Burns, Inhalation ; Humans ; Respiratory System ; injuries

Though Runx2 and Osterix are both key transcription factors in the pathway of osteoblast differentiation, whether Runx2 positively regulates Osterix being unknown. It was showed that Runx2 induced the gene expression of Osterix both in the non-osteoblastic cell lines, either pluripotent or differentiated, and in the osteoblastic cell lines. At the same time, the results also indicated that Runx2 up-regulated the activity of the 3.2 kb human Osterix promoter. Further experiments identified a highly conserved and functional Runx2 binding site "AGTGGTT" within the promoter. Thus the results support the hypothesis that Runx2 is involved in the regulation of the Osterix gene expression. Moreover, the transient transfection and dual-luciferase assay showed Osterix up-regulated the activity of the 2.3 kb type Ⅰ collagen promoter in the non-osteoblastic cells, but Runx2 did not. This difference implies that Osterix, the down stream transcription factor of Runx2 during osteoblast differentiation, is needed to stimulate the osteoblast-specific gene expression of type Ⅰ collagen.

OBJECTIVE: To examine various pneumatized extensions of the sphenoid sinus of Chinese people. METHOD: The sphenoid sinus and its surrounding structures were examined from 100 computed tomography images of the sinus. The type of the sphenoid sinus was classified according to the various extensions of the sinus. RESULT: The type of the sphenoid sinus was classified into the following 6 basic types based on the direction of pneumatization: sphenoid body, lateral, clival, lesser wing, anterior, and combined. CONCLUSION The variations in the extensions of pneumatization of the sphenoid sinus may facilitate entry into areas bordering the sphenoid sinus.
Adult ; Asian Continental Ancestry Group ; Humans ; Sphenoid Bone ; anatomy & histology ; Sphenoid Sinus ; anatomy & histology ; Tomography, X-Ray Computed

To assess the value of CD34+ cells transferred exogenous Fas ligand (FasL) in inducing apoptosis of human leukemic cells, the CD34+ cells transfected with FasL or without, pretreated with mitomycin C, was mixed with leukemic cell line U937 cells in presence or absence of daunorubicin (DNR) or cytosine arabinoside (Ara-C). After 18 h, apoptosis of cells was detected by FCM and TUNEL. Induced for 18 h by CD34+ cells transfected with FasL or without, the ratio of apoptosis of U937 cells was (5.0 +/- 1.3)%, (10.8 +/- 0.6)% (P < 0.01), respectively. Induced by FasL+ CD34+ + DNR, FasL+ CD34+ + Ara-C, the ratio was (13.4 +/- 1.0)% (P < 0.05), (17.9 +/- 1.3)% (P < 0.01), respectively. The result demonstrated that CD34+ cells transfected with exogenous FasL could induce apoptosis of human leukemic cells and showed a cytotoxic synergistic effect when used in combination with chemotherapeutic drugs, suggesting that it was possible to develop a new method in treatment of leukemia.
Antigens, CD34 ; analysis ; Apoptosis ; drug effects ; Cell Communication ; physiology ; Cytarabine ; pharmacology ; DNA, Complementary ; genetics ; Daunorubicin ; pharmacology ; Fas Ligand Protein ; Humans ; Membrane Glycoproteins ; genetics ; Mitomycin ; pharmacology ; Transfection ; U937 Cells ; fas Receptor ; genetics ; metabolism

To investigate the value of apoptosis of the allo-antigen specific T cells induced by Fas/FasL pathway in preventing graft-versus-host disease (GVHD), the CD34+ cells transfected with FasL or not, used as stimulus cells, were mixed with allo-antigen specific T lymphocytes in presence or absence of IFN-gamma and IL-2. After 5 days, apoptosis of T cells was detected by TdT nick end mediated dUTP labeling (TUNEL) and flow cytometry (FCM). The affects of these two cytokines on CD34+ cells in the graft were also compared. The ratio of apoptosis of T cells was 12.1+/-1.5% when CD34+ cells transfected with FasL was used as stimulus cells, much higher than that of CD34+ cells non-transfected (3.2+/-1.1%, P<0.01). And in presence of IFN-gamma or IL-2, the ratio reached 20.1+/-2.3%, 17.6+/-1.3% respectively (P<0.01). However, IFN-gamma up-regulated Fas expression of CD34+ cells and increased the sensibility of CD34+ cells to soluble FasL (sFasL); IL-2 showed no such effect. It is possible to induce apoptosis of the allo-antigen specific T cells of grafts activated by allo-antigen by exogenous Fas ligand expressed on recipient cells and this might provide a new approach for preventing GVHD and IL-2 may be more suitable for clinical application.
Antigens, CD34 ; biosynthesis ; immunology ; Apoptosis ; Cytotoxicity, Immunologic ; DNA, Complementary ; genetics ; Fas Ligand Protein ; Graft vs Host Disease ; prevention & control ; Interferon-gamma ; biosynthesis ; immunology ; Interleukin-2 ; biosynthesis ; immunology ; Membrane Glycoproteins ; biosynthesis ; immunology ; T-Lymphocytes ; cytology ; physiology ; fas Receptor ; biosynthesis ; immunology

OBJECTIVETo investigate the clinical characteristics of a Chinese pedigree with cardiac conductive disease complicated by atrial fibrillation and the therapeutic effect of the treatments.

METHODSAll the family members including the proband were screened with routine examination, electrocardiography, echocardiograpy, Holter recording, chest X-ray, blood biochemistry tests and autoantibody test. The proband received dual chamber pacemaker implantation combined with oral amiodarone treatment for 3 months. The patient was monitored for thyreoid function and chest X-ray during the treatments, and was followed up for another 3 months.

RESULTSClinical evidence of organic heart disease was found in none of the family members. The proband showed recurrent dizziness and chest distress, which exacerbated after exercise, and ECG showed atrial fibrillation and severe A-V block. The proband's uncle was found to have atrial fibrillation and III degree A-V block after a syncope episode at the age of 30, and received a pacemaker treatment. Her grandpa died from a heart attack without detailed clinical documentations. No other family members showed abnormal ECG or a history of any heart events. The proband's condition was improved by treatments, after which ECG and Holter recording showed pace rhythm without atrial fibrillation.

CONCLUSIONCardiac conductive disease with atrial fibrillation can present in one family, and can be managed effectively and safely with implantation of dual chamber pacemaker combined with oral amiodarone.

Asian Continental Ancestry Group ; Atrial Fibrillation ; complications ; genetics ; therapy ; Atrioventricular Block ; complications ; genetics ; therapy ; Cardiac Resynchronization Therapy ; Female ; Heart Conduction System ; abnormalities ; Humans ; Pedigree ; Young Adult

OBJECTIVETo explore a new method of alleviating graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo-BMT) through selected elimination of mouse alloreactive T cells (ARTC) by Fas-Fas ligand (FasL) passway.

METHODSThe Sca-1(+) early hematopoietic cells (EHCs) were isolated from BALB/c mouse (H-2(d)) bone marrow mononuclear cells (BMMC) by using a high gradient magnetic cell sorting system (MACS), then transferred with exogenous mouse FasL (mFasL) gene by retroviral gene transfecting technique. Afterward the transduced EHCs were expanded in vitro for one week followed by coculture with the spleen cells from BAC mouse (H-2(d) x b) as one-way mixed lymphocyte culture (OWMLC) for 6 days, then the cytotoxicity of treated BAC mouse spleen cells against Na(2)(51)CrO(4) labelling spleen cells from BALB/c mouse was observed.

RESULTSThe Sca-1(+) EHCs were successfully isolated by MACS, with a purity of (89.0 +/- 6.1)%. After transferred with exogenous mFasL gene and expanded for one week, the transferred EHCs in the 6 day OWMLC with the spleen cells from BAC mouse at a ratio of five to one resulted in an obvious inhibition of the BAC mouse spleen cells cytotoxicity against the BALB/c mouse spleen cell at different effector/target ratios as compared to the control group (P < 0.01).

CONCLUSIONThe higher exogenous mFasL-expressing mouse EHCs can deplete ARTC against their own major histocompatibility complex (MHC) antigens in vitro.

Animals ; Antigens, Ly ; immunology ; Fas Ligand Protein ; Female ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; immunology ; Membrane Glycoproteins ; genetics ; immunology ; Membrane Proteins ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Signal Transduction ; Spleen ; cytology ; immunology ; T-Lymphocytes ; immunology ; Transfection

In order to regulate the apoptosis induced by Fas-FasL system, a soluble isoform of mouse Fas was cloned from thymocytes of immature mice with the primers designed according to the full-length Fas cDNA sequence in the GeneBank. It was directionally inserted into the intermedium vector pUC19. DNA sequencing proved that it was consistent with the expected sequence. Then it was subcloned into the eukaryotic expression vector pCA13, which was used to construct the recombinant vector pCA13-FasC. By lipofectamine (LF2000)-mediated transfection, pCA13-FasC was transfected into the 293 cells. RT-PCR and Western blot indicated that the murine soluble Fas C protein was expressed in the 293 cells. Apoptosis inducing test showed that the expression of this murine Fas C could block the Fas-induced apoptosis, which confirmed the biological activity of the recombinant Fas C.
Amino Acid Sequence ; Animals ; Animals, Newborn ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Fas Ligand Protein ; Gene Expression ; Membrane Glycoproteins ; biosynthesis ; genetics ; Mice ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Transfection ; fas Receptor ; biosynthesis ; genetics

Objective To investigate the effect of ibuprofen on autophagy and astrocyte proliferation and their significances in rats with pentylenetetrazol (PTZ)-induced epilepsy. Methods Sixty male sprague-dawley rats were randomly divided into control group, PTZ group, 3-MA+PTZ group, ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group (n=12); activity of gamma-aminobutyric acid was blocked by PTZ to ignite epileptic rats in the latter 4 groups, and rats in the control group were intraperitoneally injected with normal saline once every one day; rats in the 3-MA+PTZ group and ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg) or ibuprofen (30 mg/kg) 30 min before PTZ injection;rats in the 3-MA+ibuprofen+PTZ group were given intraperitoneal injection of 3-MA (2.5 mg/kg)+ibuprofen (30 mg/kg) at the same time. The behavior and EEG features of rats were observed. Immunofluorescence staining and Western blotting were used to detect the expressions of microtubule-associated protein 1 light chain 3 (LC3) and glial fibrillary acidic protein (GFAP). Results (1) As compared with rats in the PTZ group and 3-MA+PTZ group, rats in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group had significantly decreased seizure grading and incidence of complete ignition, and significantly increased latency period (P<0.05). (2) The EEG waveform of the control group was normal; electroencephalogram of PTZ group and 3-MA+PTZ group showed sharp waves of high amplitude and spike waves; EEG wave peaks in the ibuprofen+PTZ group and 3-MA+ibuprofen+PTZ group decreased significantly, presenting frequent small spike waves and slow spike waves, among which ibuprofen+PTZ group showed more obvious changes. (3) The results of immunofluorescence staining and Western blotting showed that as compared with the PTZ group, 3-MA+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05); as compared with the PTZ group, ibuprofen+PTZ group had statistically increased LC3 expression and significantly decreased GFAP expression (P<0.05), however, 3-MA+ibuprofen+PTZ group had statistically decreased LC3 expression and significantly increased GFAP expression (P<0.05). Conclusion Ibuprofen can reduce astrocyte proliferation by promoting autophagy to affect seizures.