significantly higher than that of normal gastric tissues (P<0.01), the expression of VEGF-C and VEGF-D in the gastric cancer group and the lymph node metastasis group were significantly different (P<0.05). The expression of VEGF-C and VEGF-D in gastric carcinoma was positively correlated with lymph node metastasis (P<0.01), the expression of VEGF-C and VEGF-D in well-differentiated carcinoma, moderately differentiated carcinoma and poorly differentiated carcinoma was statistically different (P<0.05). VEGF-C and VEGF-D expressions in gastric cancer cells were not related to the patient's age, sex, and lymph node distant metastasis (P>0.05). CONCLUSION The non-intake high expression of VEGF-C and VEGF-D in gastric cancer cells is closely related to lymph node metastasis. They serve as the important reference indicator to assess the prognosis in gastric cancer patients.
Adult ; Aged ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Vascular Endothelial Growth Factor D ; genetics ; metabolism

AIM:To investigate the inhibitory effect of the"decoction for soothing liver and removing stasis and toxicity(SGQYJDF)"on hepatocellular carcinoma(HCC)proliferation in nude mice by inducing ferroptosis via the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11/xCT)/glutathione peroxidase 4(GPX4)pathway.METHODS:An ectopic subcutaneous tumor model was established by injecting SK-Hep-1 cells subcutaneously into the right axilla of nude mice.Upon formation of tumor,the mice were randomly divided into five groups(i.e.,control group,low-,medium-and high-dose SGQYJDF groups and medium-dose SGQYJDF plus Sorafenib group).Each group of mice was orally administered with the corresponding therapy for 14 consecutive days,during which the tumor size was observed regularly.At the end of treatment,the tumor growth inhibition rate was calculated based on tumor mass,and histopatho-logical changes were observed by HE staining.Then,the levels of malondialdehyde(MDA),glutathione(GSH)and fer-rous ions(Fe2+)were detected by colorimetric assays.The expression of the proliferation markers Ki-67 and GPX4 was de-tected by immunohistochemistry(IHC).The expression of p53 and xCT was detected by Immunofluorescence(IF).And the expression of p53,xCT and GPX4 was determined by Western blot.RESULTS:(1)SGQYJDF was found to dose-de-pendently decrease tumor volume(P＜0.01)and inhibit tumor mass growth(P＜0.01),and meanwhile,reduce the per-centage of Ki-67-positive cells(P＜0.01)and their proliferation ability in tumor tissues,as compared to the control group.(2)In terms of Ferroptosis-related indicators,SGQYJDF was found to dose-dependently increase the levels of Fe2+ and MDA but decrease the level of GSH in tumor tissues(P＜0.01),as compared to the control group.(3)In terms of protein expression,SGQYJDF was found to dose-dependently upregulate the expression of p53(P＜0.05)but inhibit the expres-sion of xCT(P＜0.05)and GPX4(P＜0.01).Notably,medium-dose SGQYJDF plus sorafenib showed a stronger effect than high-dose SGQYJDF.CONCLUSION:Our findings suggest that SGQYJDF can induce Ferroptosis to inhibit the proliferation of subcutaneously transplanted tumor tissues in nude mice by upregulating the expression of p53,and inhibit-ing the expression of xCT and GPX4.