Aged ; Antineoplastic Agents ; therapeutic use ; Benzamides ; therapeutic use ; Drug Resistance, Neoplasm ; Exons ; Gastrectomy ; methods ; Gastrointestinal Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Gastrointestinal Stromal Tumors ; drug therapy ; metabolism ; pathology ; surgery ; Humans ; Imatinib Mesylate ; Liver Neoplasms ; drug therapy ; secondary ; Male ; Piperazines ; therapeutic use ; Point Mutation ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Pyrimidines ; therapeutic use

Objective To investigate the morphological changes in the sciatic nerve and the dorsal root ganglions (DRGs) and also gene expression in DRGs after non-freezing cold injury, and to explore the molecular mechanism of peripheral nerve cold injury and regeneration. Methods Twenty-four male Wistar rats were used. The sciatic nerve on one side was cooled to 4℃ for 2 h, and the sciatic nerve on the opposite side was exposed, but without cooling. Sciatic nerves and L4, L5 and L6 DRGs from both sides were harvested at the 1st, 2nd and 3rd week after cooling. Any pathological changes were observed using light and electron microscopy. Laser capture microdissection (LCM) was used to investigate the DRG neurons' gene expression. The array result was verified with RT-PCR for eight genes. Results Large fiber degeneration was obvious by the 7th day after cooling. Myelinated fiber regeneration had begun by the 14th day, so this time was chosen to explore the neurons' gene expression. Ninety-six genes and expressed sequence tags (ESTs) were up-regulated greater than 2 fold. Their proteins' functions were classified as adaptive response to external stimulus, apoptosis regulation, cell adhesion, immune and inflammation response,nerve regeneration, pain associated molecules, microtubule cytoskeleton, ion-channels, neurotransmitters and receptors, and neuropeptides. Conclusions A complex molecular mechanism is involved in cold injury and regeneration of the sciatic nerve, and many genes are involved. Large scale microarray analysis is a potent means to screen out related genes, thus suggesting future repair strategies.

OBJECTIVETo investigate the phenotypic characteristics of human fetal liver cells (FLCs) and to obtain the homogenous hepatic progenitors with cloning.

METHODSImmunofluorescence and flow cytometry were used to determine the phenotypes of the FLCs. The proliferating colonies were isolated using clone ring in different culture conditions. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the mRNA expression after further cultivation.

RESULTSThe cultured FLCs showed a non-typical epithelial morphology. The positive rate for hepatic cell specific markers alpha-fetoprotein (AFP), albumin (Alb), cytokeratin 8 (CK8) and CK19 were approximately 28.1%, 84.7%, 55.1% and 9.1% respectively. Furthermore, the FLCs expressed the hematopoietic stem cell markers CD34 and CD45 with percentages of 0.04% and 0.09%. 71.8% and 75.3% of the FLCs were positive for the mesenchymal cell marker CD105 and CD166. Most of the colonies showed an elongated morphology, some with an unregular spreading-out morphology, only a small number of colonies with an epithelial-like morphology. RT-PCR results showed that among the 19 colonies obtained after further cultivation and the percentages of epithelial cell adhesion molecule (EpCAM), AFP, Alb and CK19 were 52.6%, 21.1%, 52.6% and 84.2%, respectively.

CONCLUSIONSThe clonal culture system is beneficial to obtain the homogenous hepatic progenitor cells from the heterogeneous culture of FLCs.

Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Fetus ; cytology ; Hepatocytes ; cytology ; Humans ; Stem Cells ; cytology

OBJECTIVESTo evaluate efficacy of microsurgical hemilaminectomy approach and use of intraoperative indocyanine green videoangiography for patients with intradural dorsal arteriovenous fistula.

METHODSMedical records and follow-up data of 24 patients who were microsurgically treated at a single institution for intradural dorsal arteriovenous fistula between January 2004 and August 2008 were retrospectively reviewed. Preoperatively DSA was performed for definite diagnosis. All patients were evaluated with the Aminoff and Logue scale. Preoperative, 4 patients had excellent spinal condition having mean score of 1.0; 8 cases had good spinal condition with mean score of 3.4; 9 cases had average spinal condition with mean score of 6.9; 3 cases had poor spinal condition with mean score of 10.0. Twenty two cases had one feeder while 2 cases had two feeding arteries. All the patients underwent microsurgical hemilaminectomy via a posterior approach. Two patients received complemented surgery because of the recurrence of the lesion after embolisation failed. Three patients received intraoperative indocyanine green videoangiography. Combined followed-up imaging and myelonic function were used for evaluating followed-up data.

RESULTSMean follow up was done up to 36 months. Followed-up imaging didn't reveal any residual lesion or its recurrence. Spinal functional assessment using Aminoff and Logue scale showed 16 patients of excellent outcome and had mean score of 0.7; 4 had good outcome having mean score of 4.8; 3 was of average nature with mean score of 6.7; 1 had poor outcome with 9.0 scores. With the surgical treatment, 16 cases were cured, 6 were improved and 2 cases had no change according to synthetic curative effect.

CONCLUSIONSMicrosurgical treatment, especially the microsurgical hemilaminectomy via a posterior approach, is effective treatment intradural dorsal arteriovenous fistula. Intraoperative indocyanine green videoangiography is a simple auxiliary tool for intraoperative quality control and favorable surgical outcomes.

Adult ; Aged ; Arteriovenous Fistula ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; Middle Aged ; Retrospective Studies ; Spinal Cord ; blood supply ; Treatment Outcome ; Young Adult

OBJECTIVETo compare the tubulogenesis capability of malignant glioma-derived microvessel endothelial cells (GDMEC) from human brain with that of ECV304 cells in a three dimentional model and to explore the significance of GDMEC in the study on angiogenesis.

METHODSThe GDMEC were isolated from malignant gliomas of human brain and purified by selective binding to the monoclonal antibody against CD105 bound to the magnetic MACS MicroBeads. GDMEC and endothelial-like cell line ECV304 were compared with their capabilities of formatting tubule-like structure (TLS) in the three dimentional collagen matrix, with or without inducement by various concentration of vascular endothelial growth factor (VEGF).

RESULTSThe obtained GDMEC had a high purification (98%) and could be successfully cultured in vitro. GDMECs formed more TLS than ECV304 cells of the same number and at the same time points. VEGF could induce rapid formation of TLS in a dose-dependent manner, however, ECV304 cells were less response to VEGF stimulation.

CONCLUSIONSGDMEC could maintain their endothelial characteristics and potential capability of angiogenesis. They were more response to VEGF than ECV304, therefore, more suitable for in vitro studies on tumor angiogenesis.

Brain Neoplasms ; blood supply ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; Endothelium, Vascular ; cytology ; Glioma ; blood supply ; Humans ; Immunomagnetic Separation ; Microcirculation ; pathology ; Neovascularization, Pathologic ; Vascular Endothelial Growth Factors ; administration & dosage ; pharmacology

OBJECTIVETo investigate the feasibility that transforming growth factor-beta1 (TGF-beta1) -loaded fibrin sealant (FS) promotes bone marrow mesenchymal stem cells (BMSCs) to create tissue engineering cartilage in vivo.

METHODSThe BMSCs were isolated from healthy human and amplified in vitro, and then induced by defined medium containing TGF-beta1 and dexamethasone. After 7 days the induced BMSCs were collected and mixed with TGF-beta1-loaded FS or FS as BMSCs+ FS-TGF-beta1 group and BMSCs+ FS experimental group. Then the mixture was injected by a needle into the dorsum of nude mice. In control group, only FS or BMSCs were injected. The tissue engineering specimens were harvested from nude mice 12 weeks later. Gross observation, average wet weight measurement, glycosaminoglycan (GAG) quantification, histology and immunohistochemistry were used to evaluate the results.

RESULTSThe BMSCs have possessed the shape and functional characters of chondrocyte when transferred to a defined medium. After injection of the mixture, the cartilage-like tissue were formed in two experimental groups. Compared with BMSC+ FS group, the specimens of BMSCs +FS-TGF-beta1 group were larger and firmer. Alcian staining showed better metachromatic matrix formation. The GAG contents were significantly higher. Immunohistochemical staining of collagen type II was stronger. However, no cartilage-like tissue was formed in two control groups.

CONCLUSIONTGF-beta1-loaded FS can promote BMSCs to contract injectable tissue engineering cartilage in vivo.

Animals ; Biocompatible Materials ; Cell Differentiation ; drug effects ; Cells, Cultured ; Chondrogenesis ; drug effects ; Dexamethasone ; pharmacology ; Fibrin Tissue Adhesive ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Tissue Engineering ; methods ; Transforming Growth Factor beta ; pharmacology

BACKGROUNDIt is generally accepted that gliomas are the most common primary brain tumors with poor prognosis. We aimed to explore the relationship of the immunity of the central nervous system and the genesis and development of glioma.

METHODSG422 glioma was implanted in the brain of BALB/c mice (immuno-competent mice), nude mice (T cell related immuno-deficient) and complement C3 knock-out mice (complement C3 related immunodeficient). The survival time of the host, growth and histopathology of the tumor, and concentrations of tumor necrosis factor-α (TNF-α) and interferon-γ (INF-γ) in tumor tissues were assessed.

RESULTSTumor spheres were formed in all mice after injection, and glial fibrillary acidic protein (GFAP) positive staining of the cells declared their glioma origin. The longest median survival time of (44.3 ± 6.0) days was found in BALB/c mice, followed by (24.8 ± 5.2) days in nude mice and the shortest (18.6 ± 5.8) days in complement C3 knock-out mice. Accordingly, the growth of the tumor was fastest in complement C3 knock-out mice, followed by the nude mice and slowest in the BALB/c mice. Although the proportions of infiltrating CD68(+) lymphocytes in tumor tissues showed no significant difference (P > 0.05), TNF-α level in the nude and C3 knock-out mice, (28.11 ± 4.86) µmol/L and (22.87 ± 6.36) µmol/L respectively, were significantly lower (P < 0.01) than that in the BALB/c mice, which was (230.21 ± 39.17) µmol/L. The INF-γ level was highest in the BALB/c mice ((180.76 ± 29.19) µmol/L), followed by the nude mice ((113.46 ± 23.76) µmol/L) and then the C3 knock-out mice ((16.84 ± 4.45) µmol/L).

CONCLUSIONSThe G422 glioma implanted in the brains of mice with different immune ability would be a useful model for studying the relationship of the immune system and tumor in the central nervous system. Furthermore, the T cells and complement C3 compartments of the immune response may affect the growth of implanted tumors and inflammatory factors such as TNF-α and INF-γ.

Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; Complement C3 ; genetics ; metabolism ; Glioma ; metabolism ; pathology ; Interferon-gamma ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Mice, Nude ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo construct a recombinant adenoviral vector harboring human transforming growth factor-beta type II receptor-IgG1Fc (TbetaRII-IgG1Fc) fusion gene.

METHODSThe cDNA fragments of human TbetaRII and IgG1Fc genes were amplified by RT-PCR and fused with overlap PCR to obtain the fusion gene TbetaRKK-IgG1Fc. The TbetaRII-IgG1Fc gene was cloned into the shuttle plasmid pAdTrack-CMV, which was linearized and transfected into E.coli BJ 5183 strain containing the adenoviral backbone vector. The recombinant adenovirus vector was constructed by homologous recombination. The recombinant adenoviral plasmid was linearized and transfected into 293 cells, followed by amplification and purification of the virus and detection of TbetaRII-IgG1Fc mRNA expression by RT-PCR. The functional activity of the recombinant adenoviral plasmid was assessed using enzyme-linked immunosorbent assay (ELISA).

RESULTSThe results of restriction endonuclease digestion and DNA sequencing indicated correct sequence of the target TbetaRII-IgG1Fc fusion gene. The recombinant adenoviral plasmid expressed hTbetaRII-IgG1Fc and neutralized TGF-beta1 in vitro after infection of the human lung fibroblasts (HLF), as confirmed by RT-PCR and ELISA.

CONCLUSIONSThe recombinant adenoviral plasmid capable of neutralizing TGF-beta1 in vitro is constructed successfully.

Adenoviridae ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Fibroblasts ; cytology ; Genetic Vectors ; genetics ; Humans ; Immunoglobulin Fc Fragments ; genetics ; metabolism ; Immunoglobulin G ; genetics ; metabolism ; Lung ; cytology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Receptors, Transforming Growth Factor beta ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Humans ; Infant, Newborn ; Male ; Sleep Apnea, Central ; diagnosis ; therapy

The fruiting body pattern is an important agronomic trait of the edible fungus Auricularia auricula-judae, and an important breeding target. There are two types of fruiting body pattern: the cluster type and the chrysanthemum type. We identified the fruiting body pattern of 26 test strains, and then constructed two different near-isogenic pools. Then, we developed sequence characterized amplified region (SCAR) molecular markers associated with the fruiting body pattern based on sequence-related amplified polymorphism (SRAP) markers. Ten different bands (189–522 bp) were amplified using 153 pairs of SRAP primers. The SCAR marker “SCL-18” consisted of a single 522-bp band amplified from the cluster-type strains, but not the chrysanthemum strains. This SCAR marker was closely associated with the cluster-type fruiting body trait of A. auricula-judae. These results lay the foundation for further research to locate and clone genes controlling the fruiting body pattern of A. auricula-judae.
Breeding ; Chrysanthemum ; Cicatrix ; Clone Cells ; Fruit* ; Fungi