OBJECTIVETo investigate the clinical effects of anterior cervical intervertebral space decompression under microscope in treating cervical spondylotic myelopathy in elderly patients.

METHODSFrom June 2009 to March 2012, 43 patients with cervical spondylotic myelopathy were treated with anterior cervical intervertebral space decompression and intervertebral fusion under microscope. There were 26 males and 17 females, aged from 60 to 72 years old with an average of (64.9±3.7) years. Japanese Orthopaedic Association System (JOA) score was from 7 to 12 points with an average of (9.5±1.8) points before operation. The function of nerves was assessed before and after operation according to JOA.

RESULTSAll patients were followed up from 10 to 18 months with an average of (14.7±1.6) months. Postoperative JOA score was (13.81±1.44) points (ranged, 10 to 16), had significantly higher than preoperative (P<0.01). According to the rate of the improved JOA score, 9 cases got excellent results, 26 good, 7 fair, 1 poor.

CONCLUSIONAnterior cervical intervertebral space decompression under microscope for cervical spondylotic myelopathy in elderly patients is safe and effective.

Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Humans ; Male ; Microscopy ; Middle Aged ; Spondylosis ; surgery

Content of osteocyte in bone tissue is the most abundant, the most widely distributed, and embedding the cells in the mineralized bone matrix, the life can be close to the life of the body. Osteocyte formed by the cytoplasm dendritic communication network system between osteocyte and bone surface, is of great significance to maintaining the normal physiological function of bone tissue. Bone cells as the direct receptor of bone mechanical stress, through the release of bioactive factors such as PEG2, NO, ATP and classic Wnt/beta-catenin signaling pathway mechanical stress signal can be converted to bone formation and bone resorption of biochemical signals, and the biochemical signals were transfer to the other type cells of the tissue to regulate its function activities and cause bone reconstruction function. The microcracks surrounding osteocyte can disrupt the microtubule network system,cause surrounding osteocyte autophagy. In addition, osteocyte is very important for regulation of the body mineral balance, fat metabolism, and hematopoietic function.
Animals ; Autophagy ; Hematopoiesis ; Humans ; Minerals ; metabolism ; Osteocytes ; physiology ; Signal Transduction ; Stress, Mechanical

Objectives To synthesize hepatitis B virus e gene which was suitable for yeast protein expression and express hepatitis B virus e antigen (HBeAg) in Pichia pastoris.Methods Using synonymous codons preferred by yeast usage on protein expression to replace some native codons of wild-type HBV e gene,the synthetic e gene (syneAg-gene) was achieved by a recursive PCR (rPCR).The syneAg- gene was inserted into the yeast expression vector pPICZ?A.The recombinant plasmid was transformed into GS115 yeast by electroporation.The yeast transformant induced by methanol expressed the HBeAg.Results The restriction analysis and DNA sequencing confirmed that the syneAg-gene was inserted to yeast pPICZ?A in correct orientation.SDS-PAGE,immunoblot and ELISA indicated that the secreted form of HBeAg was expressed by the yeast transformant.The expression level of HBeAg in the strain containing syneAg-gene was 63 mg/L.The titer of the recombinant HBeAg in culture superuatant was 1 :81 920 and the maximum OD (optical density) value of absorbance was 2.8 by ELISA.No cross reactivity between Pichia pastoris-derived HBeAg and anti-HBc antibody was found.Conclusion The recombinant HBeAg with high degree of specificity and immunoreactivity is expressed efficiently in Pichia pastoris.

Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1x10(6) ml(-1), the percentages of conidium germination and appressorium formation were (97.98+/-0.67)% and (97.88+/-0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 microg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.
DNA, Complementary ; genetics ; isolation & purification ; DNA, Fungal ; genetics ; isolation & purification ; Gene Library ; Genes, Fungal ; Magnaporthe ; genetics ; growth & development ; pathogenicity ; Oryza ; microbiology ; Plant Diseases ; microbiology ; RNA, Fungal ; genetics ; isolation & purification

