Objective To investigate the effect of a tissue engineered trachea for replacement fabricated using three dimensional scaffold and chondrocytes by in vitro and in vivo culturing. Methods Rib chondrocytes were isolated and expanded to two passages, then seeded in PLGA or Dacron scaffold at density of 5 × 107/ml. Cultured in vitro for two weeks, the chondrocytes-scaffold model was planted under dorsal skin between nude mice's spine. Histology of cartilage, neovascularization and organizational structure were observed with HE staining, PAS staining and electron microscopic scan were performed after 4,6,8 weeks in vivo. Results Organized structure were observed in both PLGA-chondrocyte model and dacron-chondrocyte model with cartilage formation, neovascularization and tight fibrous connective tissue between scaffold and skin after in vitro and in vivo culture. Conclusion Tissue engineered trachea fabricated using rib chondrocytes and PLGA or dacron scaffold with in vitro and in vivo culture meets the requirement of trachea replacement.

Carrier State ; virology ; Female ; Hepatitis B ; complications ; Humans ; Lymphoma, Large B-Cell, Diffuse ; virology ; Male ; Middle Aged

Objective To investigate the effect of glucocortieoids on tenasein C(TNC) expression in nasal polyp tissues and airway epithelia,and explore the possible mechanism of glucocorticoids inhibiting remodeling of nasal polyp tissue.Methods The enzyme linked immunosorbent assay(ELISA) was used to detect the protein levels of tenascin C and transforming growth factor-β1(TGF-β1) in nasal polyp tissues from intranasal glucocorticoids (Budesonide,BUD) treated group and untreated group.The cell culture,real-time RT-PCR and in situ ELISA were employed to investigate the regulatory effects of budesonide on the TNC mRNA and protein expression in BEAS-2B immortalized human bronchial epithelial cells.Results The protein levels of TNC and TGF-β1 were decreased in nasal polyp tissues from intranasal BUD-treated group as compared with untreated group(P<0.01).There was a significantly positive correlation between TNC and TGF-β1 protein levels in nasal polyp tissues (r =0.68,P<0.01).After preincubation with BUD,TNC mRNA and protein expression induced by TGF-β1 in BEAS-2B cells was significantly decreased(P<0.01).Conclusion Glucocorticoids might participate in the regulation of tissue remodeling in nasal polyp by inhibiting the TGF-β1 expression in nasal polyp tissue and suppressing the induction of TGF-β1 to up-regulatie the TNC expression in airway epithelia.

A new eremophilane derivative, (3aR,4R,5S,7S,7aS)-2-acetyl-7,7a-dihydroxy-3a,4-dimethyl-3a,4,5,6,7,7a-hexahydro-3H-inden-5-yl acetate (1) and three known compounds, 10beta-hydroxy-eremophil-7 (11)-en-12,8alpha-olide(2), beta-sitosterol (3) and beta-daucosterol(4) have been isolated from Ligularia intermedia. The compounds were isolated by column chromatography on silica gel and Sephadex LH-20,and identified on the basis of spectral analyses (MS, 1H-NMR, 13C-NMR).
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Naphthalenes ; chemistry ; isolation & purification ; Sesquiterpenes

Objective To investigate the current status of endemic fluorosis in Shandong province, and to provide scientific evidence for the development of control strategies. Methods According to "The National Technical Scheme for Endemic Disease Control in 2007", 19 counties were chosen to carry out the epidemiological investigation in 2008. Water and urinary fluoride were determined by F-ion selective electrode, dental fluorosis of children aged 8 to 12 were diagnosed by Dean method and skeletal fluorosis of adults over the age of 16 were examined clinically and by X-rays. Results In 19 counties, 186 villages were surveyed, 44 villages were found with mean water fluoride ≤ 1.00 mg/L, accounting for 23.66%(44/186);the value ＞ 1.00 mg/L in 142 villages,accounting for 76.34% (142/186);maximum water fluoride 8.88 mg/L. Total detection rate of dental fluorosis of children aged 8 to 12 was 66.35% (4518/6809), dental fluorosis index was 1.55, and defect rate was 15.39%(1048/6809). Children with urinary fluoride ＞ 1.40 mg/L was 83.29%(2149/2580), and the maximum value was 31.92 mg/L. Detection rates of skeletal fluorosis clinically and by X ray among adults over 16 years were 6.37%(5577/87 607) and 20.23% (229/1132), respectively. Conclusions Endemic fluorosis in Shandong province is still serious, prevention efforts need to be further increased.

