Objective To use detergents and nucleic acid enzyme to prepare scaffold of extracellular ma-trix , then assess the morphological and cytotoxic changes in vitro , and explore the feasibility of this type of scaffold as an ideal tissue-engineering scaffold. Methods Fifty pieces of fresh nucleus pulposus were randomly divided into a fresh control group and a decellularized group. The specimens in decellularized group were treated with 0.3%Tri-ton X-100, 0.5%sodium deoxycholate, and nuclease for 24 h. Morphological changes were studied by macroscopy, pathological staining and scanning electron microscopy. Cytotoxicity was determined by CCK-8 and LIVE/DEAD Viability/Cytotoxicity Assay Kit in vitro. Results The shape of scaffold was maintained,and the extracellular ma-trix was presented while the cells disappeared after decellularization. As compared with the fresh tissue , the scaffold and its extracts had no cytotoxicity to rabbit bone marrow stem cells. Conclusions Almost all the cells have been removed while the extracellular matrix is reserved , and the scaffold has no cytotoxicity to the seed cells. The decel-lularized scaffold can be used as an ideal substance to fabricate tissue-engineering nucleus pulposus.

Acupuncture Points ; Adult ; Aged ; Combined Modality Therapy ; Electroacupuncture ; Exercise Therapy ; Female ; Headache ; physiopathology ; therapy ; Humans ; Male ; Middle Aged ; Neck Pain ; physiopathology ; therapy ; Range of Motion, Articular ; Treatment Outcome

Objective To observe the therapeutic effects of the co-administration of γ-aminobutyric acid (CABA), sodium dimercaptopropane sulfonate (Na-DMPS) and vitamin B6 in large doses on liver and heart of rats with acute tetramine intoxication, and compare their separate effects of either GABA or Na-DMPS alone with those of the triad combination. Method Thirty rats were randomized into control group (n = 6), tetramine intoxication without treatment group (n = 6), tetramine intoxication treated with GABA group (n = 6), tetramine intoxication treated with Na-DMPS group (n = 6) and tetramine intoxication treated with triad combination (GABA + Na-DMPS + vitamin B6, GNDV n = 6) group. Samples of blood, liver tissue and heart tissue of rats with acute tetramine intoxication were collected immediately two hours after medication with different drugs. Serum alanine aminotrasferase (ALT), aspartate aminotransferase (AST), creatine kinase (CK) and creatine kinase isoenzyme (CK-MB) were measured, and the pathological changes of liver tissue and heart tissue were observed under microscope. Results The symptoms of poisoning were apparently relieved and the latency for convulsion/muscular twitch were obviously delayed in poisoned rats treated with GABA, Na-DMPS and GNDV separately. Furthermore, combination group showed the latent period delayed longer than either GABA or Na-DMPS groups The GABA, Na-DMPS and GNDV significantly lowered the serum levels of ALT, AST, CK and CK-MB in rats with tetramine intoxication, and those serum levels of enzymes were lower in GNDV group than those in either GABA group or Na-DMPS group. However, there were no difference in those serum enzymes between GABA group and Na-DMPS group. Moreover, the intoxicated rats treated with combination treatment had the slightest pathological changes in liver and heart (GNDV < GABA or Na-DMPS). Conclusions The co-administration of γ-aminobutyric acid, sodium demercaptopropane sulfonate and vitamin B6 in large doses for the treatment of tetramine intoxication is a method of choice.

Objective To construct tissue engineering nucleus pulposus by culture of rabbit bone marrow mesenchymal stem cells (rBMSCs)-nucleus pulposus acellular matrix scaffold (NPAMS)complexes (rBMSCs-NPAMS).Methods Several NPAMS were prepared,and rBMSCs was seeded into NPAMS.The scaffolds and complex were detected by general observation,HE stai-ning,immunohistochemical,qRT-PCR,scanning electron microscopy.Results The scanning electron microscopy showed the seed cell in nucleus pulposus ECM-derived scaffold could adhesion and growth.The cell attachment and proliferation were observed by HE staining.Immunohistochemical examination with typeⅡ collagen showed positive results.qRT-PCR revealed the time-depend-ent of the mRNA expression of collagen Ⅱ,and which was smaller than positive control(P<0.01).The relative expression of Ag-grecan mRNA grew in a time dependent fashion,which was still lower than positive control(P<0.01).Scaffolds and normal nucle-us pulposus had no statistical significance of the compression load under the same displacement of the compression(P>0.05).Con-clusion Natural nucleus pulposus acellular matrix scaffold composite allogeneic bone marrow mesenchymal stem cells can be suc-cessfully built into tissue engineering nucleus pulposus.

