Objective To research the 50% effective anesthetic volume ( EAV50) of 0.5% ropivacaine in nerve stimulator-guided vertical infraclavicular brachial plexus block. Methods Thirty patients scheduled for forearm or hand surgery were blocked using 0. 5% ropivacaine in nerve stimulator-guided vertical infraclavicular brachial plexus block. The EAV50, which is the anesthetic volume corresponding to 50% success and 50% failure was determined by up-and-down sequential test. The starting dose of 0. 5% ropivacaine was 0. 55ml/kg. Block failure resulted in a dose increase 110% , and block success in a reduction 110%. The sensory and motor blockade were accessed at 5- min intervals (up to 60 min). Results In nerve stimulator-guided vertical infraclavicular brachial plexus block, EAV50 for 0.5% ropivacaine was 0.417ml/kg. In up-and-down sequential test, there were significantly different in the block success rates of lateral, medial and posterior cord ( P <0.01). Conclusion In nerve stimulator-guided vertical infraclavicular brachial plexus block, enough anesthetic volume must be given to increase block success rates of all the cords of brachial plexus, and EAV50 for 0. 5% ropivacaine was 0.417ml/kg.

Objective To observe the process of Vibrio vulnificus inducing dendritic cell strain apoptosis.Methods We established the mixed culture model of mouse dendritic cell ( DC 2.4 strain) and Vibrio vulnificus( Vv1.1758 strain ),analyzed morphological characteristics of cell apoptosis by DAPI fluorescence staining,detect DNA fragmentation level of apoptosis cells by DNA Ladder assay,analyze DC2.4 apoptosis rate by Annexin V FITC/PI staining,determine activities of caspase-3 and caspase-8 by means of spectrophotometric method and detect changes of mitochondrial transmembrane potential ( △ Ψm ) by JC-1 fluorescence labeling.Results After Vv1.1758 strain and DC2.4 cell were mixed and cultured for 4 h,DAPI fluorescence staining showed typical apoptosis characteristics-chromatin condensation and marginalization; DNA agarose gel electrophoresis showed apoptosis band; apoptosis rates at 2,4 and 6 h were respectively (37.8±9.8) ％,(54.3 ± 12.7 ) ％ and ( 68.2± 14.6 ) ％ ; Mitochondrial transmembrane potentials (△Ψm) at 1 h,2 h and 4 h reduced by 7.1％,16.1％ and 46.7％ respectively; caspase-8 activity increased at 1.5 h and reached the peak at 2 h [ (2.48±0.19) U/μg],while caspase-3 activity started to increase at 3h and reached the peak at 4 h [ ( 1.91 ±0.16) U/μg ].Conclusion Vibrio vulnificus could induce dendritic cells by two pathways: reducing mitochondrial transmembrane potential and activating caspase-8 promoter and finally activate effector caspase-3 to promote apoptosis.

Objective To investigate the infection status and drug suscepetibility of mycoplasma from 6 573 patients with non-gonococcal urethritis ,and to provide the scientific bases for the clinical application of antibiotics .Methods Mycoplasma detection kit was used to detect ureaplasma urealyticum (Uu) and mycoplasma hominis(Mh) and the drug susceptibility .All the patients were divided into two groups :Chinese group and foreigner group .Results Among 5 675 Chinese patients ,2 985 patients were infected by mycoplasma(52 .6% ) .The infection rate of Uu was 2 312(40 .7% ) .35 .2% patients were male ,and 61 .4% patients were female .In 898 foreign patients ,440 patients were infected by mycoplasma(49 .0% ) .The infection rate of Uu was 327(36 .4% ) .32 .2% pa-tients were male ,and 59 .5% patients were female .In Chinese patients infected by Uu ,the susceptibility rates to MIN ,DOX ,JOS and CLA were 96 .7% ,96 .2% ,93 .7% ,89 .7% ,respectively .In foreign patients ,the susceptibility rates to MIN ,DOX ,JOS ,and CLA were 98 .9% ,98 .4% ,95 .8% ,92 .1% .Conclusion The mycoplasma infection rate of Chinese patients is higher than foreign patients .In both groups ,Uu infection is the main type .Female patients are more than male patients .The drug sensitivity rate in for-eign group is higher than that in Chinese group .mycoplasma are sensitivity to MIN ,DOX ,JOS .

Objective To investigate the invasion,internalization and the organelles damage of the cultured dendritic cells ( DC2.4 strain) during Vibrio vulnificus (Vv) infection.Methods The study model was the cultured DCs infected by Vv 1.1758 strain.Electron microscopy was used to observe the localization of bacteria in different time point of infection,cell morphology and the process of organelles changes.The cytoskeleton structure including the microfilaments and the microtubules rearrangement was examined by the fluorescence microscope.Results The Vv were pinocytosed into the DC cells through double-sides,and localized at 1-2 μm of the inner side membrane.It cost 1.27,1.87,3.43 hours reaching the infection ratio of 25％,50％,75％,respectively.Using electron microscopy,the DCs had been observed the phagosome formation within 1h,chromatin activation within 2 h,chromatin aggregation 4 h,and the significant cytoskeleton structure disruption within 6 h.Endoplasmic reticulum,mitochondria and lysosomes became swollen.In DCs,the protruding filaments gradually reduced,and their shape changed from the point-like to the linearlike aggregation at the inner side of the plasma membrane,extended microtubules disappeared,the microtubules at the outside nuclear membrane striking rearranged.Conclusion After DC was infected by Vv,the bacteria were pinocytosed into the inner side of DC membrane,and the microfilaments were observed to move from the cytoplasm to cell membrane.In addition,the microtubules moved from the synapse and the cell membrane to the nuclear membrane.The high lethality of Vv could provoke to the DCs cytoskeleton rearrangements.

Objective To research the distribution and the characteristics of the plasmid mediated quinolone resistant genes in Shewanella algae. Methods The qnr, qepA, aac(6')-Ib-cr genes were amplified by PCR, then the positive PCR products were sequenced to determine the gene type. The transferability of plasmid mediated quinolone resistance was ensured by conjugation experiment. MICs were measured by E-test. qnrA gene was mapped to plasmids to locate it. Results The qnrA gene were detected in the Shewanella algae, this is a newfound subgroup qnrA7, the GenBank accession no. was GQ463707, qnrB, qnrS,qnrC, qnrD, qepA and aac(6')-Ib-cr genes were not detected. qnrA7 reside in a plasmid about 33 kb, conjugation experiment was unsuccessful. The strain was susceptible to quinolones. Conclusion It deserves paying close attention to the report of an original qnrA subgroup in an isolate of water-borne species of Shewanella algae.

Objective To elucidate the expression of gankyrin in human gastric cancer cells and it's role in nimesulide induced apoptosis. Methods Four human gastric cancer cell lines including MKN28 (well differentiated), AGS (poorly differentiated), MKN45 (poorly differentiated), and SGC7901(moderately differentiated) were cultured and treated with nimesulide. Nimesulide induced growth inhibition and apoptosis of the cells were detected by methyl thiazolyl tetrazolium assay, and confirmed by flow cytometry. The expressions of gankyrin gene and protein were further assessed by real-time PCR and Western blotting. Results Gankyrin mRNA and protein were detected in all four human gastric cancer cell lines. The proliferations of AGS and SGC7901 cell lines were significantly suppressed by nimesulide in a time-dose dependent manner. When treated with 400 μmol/L of nimesulide for 48 hours, the significant apoptosis was found in AGS cells (23.30%±2.50%) and SGC7901 cells (16.80%±1.55% ) in comparison with controls (0.57%±0.19% and 0.88%± 0.17%, respectively, all P values <0.01). Apoptosis of AGS cells induced by nimesulide was accompanied by a considerably decreased gankyrin expression that was more significant at 24 hours (0.0035±0.0014) and 36 hours (0.0980±0.0160) in comparison with controls (0.4690±0.1190, all P values<0.01). Conclusion Gankyrin expresses in human gastric cancer cell lines and may be involved in nimesulide induced apoptosis of AGS cells.

Objective To apply electrical impedance tnmography that is a new evaluation ap-proach to monitor the development of retroperitoneal injury. We used retroperitoneal inject blood model in pigs to study the feasibility on monitoring retroperitoneal bleeding and to provide premise in theory and practice for clinical application. Methods Five pigs were used on the experiment. We insert a vessel into the retroperitoneal and inject blood to simulate retroperitoneal bleeding. Sixteen electrodes were atta-ched on the abdominal region circumference of pigs and used for electrical current injection and surface voltage measurement. Then the monitoring images were performed by electrical impedance tomography. Results The images of electrical impedance tomography retroperitoneal inject blood model of five pigs were clear, the minimal impedance scale was decreasing significantly as the bleeding volume increasing and the images were changed significantly too. The computerized tomography and the dissecting results confirmed the blood was limited in retroperitoneal. Conclusions The establishments of pigs retroper-itoneal inject blood model was successful. The images of electrical impedance tomography retroperitoneal inject blood model were clear with significant contrast. It's feasible to use electrical impedance tomography system to monitor the retroperitoneal bleeding. This technique may become a useful tool for monitoring ret-roperitoneal injury in intensive care patients.

Objective To investigate the formation of biofilm in clinical isolates of Staphylococcus epidermidis ,and to analyse the correlation between biofilm formation and antibacterial resistance of Staphylococcus epidermidis .Methods A total of 62 strains of Staphylococcus epidermidis isolated from blood specimens of inpatients with bloodstream infection ,from January 2014 to February 2015 ,were collected .The biofilm formation of Staphylococcus epidermidis was detected by using the semi‐quantitative adherence as‐say and polymerase chain reaction(PCR) amplification experiment .The antibacterial susceptibility test was carried out according to K‐B method .Results The positive rate of biofilm formation detected by using the semi‐quantitative adherence assay and PCR for icaA gene were 37 .1% (23 strains) and 43 .5% (27 strains) respectively ,and there was no statistically significant difference(P>0 .05) .There were 14 positive strains detected by both methods .The resistance rates of strains producing biofilm to antibacterial a‐gents were generally higher than those of non‐producing biofilm strains ,and there were statistically significant differences in resist‐ance rates of strains to gentamicin ,penicillin ,oxacillin ,levofloxacin and cefoxitin(P<0 .05) .All bacteria were sensitive to vancomy‐cin ,linezolid and quinupristin/dalfopristin .Conclusion There is no significant difference between the two methods in detecing bio‐film formation .The resistance rates of strains producing biofilm to antibacterial agents were generally higher than those of non‐pro‐ducing biofilm strains .

OBJECTIVETo observe the effect of copper on bone fracture healing.

METHODS160 growing chickens aged 70 days were fed in 32 cages, each consisting of five. The fracture with 1 approximately 2 mm defects at the bilateral radius of wings was created by osteotomy. The chickens were divided into four groups, 40 each. Every morning, chickens of the first group, served as controls, were fed orally 0.8% Na-CMC solution (sodium carboxymethyl cellulose), those of the other three groups were fed with copper-Na-CMC suspension (copper powder suspended in 0.8% Na-CMC) with different doses of 20, 40 and 80 mg Cu/kg body weight. Each group was sacrificed 14, 21, 28 and 35 days postoperatively. Liver was taken for analysis of Cu and Zn. Both radius were removed by dissection. CT was performed quantitatively for the ment of the gray values of the callus. The biomechanical properties of the healing radius were analyzed by a three point bending test. Afterwards, the contents of calcium, magnesium, phosphorus, copper, zinc, iron and hydroxyproline in the callus were determined.

RESULTSThe gray values of the callus increased along with the increase of copper dose and the duration of observation. 21 and 35 days after operation, the gray values of the callus were significantly higher in the high-Cu group than in the controls respectively (909 +/- 220 vs. 597 +/- 155; 973 +/- 100 vs. 763 +/- 179 HU, P < 0.05). The level of calcium and magnesium in the callus 35 days after operation was much higher in the mid and high Cu groups than in the controls respectively (Ca: 177 +/- 26.7, 176 +/- 20.5 and 137 +/- 34.7 mg/g; Mg: 2.98 +/- 0.57, 3.06 +/- 0.46 and 2.43 +/- 0.53 mg/g P < 0.05). The contents of hydroxyproline in the callus 35 days after operation were significantly higher in the high-Cu group than in the controls (34.23 +/- 1.96 vs. 32.17 +/- 1.93 mg/g respectively). The biomechanical properties of the repaired radius had a better improvement tendency in the three Cu-fed groups than in the controls.

CONCLUSIONCopper is helpful in fracture healing; however, the effect of copper on fracture healing needs further study.

Animals ; Bony Callus ; Chickens ; Copper ; Fracture Healing ; Zinc

OBJECTIVEDevelop an LC-MS method to determine evodiamine and rutaecarpine in rats plasma simultaneously. The method was employed to investigate pharmacokinetics of evodiamine and rutaecarpine.

METHODBlood samples were collected in different time after oral administrated with the extracts of Euodiae Fructus, the plasma concentration of evodiamine and rutaecarpine was determined by LC-MS, pharmacokinetic parameters were calculated by WinNonlin 5.1 software.

RESULTThe linear ranges of evodiamine and rutaecarpine were 0.5-100 microg x L(-1) (r = 0.995 9), 1-200 microg x L(-1) (r = 0.999 3) respectively. The average recovery were exceeded 76% (n = 5), the precision of inner-day and inter-day were less than 15%. The pharmacokinetics parameters AUC, t1/2, CL _F of evodiamine were: (2 215.24 +/- 414.49), (4 230.62 +/- 753.77), (13 219.21 +/- 3 740.95) min x ng(-1) x mL(-1); (146.57 +/- 38.38), (114.38 +/- 14.65), (163.37 +/- 8.83) min; (184 607.29 +/- 32 502.21), (192 878.22 +/- 31 897.37), (19 3224.63 +/- 62 278.74) mL x min(-1). The pharmacokinetics parameters AUC, t1/2, CL_F of rutaecarpine were (2 283.53 +/- 298.51), (4 424.84 +/- 276.95), (14 239.93 +/- 3648.27) min x ng(-1) x mL(-1); (167.10 +/- 15.82), (131.58 +/- 20.07), (144.41 +/- 13.65) min; (1 177 340.54 +/- 2 4942.21), (181 262.92 +/- 11 162.22), (177 508.10 +/- 52 611.80) mL x min(-1).

CONCLUSIONThe method described in this report has high sensitivity and selectivity, and was suitable for pharmacokinetic studies of evodiamine and rutaecarpine. The kinetic process of evodiamine and rutaecarpine in rats in vivo were all yielded to be one-compartment model.

Administration, Oral ; Animals ; Evodia ; Indole Alkaloids ; pharmacokinetics ; Male ; Plant Extracts ; pharmacokinetics ; Quinazolines ; pharmacokinetics ; Rats ; Rats, Wistar