1.Effect of mutant ?-catenin gene on the proliferation of hepatocytes
Xianzhang SHANG ; Jun MIN ; Zhonghua CHU ; Jishen CHEN
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: The ?-catenin is a key molecule in the Wnt signal pathway, which plays a critical role in normal development and tumorigenesis. However, the mechanisms of the ?-catenin on the cell growth control are still not completely defined. The aim of this study was to test the hypothesis that the mutant ?-catenin may regulate the hepatocyte proliferation. METHODS: The immortalized murine hepatocyte cell line, AML12, was used for this study. A plasmid that contain mutant ?-catenin S33Y was transfected into the AML12 cells and a stable cell line AML12S33Y was established. The cell growth property of this cell line and the parental cell were compared by flow cytometry analysis and direct cell count. The cells were also tested for the ability to form soft agar colonies, and the ability to form tumors in the severe immune deficient mice (SCID). RESULTS: 1. The mutant ?-catenin containing cell line AML12S33Y has higher proliferating index compared with the parental AML12 cells ( P
2.Effect of Surgical Treatment on Primary Gastrointestinal Non-Hodgking Lymphoma in Children
zheng-yun, ZHANG ; min, XU ; jun, CHU ; qi-min, CHEN ; jing-yan, TANG ; ci, PAN
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To explore the effect of surgical treatment on primary gastrointestinal non-Hodgking lymphoma(NHL) in children.Methods Nine cases of clinical and follow-up data of primary gastrointestinal NHL were studied retrospectively to evaluate the effect of surgical treatment on primary gastrointestinal NHL in children.Results Seven cases were male and 2 cases were female.The mean age was(5.59?3.27)years old.The clinical manifestation included abdominal mass (7 cases),abdominal pain (5 cases),fever (2 cases),haematemesis and melena (2 cases),constipation (1 case) and paroxysmal abdominal pain with vomiting (1 case).Nine cases were diagnosed as primary gastrointestinal NHL,including 1 case of intussusception,1 case of acute appendicitis,2 cases of gastrointestinal obstruction,2 cases of gastrointestinal bleeding and 3 cases of abdominal mass.One case received the operation of intussusception reduction,tumor resection and intestinal anastomosis.One case received appendectomy.One case received the operation of tumor biopsy and transverse colon colostomy.Six cases received laparotomy.Six cases were diagnosed as Burkitt lymphoma.One case was anaplastic large cell lymphoma and 2 cases were diffuse large B-cell lymphoma.One case was at stage Ⅰ,1 case was at stage ⅠE,2 cases were at stage Ⅱ,3 cases were at stage ⅡE and 2 cases were at stage Ⅲ.Nine patients had received operation.One case died after operation and 8 cases had received combined chemotherapy.The 1 and 3 years survival rates were 75.0% and 37.5%,respectively.Conclusions Acute abdomen is often the first symptom of primary gastrointestinal NHL in children and comprehensive surgical treatment is an effective procedure for it.
3.The effect of somatostatin on expression of metalloproteinase-2 and tissue inhibitor matrix metalloproteinase-2 of implanted liver tumor after partial hepatectomy in nude mice
Shiwen GONG ; Jun MIN ; Yajin CHEN ; Qingjia OU ; Tao CHEN ; Zhonghua CHU ; Shikun QIAN ; Huizhen LIANG
Chinese Journal of General Surgery 1994;0(05):-
Objective To observe the effect of somatostatin (SST) on the expression of metalloproteinase-2(MMP-2) and tissue inhibitor matrix metalloproteinase-2(TIMP-2) of implanted tumor in nude mice after partial hepatectomy.Method Nude mice were divided into group A(n=10) implanted by human hepatocellular carcinoma(HCC),group B(n=10, partial hepatectomy) and C(n=10,HCC implantation after liver resection). SST(250 ?g/kg,b.i.d) was given intraperitoneally in group C. Mice were sacrificed on 35th d, tumor was measured, the expression of MVD.MMP-2 and TIMP-2 was detected by immunochemical staining and quantitative image computer-analysis. Results In group B, the weight and volume and expression of MVD.MMP-2 of tumor tissue was markedly incressed than in group A(P
4.A pilot study of molecular mechanism of salmon milt DNA (SMD) retards mouse insenescence
Min CHENG ; Boyu DU ; Wenfeng CHU ; Wenhua CHEN ; Ping YU ; Rong LI
Chinese Journal of Tissue Engineering Research 2003;7(20):2801-2803
Aim To iuvestigate the effect and the mechanism of salmonmilt DNA (SMD) on age-related involutions in mouse thymus. MethodsFemale BALB/c of 10 months were divided randomly into three groupsaccording to their weights: high dosage group 333.33 mg/(kg @ d), lowdosage group 166. 67 mg/(kg @ d) and control group 0 mg/(kg @ d) .After five weeks, with Image-Pro Plus (version. 4.0) software, the thymusindexes and the thymoctytes in the thymus section were measured, as wellas the thymus cortex thickness. All the data were analyzed by SAS statisticsoftware. Mieroarray technique was applied to screen the gene fragments,which were differently expressed between the high dosage group and thecontrol group, together with RT-PCR to further confirm some of them.Results No significant differences of the variables including bodyweight, thymus weight and thymus indexes among the three groups werefound (F < 3.0 and P > 0.05, respectively). The thymocytes quanti-ties of thymus cortex and medulla in the high dosage group were significantlyhigher than those of the control group [cortex D(H, C) = 9.46, P < 0.01;medulla t( H.C) = 2.53, P < 0.05]. The thymus cortex thicknesses of bothSMD supplement groups were significantly higher than that of the control group[cortex D(L,C)=3.65, P> 0.05; medulla t(L, C)=0.8, P> 0.05] .112differently expressed gene fragments were isolated. Furthermore, we foundthe fragments with the logged number of U23789, X80232 and Aw209102were highly expressed in the high dosage group when RT-PCR techniquewas used. Conclusion SMD may reverse the age-related involutions inmouse thymus via up-regulation the expression of proliferation related genesand development and differentiation related genes simultaneously.
5.Analysis on pulse diagram characteristics of subjects with subhealth state.
Guixiang CHU ; Qingguang CHEN ; Jiatuo XU ; Bo YU ; Min ZHANG ; Longtao CUI ; Hongjin WU ; Zhaofu FEI
Journal of Integrative Medicine 2012;10(10):1099-105
To study the pulse diagram parameters of subjects with subhealth state and to find the pulse parameters for subhealth state evaluation.
6.The biological characteristics of cytokine-induced killer cells
Weishi GAO ; Jun MIN ; Zhonghua CHU ; Tao CHEN ; Qing WEI ; Qingji OU
Chinese Journal of Pathophysiology 2000;0(07):-
AIM: To investigate the biological characteristics of cytokine-induced killer (CIK) cells in vitro METHODS: The non-adhere peripheral blood monoclear cells from healthy donors were induced into CIK cells in the presence of IFN-?, IL-1?, IL-2 and anti-CD3 antibody. LAK (lymphokine activated killer) cells were prepared as a control. The cellular phenotype were detected by FCM and immunocytochemistry and the cytotoxicity was measured by LDH release assay. RESULTS: After 2 weeks of induction, the proliferation rate of CIK cells reached a peak and the proportion of CD3 + population was above 95%, and then the cells growth entered to plateau phase at week 3. The proportion of CD3 +CD56 + NKT subset cells was 16 5% on day 15 and it had no obvious variety between 2 and 4 weeks. Correspondingly, LAK cells grew slowly and had lower proliferation rate compared with the CIK cells ( P
7.Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro
Yunfeng CAI ; Jun MIN ; Tianling FANG ; Zhonghua CHU ; Xiaogeng DENG ; Jingsong HE ; Jishen CHEN
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro . METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.
