Acute Disease ; Adult ; Benzene ; poisoning ; Female ; Humans ; Leukemia ; chemically induced ; Occupational Exposure ; adverse effects

Objective:To investigate the influence of sCD80-Linker-sCD40L fusion protein on the unspecific anti- tumor immunity in vitro.Methods:Ovarian cancer SKOV3 cells were separately transfected with recombinant adenoviral vectors containing sCD80-Linker-sCD40L fusion gene,sCD80 gene,sCD40L gene or with control adenovirus.The expres- sion of the sCD80-Linker-sCD40L fusion protein,sCD80 protein and sCD40L protein in the supernatants of SKOV3 cells was determined by ELISA.Dendritic cells(DCs)were cultured with peripheral blood mononuclear cells from a patient with ovarian carcinoma.DCs and autologous T cells were co-cuhured and were exposed to different supernatants for 48 h. The allostimulatory effects of DCs on T cells were determined by mixed lymphocyte reaction(MLR).The unspecific kill- ing activities of induced T cells against SKOV3/K562 cells were measured by LDH-releasing assay.Results:ELISA assay showed that levels of the sCD80-Linker-sCD40L fusion protein,sCD80 protein and sCD40L protein in the supernatants of transfeced SKOV3 cells were 2.791 ng/ml,1.956 ng/ml and 1.407 ng/ml,respectively.The fusion protein-exposed DCs ([0.382?0.053]vs[0.167?0.028],P

OBJECTIVETo evaluate the feasibility and efficacy of unilateral pedicle screw fixation in treating thoracolumbar fractures through paraspinal approach.

METHODSFrom January 2006 to January 2009,21 patients with single level thoracolumbar fracture without neurological symptoms were treated with unilateral pedicle screw fixation through paraspinal approach. There were 14 males and 7 females,aged from 21 to 65 years old with a mean of 36.4 years. The duration from injury to operation ranged from 6 h to 5 d with an average of 3 d. According to the classification of Denis fracture, compression fractures happedned in 12 cases and burst fractures happened in 9 cases,including 1 case with T5 fracture, 2 cases with T7 fracture, 2 cases with T10 fracture, 3 cases with T11 fracture, 8 cases with T12 fracture, and 5 cases with L1 fracture. Based on the Flankel grade, all patients were classified as grade E. Anterior vertebral body height ratio, sagittal Cobb angle, condition of internal fixation failure, visual analogue score (VAS) were evaluated.

RESULTSAll patients were followed up from 12 to 36 months with an average of 20.5 months. No internal fixation failure was found. Anterior vertebral body height ratios at preoperative 3 days after operation and last follow-up were 54.3 +/- 2.8, 92.9 +/- 1.5, 93.8 +/- 1.7, respectively;sagittal Cobb angle at the three timepoints were (27.8 +/- 2.5) degrees, (5.3 +/- 0.8) degrees, (6.3 +/- 1.4) degrees, respectively; the difference was statistical significant (P < 0.05). VAS was (1.2 +/- 0.4) points at last follow-up and had obviously improved (P < 0.05).

CONCLUSIONTreatment of thoracolumbar fractures with unilateral pedicle screw fixation through paraspinal approach is safe with the advantages of micro-trauma and less blood loss,which can not only completely retain the posterior spinal complex structure, reinforce the spinal stability, raise the reductional quality, but also improve the strength of fixation and the distribution of stress force.

Adult ; Bone Screws ; Feasibility Studies ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Spinal Fractures ; diagnostic imaging ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Young Adult

Acid Anhydride Hydrolases ; Carcinoma, Hepatocellular ; etiology ; genetics ; Humans ; Liver Neoplasms ; etiology ; genetics ; Mutation ; Neoplasm Proteins ; genetics ; RNA, Messenger ; analysis

Objective To investigate the clinical,neuroimaging and myopathological features of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes(MELAS).Methods The clinical manifestations,neuroimaging and myopathological features of 31 patients with MELAS diagnosed in our Neuromuscular Center in the recent 7 years were retrospectively analyzed.A3243G point mutations were analyzed by RFLP method in 10 patients.Results ①Clinical features:There were 18 male patients and 13 female patients.The age of onset ranged from 3 to 43 years,averaging 21.9 years.The averaged duration was 4.9 years.Thirteen patients in this group had family history of maternal inheritance pattern.The main clinical manifestations included short stature(26 patients),recurrent headache and vomiting(24 patients), muscle weakness(22 patients),epileptic seizure(21 patients),cognitive decline(19 patients),visual disturbance(17 patients),sensorineural deafness(16 patients),ataxia(6 patients),psychiatric symptom (8 patients),external ophathalmoplegia(2 patients)and diabetes mellitus(9 patients).The serum CK level was slightly elevated in 6 patients,and the fasting blood lactic acid was increased in 15 of the 18 detected patients.②Neuroimaging features:The stroke-like lesions were mostly confined to cerebral cortex, including temporal lobe(24 patients),occipital lobe(21 patients),parietal lobe(12 patients)and frontal lobe(4 patients).Three patients had deep white matter involvement.Migrating stroke-like lesions were confirmed in 4 patients by repeated cranial CT/MRI examination.In addition,cerebral atrophy(17 patients)and bilateral basilar ganglion calcification(11 patients)were found.③Myopathological features: Scattered ragged red fibers(RRF)in various number were found in all the patients by MGT staining.Other founding included strongly SDH-reactive blood vessel(27 patients),COX enzyme deficiency(19 patients), and mild to moderate lipid storage in RRF(20 patients).④MtDNA analysis showed 9 patients with A3243G point mutation in all the detected 13 patients.Conclusion The clinical and neuroimaging features may offer important clue to the diagnosis of MELAS,but a definite diagnosis of MELAS relies on the myopathology and mtDNA mutation analysis.