The study was aimed to explore the possible association between HLA-DRB1 allele polymorphism and the susceptibility to leukemia in Gansu Chinese Han nationality and to find the genes susceptible to leukemia. HLA-DRB1 genes in 74 patients with leukemia from northwestern China and 82 healthy Chinese controls were determined by polymerase chain reaction and sequence-specific oligonucleotide probe hybridizations (PCR/SSO) DNA analysis. The results showed that as compared with control, the allele frequencies of HLA-DRB1*03 (chi(2) = 8.125, P = 0.004), HLA-DRB1*07 (chi(2) = 13.526, P = 0.000), HLA-DRB1*08 (chi(2) = 18.855, P = 0.000), HLA-DRB1*13 (chi(2) = 7.039, P = 0.008) significantly increased in AML group. The allele frequencies of HLA-DRB1*07 (chi(2) = 5.689, P = 0.017), HLA-DRB1*11 (chi(2) = 7.73, P = 0.005), HLA-DRB1*12 (chi(2) = 4.234, P = 0.040), HLA-DRB1*13 (chi(2) = 38.333, P = 0.000) significantly increased in CML group. The allele frequency of HLA-DRB1*01 (chi(2) = 5.294, P = 0.021) significantly increased in ALL group. It is concluded that the susceptibility to AML in Gansu Han nationality is positively related to HLA-DRB1*03. 1*07.1*08.1*13. CML positively correlates with HLA-DRB1*07. 1*11.1*12.1*13, and ALL may be positively in relation with HLA-DRB1*01. Allele polymorphism is associated with the leukemia occurrence.
Acute Disease ; Alleles ; Asian Continental Ancestry Group ; genetics ; China ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Leukemia ; ethnology ; genetics ; Polymorphism, Genetic

Background Monosodium L-glutamate (MSG) is a food flavour enhancer and its potential harmfulness to the heart remains controversial.We investigated whether MSG could induce cardiac arrhythmias and apoptosis via the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor.Methods Myocardial infarction (MI) was created by ligating the coronary artery and ventricular arrhythmias were monitored by electrocardiogram in the rat in vivo.Neonatal rat cardiomyocytes were isolated and cultured.Cell viability was estimated by 3-(4,5)-dimethylthiahiazo(-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay.Calcium mobilization was monitored by confocal microscopy.Cardiomyocyte apoptosis was evaluated by acridine orange staining,flow cytometry,DNA laddering,reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Results MSG (i.v.) decreased the heart rate at 0.5 g/kg and serious bradycardia at 1.5 g/kg,but could not induce ventricular tachyarrhythmias in normal rats in vivo.In rats with acute MI in vivo,however,MSG (1.5 g/kg,i.v.) induced ventricular tachyarrhythmias and these arrhythmias could be prevented by blocking the AMPA and N-methyl-d-aspartate (NMDA) receptors.Selectively activating the AMPA or NMDA receptor induced ventricular tachyarrhythmias in MI rats.At the cellular level,AMPA induced calcium mobilization,oxidative stress,mitochondrial dysfunction and apoptosis in cultured cardiomyocytes,especially when the AMPA receptor desensitization were blocked by cyclothiazide.The above toxic cellular effects of AMPA were abolished by AMPA receptor blockade or by H2O2 scavengers.Conclusions MSG induces bradycardia in normal rats,but triggers lethal tachyarrhythmias in myocardial infarcted rats probably by hindering AMPA receptors.AMPA receptor overstimulation also induces cardiomyocyte apoptosis,which may facilitate arrhythmia.

To investigate the rat intestinal absorption of stearic acid-octaarginine (SA-R8) modified solid lipid nanoparticles containing paclitaxel (SA-R8-PTX-SLN), compared with the commercially available preparation of PTX (Taxol) and PTX-loaded solid lipid nanoparticles (PTX-SLN), the in situ intestinal absorption of SA-R8-PTX-SLN was investigated by means of single-pass rat intestinal perfusion technique. The absorptions of the preparations were investigated at different intestinal segments, different drug concentrations and in the presence of P-glycoprotein inhibitor (verapamil). The results showed that PTX could be absorbed at each intestinal segment and the three preparations all showed maximum absorptions at the duodenum. The cumulative absorptions of three preparations at each intestinal segment appeared SA-R8-PTX-SLN > PTX-SLN > Taxol (P < 0.05). SA-R8-PTX-SLN showed a liner absorption manner at the duodenum in the examined drug concentration range. The cumulative absorptions of Taxol and PTX-SLN were significantly promoted after the addition of P-glycoprotein inhibitor (verapamil) into the preparation (P < 0.05), but absorption of SA-R8-PTX-SLN existed no significantly difference compared with the preparation without verapamil (P > 0.05). SA-R8 and SLN might both effectively improve the oral absorption of PTX in the intestinal tract.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; pharmacokinetics ; Cell-Penetrating Peptides ; chemistry ; Drug Carriers ; Intestinal Absorption ; drug effects ; Lipids ; chemistry ; Male ; Nanoparticles ; Oligopeptides ; chemistry ; Paclitaxel ; administration & dosage ; chemistry ; pharmacokinetics ; Perfusion ; Rats ; Rats, Sprague-Dawley ; Stearic Acids ; chemistry ; Verapamil ; pharmacology

