Objective To study the characteristic cluster structure of good-quality natural drinking water.Methods The coupling 17 O NMR(nuclear magnetic resonance)FWHM(full width half maximum)was selected as the index to weigh the rel-ative average size of water cluster in liquid water.The 17 O NMR FWHM of water samples(nature water and artificial water)were measured by ARX400NMR spectroscopy.Results 17 O NMR FWHMs of21representative water samples were in the range of53-145Hz.Conclusion 17 O NMR FWHM of good-quality natural drinking water mainly lay from70to90Hz.

Objective To investigate the clinical and radiographic outcome of traumatic pubic symphyseal diastases fixed with the locked plate. Methods From December 2007 to December 2009,17 patients(11 males,6 females,at mean age of 45.3 years)with pubic symphysis diastasis of unstable pelvic ring injuries were treated with open reduction and fixation with the locked plate.According to Tile classification system,five patients were with type B1 fractures,one with type B2,one with type B3,six with type C1,three with type C2 and one with type C3.All operations were performed under general anesthesia.Pubic symphyseal diastasis was treated by open reduction and fixation with the locked plate.There were 13 patients with associated posterior pelvic disruption,of which seven patients were treated by open reduction via anterior approach and fixed wich the reconstruction plates,three by close reduction and fixed with percutaneoua sacroiliac cannulated screws and three by posterior approach and fixed with M type plates. Results Of all the patients,16 patients were followed up for a mean time of 16 months(6-30months),which showed mean blood loss of 200 ml(50-600 ml)and mean hospital stay of 21 days (14-62 days).The clinical outcome was measured according to Majeed scores system,which showed excellent results in seven patients,good in seven and fair in two. Conclusions The locked plate fixation takes advantages of sailsfactory clinical outcomes with less operative trauma and a lower implant failure and wound infection rate in treatment of traumatic pubis symphysis diastasis.Locked plate fixation of symphysis call be performed alone when the posterior pelvic ring is only partially disrupted (Tile B).Posterior fixation construction should be used if the pelvic ring is under complete instability(Tile C).

Objective:To investigate the immunogenicity of pORF5 plasmid protein,and further to screen and identify its im-munodominant domian.Methods: 10 different fragments of pORF5 gene including full length were amplified from the DNA of Chlamydia trachomatis serovar D by PCR and cloned into appropriate site of pGEX-6p vector to construct recombinant vectors after digested with BamHⅠand NotⅠrestriction endonucleases.After identification by PCR and sequencing,the recombinant plasmids were transformed into XL1 Blue E.coli to express the GST fusion proteins.ELISA and Western blot were carried out to identify the immunogenicity and immunoreaction of pORF5 plasmid protein.10 different fragments were reacted with sera from patients urogenitally infected with Chlamydia trachomatis, mouse polyclonal antibodies and mouse monoclonal antibodies of pORF5 plasmid protein with ELISA method.Results: pORF5 plasmid protein displayed strong immunogenicity and could induce a strong antibody response in human.The reactivity of human antibodies almost completely disappeared,when the native structure of pORF5 plasmid protein was de-stroyed.F6 that only lacked the N-terminal 66 amino acids was recognized by antibodies in ELISA as strongly as the whole pORF5 plasmid protein was.However,no other fragments were significantly recognized although there was a minimal reactivity of F2 and F3 with antibodies.Conclusion:pORF5 plasmid protein was an immunodominant antigen containing conformation-dependent epitope,and the C-terminal three quarters of pORF5 amino acid sequence was required for maintaining its immune dominance and conformation.The significance of the above findings lay a foundation for the further study on pORF5 protein function and vaccine development.

Aim To observe the influence of CCK-8 on expression of MMPs/TIMP-1 in TNF-α-induced rat fibroblast-like synovial cell line RSC-364.Methods The secretion levels of MMP-1, MMP-3, MMP-9 and TIMP-1 were determined using ELISA;MMP-3 and MMP-9 mRNA expressions were detected by RT-PCR.Results MMP-3 and MMP-9 could not be examined in RSC-364 incubated with CCK-8 and unstimulated RSC-364, which was able to product a little MMP-1, TIMP-1 and express even less MMP-3,-9 mRNA.CCK-8 inhibited the increase in MMP-1, MMP-3, MMP-9 secretion and MMP-3,-9 mRNA expression in TNF-α-induced RSC-364.TIMP-1 production was also increased in TNF-α-induced RSC-364.CCK-8 had no effect on TIMP-1 production in TNF-α-induced RSC-364, but was able to reduce the ratios of MMP-1, MMP-3, MMP-9 to TIMP-1.Conclusion The inhibitory effect of CCK-8 on MMPs activity may be related to the decrease of MMPs mRNA expression, MMPs secretion and the ratios of MMPs to TIMP-1 in TNF-α-induced RSC-364, which indicates that CCK-8 might be a possible regulator in the pathogenesis of rheumatoid arthritis.

Objective To perform comparative study on database performance before and after upgrading hardware & software of database. Methods The comparison was carried out from the aspects of hardware expansibility, processing ability, system stability and database performance. Results The reasons for the change of performance and the change law were analyzed. Conclusion After upgrading, the system performance is enhanced greatly.

Objective To investigate the correlation between large platelet ratio (P-LCR) and stroke in patients with atrial fibrillation. Methods 152 consecutive patients with atrial fibrillation were selected and divided into two groups according to whether accompanying stoke or not. Clinical features of patients and blood test indicator P-LCR were analyzed by Logistic regression analysis. Results In total, 152 subjects, including 47 patients with stoke and 105 patients without stoke,were included in the study. Univariate analysis showed that the difference in age, diabetes, hypertension, lipid-lowering therapy and P-LCR between two groups was statistically significant (P < 0.05). Logistic regression analysis showed that age, diabetes and P-LCR were independent risk factors of stroke in patients with atrial fibrillation. Conclusions Age, diabetes and P-LCR are independent risk factors of stroke in patients with atrial fibrillation.

Objective To observe the morphological characteristics of HCV particles and intracel-lular ultrastructure changes in Huh7. 5 cells which was infected with chimeric HCV via transmission electron microscopy. Methods Plasmid J6/JFH encoding the full length HCV chimeric genome was transcribed to HCV RNA in vitro and the RNA was transfected into Huh7.5 cells by electroporation. Quantitative real-time PCR(qRT-PCR) was used to assay HCV copies of the supernatant of transfected cells. Indirect immunofluo-rescence was used to detect the expression of HCV proteins. The cell culture superoatant were used to infect narve Huh7.5 cells, transmission electron microscopy was used to observe morphological characteristics of vi-rus particles and intracellular ultrastructure changes in infected Huh7. 5 cells. Results qRT-PCR showed high level virus copies in supernatant of transfected cells collected in different times, indirect immuno-fluo-rescencc proved high expression of HCV NS5A proteins in the transfected cells. Large numbers of enveloped or unenveloped virus-like particles (VLPs) were observed in infected Huh7. 5 cells via transmission electronmicroscopy. We also found hyperplasia of some membrane-enclosed organelles in the cytoplasm. Several fea-tures characterizing flavivirus infected cells and a cytoplasmic inclusion of unknown origin were observed. Conclusion The chimeric HCV from in vitro cell culture system is proved to be intact virus particles which can efficiently infect Huh7.5 cells.

BACKGROUND:Recently,researches have found that insulin-like growth factor-1(IGF-1)can induce the differentiation of bone marrow-derived mesenchymal stem cells(BMSCs)into chondrocytes,but there are no reports concerning the differentiation of adipose-derived mesenchymal stem cells(ADMSCs)into chondrocytes induced by IGF-1,as well as interaction with transforming growth factor-β1(TGF-β1)during this process.OBJECTIVE:To explore the possibility of inducing ADMSCs chondrogenic differentiation by using IGF-1 and the interaction with TGF-β1 in induction.METHODS:ADMSCs were obtained,and seeded at 2×10~5 cells/cm~2 in culture flask.Insulin-free chondrogenic media containing IGF-1 or(and)TGF-β1 were used to induce ADMSCs.2 weeks later,cells were harvested and stained by using toluidine blue and collagen Ⅱ antibody immunohistochemistry.Intracellular sulfated proteoglycan and collagen Ⅱ coloring were observed.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of collagen Ⅱ,aggrecan and Sox9 mRNA.RESULTS AND CONCLUSION:After induced,toluidine blue stain exhibited that the cells in the three induction groups were polygonal,with cytoplasm and cell membrane of blue different dyeing.Immunohistochemistry for type Ⅱ collagen demonstrated that cytoplasm and cell membrane were stained brown in three induction groups.RT-PCR revealed that the expression of collagen Ⅱ,aggrecan,Sox9 mRNA of IGF + TGF group were significantly greater than the IGF and TGF groups,and IGF and TGF groups were significantly stronger than the control group.No significant difference was determined between the IGF and TGF groups.These results indicated that IGF-1 can induce chondrogenic differentiation from ADMSCs,expressing chondrocyte specific cell phenotype.There is synergism of IGF-1 and TGF-01 to induce the differentiation of ADMSCs into chondrocytes.

BACKGROUND: Adipose-derived mesenchymal stem cells (ADSCs) can be induced toward the chondrogenic lineages with chondrogenic medium contained transforming growth factor-β_1(TGF-β_1). However, it remains concerned about the disadvantage with use of TGF-β_1 in vitro, which can induce chemotaxis, activate inflammatory cells, cause local defect and want an ideal matrix to deliver proteins. Therefore, with advanced gene-delivery techniques, TGF-β_1 can be long-lasting expressed by transduced stem cells, which is very useful for Chondrogenic Tissue Engineering.OBJECTIVE: To master the method of construction and transfection of eukaryotic expression vector for rat transforming growth factor-β_1 and to study the possibility of gene transfection to ADSCs with this vector.METHODS: Recombining DNA techniques were applied to construct recombinant plasmid pcDNA3.1-TGF-β_1. And this plasmid was verified by restriction endonuclease mapping and DNA sequencing; Then the plasmid with TGF-β_1 gene or not was transfected into ADSCs by use of LipofectamineTM2000. After infection, transduced ADSCs were diluted and cultured with neomycin (G418). Gene transfer efficiency compared on the basis of green fluorescent protein expression was assessed.RESULTS AND CONCLUSION: Digestion of the plasmid with double restriction endo- nuclease XboⅠand Hind Ⅲ showed about two specific electrophoretic strips (1.35 bp and 5.4 kb), and the sequence of the rat TGF-β_1 gene in recombinant was concorded with that reported in Genbank. There were 80 percent of the cells which were transduced, and the expressions of mRNA and protein of TGF-β_1 in ADSCs were discovered positively. These indicated that the eukaryotic expression vector for rat TGF-β_1 (i.e. pcDNA3.1-TGF-β_1) , which is able to transfect the ADSCs, can be constructed through genetic recombination.

AIM: To investigate the effect of sulfated cholecystokinin octapeptide (CCK -8 ) on TNF -α induced IL - 6 mRNA expression, NF - κB activation in the rat fibroblast - like synovial cell strain RSC - 364 and its possible receptor mechanisms. METHODS: RSC -364 cells were stimulated with TNF - α( 10 μg/L) in the presence or absence of sCCK- 8( 10-8 - 10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L). IL -6 and CCK receptor A/B (CCK- AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction (RT- PCR) at 3 h after stimulation, and nuclear factor - κB (NF - κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at lh after stimulation. At 30 min of stimulation the IκB protein level in cytoplasma was measured by Western blotting. RESULTS: Both CCK - AR and CCK - BR were constitutively expressed on RSC - 364. sCCK - 8, at concentrations from 10-8 mol/L to 10 -6 mol/L, significantly increased IL - 6 mRNA expression, CCK - AR and CCK - BR mRNA expression, NF - κB binding activity and IκB protein degradation. The effects of sCCK - 8 on NF - κB activity and IκB degradation level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: sCCK - 8 upregulats TNF - α- induced IL - 6 mRNA expression by NF - κB pathway through its receptor on rat synoviocytes, suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.