In the present study, the influence of essential free fatty acids on AFP secretion and cell growth of BEL-7402 human hepatocellular cacinoma cell line was investigated by radiommunoassay. The results demonstrated that 40 - 50?g/ ml concentration of linolenic acid could inhibit the AFP secretion obviously( P 0.05) . All of these studies about anti-cancer effect of linolenic acid will provide the principle for the patient for health care and therapy.

Objective To investigate the expression changes of translation elongation factor 1?1 in anti-BPDE transformed and carcinoma 16 HBEs. Methods Suppression subtractive hybridization (SSH), bioinformatics and semi-quantitative RT-PCR were applied. The cDNA of anti-BPDE transformed and carcinoma 16 HBE cells were used as tester respectively, and the cDNA of normal 16 HBE was used as driver, the library of subtractive hybridization were profiled and inverted into TA cloning vector after two times of hybridization and two times of PCR. After the screening, sequencing and analysis of sequences, semi-quantitative RT-PCR was performed accompanying to the inner reference of ?-actin. Results 9 differentially expressed fragments were consistent with translation elongation factor 1?1 in different regions in Genbank, and the expressions were up regulated in BPDE transformed and carcinoma 16 HBE cells. Conclusion Translation elongation factor 1?1 may be related to the transforming effect and carcinogenesis of anti-BPDE.

Objective　To findout a good method for malignant transformation of human bronchial epithelial cells 16HBE cell induced by anti-benzo (a) pyrene-trans-7,8-dihydrodiol-9,10-epoxide (anti-BPDE) using the objective and simple index:colony forming frequency of cells in semisolid agar culture medium,and to establish a best model of malignant transformation of 16HBE cells.Methods　The tests on viable rate and colony formation of 16HBE cells were carried out to determine the exposure doses of anti-BPDE.The l6HBE cells were treated once or several times by anti BPDE at different doses and the transformed foci were observed and assessed at the different stages during the period of the whole experiment.The malignant features of transformed 16HBE cells with a good dose-response relationship were identified by the method of semisolid agar culture.The anti-BPDE induced colony forming frequencies of 16HBE cells in semisolid agar medium in each dose group were statistically compared.Results　The best method for malignant transformation of 16HBE cells induced by anti-BPDE was that the 16HBE cells were intermittently treated several times with anti-BPDE at doses of 0.1,0.5,1.0 and 2.0 μmol/L respectively and were inoculated for 15 generations,the related colony forming frequencies were 2.0‰,5.5‰,7.0‰ and 10.5‰respectively with a significant dose-response relation ship and a good linear correlation (r=0.9741,P＜0.05).Conclusion　The research infered that the malignant transformation of human bronchial epithelial cell-16HBE cells could be induced successfully by anti-BPDE,a suitable model could be established by anti-BPDE also.

Objective To find a ideal method for inguinal herniorrhaphy. Methods One hundred patients with inguinal hernia were randomized to receive respectively PHS and Mesh Plug & Patch tension-free herniorrhaphy. Results In PHS group, the average operative time was 35 min. The postoperative complication rate was 2% (1/50), the average hospital stay was 2 days. Follow-up of 6～10 months has found no recurrence. In Plug group, the average operative time was 40 min,complication rate was 8% (4/50), the average hospital stay were 3 days. Postoperatively, 6 patients complained of temporary foreign body feeling. Follow-up of 6～10 months has found no recurrence in either group. Conclusions Both PHS and Mesh Plug & Patch are ideal materials for herniorrhaphy. PHS is especially suitable for lean inguinal hernia patients with large defect on transversalis fascia. Plug & Patch has an advantage in fat patients with small defect on transversalis fascia.

Objective: To investigate the optimal culture conditio ns for adipose tissue-derived stromal cells(ADSCs) and for the induction of these cells to differentiate into osteogenic cells. Methods: ADSCs were cultured with routine methods,bFGF at 20 ng/ml was added into the medium and the proliferative of ADSCs was examined by cell counting. 0.1 ?mol /L of dexamethasone,10 mmol/L of ?-glycerophosphate and 50 ?mol/L of ascorbic acid were adapted to induce the cells to differentiate into osteogenic cells, ADSCs were identified by immunocytochemistry and differentiated osteogenic cells were identified by alkaline phosphatase(AP) staining and immunocytochemistry. Result: A population of ADSCs could be isolated from adul t human adipose tissue,the cells were fibroblast-like and could be maintaine d in vitro for extended periods with stable population doubling.The cells w ere expanded as undifferentiated in culture for more than 10 passages, indicati ng their proliferative capacity.bFGF stimulated the cell proliferation.Dexameth asone,?-glycerophosphate and ascorbic acid induced (40?8.6)% of ADSCs to ex press alkaline phosphatase(AP) ,(35?10.6)% of AP positive ADSCs were found to be collagen I positive. Calcification plaques were occasionally found in the cul tures. Conclusion:The data support the hypothesis that adu lt human adipose tissue contains stem cells capable of diffferentiating into ost eogenic cells.

BACKGROUND:There are myoblasts in human embryonic skeletal muscle. It remains poorly understand whether myoblasts in vitro can form myotube and what are the corresponding markers for identifying myoblasts and myotubes. OBJECTIVE:To investigate whether in vitro cultured myoblasts from human embryonic skeletal muscle can form myotube and whether they can express neural markers. METHODS:Human embryonic muscle-derived myoblasts were cultured in serum-containing medium. When the primary culture was established, cultured cel s were identified with immunocytochemistry for neural markers, such asβ-tubulin markers (desmin, myogenin, smooth muscle actin and myosin). RESULTS AND CONCLUSION:A population of myoblasts could migrate from human embryonic muscle tissues. They could express the markers for skeletal muscle such as desmin and myogenin, and they could express neuron specific enolase, nestin and neurofilament 200. They could form myotubes in vitro, and myotubes expressedβⅢ-tubulin, neurofilament 200 and glial fibril ary acidic protein. The data support the hypothesis that myoblasts from human embryonic muscle express neural markers and muscle markers, and cultured myoblasts and myotubes expressed neuron specific enolase,β-tubulin Ⅲ, nestin, neurofilament 200 and glial fibrillary acidic protein. This indicates that these markers could not be used for cel identification of trans-differentiation study from muscle origin to nervous system.

BACKGROUND:At present,the most frequently agents used for neural induction of bone marrow stromal cells(BMSCs)in vitro are anti-oxidants,such as beta-mercaptoethanol and all trans-retinoids.The majorities of induction from BMSCs are neuron-like cells in these protocols;however,whether it has neuronal function or not should be further studied.OBJECTIVE:TO investigate the differentiated characteristics of inducing human BMSCs into neural cells in serum-free medium.DESIGN:Observational study.SETTING:Chaozhou Central Hospital.MATERIALS:The experiment was carried out in the Chaozhou Central Hospital from April 2004 to December 2005.Adult bone marrows were derived from femoral and tibial bone marrow of three patients with fracture.All patients provided the confirmed consent and were approved by the Ethics Committee of Chaozhou Central Hospital.DMEMIF12 medium(1:1),fetal bovine serum (FBS), glutamine, N2 supplements and B27 Supplements were from GIBCO/BRL Company;recombinant basic fibroblast growth factor(bFGF)and recombinant epidermal growth factor(EGF)from Sigma Company;monoclonal antibody for vimentin(1:100),monoclonal antibody for myelin basic protein(MBP) (1:100),monoclonal antibody for S1 00(1:1 00),monoclonal antibody for neuron specific enolase(NSE)(1:1 00),and monoclonal antibody for neurofilament 200(NF200)(1:1 00)from Beijing Zhongshan Company;monoclonal antibody for glial fibrillary acidic protein (GFAP)(1:200)and polyclonal antibody for nestin(1:100)from Boster Company(Wuhan);mouse monoclonal antibody for beta-tubulin 3(1:1 000)from Sigma Company;SP-9000 kits and quick AEC from Beijing Zhongshan Company; culture dishes and flasks from Coming Company.METHODS:BMSCs from human bone marrow were cultured in serum-containing medium.When the primary culture was established, BMSCs were transferred into serum-free medium containing N2 or B27 supplement with 20 μg/L basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF),and cells were cultured in an incubator containing C02of 0.05 volume fraction at 37℃. Morphological changes of BMSCs in serum-free medium were observed under phase contrast microscope. And two days after culture. Expression of relative markers of BMSCs was detected withimmunocytochemistry.MAIN OUTCOME MEASURES:Morphological changes of BMSCs and expression of relative markers of nerve cells.RESULTS:A population of BMSCs could be isolated from adult human bone marrow,and they were processed to obtain a fibroblast-like population and were expanded as undifferentiated cells in culture for more than 10 passages.indicating their proliferative capacity.They could form spheroid state when they were sub-cultured in serum-free media supplemented with bFGF and EGF.these cells could express the markers for neural stem cells such as vimentin and nestin;they could expressed neuron specific enolase(NSE),beta-tubulin 3,TrkC and neurofilament 200(NF200)when they were plated on dishes with serum-containing medium; some cells exhibited the phenotypes for astrocytes.expressing gilal fibrillary acidic protein(GFAP)and S100 protein.CONCLUSION:The morphology,protein expression and differentiation ability of BMSCs in serum-free medium was similar to those of neural stem cells.The data support the hypothesis that adult bone marrow contains stem cells capable of differentiating into neural cells,the serum-free media make BMSCs overcome their mesenchymal commitment,showing the phenotypes for neural stem cells.

OBJECTIVETo clone differentially expressed cDNA sequences involved in malignant transformation induced by benzo(a)pyrene metabolite dihydroxyepoxy benzo pyrene (BPDE).

METHODThe malignant transformation of human bronchial epithelial cell line 16HBE induced by BPDE in vitro was used as a model for comparing gene expression between the transformed cells and controls. cDNA representational difference analysis (cDNA-RDA) was performed to isolate differentially expressed cDNA fragment in transformed cells. The cDNA fragments were ligated to pGEM-T vector and transformed into JM109 bacteria. The plasmid DNA were sequenced and compared with data in GenBank by BLASTN.

RESULTSFive cDNA sequences were found to be novel ones and were registered in dbest database, which assigned accession numbers in GenBank are BG354691, BG354692, BG354693, BG354694 and BG354695, respectively. Eight of the remaining cDNA sequences showed sequence homology to those previously reported such as ribosomal protein S23, MLN137, ACTN4, transforming growth factor and G protein gene.

CONCLUSIONSThese 13 genes may be involved in BPDE-induced malignant transformation, but their biological characteristics and functions are left to further studies.

Benzopyrenes ; metabolism ; pharmacology ; Carcinogens ; pharmacology ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; Cells, Cultured ; Cloning, Molecular ; DNA, Complementary ; analysis ; drug effects ; DNA, Neoplasm ; analysis ; Gene Expression ; drug effects ; Humans

OBJECTIVE: To construct and obtain ideal protein delivery vectors by researching the delivery efficiency and cytotoxicity to Hela cells using mPEG-PLGA-BSA-FITC-NPs. METHOD: The mPEG-PLGA nanoparticle was obtained through surface modification of PLGA with PEG, and deliver BSA-FITC into Hela cells in vitro. The positive cells were counted by Laser scanning confocal microscopy and the survival rate of Hela cells was calculated by MTT assay at different time points. RESULT: mPEG-PLGA-BSA-FITC-NPs shows the classic nanometer size, and the encapsulation efficiency reached 51. 2%. At the same time, the nanoparticles possess characteristics of slow release. By optimizing the delivery conditions, the highest efficiency of mPEG-PLGA-BSA-FITC-NPs was above 65.2%, and the cellular viability was about 85.7%. CONCLUSION mPEG-PLGA-BSA-FITC-NPs nanoparticles can successfully carry the target protein into cells as safe and effective as novel delivery materials of protein in vitro, and has shown slow release characteristics. The mPEG-PLGA-BSA-FITC-NPs provide ideal delivery vector for future application in clinical treatment of disease using nano-materials.
Drug Carriers ; Fluorescein-5-isothiocyanate ; analogs & derivatives ; HeLa Cells ; Humans ; Nanoparticles ; Particle Size ; Polyesters ; Polyethylene Glycols ; Serum Albumin, Bovine

Objective To study the effects of thymus transplantation(TT)combined with CD4--DLI on T cell reconstitution after allogeneic hematopoietic stem cell transplantation(allo-HSCT). Methods BALB/c mice were randomly divided into three groups:hematopoietic stem cell transplantation (HSCT group),hematopoietic stem cell transplantation combined with thymus transplantation(TT group)and hematopoietic stem cell transplanta-tion combined with thymus transplantation plus CD4+ T cell-depleted lymphocyte infusion(CD4--DLI group). On day-1,the mice were treated with the lethal dose of radiotherapy. On day 0,C57BL/6 mice were used as donor for hematopoietic stem cell transplantation. The mice were sacrificed on 5 days,2 weeks,4 weeks and 3 months after transplantation,respectively. The peripheral blood and spleen cells of mice were collected for determinations of T cell surface antigen,T cell receptor,naive T cells and intracellular cytokines. HE staining was used to assess the development of donor thymus. Results TT and CD4--DLI did not impair each other′s effects on T cell reconstitu-tion. TT combined with CD4--DLI increased the number of T cell reconstruction. CD4--DLI promoted the effect of TT on enlargement naive CD4+and CD8+T cell pool. Combination of TT and CD4--DLI enhanced the cytokine pro-duction of T cells. Conclusion TT combined with CD4--DLI had no side effects on TCR repertoire and thymus. Conclusion TT combined with CD4--DLI can enhance the reconstitution of T cell number and function via thymus dependent and thymus independent mechanism.