Arthritis, Juvenile ; diagnosis ; pathology ; Biomarkers, Tumor ; blood ; Biopsy ; Bone Neoplasms ; diagnosis ; pathology ; Child, Preschool ; Diagnostic Errors ; Female ; Hip Joint ; diagnostic imaging ; pathology ; Humans ; Ilium ; diagnostic imaging ; pathology ; Magnetic Resonance Imaging ; Radiography ; Sarcoma, Ewing ; diagnosis ; pathology

BACKGROUND:Previous in vitro studies mainly focused on morphological and nerve marker aspects in the study of bone marrow mesenchymal stem cells (BMSCs)-derived neuron-like cells,but less focused on the olectrophysiological properties of neuron-like cells following differentiation.OBJECTIVE:To study the electrophysiological changes of differentiation from BMSCs into neuron-like cells after induction by brain-derived neurotrophic factor (BDNF)/basic fibroblast growth factor (bFGF)/alltrans-retinolc acid (AT-RA).DESIGN,TIME AND SETTING:The cytological comparative in vitro study was performed at the Department of Neural Cell Laboratory,Tianjin Huanhu Hospital and College of life Science of Nankai University from June 2005 to October 2007.MATERIALS:Totally 3 male Wistar rats (6-week old,weighing about 160 g) were used in this study.METHODS:BMSCs were cultured and purified by their characteristic of plastic adhesion in vitro,and then induced by BDNF,bFGF and AT-RA,and differentiate into neuron-like cells.Whole-cell patch damp technique was used to detect cell membrane current prior to and 3 days following induction.MAIN OUTCOME MEASURES:MSC phenotype was determined by flow cytometry.Cell morphology was observed under the inverted microscope before and after differentiation.Neuron specific enolase expression was assessed by immunocytochemistry.Whole-cell current results were measured.RESULTS:Flow cytometry results demonstrated that CD90 positive rate (99±3)%,CD31 (3.4±0.8)%,and CD34 (0.3±0.1)%.This indicated that these cells were undifferentiated stem cells,with purity of 95%.Undifferentiated MSCs under the optical microscope were mostly fiat cells with processes,similar to fibroblast-like cells.Neuron-like cells appeared 3 days following induction.Immunocytochemistry results showed that MSCs before induction were weakly positive for neuron specific enolase,but strongly positive for neuron specific enolase.At 72 hours,the differentiated rate was (24.01±3.76)%.The peak currents of outward currents in neuron-like cells were significantly higher in the induction group compared with the control group (P ＜ 0.05),but no inward Na~+ current was detected.CONCLUSION:(bFGF & BDNF)/AT-RA could induce the differentiation of MSCs into neuron-like cells.These cells showed the tendency to differentiate into mature neurons,though having no electrophysiological properties of mature neurons.

OBJECTIVETo evaluate the inflicted by radar electromagnetic radiation to the sperm DNA of radar operators.

METHODSSperm concentration, viability, motility, sperm abnormality were determined by routine sperm analysis and sperm chromatin structure assay (SCSA) in the highly exposed group(n = 88), lowly exposed group(n = 143) and control group(n = 39).

RESULTSSperm motility, viability of the highly exposed group reduced compared with that of the lowly exposed group and control group, while sperm abnormality increased. The COMP alpha reduction of the highly exposed group indicated that the highly exposed group had a medium fertility potential. The multifactor variable analysis showed that daily working time was a dangerous factor in sperm abnormality and abstinence time was a dangerous factor in the parameter of SCSA.

CONCLUSIONSRadar radiation inflicts damage to male reproduction system and it is important to take protective measures.

Adolescent ; Adult ; Chromatin ; ultrastructure ; DNA Damage ; Dose-Response Relationship, Radiation ; Electromagnetic Phenomena ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Radar ; Spermatozoa ; radiation effects ; ultrastructure

Action Potentials ; Animals ; Electromyography ; Environmental Exposure ; Hypothermia ; Rats ; Rats, Wistar ; Seawater ; Thigh ; physiology ; physiopathology

Though mesenchymal stem cells (MSC) have been clinically used to repair a variety of damaged tissues, the underlying mechanisms remain elusively as the majority of the ex vivo expanded MSC die shortly after transplantation. To explore the mechanism in which the death cells play tissue repair effect, apoptosis of rat bone marrow MSC was induced by culturing cells in the conditions of hypoxia or/and serum-free medium, and the subcellular structures in the supernatants were analyzed. The results showed that apoptosis occurred in the presence of either hypoxia or serum-free condition as well, and the apoptotic proportion reached up to (17.44 ± 2.15) after the cells were treated by hypoxia plus serum free culture for 72 hours. The flow cytometric analysis of the sub-cellular substances harvested by ultracentrifugation of the supernatants found that the MSC released substantial amount of membrane microparticles into the supernatants, which expressed CD29, CD44A and Annexin-V-binding phosphatidylserine. It is concluded that the MSC can release membrane microparticles after induction, the amount of these membrane microparticles was around 15-fold of the parent cell numbers. The membrane microparticles is the mediators in the cross-talk between the transplanted cells and their surrounding tissues. This study provides some novel information for the mechanisms of MSC therapy.
Animals ; Apoptosis ; Cell Count ; Cell Hypoxia ; Cell-Derived Microparticles ; metabolism ; Cells, Cultured ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Rats ; Rats, Wistar

Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.
Chromatography, Affinity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Hepatitis B Antibodies ; biosynthesis ; genetics ; isolation & purification ; Hepatitis B Surface Antigens ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; genetics ; isolation & purification ; Pichia ; genetics ; Recombinant Proteins ; biosynthesis ; isolation & purification

OBJECTIVETo evaluate the inhibitory effect of small interfering RNA (siRNA) targeting HBV C gene region on hepatitis B virus (HBV) in vivo.

METHODSAn animal model of HBV infection was developed hydrodynamically, and pcDNA3.1-HBV and siRNA were together injected into the tail vein of the BALB/c mice. HBsAg was analyzed by time-resolved immunofluorometric assay, HBV DNA was analyzed by fluorogenic quantitative PCR (FQ-PCR), HBV C-mRNA was detected by semi-quantitative RT-PCR, and viral specific proteins (HBsAg and HBcAg) in the mice livers were assayed using immunohistochemical staining.

RESULTSIn the mice, the siRNA effectively inhibited HBV replication and expression compared with the controls. The inhibitive effect of siRNA on HBV lasted at least 3 days.

CONCLUSIONThese results demonstrate that RNAi can substantially inhibit HBV replication and expression in vivo.

Animals ; Female ; Hepatitis B ; therapy ; Hepatitis B virus ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; RNA, Small Interfering ; physiology ; RNA-Induced Silencing Complex ; Random Allocation ; Virus Replication ; genetics

OBJECTIVEThrombin and factor Xa are key players in the process of arterial thrombi formation and lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) is a cell surface endocytosis receptor for atherogenic oxidized LDL (ox-LDL). Here we investigate whether thrombin and factor Xa can induce LOX-1 protein expressions in cell-associated forms and soluble forms in cultured bovine aortic smooth muscle cells (BSMCs).

METHODSBSMCs were treated with thrombin or factor Xa in the presence or absence of AG1478, an epidermal growth factor (EGF) receptor-associated tyrosine kinase inhibitor. Total cell lysates and concentrated culture medium were then analyzed by Western blot using a mouse anti-LOX-1 monoclonal antibody.

RESULTSLOX-1 protein levels in cell lysates and culture medium were significantly increased by thrombin and factor Xa in a concentration- and time-dependent manner. Upregulation of LOX-1 protein expressions in cell lysates and concentrated culture medium was observed at concentrations above 2.0 and 3.0 U/ml of thrombin and 50 and 100 nmol/L of factor Xa, respectively. Increased LOX-1 protein expressions in cell lysates and cell culture medium were detectable as early as 4 h and peaked at 12 h after treatment with thrombin or factor Xa. LOX-1 expression induced by thrombin and factor Xa could be blocked by pretreatment with AG1478.

CONCLUSIONSThrombin and factor Xa can act as LOX-1 inducers via tyrosine kinase activation.

Animals ; Atherosclerosis ; pathology ; Cattle ; Cells, Cultured ; Factor Xa ; pharmacology ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Scavenger Receptors, Class E ; biosynthesis ; Thrombin ; pharmacology

Objective To evaluate the efficacy of Epipolis laser in situ keratomileusis(Epi-LASIK)and laser in situ keratomileu- sis(LASIK)on high myopia patients.Design Prospective,case-controlled study.Participants 62 patients(123 eyes)with high myopia. Methods 62 patients(123 eyes)underwent Epi-LASIK or LASIK surgery for high myopia:28 patients(56 eyes)underwent Epi-LASIK and 34 patients(67 eyes)underwent LASIK.The differences in postoperative uncorrected visual acuity(UCVA),best corrected visual acuity(BCVA),refraction,and root mean square(RMS)of high-range wavefront aberration were compared one-week,one-month, three-months and six-months postoperatively.Main Outcome Measures UCVA,BCVA,refraction,and RMS of high-range wavefront aberration.Results There was no serious complication during and after the operation.The recovery of postoperative UCVA after Epi-LASIK was slower than that of LASIK.One week postoperatively,the proportion of UCVA≥0.8 of Epi-LASIK(46.4%)was less than that of LASIK(77.6%)(P=0.0003).No significant differences were found in those of Epi-LASIK(85.7%,94.6%,91.1%)and those of LASIK(92.5%,95.5%,94.0%)one-month,three-months and six-months postoperatively(P=0.590,0.822,0.530).BCVA was same after Epi-LASIK and LASIK.The proportion of mean spherical equivalents within?0.50D for Epi-LASIK(42.9%,51.8%,60.7%,64.3%)had no difference with those for LASIK(53.7%,59.7%,71.6%,73.1%)one-week,one-month,three-months and six-months postoperatively (P=0.230,0.378,0.200,0.290).The postoperative RMS increased after both surgeries,especially after LASIK.At postoperative one-month,three-months and six-months RMS of Epi-LASIK(1.51?0.77)?m,(1.32?0.76)?m,(1.18?0.71)?m were much higher than the (0.87?0.27)?m preoperative ones(P=0.016,0.019,0.026).At postoperative one-month,three-months and six-months RMS of LASIK (2.41?0.81)?m,(2.17?0.63)?m,(1.89?0.87)?m were also much higher than the preoperative(0.91?0.22)?m(P=0.011,0.008,0.006). There were significant differences between the RMS of Epi-LASIK and LASIK one-month,three-months and six-months postoperatively (P=0.039,0.035,0.033).The I grade haze was found in two eyes after Epi-LASIK.Conclusion Epi-LASIK has better visual quality re- sult than LASIK on correcting high myopia.(Ophthalmol CHN,2007,16:336-339)

Objective: To explore the correlation between serum vitamin D level and glucose and lipid metabolism among primary and middle school students in Inner Mongolia, so as to providing a scientific basis for guiding rational vitamin D supplementation among primary and middle school students in Inner Mongolia Region. Methods: A total of 2 206 students of primary and middle school students from 8 counties (districts) in Inner Mongolia were enrolled for a questionnaire survey, physical examination, and laboratory test by a multi stage stratified random sampling approach.The levels of fasting blood glucose, blood lipid and other related indexes with different vitamin D levels were analyzed, and their correlation was further discussed. Results: The deficiency and insufficiency rates of vitamin D among primary and middle school students were 33.2% and 42.8%, respectively, and the suitable rate was 24.0%. The body mass index (BMI) and waist height ratio (WHtR) of the vitamin D deficiency group [(22.87±7.41) kg/m 2 , 0.46±0.08)]were higher than the insufficiency [(20.59±8.00)kg/m 2, 0.44±0.09)] and suitable group [(18.01±7.38)kg/m 2, 0.43 ±0.08)] ( P <0.05). Compared with the vitamin D suitable group [(3.87±0.85) mmol/L], the total cholesterol (TC) level in the deficiency group [(3.73±0.67)mmol/L] decreased, with a statistically significant difference ( P <0.05). The levels of high density lipoproteins (HDL) in the vitamin D insufficiency[(1.40±0.33)mmol/L] and deficiency groups [(1.34±0.31)mmol/L] were lower than the suitable group [(1.48±0.34)mmol/L], with a statistically significant difference ( P <0.05). The detection rate of hyperuricemia (HUA) in the vitamin D deficiency group (22.95%) was higher than the suitable group (17.20%), with a statistically significant difference ( P <0.016 7). Pearson correlation analysis showed that vitamin D was negatively correlated with BMI and WHtR ( r =-0.23, -0.11), and positively correlated with TC and HDL level ( r =0.06, 0.16) ( P <0.05). Conclusions The deficiency and insufficiency rates of serum vitamin D among primary and middle school students in Inner Mongolia are high which are associated with several indicators of blood lipids. Therefore, it is recommended to supplement vitamin D preparations in moderation in daily life.