Objective To establish a method for the determination of eight heavy metal elements ofPb,Cd,As,Hg,Co,V,Se,Mo in iron dextran by inductively coupled plasma optical emission spectrometry (ICP-OES).Method Through selection of detection wavelengths,optimization of instrument parameters,correction of spectral interference,verification of ionization interference and investigation of suppression methods,the eight heavy metal elements were analyzed by ICP-OES.Results The recovery rate of the detected elements increased 5％ to 10％ by suppression of ionization interference.The accuracy of the method was good.The spiked recovery rates of the detected elements were from 95.7％ to 101.1％.The precision of the method were good (RSD ＜ 3.6％,n =6).The linearities of the detected elements were good,and the correlation coefficients were all greater than 0.999.The detection limits were from 0.15 to 8.09 ng/mL.The quantization limits were from 0.46 to 24.26 ng/mL.Conclusion The method was accurate,sensitivity,rapid and reliable,which can be applied to the determination of contents of eight heavy metal elements in iron dextran.

Objective To evaluate the effects of isoflurane anesthesia on NR1 expression and neuronal apoptosis in the hippocampus and cortex of adult rats .Methods Thirty-six adult male Sprague-Dawley rats , weighing 250-280 g ,were randomly assigned into 3 groups using a random number table :control group (group C , n=6 ) ,O2 inhalation group (group O , n=6 ) and isoflurane anesthesia group (group I , n=24 ) .The rats were exposed to 2% isoflurane (group I) ,to pure oxygen (group O) ,or to air (group C) for 2 h .At 2 h ,and 1 ,7 and 14 days after the rats were awake (T1-4 ) ,Morris water maze test was performed .The rats were then sacrificed and brains were removed for isolation of the hippocampus and cortex .NR1 expression was detected using SABC immuno-histochemical technique and neuronal apoptosis was determined using TUNEL .Results Compared with group C , the escape latency at T2 and total swimming distance at T1 ,2 were significantly prolonged , and the expression of NR1 in hippocampi was down-regulated at T1 ,2 in group I ,and the expression of NR1 in the cortex was down-regulated in O and I groups ( P<0.05) .There was no significant difference in the apoptosis index between the three groups ( P> 0.05 ) .Conclusion Isoflurane anesthesia can decrease the cognitive function transiently ,which is related to inhibition of up-regulation of NR1 expression in the hippocampi ,but not related to neuronal apoptosis in adult rats .

Objective To establish a method for the determination of eight heavy metal elements (Pb,Cd,As,Hg,Co,V,Se,and Mo) in iron dextran by inductively coupled plasma optical emission spectrometry (ICP-OES).Methods Through detection wavelength selection,optimization of instrument parameters and applying interference element correction (IEC) method to correction of spectral interference,the eight heavy metal elements were analyzed by ICP-OES.Results The elements recoveries were from 88.7％ to 101.3％ by correction of spectral interference with IEC method.The accuracy of the method was good.The linearities of the detected elements were good,and the correlation coefficients were all greater than 0.999.The detection limits were from 0.12 to 7.26 ng/rnL.The quantization limits were from 0.40 to 23.96 ng/mL.The precision of the method was good (RSD＜3.5％,n=6).Conclusion The results of the spectral interference correction by IEC method are greatly superior to the results of conventional method.The established method is accurate,sensitive,and rapid,which can be applied to the determination of contents of eight heavy metal elements in iron dextran.

Objective To compare the dissolution curves of reference preparation and self-prepared Iloperidone Tablets in four different pH dissolution media (0.1 mol/L HC1 solution,pH 4.5 acetate buffer solution,pH 6.8 phosphate buffer solution,and water).Methods The solubility of Iloperidone in different pH solutions was measured,the dissolution curves of two preparations in four different pH dissolution media were determined by HPLC,and the similarity was investigated according to the f2 factor method.Results The f2 factors between reference preparation and self-prepared Iloperidone Tablets in four different media were more than 50.Conclusion The two preparations are equivalent in four different pH dissolution media in vitro.

Objective To evaluate the effect of emulsified isoflurane on cognitive function in rats.Methods Seventy-two adult male SD rats,aged 8 weeks,weighing 250-300 g,were randomly divided into 3 groups:normal control group (group C,n =12),intralipid group(group E,n =12),and 8％ emulsified isoflurane group ( group EI,n =48).Morris water maze test was performed at 2 h after administration in group E and at 2 h,1,7,14 d after administration in 12 rats at each time point in group EI.The escape latency,staying time at the original platform quadrant,frequency of crossing the original platform and swimming speed were recorded.Orbital blood samples were taken from 6 rats in each group after water maze test for determination of the plasma corticosterone concentration,and then the animals were sacrificed and their hippocampi were removed for determination of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) contents.Brains were removed from another 6 rats in each group after water maze test for determination of the expression of BDNF and NGF in DG,CA3,CA2,CA1 of hippocampus.Results Compared with group C,in group EI the escape latency at 2 h after administration was prolonged,staying time at the original platform quadrant was shortened,the expression of BDNF in DG and CA3 of hippocampus was down-regulated and the BDNF content in hippocampus was decreased at 2 h and 1 d after administration( P ＜ 0.05 or 0.01).The escape latency was shortened and staying time at the original platform quadrant was prolonged at 7 and 14 d after administration,the content of NGF in hippocampus was increased at 1,7 and 14 d after administration and the expression of BDNF in DG and CA3 of hippocampus was up-regulated at 1d after administration as compared with those at 2 h after administration in group E1( P ＜ 0.05 or 0.01).There was no significant difference in the variables mentioned above between groups E and C( P ＞ 0.05 ).There was no significant difference in plasma corticosterone concentration among the 3 groups ( P ＞ 0.01 ).Conclusion The mechanism by which emulsified isoflurane results in transient cognitive impairment in rats is related to down-regulating the expression of BDNF in hippocampus,but not related to corticosterone and NGF.

Objective To investigate the possibility of establishing a Modic changes (MCs) animal model,and explore the pathogenesis of MCs through imaging,histology and molecular biology experiments.Methods Fifty four New Zealand rabbits (weight 2.5-3.0 kg,half male and half female) were randomly divided into 3 groups:sham group (n=l8),muscle embedment group (n=18) and NP embedment group (n=18).In NP embedment group,the L4-5 and L5-6 discs were exposed by the lumbar anterolateral surgical approach.A 16 G needle was used to puncture the L5-6 vertebral body close to the epiphyseal plate.The depth of the drilling was approximately 3 mm.A 5 ml syringe was then put into the L4-5 intervertebral disc and extracted the NP,which was injected into the drilled hole of the vertebral body.The muscle embedment group and sham group shared the same operating procedures and drilling methods with the NP embedment group.Some pieces of muscle acquired from paraspinal muscles were put into the drilled hole in muscle embedment group,while nothing was put into the drilled hole in sham group.After that,the bleeding stopping,tissue washing and suture were done in all groups.12 weeks,16 weeks and 20 weeks after the surgery,MRI scan was applied to each group.All the specimens were tested by HE staining,real-time fluorescence quantitative PCR and Western blot to observe the expression of inflammatory cytokines.Results After modeling for 12 weeks,16 weeks and 20 weeks,MRI showed low signal changes on T1WI and mixed high signal in the context of low signal changes on T2WI in the NP embedment group.However,the muscle embedment and sham group showed no significant signal changes.Gross observation and HE staining confirmed that there was abnormal tissue proliferation in the imbed site of the NP embedment group.RT-PCR and Western blot showed high expression of IL-4,IL-17 and IFN-γin the NP embedment group,which were positively correlated with the length of the postoperative period.There was no significant difference between the muscle embedment group and sham group.Conclusion The puncturing of vertebral body close to endplate and putting nucleus into it can create an animal model of MCs.Autoimmune factors may play an important role in MCs.

Objective To observe the expression of Interleukin-17 (IL-17) in different types of herniated disc tissue,and to investigate Modic changes in corresponding injured segment and relationship between Modic changes and herniated types.Methods Fifty patients with single level disc herniation were enrolled in this study.According to clinical diagnosis criteria,imaging (CT and MR) examination results,and the view in surgery process the patients were divided into 2 groups:herniation group(35 patients),degeneration group(15 patients).According to MRI,patients were divided into 2 groups:Modic changes group(14 patients),non-Modic changes group (36 patients).The disc tissues were observed by HE staining and immunohistochemical staining,and the expression of IL-17 was detected by using real-time quantitative PCR.Results Pathological examination revealed that there were more granulation tissue and neovascularization in herniation group than in degeneration group.Immunohistochemical result showed that there were more IL-17-positive cells in herniation group than in degeneration group; there were more IL-17-postive cells in Modic changes group than in non-Modic changes group.The expression of IL-17 mRNA in herniation group was 6.7701 times more than that in degeneration group.The expression of IL-17mRNA in Modic changes group was 4.2836 times more than that in non-Modic changes group. Conclusion The herniated disc nucleus pulposus can cause more prominent local inflammation.Autoimmune responses maybe play an important role in pathophysiological mechanisms of Modic changes.

Objective To screen and validate differentially expressed microRNAs in human degenerative nucleus pulposus cells (NPCs) and to predict their target genes,and to investigate the role of JNK pathway in degenerative intervertebral disc disease.Methods Eight degenerative nucleus pulposus tissues were harvested from 8 patients (6 males and 2 females) with lumbar degenerative disease,and three normal nucleus pulposus tissues were harvested from 3 patients (3 males) with lumbar fracture,intraoperatively.Differentially expressed microRNAs were screened by microRNA microarray analysis and validated by real-time qPCR.Target genes of highly expressed microRNAs were predicted by analyzing information from data bases:MicroCosm v5,TargetScan 5.1 and microRNA.org.Signal pathways associated with the target genes were analyzed,and qPCR was used to validate the screening results.Results Twenty differentially expressed microRNAs were identified.The microRNA-513a-5p and microRNA-494 were highly expressed in degenerative nucleus pulposus tissue,which was corresponding to the verification results,and the ratio was 2.222 2 and 2.948 5,respectively.Target genes of microRNA-513a-5p and microRNA-494 were MKK4 and JunD,respectively.MKK4 and JunD were both involved in JNK signaling pathway and located in upstream and downstream of this pathway,respectively.Conclusion In human degenerative nucleus pulposus cells,microRNA-513a-5p and microRNA-494 are highly expressed microRNAs,whose target genes are MKK4 and JunD,respectively.Both MKK4 and JunD are involved in JNK signaling pathway.These results indicate that JNK pathway may play important role in the pathogenesis of degenerative intervertebral disc disease.

Objective To investigate the effect of tension stress on the changes of mRNA expression of interleukin-17B (IL-17) and inducible nitric oxide synthase (iNOS) in rat discs.Methods Sixty-four female Sprague-Dawley rats were randomly divided into 4 groups:sham (S),tail-suspended (TS),tail-suspended with needle puncture (TSNP) and single needle puncture (SNP) groups.Tail-suspension device made rats of TS group and TSNP group hindlimb suspended,which exerted tension force on the coccygeal vertebrae disc;and aseptic needle puncture on the tail disc between the seventh and eighth coccygeal vertebrae (Co7/Co8)of TSNP,SNP group induced the inflammatory environment.The discs were harvested 4 weeks later.Measurements included gross observation,histological observation and real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) analysis.Results Gross and histological examination found that varying degrees of degenerative changes in the annulus and nucleus in TS,SNP and TSNP groups,but not in the S group.RT-qPCR results showed that the mRNA of IL-17B and iNOS were significantly higher in the TSNP and SNP groups than that in the S group.The values of 2-△△Q were 12.99 and 8.12 in TSNP groups,and the 2-△△Q values of SNP group demonstrated 22.49 and 14.98 compared to S groups.Sole application of tension stress brought by the tail-suspension slightly enhanced the expression of IL-17B and iNOS,the values of 2-△△Q were 1.22 and 1.49,which didn't reach a statistically significant change.Compared with SNP group,the mRNA expression of IL-17B and iNOS in TSNP groups were shapely decreased,the values of 2-△△Q were-9.50 and -6.86 respectively.Conclusion Relatively low magnitude tension stress might play a key role in anti-inflammatory process and the relief of discogenic pain.

Objective To develop and evaluate a porcine model for laparoscopic ureterovesical reimplantation (LUR) training. Methods Ten female pigs with a mean weight of 30 kg were used and the animals were placed on supine position after anesthesia. One 10 mm port and two 5 mm ports were placed after the establishment of pneumoperitoneum. The horn of uterus was used as "ureter". A model simulating the performance of LUR was then established on the mini-pigs. Four trainees per-formed the LUR procedures on the animal models during an advanced laparoscopic urology training course, following the technique criteria exactly used in LUR. The learning curve was analyzed in terms of operation time. Results The porcine model for laparoscopic training was established suc-cessfully and 4 LUR trainings was performed on each porcine. Each trainee performed 10 LURs on the models during the training course. The operation time declined from 170±10 rain initially to 90±4 rain after the training course (P<0.01). At the end of this training, all trainees could accomplish a watertight LUR procedure on the model. Conclusions The establishment of the training model is feasible. The trainees could acquire skills needed to perform LUR in vivo based on this simple model and to develop dexterity and facility in laparoscopic manipulation of needles and sutures as well. The model provides a platform for basic techniques training of the ureteral reconstruction procedures.