A new sesquiterpenoid, 8α-hydroxy-6β-methoxy-1-oxoeremophila-7 (11), 9 (10) -diene-12, 8-olide (1) and five known compounds, petasin (2), caffeic acid (3), hepta-cosanol (4), β-sitosterol (5) and β-daucosterol (6) have been isolated from the roots of Ligularia intermedia. The compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR).
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Imaging ; Molecular Structure ; Plant Roots ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo investigate the osteogenesis mechanism by analysis of the expression of insulin-like growth factor-I (IGF-1) and alkaline phosphatase (ALP) in the reconstruction of cleft palate (CP) with distraction osteogenesis (DO) in rhesus.

METHODSThe CP animal models were established surgically. 21 rhesus in experimental group underwent DO to close the soft and bony defect, followed by consolidations. Every 3 animals were killed and the specimen were taken out after consolidation of 1, 2, 4, 6, 8, 12, 24 weeks. The mRNA of IGF-1 and ALP were detected with Real-time RT-PCR technique. The expression of IGF-1 and ALP was quantitatively analyzed by ELISA. The results were compared with those in control and sham groups (each of 2 animals), respectively.

RESULTSSince consolidation, the mRNA of IGF-1 and ALP increased significantly at one week and reached the peak at two weeks, but decrease to control level after 12 weeks of consolidation. The expression of IGF-1 also increased to peak level after two weeks of consolidation. The expression of ALT increased significantly since consolidation and reach the peak value after six weeks. They all decreased to nearly control level after 8-12 weeks.

CONCLUSIONSThe palate cleft can be successfully closed with new formed bone after DO. The mechanism of bone consolidation is intramembranous bone formation.

Alkaline Phosphatase ; metabolism ; Animals ; Bone Regeneration ; Cleft Palate ; metabolism ; surgery ; Insulin-Like Growth Factor I ; metabolism ; Macaca ; Osteogenesis, Distraction ; Postoperative Period

OBJECTIVEAllogeneic bone marrow transplantation (Allo-BMT) has improved long-term survival in children patients with refractory leukemia. For patients who do not have an HLA-identical sibling, the treatment option is limited. Using related mismatch donors or parental donors for Allo-BMT was at high risk for acute severe GVHD. The purpose of this study was to explore the effects of CD25 monoclonal antibody on reducing the incidence of severe acute GVHD in haploidentical bone marrow transplantation for the treatment of childhood leukemia.

METHODSTen children with leukemia received haplotype Allo-BMT from HLA two or three loci mismatched related donors. Most patients were classified as in high risk category. The donors of patients were given G-CSF (Lenograstim Chugai) 250 microg/day for seven doses prior to marrow harvest. The non-T-cell depleted haploidentical bone marrow was infused. In addition to combination of CSA, MTX, ATG (Fresenius Hemocare, Germany) and mycophenolate mofetil (MMF) for GVHD prophylaxis, CD(25) monoclonal antibody (Simulect, Novartis Pharma Switzerland) was administered to prevent acute severe GVHD.A total of 40 mg Simulect was given in two doses of 20 mg each by 30 min intravenous infusion 2 h before transplantation and day 4 after transplantation. The outcomes were compared with those of 8 children patients with leukemia who received haploidentical bone marrow transplantation without CD(25) antibody for GVHD prophylaxis.

RESULTSAll patients were engrafted and sustained full donor-type engraftment. 100% donors hematopoietic cells after transplantation was determined by cytogenetic evidence analysis. The median days of granulocyte exceeding 0.5 x 10(9)/L and pallets exceeding 20 x 10(9)/L were 19 and 22 days, respectively. Patients were monitored till up to days 100 for the sign of aGVHD. None developed the grade II-IV acute GVHD. However, the incidence of the grade II-IV acute GVHD in control group was 50% (P = 0.0147). Eight patients could be evaluated for chronic GVHD. All experienced chronic GVHD confined to the skin. The median follow-up duration was 12 months (range 9 - 24 months). Two patients died from transplant related mortality, one had patient relapsed disease and died. The remaining seven patients were alive in disease-free situation.

CONCLUSIONThe use of Simulect in haploidentical bone marrow transplantation for the treatment of children patients with leukemia is effective for preventing acute severe GVHD and may reduce transplant-related mortality.

Acute Disease ; Adolescent ; Adult ; Antibodies, Monoclonal ; therapeutic use ; Bone Marrow Transplantation ; adverse effects ; methods ; Child ; Family ; Female ; Graft vs Host Disease ; blood ; etiology ; prevention & control ; Humans ; Immunosuppressive Agents ; therapeutic use ; Leukemia ; mortality ; therapy ; Male ; Middle Aged ; Receptors, Interleukin-2 ; immunology ; Tissue Donors ; Transplantation, Homologous ; Treatment Outcome

To investigate the properties of haploidentical donor-derived bone marrow engraftment and hematopoietic reconstitution in patients received bone marrow transplantation, 15 patients with leukemia received bone marrow grafts without T cell depletion from their family donors of those with 2-3 mismatched loci of HLA antigens. The donors were given G-CSF 250 micro g/day for 7 days prior to marrow harvest. All patients were treated with conditioning regimens consisting of high-dose of Ara-C, cyclophosphamide, and total body irradiation. A four-agent based GVHD prophylaxis was used as cyclosporine A, MTX, ATG and mycophenolate mofetile (MMF). Donor engraftment was evaluated as identification of HLA locus, chromosome karyotype, DNA fingerprinting, blood type and other parameters such as occurrence of GVHD, recovery of peripheral blood cell counts as well as normal myelogram. The results showed that successful and stable engraftment was established in all patients. The median time of granulocyte counts > 0.5 x 10(9)/L and platelet > 20 x 10(9)/L was 18 (13-23) and 22 (16-32) days, respectively. One of the patients relapsed despite the bone marrow chimerism appearing after transplantation. The grade I acute GVHD occurred in 8 and grade II-IV in 5 of the 15 patients. Of the patients, 7 received marrow grafts from donors of opposite sex were identified for donor engraftment by chromosome analysis, 4 by blood typing, 7 with HLA locus analysis and 1 with DNA fingerprinting. In conclusion, HLA haploidentical bone marrow transplantation is feasible with a series of management including mobilization with G-CSF in donors, intensive conditioning and proper immunosuppressants, which enable the allo-transplants to stride across the immunological barrier.
Adolescent ; Adult ; Bone Marrow Transplantation ; Child ; Female ; Graft vs Host Disease ; etiology ; Haplotypes ; Hematopoiesis ; Histocompatibility Testing ; Humans ; Leukemia ; blood ; therapy ; Male

This study was aimed to detect the changes of bcr/able gene level in ph+ CML patients at different stages after allo-HSCT by real-time quantitative PCR and to evaluate the significance of this detection. The serial detection of bcr/abl fusion gene levels in 21 cases of CML treated with allo-HSCT was performed by RQ-PCR. The results showed that the bcr/able fusion gene could not be detected in 7 out 21 CML cases with positive fusion gene after allo-HSCT, while the bcr/abl fusion gene of different levels could be detected in 14 cases within 1-6 months. Dynamic detection indicated that the bcr/abl fusion gene levels in 9 cases were lower with relative value 0.0074%-0.088% and then could not be detected within 3-7 months after allo-HSCT. The bcr/abl fusion gene levels in 5 cases diagnosed as molecular relapse were between 0.077%-75%. The bcr/abl fusion gene levels in 1 out of 5 cases were 0.95%, 1.5%, and 0.16% in month 1, 2 and 3, respectively, and turned to negative in the month 4 without any treatment after allo-HSCT. 2 cases received the donor peripheral blood stem cell infusion, and then their bcr/abl mRNA levels could not be detected in bone marrow. Another 2 cases developed to the hematologic relapse, 1 out of 2 cases reached CR again after infusion of donor peripheral blood stem cells and chemotherapy, the other one died. It is concluded that serial quantifications of bcr/abl mRNA levels by RQ-PCR are reliable and can be used to detect the MRD, to monitor the outcome and to predict the relapse.
Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genes, abl ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; surgery ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult