OBJECTIVETo investigate the enhancive effect of N,N'-dinitrosopiperazine (DNP) on induced carcinogenesis in nasal and/or nasopharyngeal epithelia among TgN(p53mt-LMP1)/HT transgenic mice to examine the underlying mechanism for the development of nasopharyngeal carcinoma (NPC).

METHODSTgN(p53mt-LMP1)/HT transgenic mice and the same strain of C(57)BL/6J wild-type mice both at the age of 5 months were randomly divided into 2 groups in parallel, respectively, i.e., TgN(p53mt-LMP1)/HT cancerous lesion-inducing group (TI), TgN(p53mt-LMP1)/HT control group (TC), C57BL/6J cancerous lesion-inducing group (CI), and C57BL/6J control group (CC). TI and CI mice were treated only with DNP for 16 weeks, twice each week, while TC and CC mice were given the same volume of saline as controls. At the end of treatment, animals were sacrificed to collect epithelial tissue samples from nasal cavity and nasopharynx for pathohistological evaluation by haematoxylin and eosin (HE) staining and for determination on the expression of TRAF2, c-Jun, and p16 by immunohistochemistry.

RESULTSAtypical hyperplasia was more significant in the samples of TI than in those of TC, CI, and CC, with the rates of lesions being 90%, 10%, 0, and 0 (P<0.01) respectively, though DNP was used alone in a much shortened inducing period at less dosage and without the use of carcinogenic promoter 12-O-tetradecanoylphorbol-13-acetate as usual. The expressions of tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2) and c-Jun in these samples were significantly up-regulated in TI (P<0.01), while the expression of p16 was significantly lower in TI than in the other groups (P<0.01).

CONCLUSIONTgN(p53mt-LMP1)/HT mice hold inherited constitutional defect in immune surveillance function, which can be aggravated by environmental carcinogens, such as DNP used even though in a much less strength. The enhanced carcinogenesis-inducing effect of DNP on TgN(p53mt-LMP1)/HT mice should be closely associated with abnormal signaling of activator protein-1 (AP-1) pathway, especially up-regulated expressions of TRAF2 and c-Jun, and down-regulated expression of p16.

Animals ; Epithelial Cells ; drug effects ; metabolism ; pathology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Mice ; Mice, Transgenic ; Mutation ; genetics ; Nasopharyngeal Neoplasms ; chemically induced ; genetics ; metabolism ; pathology ; Nitrosamines ; pharmacology ; Nose Neoplasms ; chemically induced ; genetics ; metabolism ; pathology ; Precancerous Conditions ; chemically induced ; genetics ; pathology ; TNF Receptor-Associated Factor 2 ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism

OBJECTIVETo investigate the feasibility of using natural poritos as scaffolds in bone tissue engineering (TE) and repair of caprine mandibular segmental defect with titanium reticulum reinforced.

METHODSNatural poritos with a pore of 190-230 microm in size and porosity of about 50percent-65percent was molded into the shape of granules 5 mm x 5 mm x 5 mm in size. Expanded autologous caprine marrow mesenchymal stem cells were induced by recombinant human morphogenetic protein-2 (rhBMP2) to improve osteoblastic phenotype. Then marrow derived osteoblasts were seeded into poritos in density of 4 x 10(7)/ml and incubated in vitro for 48 hours prior to implantation. Then osteoblastic cells/poritos complexes were implanted into mandibular defect and the defect was reinforced by titanium reticulum. Implantation of poritos alone acted as the control. Bone regeneration was assessed 4, 8, 16 weeks after implantation using roentgenographic analysis and histological observation was done after 16 weeks.

RESULTSNew bone could be observed histologically on the surface and in the pores of natural coral in all specimens in the cell-seeding group, whereas in the control group there was no evidence of osteogenesis process in the center of the construction. The results showed that new bone grafts were successfully restored 16 weeks after implantation.

CONCLUSIONSThis study suggests the feasibility of using porous coral as scaffold material transplanted with marrow derived osteoblasts by TE method. By means of titanium reticulum reinforcement, mandibular defect could be successfully restored. It shows the potentiality of using this method for the reconstruction of bone defect in clinic.

Animals ; Anthozoa ; Bone Marrow Cells ; Bone Morphogenetic Proteins ; Cell Culture Techniques ; Chondrogenesis ; Goats ; Mandible ; diagnostic imaging ; pathology ; surgery ; Mice ; Osteoblasts ; transplantation ; Osteogenesis ; Porosity ; Radiography ; Reconstructive Surgical Procedures ; Stents ; Tissue Engineering ; Titanium

OBJECTIVETo discuss the clinical effectiveness in treating thoracolumbar fractures adopting the rehabilitation exercise utilizing knee pads on the orthopedic traction bed.

METHODSFrom June 1996 to June 2006, we studied the clinical effectiveness of thoracolumbar fractures utilizing knee pads on the orthopedic traction bed for rehabilitation exercise. The cases surveyed total 209, 163 of which had full data. There were 98 males and 65 females with the age from 17 to 74 years (mean, 14.5 years). Consulting time after injury from 30 min to 7 days. Fracture site in T11 had 8 cases, in T12 24 cases, in L1 73 cases, in L2 33 cases, in L3 8 cases, in L4 3 cases, in T12 and L1 14 cases. Compression degree of vertebral anterior border: full compression had 1 case,more than 4/5 had 23, more than 2/3 had 67, more than 1/2 had 40, in 1/3 had 46.

RESULTSAmong them, 8 cases with legs paresis no recovery in nerval function or stopping recovery changed methods, and underwent surgical treatment. Others 155 cases were followed up from 2 to 12 years with an average of 3 years and 4 months. The average height of vertebral anterior borders of the 169 injured compressed had increased from 1.55 cm before treatment to 2.70 cm after treatment with an average of 1.15 cm. The height of the injured vertebral anterior borders had recovered from 50.5% (1.55/3.07) before treatment to 89.4% (2.70/3.02) after treatment. Kyphosis angle of the injured vertebral bodies had recovered from 13.25 degrees to -1.6 degrees in average. Twenty-three cases associated with dislocation basic reduction.

CONCLUSIONRehabilitation exercise using knee pads on the orthopedic traction bed can obtain satisfactory clinical effect in treating thoracolumbar fractures, the method is easy. At 3, 7, 10 days after treatment, the height of bed should be adjusted according X-ray.

Adolescent ; Adult ; Aged ; Exercise Therapy ; instrumentation ; methods ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; injuries ; physiopathology ; surgery ; Male ; Middle Aged ; Orthopedic Equipment ; Recovery of Function ; Spinal Fractures ; physiopathology ; rehabilitation ; surgery ; Thoracic Vertebrae ; injuries ; physiopathology ; surgery ; Treatment Outcome ; Young Adult

Objective To study the proliferation of CRTH2 (CD4+ CD294+ Th2)cells promoted by pneumococcal vaccine through antigen presentation of dendritic cells (DCs),so as to provide a new approach for amplification and sorting of Th2 cells.Methods CDs induced from peripheral blood mononuclear cells were cocultured with T lym-phocytes after loading pneumococcal vaccine antigen,mixed lymphocyte reaction (MLR)was detected by cell count-ing kit-8(CCK8),DCs and CRTH2 cells were analyzed by flow cytometry.Results Pneumococcal vaccine could promote the maturation of DCs,together with TNF-a,it was adjuvant for maturation of DCs.Pneumococcal vaccine antigen-loaded DCs could increase the rate of subsets of CRTH2 cells on day 5([0.93±0.10]%)compared with day 1([0.70±0.02]%),and absolute number also increased (both P <0.05).Conclusion Amplification of CRTH2 cells can be greatly promoted by pneumococcal vaccine antigen-loaded DCs,which might be one of the effective way to induce amplification of Th2 cells.