To find anti-hypertensive lead drug, angiotensin converting enzyme (ACE) inhibitory peptides were synthesized and their effects on inhibiting ACE activity were investigated. ACE inhibitory peptides were synthesized via Fmoc solid-phase synthesis, isolated and purified through reversed phase high-performance liquid chromatography (RP-HPLC), and identified by mass spectrometry. A RP-HPLC analysis method was used to test ACE inhibitory activity in vitro of these ACE inhibitory peptides. Six octapeptides were successfully synthesized, and the analytical results of mass spectrum were consistent with their theoretically calculated data. Among these synthetic octapeptides, the anti-SARS (severe acute respiratory syndromes) octapeptide had the most obvious ACE inhibitory activity with an IC50 value of 3.4 x 10(-5) mol x L(-1). So octapeptide AVLQSGFR-OH (anti-SARS peptide) was found to be the strongest candidate for potential development as an anti-hypertensive drug and had the implication of further study.
Angiotensin-Converting Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Antihypertensive Agents ; chemical synthesis ; chemistry ; pharmacology ; Chromatography, High Pressure Liquid ; methods ; Mass Spectrometry ; Molecular Structure ; Oligopeptides ; chemical synthesis ; chemistry ; pharmacology ; Peptidyl-Dipeptidase A ; drug effects ; Solid-Phase Synthesis Techniques

The study aims to screen the ability of the bacteria to metabolize ononin and assess the effect of ononin on the intestinal bacteria. Fresh human fecal sample was obtained from a healthy volunteer, diluted serially in sterile water and sixty-nine different bacterial colonies were picked out ultimately. UPLC-Q-TOF/MS with automated data analysis software (MetaboLynx) was applied to fast analysis of ononin metabolites. Furthermore, an E(max) precision microplate reader was employed to determine the growth situation of Enterococcous sp., Enterobacter sp., Lactobacilli sp., and Bifidobacteria sp. Results indicated that hydrogenation, demethylation, hydroxylation and deglycosylation were the major metabolic pathways of ononin by human intestinal bacteria in vitro. Ononin can inhibit the growth of pathogen such as Enterococcus sp., Enterobacter sp. and can promote the growth of probiotics such as Bifidobacteria sp. and Lactobacilli sp. This study suggested that intestinal bacteria have the metabolic effects of ononin and the biotransformation was completed by different bacteria. And ononin can affect the balance of intestinal flora and the degree of influence varies depending on the bacterial species and the concentration of ononin.
Bacteria ; metabolism ; Biotransformation ; Feces ; microbiology ; Glucosides ; metabolism ; Humans ; Intestines ; microbiology ; Isoflavones ; metabolism ; Metabolic Networks and Pathways

AIM:To study the regulation of human chromosome 1 open reading frame 109(c1orf109)gene by transcription factor Myc-associated zinc-finger protein(MAZ)in vitro.METHODS:In vitro study,electrophoretic mobili-ty shift assay was performed to screen the binding sites of MAZ in the promoter region of c1orf109 gene.The HeLa cells were co-transfected with the enhanced green fluorescent protein reporter vector driven by c1orf109 promoter,and MAZ and transcription factor specificity protein 1(Sp1)expression plasmids.After 24 h,the transcriptional expression of c1orf109 gene in the co-transfected cells was determined by confocal scanning microscopy and flow cytometry.RESULTS:The c1orf109 promoter could bind to MAZ ,and shared the binding sites with Sp 1.MAZ and Sp1 both inhibited the transcrip-tional expression of c1orf109 gene and the inhibition effect of Sp1 was greater than MAZ(P<0.05).CONCLUSION:Both MAZ and Sp1 regulate the expression of c1orf109 gene in physiological and pathological conditions ,and this regulation is redundancy with the same direction.The existence of redundancy transcriptional regulation manner of this gene suggests that precise regulation of c1orf109 gene is vital important for cell biological processing.

Naringin has been reported to possess a wild range of biological activities. However, the route and metabolites of naringin produced by intestinal bacteria are not well understood. In this paper, different bacteria were isolated from human feces and their abilities to convert naringin to different metabolites were studied. Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) with automated data analysis software (MetaboLynx) was applied to fast analysis of naringin metabolites. Using MSE and mass defect filter techniques, three metabolites were detected and tentatively identified. The results indicated that acetylation, hydrolyzation and hydrolyzation with hydrogenation were the major metabolic pathways of naringin in vitro. Then, we studied the gene sequence of the 16S rRNA of the bacteria by extraction of genomic DNA of the strain, PCR amplification and clone of the 16S rRNA. The consequence proved that Enterococcus sp.30, Bacillus sp.46, Escherichia sp.54 and Escherichia sp.63 have the peculiar metabolism characteristic of naringin.
Bacillus ; genetics ; isolation & purification ; metabolism ; Chromatography, High Pressure Liquid ; Enterococcus ; genetics ; isolation & purification ; metabolism ; Escherichia ; genetics ; isolation & purification ; metabolism ; Feces ; microbiology ; Female ; Flavanones ; metabolism ; Humans ; Metabolic Networks and Pathways ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

BACKGROUNDIn our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA (cDNA) chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma (MM) patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM.

METHODSThe full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples.

RESULTSDAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C (PKC) phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients (P < 0.001) that matched with the results of the RT-PCR.

CONCLUSIONSDAZAP2 is downregulated in MM samples and it may be a signal molecule in MM cells. DAZAP2 is involved in the pathogenesis of MM and could be used as a genetic marker for MM.

Adult ; Aged ; Amino Acid Sequence ; Blotting, Western ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; etiology ; metabolism ; RNA, Messenger ; analysis ; RNA-Binding Proteins ; analysis ; chemistry ; genetics