Objective To establish embryonic stem cells (ESC) that can express green fluorescent protein (CFP) and differentiate them into neurons. It would provide tagging neurons for clinical transplantation to cure neural system diseases. Methods ESC (R1) was transfeeted with a plasmid containing the GFP by electroporation. A transgeuic cell line was obtained after selection with G418. The ESCs were characterized by AKP staining. Monolayer differentiation method was used to induce neural differentiation derived from GFP-ESC and immunofluorescence method was used to identify Tuj1 positive cells. Results There was no significant difference(X2=3.14,P0.05) in transfect rates between liposome and electroporation (65% vs 79%). The AKP staining of GFP-ESC was positive. GFP-ESC could be differentiated into neural cells. Conclusions These results show that ESC expressing GFP has been estabhshed, which can be differemiated into neurons.

Objective To investigate the effect of Gout Gra nule(GG)for gout.Methods Gout -relieving effect of GG was observed in rats with adjuvant arthritis induced by sodium urate and i n mice with hyperuricemia induced by hypoxan-thine.Anti -inflammation of GG was i nvestigated in mice with xylene -ind uced auricular swelling and in rats with edema of hind paw induced by injection of carrageenin.WBC migratory response c aused by carboxymethylcellulose(CMC)in mice was also observed.Results GG could reduce the blood level of uric acid and inhibit acute inflammatio n in model ani-mals and also tended to promote the elimination of uric acid.Conclusion GG had a remarkable effect on the acut e attack of gout.

Objective: To prepare a blood deficient model and study the effect of Tangkuei Blood Supplementing Decoction on hemopoiesis in this model. Methods: This model was made in mice by i.p. an accumulate doses of 60mg/kg acetylphenylhydrazine (APH) and 160mg/kg cyclophosphamidum (CY). The RBCs, WBCs, reticulocytes and bone marrow nucleated cells (BMNC) were counted, the micro structure of bone marrow was observed. The swimming time, the body temperature and the plasma cAMP, cGMP levels were measured. The effects of Tangkuei Blood Supplementing Decoction by p.o. administration on promoting the hemopioetic function were observed with the model mice. Results: Tangkuei Blood Supplementing Decoction could remarkably increase RBC, WBC, BMNC, improve the proportion of reticulolytes in peripheral blood and the micro struture of bone marrow, prolony the swimming time, raise the body temperature and the specific value of cAMP/cGMP. Conclusion: The model exhibits the main features of blood deficiency, and can be used as one of models of blood deficiency. Tangkuei Blood Supplementing Decoction can obviously improve the dual deficiention of qi and blood of model mice by supplementing qi and blood.

OBJECTIVETo study 3 different strategies of urine drainage following hypospadias urethroplasty, the clinical nursing in their application, and their effects.

METHODSWe retrospectively analyzed the clinical data of 595 cases of hypospadias treated by urethroplasty. After surgery, 133 of the patients underwent urine drainage by suprapubic cystostomy (group A), 202 by urethral stent- tube indwelling (group B), and 260 by early initiative micturition with the urethral stent-tube (group C). All the patients received routine postoperative nursing care required for hypospadias repair.

RESULTSOperations were successfully completed in all the cases. Group C showed a remarkably shorter hospital stay and lower incidence rates of urinary fistula and urethral stricture than groups A and B (P<0.05), but there were no significant differences in the three indexes between A and B (P<0.05).

CONCLUSIONFor urine drainage following hypospadias repair, early initiative micturition with the urethral stent-tube can significantly reduce postoperative complications, decrease difficulties and workload of nursing care, and shorten the hospital stay of the patient.

Cystostomy ; Drainage ; methods ; Humans ; Hypospadias ; surgery ; Length of Stay ; Male ; Postoperative Complications ; prevention & control ; Reconstructive Surgical Procedures ; Retrospective Studies ; Stents ; Urethra ; surgery ; Urethral Stricture ; prevention & control ; Urinary Fistula ; prevention & control ; Urine ; Urologic Surgical Procedures, Male

White-rot fungi is a kind of basidiomycetes making wood rotten. For their particular metabolism and extracellular degrading ability, they can degrade a lot of organic pollutants, and then become the hot point of international academic research. This paper reviews the recent research progress in many aspects,such as the sort and degradation mechanism of white rot fungus, advances in applied research for white rot fungi on industry and environmental pollution disposal and so on. In addition, some suggestions on the prospective application in the composting of municipal solid waste are presented in the end.

OBJECTIVENucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes.

METHODSEukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results.

RESULTThe recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours.

CONCLUSIONThe nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.

Cancer Vaccines ; biosynthesis ; Gene Expression ; Genetic Vectors ; HN Protein ; genetics ; Hep G2 Cells ; Humans ; Interleukin-18 ; genetics ; Laryngeal Neoplasms ; immunology ; prevention & control ; Newcastle disease virus ; immunology ; Plasmids ; Transfection ; Vaccines, DNA ; biosynthesis

OBJECTIVETo analyze the phenotypic and functional characteristics of endothelial (T3A) cells derived from human hepatocellular cell carcinoma.

METHODSEndothelial cells were isolated from human hepatocellular carcinoma specimens. The identification of T3A cells was performed by checking von Willebrand Factor (vWF), CD31, CD34 and Dil-Ac-LDL uptake. The cell surface fenestrations, a specific morphological feature of tumor derived EC, were investigated by scanning and transmission electron microscopy. The phenotypic characteristics of T3A cells were analyzed by fluorescence-activated cell sorter (FACS) and were further conformed by real-time PCR at transcription level. Furthermore, tumor necrosis factor alpha (TNFalpha)-induced cytotoxicity was evaluated by 3-(4, 5-dimethythiazolyl) -2, -diphenyl-2H-tetrazolium-bromide (MTT) assay; Matrix metalloproteinase secretion was detected by zymography; Angiogenic ability in vitro was analyzed by culturing T3A cells in three-dimensional Matrigel plug. Coagulant and fibrinolytic activities were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe isolated T3A cells exhibited classic "spindle-shape" morphology and monolayer growth and contact inhibition properties. Immunofluorescent staining showed that T3A cells expressed vWF, CD31, CD34, and uptake of Dil-Ac-LDL at a high level. The cell surface fenestrations were observed on T3A cells by scanning and transmission electron microscopy. By FACS and real-time PCR, T3A cells were found to express alphav3, alphavbeta5 and TNF receptor p75 at high levels, and TNF receptor p55 and ICAM-1 at low levels, as compared with those in human liver sinusoidal endothelial cells (LSEC). In response to TNFalpha, LSEC exhibited a dose-dependent cytotoxicity, while T3A cells were resistant. Gelatin zymography showed that MMP-2 activity was higher in T3A cells than that in LSEC. In a three-dimensional plug of Matrigel, T3A cells exhibited stronger angiogenic ability as compared with LSEC. In addition, T3A cells released more tissue factor (TF), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI-1) and urine plasminogen activator (u-PA) than LSEC in response to TNFalpha.

CONCLUSIONTumor-derived endothelial cells are phenotypically and functionally different from those derived from normal liver tissue.

Antigens, CD34 ; metabolism ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Cell Shape ; Cells, Cultured ; Endothelial Cells ; metabolism ; pathology ; ultrastructure ; Gene Expression ; Humans ; Integrin alphaVbeta3 ; metabolism ; Integrins ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Lung ; blood supply ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; metabolism ; Microscopy, Electron, Scanning ; Neovascularization, Pathologic ; metabolism ; pathology ; Phenotype ; Plasminogen Activator Inhibitor 1 ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Tumor Necrosis Factor, Type I ; metabolism ; Receptors, Vitronectin ; metabolism ; Tissue Plasminogen Activator ; metabolism ; Tumor Cells, Cultured ; Tumor Necrosis Factor Decoy Receptors ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; von Willebrand Factor ; metabolism

OBJECTIVETo investigate the role of mutation and mRNA expression of tumor suppressor gene phosphatase and tensin homolog deleted in chromosome 10 (PTEN) in tumorigenesis and progression of oral squamous cell carcinoma (OSCC).