Programmed death-1 ligand-1(PD-L1) is a recently identified member of the B7 family molecules and is shown to mediate the inhibition of immune responses. This study was purposed to enhance the weak immunological function of dendritic cells (DCs) derived from the patients with chronic myelocytic leukemia (CML) by blockade of the expression of PD-L1. Bone marrow mononuclear cells (BMMNCs) of CML patients were induced into DCs in the presence of cytokine cocktail of rhGM-CSF, rhIL-4 and TNF-alpha. The phenotypes of DCs were detected by flow cytometry, mixed lymphocyte reaction was analyzed by MTT assay and IFN-gamma, IL-2 and IL-10 in the cell culture supernatant were detected by ELISA. The results showed that the expression of PD-L1 on CML-DCs was upregulated with the maturation of CML-DCs. PD-L1-blockaded DCs could enhance T lymphocyte proliferation, increase the secretion of IL-2 and IFN-gamma, and inhibit the production of IL-10. Taken together, PD-L1-blockaded DCs originated from CML cells had more potent immunostimulatory capability. It is concluded that PD-L1 blockaded can enhance the function of CML-DCs. This approach presents new possibilities for achieving anti-tumor immunity by DC-based vaccination.
Antigens, CD ; metabolism ; B7-H1 Antigen ; Dendritic Cells ; cytology ; immunology ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; metabolism ; Recombinant Proteins ; Tumor Necrosis Factor-alpha ; pharmacology

This study was purposed to investigate the possibility of differentiating the acute promyelocytic leukemia (APL) cells into dendritic cells (DCs) induced by all-trans retinoic acid (ATRA) combined with classic cytokines so as to provide a new approach for development of APL-DC vaccine. The bone marrow mononuclear cells from a new diagnosed patient with APL and HL-60 cells were separately cultured in complete culture medium. The cells were treated by ATRA, GM-CSF, IL-4 and TNFalpha in experimental groups and no ATRA was added in control and blank control groups. The cell morphology was observed by light microscopy, the phenotypes of DCs were detected by flow cytometry, the level of IL-12 was measured by using ELISA, the mixed lymphocyte reaction (MLR) and effect of cytotoxic T-lymphocyte (CTL) were assayed by MTT method. The results indicated that in experiment groups, the cells had dendritic appearance and cytogenetic characteristics of APL; expression of CD1a, CD83, CD80, CD86, HLA-DR and CD1d as well as level of IL-12 obviously increased; the MLR and CTL effects were significant, but increase of CD1a expression in HL60-DCs did not show statistical difference from control and blank control groups. It is concluded that ATRA can successfully induce APL cells to differentiate into functionally mature DSs which obviously mediate MLR and CTL effects. The APL-DCs derived by ATRA can notably express CD1d that may activate CD1d-restricted NKT cells and promote proliferation of NRT cells. The exact mechanism of which should be further studied.
Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Dendritic Cells ; cytology ; drug effects ; metabolism ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Tretinoin ; pharmacology

Using cDNA from Rehmannia glutinosa leaf as template, a 972 bp fragment of expansin gene which containing a 762 bp ORF that encoded 253 amino acids, was cloned, named RgEXPA10, which GenBank accession number for this gene is KF011918. A 1 207 bp genomic sequence of RgEXPA10 was amplified by PCR with leaf DNA as template, sequencing analysis revealed that three exons and two introns in RgEXPA10 genomic sequence, and which GenBank accession number is KF011919. Molecular and bioinformatic analyses indicated that RgEXPA10 protein have DPBB_1 and Pollen_allerg_1 domain, also including a 26 aa nuclear localization signal and a 19 aa transmembrane region. Phylogenetic analysis revealed that RgEXPA10 showed the highest homology with AtEXPA8 among the 26 α-expansins in Arabidopsis thaliana. However, the RgEXPA10 indicated the highest homology with the expansin from Solanum lycopersicum among 22 plant species. Expression patterns using qRT-PCR analysis showed that RgEXPA10 mainly expressed in unfolded leaf, followed by the tuberous root at stage of expanding period, and rarely expressed in senescing leaf. And RgEXPA10 showed higher expression level in tuberous root at 60 and 90 days after emergence. The transcription level of RgEXPA10 significantly reduced under all the three stresses including continuous cropping conditions, salinity and waterlogging. This study will lay foundations for molecular function in development and regulation of different stresses for R. glutinosa.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; Gene Expression Regulation, Plant ; Genes, Plant ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; Plant Proteins ; genetics ; Plant Roots ; Rehmannia ; genetics

This study was aimed to explore the role and mechanism of AML1a in abnormal hematopoiesis in mice. Plasmids pMSCV-FLAG-AML1a-IRES-YFP and pMSCV-IRES-YFP together with envelope-encoding plasmid pECO and packaging plasmid pGP were respectively transfected into 293T cells by using a method of calcium phosphate precipitation to produce retrovirus. Bone marrow mononuclear cells (BMMNC) from male C57BL/6J mice were transfected with the retroviral vector MSCV expressing FLAG-AML1a fusion protein and yellow fluorescent protein (YFP). The cells were cultured in M3434 semi-solid medium for colony formation assay and in M5300 fluid medium containing murine IL-3 (mIL-3), IL-6 (mIL-6) and SCF (mSCF) for long-term culture. The results showed that transfection of AML1a into BMMNC enhanced colony formation, colony size of the AML1a group was significantly larger than that of the control group, and the colonies were mainly composed of CFU-E and CFU-GEMM. In the long-term culture, AML1a-transfected BMMNC showed differentiation block, while the control cells were in a more mature stage. It is concluded that AML1a may block the normal hematopoiesis at the stage of primitive progenitors. At the same time, AML1a also enhances the proliferation activity of primitive progenitor cells.
Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cell Proliferation ; Colony-Forming Units Assay ; Core Binding Factor Alpha 2 Subunit ; genetics ; Genetic Vectors ; Hematopoietic Stem Cells ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Retroviridae ; genetics ; Transfection

OBJECTIVETo evaluate the effect of HSGJ on chronic allograft nephropathy (CAN) using standard rat model of CAN.

METHODRenal transplantation was performed with Fisher rats as donors and Lewis rats as recipients. All the recipients were randomly divided into control group and medication groups (high and low dosage of HSGJ, fed every other day). After 16 weeks of treatment, renal function and the histological alteration of CAN were measured. The expression of the TGFbeta1 mRNA in the allograft was evaluated by real-time PCR.

RESULTThe content of 24 h urine protein and the level of serum creatinine in the medication groups were significantly decreased (P < 0.01) as compared with control group, whereas the creatinine clearance was increased (P < 0.01). The degree of glomerular sclerosis and the Banff score of medication groups were lower than the control group respectively (P < 0.01), in consistent with decreased expression of the TGF 1mRNA.

CONCLUSIONHSGJ can prevent the chronic allograft nephropathy and the mechanism may be related with its influence on the expression of the TGFbeta1.

Animals ; Chronic Disease ; Drugs, Chinese Herbal ; therapeutic use ; Glomerulonephritis ; etiology ; immunology ; prevention & control ; Graft Rejection ; drug therapy ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; adverse effects ; Random Allocation ; Rats ; Rats, Inbred F344 ; Rats, Inbred Lew ; Transplantation, Homologous

OBJECTIVETo investigate the feasibility and safety of video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity.

METHODSThe clinical data of 120 patients who underwent esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity from March to December 2011 was analyzed retrospectively. In the video-assisted thoracoscopic surgery group, there were 60 patients [41 male and 19 female patients with aver age of (62 ± 7) years old] who underwent video-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. In the routine thoracotomy group, there were 60 patients [39 male and 21 female patients with aver age of (62 ± 9) years old] who underwent routine thoracotomy esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity. Operation time, intra-operative blood loss, postoperative total thoracic drainage in 3 days, total number of harvested lymph nodes, hospitalization, cost of hospitalization and complications were compared between the two groups.

RESULTThe operations were carried out successfully in two groups. There was no perioperative death in all patients. There was no statistical difference in intra-operative blood loss, postoperative total thoracic drainage and cost of hospitalization between the two groups. Operation time of rideo-assisted thoracoscopic surgery group was significantly longer than that of thoracotomy group ((188 ± 38) minutes vs. (138 ± 50) minutes, t = 6.171, P = 0.000), but postoperative hospitalization was significantly lower ((14 ± 3) d vs. (18 ± 6) d, t = -4.093, P = 0.000) and total number of harvested lymph nodes was lower (17 ± 9 vs. 21 ± 11, t = -2.058, P = 0.042). There was significantly statistical difference in total postoperative main complication (25.0% vs. 48.3%, χ(2) = 7.033, P = 0.008). And postoperative incisional infection of VATE group patients was significantly lower than that of thoracotomy group patients (6.7% vs. 25.0%, χ(2) = 7.566, P = 0.006).

CONCLUSIONSVideo-assisted thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity is technically feasible and safe, with minimized trauma and quick recovery. The recent result is satisfactory.

Aged ; Aged, 80 and over ; Anastomosis, Surgical ; methods ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Humans ; Lymph Node Excision ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Surgery, Video-Assisted ; Thoracotomy