In this study the MTP1 gene, encoding a type III integral transmembrane protein, was isolated from the rice blast fungus Magnaporthe oryzae. The Mtp1 protein is 520 amino acids long and is comparable to the Ytp1 protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP (green fluorescent protein) fusion expression results indicate that Mtp1 is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily expressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for pathogenicity. The Deltamtp1 mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use.
Fungal Proteins ; genetics ; physiology ; Genes, Fungal ; Magnaporthe ; genetics ; Membrane Proteins ; genetics ; Oryza ; microbiology ; Promoter Regions, Genetic

To investigate the effects of allicin on rats by NMR-based metabonomic method, the changes of endogenous metabolites in normal rat urine and the influences on metabolism were analyzed with bio-nuclear magnetic resonance (NMR) method and partial least-squares discriminant analysis (PLS-DA) after intraperitoneal administration of allicin solution. The identified biochemical effects associated with allicin dosing included elevated then gradually recovered urinary levels of Kreb's cycle intermediates, such as citrate, alpha-ketoglutarate and succinate and increased concentrations of ketones. Meanwhile, decreased urinary concentrations of glucose, lactate, alanine, hippurate and trimethylamine oxide were observed. The PLS-DA revealed that the metabonomic profiles of allicin treated groups were obviously different from those of the control group. Allicin may change metabolism significantly in normal rats. The study of the pharmacologic mechanism of allicin by metabonomic method is practicable and it could be explored further.
Animals ; Magnetic Resonance Spectroscopy ; Male ; Metabolomics ; Rats ; Rats, Sprague-Dawley ; Sulfinic Acids ; metabolism ; urine

To analyze the chemical components and decomposition products in allicin extract of garlic, the chemical components screening and identification were made with HPLC-MS/MS method by full scan TIC MS, HPLC retention time, product MS spectra and chemical reference standards. The stability of the extract in water and alcoholic solutions was also investigated. There were five major components in allicin extract which were all identified as thiosulfinates. The extract was stable for at least 3 months when stored at -20 degrees C as water solution, but obvious decomposition was observed with the increase of alcoholic concentration. The decomposition products were also identified by HPLC-MS/MS.
Chromatography, High Pressure Liquid ; Drug Stability ; Garlic ; chemistry ; Plants, Medicinal ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Sulfinic Acids ; isolation & purification ; metabolism ; Tandem Mass Spectrometry ; Thiosulfates ; analysis

Australia ; Environmental Monitoring ; Humans ; Occupational Health ; Safety Management ; methods ; organization & administration

OBJECTIVETo study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).

METHODSThe effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.

RESULTS(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.

CONCLUSIONMSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogenes ; Receptor Protein-Tyrosine Kinases ; metabolism

OBJECTIVETo observe the effects of AML1-ETO fusion gene on the transcription activity of p21WAF1/CIP1 gene. And to explore the enhancement of leukemia pathogenesis of AML1-ETO.

METHODSThe luciferase reporter plasmids of p21WAF1/CIP1 gene promoter were constructed, and co-transfected into CV-1 cells with AML1-ETO, AML1b and AML1a expression plasmids. The trans-activity of p21WAF1/CIP1 gene promoter was assayed by luminometer.

RESULTSAML1-ETO exhibited a distinct inhibition activity of p21WAF1/CIP1 gene promoter with a sequence-specificity and dosage-dependent manner. The trans-activity of p21WAF1/CIP1 gene promoter decreased to (19 +/- 4)% compared to control group, when 1000 ng pCMV5-AML1-ETO plasmid was used. AML1b and AMLla showed less inhibition activity. The trans-activity of p21WAF1/CIP1 gene promoter decreased to (61 +/- 16)% and (59 +/- 16)% compared to control group, respectively, when 1000 ng plasmid was used.

CONCLUSIONAML1-ETO exhibits more inhibition activity of p21WAF1/CIP1 gene promoter than AML1b and AMLla, results from recruiting transcription co-repression complex efficiently by ETO. Based on previous researches, the effects of exogenous AML1-ETO on p21WAF1/CIP1 gene promote may be dependent on the type of cell lines.

Animals ; Cell Line ; Core Binding Factor Alpha 2 Subunit ; genetics ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; Haplorhini ; Oncogene Proteins, Fusion ; genetics ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics ; RUNX1 Translocation Partner 1 Protein ; Transcription, Genetic ; Transfection