METHODSThe mutation of exon 3, 5, 6 and exon 8 of PTEN gene in 42 oral squamous cell carcinoma tissue and paired normal tissue were detected by polymerase chain reaction (PCR) and DNA sequencing methods. The levels of PTEN mRNA expression in these tissues were assayed using semi-quantitative RT-PCR. Data was analyzed with SPSS 14.0 software package.

RESULTSMutated exon 5 of PTEN gene was found in 2 cases of advanced OSCC. The expression of PTEN mRNA was detected in all OSCC and paired normal tissue. The level of PTEN mRNA in OSCC tissue (0.36 +/- 0.12) was significantly lower than that of paired normal tissue (0.64 +/- 0.09, P < 0.01).

CONCLUSIONSThe decreased expression of PTEN mRNA contributes to tumorigenesis of OSCC.

Carcinoma, Squamous Cell ; genetics ; Chromosomes, Human, Pair 10 ; Exons ; Gene Expression ; Genes, Tumor Suppressor ; Humans ; Microfilament Proteins ; Mouth Neoplasms ; genetics ; Mutation ; PTEN Phosphohydrolase ; genetics ; Phosphoric Monoester Hydrolases ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; metabolism ; Tensins

OBJECTIVETo investigate the mechanisms of apoptosis induced in human hepatoma cell line SMMC7721 by plasmid pVHN constructed with Newcastle disease virus (NDV) HN gene.

METHODSTwenty-four h after transfection with liposome-plasmid pVHN complexes in vitro, the mortality rate of SMMC7721 cells was determined by MTT staining and flow cytometry (FCM) with PI staining. The alteration of mitochondrial trans-membrane potential of the cells was detected by FCM with rhodamine 123 staining. Cell genomic DNA was detected by agarose electrophoresis. The activation of caspase-3 was assayed by its substrate color reaction.

RESULTSSignificant apoptosis was induced by transfection with plasmid pVHN into the cells for 24 h and the mortality rate was 50.0% (the mortality rate of control group was 5.2%). Genomic DNA was fragmented and mitochondrial trans-membrane potential was decreased, but caspase-3 activity increased.

CONCLUSIONSignificant apoptosis in SMMC7721 cells can be induced by NDV HN gene. Apoptosis may be resulted from the decrease of mitochondrial trans-membrane potential and activation of Caspase-3.

Apoptosis ; physiology ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; HN Protein ; genetics ; Humans ; Liver Neoplasms ; pathology ; Newcastle disease virus ; genetics ; Transfection ; Vaccines, DNA ; immunology

To study the effect of mesenchymal stem cell (MSC) on immune function, MSCs were isolated and cultured from human bone marrow cells. The purity of MSCs were identified with the spindle-fibroblastic morphology characterization by microphotograph and the phenotypes were tested by flow cytometry. MSCs were plated in 96-well plates (2,000/well and 1,000/well), and cocultured for 3 days with T cells isolated from cord blood. Cord blood T cells non-cocultured with MSC acted as control group. After cord blood T cells stimulated by PHA for 60 hours, [(3)H]-thymidine was added to each well and T cell proliferation was assessed by [(3)H] thymidine incorporation. The results showed that cord blood T cell proliferation was suppressed when 2,000 MSCs were plated each well and cord blood T cell proliferation was activated when 1,000 MSCs were plated. Our results suggested that the immunomodulatory function of MSC seemed dependent on cell dose. High concentration of MSC most often resulted in inhibition, while low concentration resulted in stimulation.
Antigens, CD ; analysis ; Bone Marrow Cells ; cytology ; Cell Division ; immunology ; Cells, Cultured ; Coculture Techniques ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Lymphocyte Activation ; drug effects ; immunology ; Mesoderm ; cytology ; Phytohemagglutinins ; pharmacology ; Stem Cells ; cytology ; T-Lymphocytes ; cytology ; drug effects ; metabolism ; Thymidine ; metabolism ; Tritium