OBJECTIVE: To establish an SPE(solid phase extraction)-HPLC method for the determination of diclofenac sodium in human plasma.METHODS: The separation of diclofenac sodium was performed on ODS-2 column with a detection wavelength of 280nm.The mobile phase was composed of phosphate buffer(pH6.5)-methanol(40∶ 60) at a flow rate of 1.0mL? min-1.RESULTS: The linear range of diclofenac sodium was 16.2～ 2 024.0ng? mL-1(r=0.998 9).The methodological recovery ranged between 90% to 110% and the extraction recovery was above 90%.Both intraday RSD and inter-day RSD were less than 5%.CONCLUSION: The method is simple,accurate and reliable,and suitable for the pharmacokinetic study of diclofenac sodium.

Objective\ To evaluate the immediate effects and strength of allograft fusion cage(AFC) used for reconstructing stability of cervical spine. Methods\ Discs of C 5 and C 6 were resected on 8 fresh human cervical spine specimens, and autogenous iliac bone grafts(AIBG), and AFCs were implanted into the intervertebral spaces respectively. Compression test, pull out test and segmental motion measurement were studied.Results\ Comparing with intact and AIBG groups, the range of motions of C5-6 in AFC group were decreased in all directions except for extension; with (502?114) N compressive load,the vertebrae in AFC group were broken while the AFC were intact, but the AIBG were broken at (135?42) N load; with 300 N drawing load,no loosening was found between AFCs and vertebra, but it was found between AIBG and vertebra at 60 N load. Conclusion\ AFC could provide enough support,anti slide ability and could remain or increase the height of intervertebral spaces. It completely meets clinical and biomechanical requirements.

Objective To evaluate the effects of allograft fusion cage(AFC) on anterior cervical interbody fusion. Methods AFCs were implanted in 61 degenerative cervical intervertebral spaces of 39 cases, who needed anterior cervical interbody fusion from September 1995 to December 1999. 31 cases were diagnosed as cervical spondylolysis， 2 cases as acute protrusion of cervical intervertebral disc and 6 cases as fracture and dislocation of cervical spine. The clinical effects and complications were observed, and the postoperative presentations of X－ ray examination of cervical spine were also evaluated. Results Thirty- nine cases were followed up with a mean period of 28.6 months. No neurologic complications appeared, and no AFCs shifted or dislocated. The clinical effects were satisfactory. 61 intervertebral spaces were confirmed to be solid fused completely by constant X－ ray examination at 3.9 months in average after operation. There were no collapse or angular deformities in 59 spaces of them,the other 2 spaces lost a little height because of removal of external fixation too early. Conclusion The implantation of AFC was simple, stable, less injury with similar intervertebral osseous fusion rate compared to the conventional anterior cervical interbody fusion. Forthermore, the implantation of AFC does not need auto iliac crest graft or the use of metal fixations. Some complications caused by implanting auto iliac crest and metal fixations can be avoided.

Objective To compare the survival conditions of patients with insulinoma after enucleation of insulinoma or partial resection of pancreas.Methods The clinical data of 99 patients with insulinoma,treated with surgery from May.2003 to Aug.2015 were retrospectively analyzed.Of the 99 patients,38 received enucleation of insulinoma alone and 61 received partial resection of pancreas.The overall data were analyzed by SPSS 21.0 software.Results Average survival of patients after enucleation of insulinoma (103.3 months) was longer than that of patients after partial resection of pancreas (77.5 months),and the difference had statistical significance (P=0.006).The difference of the incidence of most chronic or temporary complications had no statistical significance between the two groups (P＞0.05),except for new-onset diabetes (P=0.004).Conclusion Enucleation of insulinoma should be firstly recommended for patients with insulinoma in suitable size,which can provide patients with better survival condition.

Objective To analyze the characteristics of toxin, the PCR-ribotyping(RT) and the multilocus sequence typing(MLST) of Clostridium difficile strains isolated from China-Japan Friendship Hospital in order to provide a basis for monitoring the outbreak of nosocomial Clostridium difficile infection.Methods A total of 321 samples were collected from the patients with suspected Clostridium difficile infection(CDI) in China-Japan Friendship Hospital(CJFH) during 2012 to 2013.All Clostridium difficile strains were isolated and identified by the standard phenotypic culture method.Cytotoxicity test was performed to detect toxin B.Toxin genes (tcdA and tcdB) and binary toxin genes (cdtA and cdtB) harbored by those strains were analyzed.RT and MLST were used for homologous analysis.Clinical data of the patients were collected to analyze the isolation rate of Clostridium difficile in different populations.Results Forty-eight strains of Clostridium difficile were isolated from 46 patients with diarrhea and three of them were isolated from the same patient.The incidence of CDI among all patients, outpatients and inpatients were 14.3%(46/321), 12.8%(5/39) and 14.5%(41/282), respectively.Toxin B was detected in all of the strains as indicated by the cytotoxicity test.Strains of sequence type 1(ST1) showed the strongest cytotoxicity of all the isolated Clostridium difficile strains.Ten out of the 48 strains (20.8%) were tcdA(-)/tcdB(+) strains, which belonged to either ST37 or ST81.The results of RT and MLST were consistent in assigning the strains into nine types, in which the predominant type was ST1/RT027 accounting for 27.1% (13/48).All of the ST1/RT027 strains presented a toxin gene profile of tcdA(+)/tcdB(+) and cdtA(+)/cdtB(+).Most of the ST1/RT027 strains were isolated from the Traditional Chinese Medicine Department of Respiratory, where smallnosocomial outbreaks of ST1/RT027 strain infection might happen.Conclusion CDI diagnosed in CJFH mainly belongs to nosocomial infection.Most of the isolated strains harbor tcdA(+)/tcdB(+) genes.Surveillance for the outbreaks of CDI caused by ST1/RT027 strains over producing toxins A and B should be strengthened in hospitals.

Objective To investigate the feasibility and effects of endoscopic surgical treatment of lumbar intervertebral disc herniation associated with veitebral osteochondrosis.Methods From June 2008 to December 2015,276 cases of lumbar intervertebral disc herniation associated with vertebral osteochondrosis were treated with endoscopic surgery,including 185 men and 91 women,with an average 39.2 years old (range,16-65 years old).The involved level included L2.3 in 2 cases,L3.4 in 9 cases,L4,5 in 126 cases and L5S1 in 139 cases.On preoperative axial CT,the diameter of ossification was more than half of the transverse or sagittal diameter of the spinal canal in 89 cases,and no more than half of the transverse and sagittal diameter of the spinal canal in 187 cases.All patients were operated on the side with serious symptom,181 cases were operated with mobile microendoscopic discectomy (MMED),and 95 cases were operated with percutaneous endoscopic surgery,including percutaneous transforaminal endoscopic discectomy (PTED) in 61 cases and the percutaneous interlaminar endoscopic discectomy (PIED) in 34 cases.The operation and complications were analyzed.Results The soft herniation,broken disc material and the periphery of compressing ossification were removed under the endoscope in all cases,until the nerve was well decompressed.However,the ossification was not complete resected.Dural sac tear occurred in 3 cases of MMED.In the early stage of PTED,2 cases converted to MMED because of intraoperative pain and difficulty,and one case had exiting nerve root injury.At the final follow-up of 12-60 months (average,20.6 months),visual analogue scale decreased from preoperative 8.5±1.2 to 1.0±0.9,Oswestry disability index decreased from preoperative 40.2±8.6 to 3.1±3.0.According to Macnab scale,the results were excellent in 89,good in 154 cases,moderate in 33 cases.Conclusion For most lumbar intervertebral disc herniation associated with vertebral osteochondrosis,good results can be achieve by removal of herniated and broken intervertebral disc and decompression of nerve with endoscope.Therefore,we speculate that the soft disc herniation and spinal stenosis are main pathogenic factors,and that the complete resection of ossification is not needed.

Objective To assess the prospect of integrated biphasic silk fibroin scaffold made by annulus fibrosus-nu?cleus pulposus tissue engineering in application as integrated intervertebral disc(IVD). Methods An integrated annulus fi brosus-nucleus pulposus(AF-NP)biphasic scaffold was made by silk fi broin using two different uncomplicated methods which were paraffin spheres-leaching method(outer AF phase)and phase separation method(inner NP phase). The scaf?fold was investigated by general observation, stereomicroscope and scanning electron microscopy(SEM). Its pore size, poros?ity, and compressive elastic modulus were determined. AF and NP cells were isolated from rabbit IVD and seeded into the corresponding phase of the scaffold respectively. The cell-scaffold complex was cultured for 48 hours. The biocompatibility of the scaffold was evaluated by SEM, live/dead staining while CCK-8 assay was used to assess cell proliferation. Results Stereomicroscope and SEM showed that AF phase and NP phase integrated perfectly without cross-linking. Both phases pos?sessed highly interconnected porous structure [pore size of AF and NP phase were(220.0±23.1)μm and(90.0±17.8)μm, re?spectively] and highly porosity(AF and NP phase were respectively 91%and 93%). In addition, this silk biphasic scaffold had impressive mechanical properties(150.7 ± 6.8)kPa. SEM revealed that disc cells attached to regions of pore walls, dis?tributed uniformly and secreted extracellular matrix. Live/Dead staining and cell count kit-8(CCK-8)analysis showed that the silk composite scaffold was non-cytotoxic to disc cells. Conclusion This silk biphasic AF-NP scaffold has satisfied pore size, porosity, biomechanical properties and biocompatibility, so it is ideal candidate for IVD tissue engineering.

Objective To establish reference values of normal fetal heart conduction time intervals by tissue Doppler imaging(TDI)and pulsed Doppler(PD)echocardiography,and to assess their correlation with gestational-age and fetal heart rate.Methods One hundred and eighty-nine pregnant women underwent detailed echocardiographic examinations.Atrio-ventricular conduction time interval(AV)and the time interval from onset of ventricular contraction to the onset of atrial contraction of next cardiac cycle(VA)were measured by TDI and PD echocardiography.Results TDI-AV was(126.56±15.33)ms(95% CI 124.10～129.03 ms),TDI-VA was(285.22±24.53)ms(95% CI 281.27～289.16 ms),PD-AV was(127.42±12.88)ms(95% CI 125.35～129.49 ms),PD-VA was(287.42±25.19)ms(95% CI 283.37～291.47 ms).A paired t test revealed no systematic difference between the two approaches used to measure AV and VA.AV and VA were significantly positively correlated with gestational age,and significantly negatively correlated with fetal heart rate.Heart conduction time intervals were altered in fetus with paroxysmal arrhythmia.Conclusions This study established the normal values of fetal heart conduction time intervals.Prenatal determination of fetal heart conductional time intervals has important potential clinical utility in assessing fetal arrhythmia.

Objective To establish an animal model of annulus fibrosus (AF) partial defect for the repairing of interver?tebral disc (IVD) defect. Methods Image J 1.46r software was used to measure the T12/L1-L6/S1 intervertebral height in ovine lumbar spine X-ray films. AF thickness was measured by axial split disc. A 11 blade was used to make a trapezoid de?fect of upper bottom 3 mm, lower bottom 5 mm, height 5 mm and thickness 3 mm, whose lower bottom toward the nucleus pulposus (NP) in the left front of ovine lumbar IVD in vitro. The minimally invasive lateral approach was used to make the same type of trapezoid defect in the left front of the ovine lumbar IVD in vivo. The trapezoidal defect length of the axial divid?ing disc was measured, AF and a small amount of NP from trapezoidal defect in IVD were weighed, and the production of trapezoidal defect in IVD was evaluated. Results The lumbar intervertebral space height of ovine was (4.45 ± 0.28) mm. There were significant differences in the thickness of AF (4.08±0.50) mm , thickness (3 mm) and height (5 mm) of trapezoidal defect (P<0.05), respectively. There were no significant differences in trapezoidal defects in ovine lumbar IVD in vitro on the upper bottom (3.03 ± 0.09)mm, the lower bottom (5.03 ± 0.09) mm, the height (4.97 ± 0.10) mm, the thickness(3.02 ± 0.06) mm and the trapezoidal defect predetermined value on the upper bottom 3 mm, the lower bottom 5 mm, the height 5 mm and the thickness 3 mm (P>0. 05). The weights of the AF and NP taken out from ovine lumbar IVD in vitro and in vivo were (0.162 ± 0.011) g and (0.166 ± 0.014) g, and there was no significant difference between them (P > 0.05). Conclusion Through the operation of minimally invasive lateral approach, the method of making a trapezoidal defect in the experiments can establish animal model of AF partial defect, which meets the requirements for the repairing of IVD defect, and is simple, safe and reliable.

Objective To determine the usefulness of real time three-dimensional transesophageal echocardiography(RT-3D TEE) in the preoperative assessment of mitral valve (MV) pathology by comparing images with surgical findings and to evaluate the function of MV postoperatively. Methods Nineteen consecutive adult patients with established diagnosis of mitral regurgitation(MR) scheduled for surgical correction were enrolled. Intraoperative 2D and 3D transesophageal echocardiography(TEE) were performed. All the 3D images were compared with findings obtained from direct surgical inspection. Postoperative RT-3D TEE was performed immediately to evaluate outcomes of mitral annuloplasty. Results Superb 3D-TEE en face views of the MV were obtained in all patients. Correct diagnoses of MV pathology in agreement with the surgical findings were made in 16 of 19 patients (84. 2%), however in the remaining 3 patients, the diagnoses were incorrect despite good image quality. In one, RT-3D TEE diagnosed prolapse of A2 segment and P2 scallop,which was not confirmed at surgery, but MV degeneration and annular dilatation were observed. In another one, RT-3D TEE revealed prolapse of P3 scallop, while at surgery prolapse involved P2 and P3 with ruptured chordae. In the rest one,surgically detected prolapse of P2 was missed by RT-3D TEE. On the other hand with 2D TEE,the diagnoses correlated poorly with surgical findings,only 10 patients were accurately diagnosed (52.6%). In some patients,2D TEE was able to identify the MVP, but it had difficulty in defining the exact location of the prolapsed segment or scallop. The severity of MR decreased significantly after surgery. MV repair was successful in all patients except one(5.1%), in whom moderate to severe MR was still present and MV replacement was conducted subsequently as an alternative.Conclusions RT-3D TEE provides excellent imaging of MV components, including the anterior and posterior leaflets, as well as annulus and subvalvular structures, which can be especially helpful in planning the most appropriate surgery strategy. RT-3D TEE offers exact anatomic characteristics of mitral annuloplasty rings and bands, providing additional information for the evaluation of surgical outcomes.