Aviation ; Humans ; Physical Fitness ; Resistance Training ; Schools ; Students

OBJECTIVETo prepare monoclonal antibody (mAb) against prM epitope.

METHODSThe gene encoding prM was isolated using RT-PCR from brain of JEV infected mouse and cloned into prokaryotic expression vector pET-32a. Recombinant plasmid was transformed into E.coli BL21/DE3/LysS, then the transformed cells were expressed with the induction of IPTG. The expression and purification of the prM protein was analyzed by SDS-PAGE. The BALB/c mice were immunized with purified prM protein. Hybridoma cell lines secreting monoclonal antibodies against prM were established after cell fusion of mouse splenic cell and P3-X63-Ag8.653 cells. The specificity of mAb was identified by ELISA, Western Blot and Immunohistochemistry assay.

RESULTSmAb against prM epitope of JEV was prepared successfully.

CONCLUSIONThe obtained prM specific mAb was valuable for the prevention and dignosis of Japanese encephalitis.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; isolation & purification ; Antibody Specificity ; BALB 3T3 Cells ; Cell Line ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Encephalitis Virus, Japanese ; genetics ; immunology ; Epitopes ; immunology ; Escherichia coli ; genetics ; Mice ; Plasmids ; genetics ; metabolism ; Prokaryotic Cells ; metabolism ; Sequence Analysis, DNA ; Viral Proteins ; biosynthesis ; genetics ; immunology ; isolation & purification

OBJECTIVETo compare differences between Crowe IV developmental dysplasia of the hip (DDH) with secondary acetabulum and Crowe IV DDH without secondary acetabulum,and determine whether it is necessary to divide Crowe IV DDH into two subtypes.

METHODSFrom June 2007 to May 2015,145 hips of 112 Crowe N patients who underwent total hip arthroplasty (THA) using S-ROM stem were divided into two groups: secondary acetabulum formaton group (group A) and no secondary acetabulum formaton group (group B). In group A,there were 12 females, 96 males,with an average age of (39.38 ± 11.19) years old. In group B, there were 2 females, 35 males, with an average age of (38.19 ± 10.92) years old. All the patients were evaluated by using Harris Hip Score. Radiographic evaluations were made preoperatively and during follow up. The differences between two groups were compared on dislocation height, canal flare index (CFI), subtrochanteric shortening osteotomy (SSTO) usage, pre- and post-operation Harris scores, complications.

RESULTSThe dislocation height for group A was (4.74 ± 1.57) cm, while the dislocation height for group B was (3.12 ± 1.15) cm. Significantly difference was detected between two groups. The CFI for group A was 2.69 ± 0.68, while the CFI for group B was 3.42 ± 0.79, and the significantly difference was detected between two groups. Harris scores were totally improved from 58.18 ± 15.67 preoperatively to 91.20 ± 3.79 post-operatively and the difference was significant. Pre-operative Harris scores was 58.1 ± 15.3 in group A, 58.3 ± 16.9 in group B. Post-operative Harris scores was 91.0 ± 4.1 in group A, 91.0 ± 5.1 in group B. No significant difference was found on Harris scores between A and B preoperatively and post-operatively. Complications of 4 cases peri-prosthesis fracture, 4 cases dislocation and 4 cases nerve injury occur in group A; While only one case dislocation and one case nerve injury occur in group B. No statistical significance was detected.

CONCLUSIONCrowe IV DDH with secondary acetabulum is significantly different from Crowe IV DDH without secondary acetabulum on dislocation height and femoral morphology, which causes the different selections of surgical techniques (SSTO usage or not). These important differences in fundamental parameters indicate the necessity to further divide Crowe IV DDH into IVA and IVB two subtypes.

Adolescent ; Adult ; Aged ; Female ; Hip Dislocation, Congenital ; classification ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy