Objective To explore the mechanism of spontaneous seizures in adaptor protein complex type 3B knockout mice(AP3M2KO mice).Methods AP3M2KO mice were generated.Seizures and electroencephalogram(EEG)were monitored using video camera and telemetry system.Glutamate and GABA releases were determined using in vivo microdialysis method.Results AP3M2KO mice began to suffer from spontaneous seizures 8 weeks after the birth,but did not show any other behavior abnormality.The onset of ictal discharge over the temporal region was synchronized with seizures.There were no significant differences in basal glutamate and GABA releases in hippocampus between AP3M2KO((0.35±0.08)pmol/20μl and(2.94±1.69)fmol/20μl,respectively)and wild-type mice.However,the 50 mmol/L K+-evoked GABA release was impaired in AP3M2KO mice((63.5±11.8)fmol/20μl vs(209.2±63.7)fmol/20 μl,t=4.405,P＜0.05),whereas no significant difference was found in K+-evoked glutamate release.Conclusions AP3M2KO mice suffer from epileptic seizures similar to the clinical features of human epilepsy.The impairment of inhibitory GABAergic transmission iS involved in the mechanism of spontaneous seizures in AP3M2KO mice.

Endothelial-to-mesenchymal transition(EndoMT)is a change in the transformation or differentiation of endothelial cells into mesenchymal cells under physiological or pathological conditions, accompanied by changes in phenotype and function, and is an important part of fiber repair. It is widely involved in the pathophysiological process of embryonic development, tumor invasion and a variety of fibrotic diseases. Research on the role of EndoMT in ocular diseases has also made some progress. This article will review the basic biological characteristics, mechanism and research results of EndoMT in ophthalmological diseases, intending to theoretically reveal its possibility as a treatment target and a key point of regenerative medicine technology in related diseases, provide a reference for clinical practice and scientific research.

OBJECTIVE: The aim of this study was to induce oral submucous fibrosis (OSF) in Sprague-Dawley(SD) rat models by arecoline and mechanical stimulation. METHODS: Two factors factorial design was used to divide 48 rats into 8 groups (n=6). Different concentrations of arecoline (0, 0.5, 2, and 8 mg·mL⁻¹) and mechanical stimulation (with or without brush) were treated. After 16 weeks of treatment, the mouth opening was measured, the pathological changes of the buccal mucosa were observed, and the expressions of type Ⅲ collagen, transforming growth factor β1 (TGF-β1), and interferon-γ (IFN-γ) were detected. RESULTS: In rats with moderate and high concentrations of arecoline, typical OSF pathological features were observed in the buccal mucosa, the mouth openings were significantly reduced, and the expression levels of type Ⅲ colla-gen and TGF-β1 were significantly increased (P<0.05). Although mechanical stimulation can increase the three indexes of mucosa (P<0.05), no pathological change and difference in the mouth opening was observed (P>0.05). CONCLUSIONS Moderate and high concentrations of arecoline can induce OSF in SD rats, but mechanical stimulation cannot induce OSF.
Animals ; Arecoline ; pharmacology ; Fibroblasts ; Mouth Mucosa ; Oral Submucous Fibrosis ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the role of spinal MAPK-ERK signal pathway in myocardial ischemia-reperfusion (I/R) injury.

METHODSSixty male Sprague-Dawley(SD) rats (80-100 g) were randomly divided into 3 groups: sham (n=10), PD98059 (n=25) and I/R groups (n=25). Three days after successful intrathecal implantation, 5 μg DMSO was injected intrathecally into the sham group, and then the left coronary arteries were separated without being tied. Rats in the I/R and PD98059 groups were injected with 5 μL DMSO and PD98059 (5 μg) 30 minutes before thoractomy respectively. Then the left coronary artery was tied for 30 minutes followed by 120 minutes of reperfusion. After the experiments, the ERK phosphorylation condition of T1-T4 spinal cord segments was detected with immunofluorescence; the myocardiac apoptosic index and infarct size were measured.

RESULTSExpression of p-ERK in the I/R group was significantly higher than in the sham and PD98059 groups (P<0.05). Myocardial apoptotic index and infarct size in the PD98059 group were significantly lower than in the I/R group (P<0.05), but higher in the PD98059 group than in the sham group (P<0.05).

CONCLUSIONSThe MAPK-ERK pathway in the superior thorathic spinal cord can be activated by myocardial ischemia-reperfusion and inhibition of the pathway can play a protective role in myocardial ischemia-reperfusion injury.

Animals ; Apoptosis ; drug effects ; Flavonoids ; pharmacology ; therapeutic use ; MAP Kinase Signaling System ; physiology ; Male ; Myocardial Infarction ; drug therapy ; Myocardial Reperfusion Injury ; etiology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; Spinal Cord ; physiology

OBJECTIVETo explore the mechanism of anti-proliferative effect of indomethacin (IN) on chronic myelogenous leukemia (CML) cells.

METHODSMTT was applied to assay CML cells viability under IN intervention. STAT1, STAT5 proteins were analyzed by Western blot, the expressions of phosphorylated STAT1 or STAT5 by immunoprecipitation combined with Western blot, the cellular localization of p-STATs proteins by indirect immunofluorescence technique, and the detection of Bcl-X(L) and COX-2 protein by Western blot.

RESULTSIN could significantly inhibit the viability of CML cells. 0 approximately 400 micromol/L of IN could down-regulate the expression of p-STAT1 or p-STAT5 in a dose-response manner, p-STATs were distributed mainly in the nucleus as scattering spots. The expression of COX-2 protein could be detected in K562 cells. Both Bcl-X(L) and COX-2 proteins could be inhibited by IN in a dose-dependent manner.

CONCLUSIONSIN could significantly inhibit the proliferation of CML cells, the mechanism of which might be related to the suppression of STATs/Bcl-X(L) signal transduction pathway. There exists COX-2 protein expression in K562 cells, the anti-leukemia effect of IN was possibly dependent on COX-2 pathway.

Blotting, Western ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Down-Regulation ; drug effects ; Humans ; Indomethacin ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Microscopy, Fluorescence ; Phosphorylation ; drug effects ; STAT1 Transcription Factor ; metabolism ; STAT5 Transcription Factor ; metabolism ; bcl-X Protein ; metabolism

OBJECTIVETo identify hemin-induced gene expression in K562 cells.

METHODSPoly A(+) RNAs were isolated from hemin-induced (tester) and non-induced K562 cells (driver) respectively, and double-strand cDNAs were synthesized by reverse transcription. The forward subtracted cDNA library was constructed by using suppression subtractive hybridization (SSH) techniques. The recombinant plasmids were extracted and the positive clones were identified by EcoR I digestion after the amplification and screening of the library. The inserts were amplified by PCR. The upregulated cDNA transcripts were identified by reverse dot blot hybridization, DNA sequencing and homology analysis with GenBank database "blast" respectively.

RESULTSFifteen upregulated clones were identified and most of them were homologous to the mRNA sequences of protein with known function, including globin epsilon1, glutathione S-transferase like glutathione transferase Omega (GSTTLp28), selenoprotein X1 (SEPX1), triosephosphate isomerase (TPI1), ribosomal protein L7 (RPL7), ribosomal protein S13 (RPS13), ferritin light polypeptide, globin A gamma, RAD 51 homolog C(RAD51C), ferritin heavy polypeptide, X-box binding protein (XBP1). A part of the hemin-induced cDNA clones exhibited sequence similarities to that of the GenBank registered mRNA with unknown function of their expressed proteins, including the cDNA clones of DKFZp434I116, hypothetical protein HSPC014 and NOL1R2 proteins.

CONCLUSIONSHemin mainly induces the genes expression related to erythroid differentiation, protein synthesis and metabolism in K562 cell. There results provide comprehensive information useful for the differential gene expression in hemin-induced erythroid differentiation and for further function study of genes involved in hematopoiesis.

Blotting, Northern ; DNA, Complementary ; drug effects ; genetics ; Gene Expression ; drug effects ; Hemin ; pharmacology ; Humans ; K562 Cells ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction

OBJECTIVE: To evaluate the protective effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) on acute hepatic failure induced by galactosamine (D-GalN) and lipopolysaccharide (LPS) in mice, and to explore its mechanism. METHODS: The mice were intraperitoneally administered D-GalN (800 mg/kg) and LPS (10 microg/kg), and then were intraperitoneally injected either saline (the control group )or rhG-CSF at 300 microg/kg body weight (the therapy group) at 4 h, 2 h and 0 h before the D-GalN/LPS injection. The survival rate of the mice was estimated at 24 h after the D-GalN/LPS injection. The degree of hepatic injury was evaluated at 6 h after the D-GalN/LPS injection, and the levels of TNF-alpha, IFN-gamma, IL-6 and IL-10 mRNA were simultaneously measured by semiquantitative RT-PCR. RESULTS: The survival rate of the therapy group was significantly higher than that of the control group (68.4% vs 20%, P<0.01). As compared with the control group, the degree of liver injury in the therapy group significantly decreased (P<0.05), and the levels of TNF-alpha and IFN-gamma mRNA in the hepatic tissue also reduced remarkably (P<0.01, respectively), while the levels of IL-6 and IL-10 mRNA increased (P<0.01, respectively) at 6 h after the D-GalN/LPS injection. CONCLUSION G-CSF can protect the mice from acute hepatic failure induced by D-GalN/LPS.
Animals ; Galactosamine ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Lipopolysaccharides ; Liver Failure, Acute ; chemically induced ; drug therapy ; Male ; Mice ; Protective Agents ; therapeutic use ; Random Allocation ; Recombinant Proteins

Because of the versatility of autologous cartilage and proven advantages, it has been widely used in all kinds of rhinoplasty. Autologous cartilage have good biocompatibility with less complications such as infection, extrusion of graft, compared to biological synthetic materials, so it is generally considered the primary graft for nasal surgery especially septal, conchal and costal cartilage. With the popular clinical application of autologous cartilage, problem also arised and considered.The purpose of this paper is to review the application of autologous cartilage and complications in rhinoplasty.