8.The correlation between adhesion molecules and chemos ensitivity in human hepatocellular carcinoma
Zhonghua CHU ; Jun MIN ; Yunbiao LING ; Shuang CHEN ; Haiyan ZHAO ; Jianping LIU ; Qingjia OU
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To investigate the expression of adhes i on molecules in human hepatocellular carcinoma (HCC), and analyze the correlatio n between the expression of adhesion molecules and chemosensitivity. METHODS: The surgical and needle specimens from 64 patients were tested by the ATP-TCA. The expression of adhesion molecules and multi-drug resi stance genes (MDR) of tumor tis sues of 64 cases and adjacent tissues of 12 cases of HCC were detected with RT-P CR. RESULTS: The expression level of E-cadherin, ICAM-1, CD 44, CD 44V, ? 5, ? 1 in liver cancer tissues was 1.24?0.54, 0.96?0.3 7, 0.62?0 .73, 0.86?0.33, 0.97?0.49, 1.41?0.24, respectively. There was a signif icant difference between CD 44 and E-cadherin, ? 1. The expression levels of MD R 1, MRP, GST-?, LRP, TOPO II mRNA in liver cancer tissues were 1.17?0.47, 1 .59?0.33, 1.18?0.48, 1.03?0.48, 1.00?0.31, respectively. The express ion level of adhesion molecule mRNA had positive Spearman correlation with the expression level of MDR mRNA. ICAM-1, ? 5 had positive Spearman correlation w ith MDR 1. E-cadherin and CD 44 had negative Spearman correlation with MDR 1. E-cadherin had negative Spearman correlation with MRP. ICAM-1 had positive Spearman correlation with LRP. E-cadherin and CD 44 had negative Spearman c orrelation with LRP. CONCLUSION: The expression levels of adhesion molecule mRNA ha ve correlation with the effect of chemotherapy and the expression of MDR genes .
9.Spleno-left adrenal vein shunt for portal cavernous transformation
Zhengjun ZHANG ; Qimin CHEN ; Min XU ; Jun CHU ; Zhilong YAN ; Li HONG ; Song GU ; Ming HU
Chinese Journal of General Surgery 2010;25(1):17-19
Objective To evaluate the effect of spleno-left adrenal vein shunt for the treatment of portal hypertensive upper GI bleeding caused by portal vein cavernous transformation in children.Methods Spleno-left adrenal vein shunt was performed in 8 children with portal hypertension due to cavernous transformation.The clinical data was reviewed.Results Portal vein pressure decreased significantly from (30±11)mm Hg to(22±7) mm Hg after shunt.There was no mortality perioperatively and during the follow-up.There were no recurrent hemorrhage nor hepatic encephalopathy occurring in the follow-up and all the children have normal intelligence and normal liver function though blood ammonia level increased significantly from(18±7)μmol/L to (60±17)μmol/L in 4 cases.In 7 cases in which preoperative whole blood cell count significantly decreased,the postoperative WBC,RBC,Hb and PLT was (7.64 ±4.46)×10~9/L,(4.54±0.97)×10~(12)/L,(133±5) g/L and (355.40±107.36)×10~9/L respectively (all P <0.05).In one case suffering from preoperative low PLT count the postop PLT reached 333×10~9/L,which was significantly higher than that preoperatively.Esophageal varices ameliorated in 6 cases.No stenosis of anastomotic stoma and thrombosis developed.Conclusion Spleno-left adrenal vein shunt is an effective procedure to treat portal vein cavernous transformation induced portal hypertension in children.
10.Comparative study on the regulatory effects on senescence related cell cycle gene expression by TCM principles of tonifying shen, invigorating pi benefiting qi, and activating blood circulation.
Fang-min CHEN ; Wei-kang ZHAO ; Pin-chu XU
Chinese Journal of Integrated Traditional and Western Medicine 2003;23(11):837-840
OBJECTIVETo compare the effects of TCM therapeutic principles of tonifying Shen (TS), benefiting Qi (BQ), invigorating Pi (IP) and activating blood circulation (ABC) herbs in regulating the gene expression in senescence related cell cycle.
METHODSDrug sera containing TCM herbs of the above-mentioned principles were used to treat the aged human diploid fibroblast cell line 2BS. The effect of TCM on the senescence related cell cycle and its related gene expression (P16INK4, Cyclin D1 and PCNA) were examined by means of cell proliferative doublings, flow cytometry, RT-PCR and Western blot analysis.
RESULTSTCM herbs of TS and BQ could improve the cell cycle, down-regulate the P16 and Cyclin D1 mRNA/protein expression, up-regulate PCNA mRNA/protein expression, while TCM herbs of IP and ABC showed insignificant effect on these indexes.
CONCLUSIONTCM herbs of TS and BQ have effect in improving cell cycle, it may be achieved through promoting the P16 pathway of gene expression.
Aging ; genetics ; Animals ; Cell Cycle ; drug effects ; Cells, Cultured ; Cellular Senescence ; genetics ; Cyclin D1 ; biosynthesis ; genetics ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Fibroblasts ; cytology ; Gene Expression Regulation ; Lung ; cytology ; Male ; Medicine, Chinese Traditional ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley