OBJECTIVE:To establish an HPLC method for the determination of dextromethorphan and dextrophan in human urine and to discriminate phenotype of CYP2D6.METHODS:The urine samples were hydrolyzed by enzyme,distilled and purified by acid and alkali before direct introduction.HPLC system was adopted using Waters Nova-Pak Phenyl column with mobile phase consisted of acetonitrile-water(60∶40)at a flow rate of 1.0 mL?min-1.The fluorescence detection was performed with excitation wavelength at 280 nm and emission wavelength at 310 nm.The column temperature was 25℃.RESULTS:The linear ranges of dextromethorphan and dextrophan were 0.065～4.148 ?g?mL-1(r=0.998 5)and 0.353～22.592 ?g?mL-1(r=0.999 7),respectively.The lower detection limits were 0.016 ?g?mL-1 and 0.018 ?g?mL-1,respectively.The average recoveries were 90.5% and 93.5%,resepctively;the intra-day RSD were no more than 3.26% and 3.47% and inter-day RSD were no more than 3.65% and 4.05%,respectively.All the 8 subjects were extensive metabolizers(EM).CONCLUSION:The method is simple and reliable,and suitable for the determination of both dextromethorphan and dextrophan in human urine and as well as the study of the phenotype of CYP2D6 polymorphisms.

Objective To study the clinical effect of the minimally invasive surgery for treating recurrent lumbar disc protrusion . Methods Thirty operative cases of recurrent lumbar disc protrusion treated with the minimally invasive surgery from July 2009 to July 2013 were retrospectively analyzed for statistically describing the age ,body mass index ,recurrent segment ,operating time ,intr-aoperative bleeding volume and postoperative painless walking time .The recurrence segments were 1 case of L34 ,10 cases of L45 and 19 cases of L5 S1 ;the visual analogue scale (VAS) was adopted to evaluate the operative effect .Results The operating time was 65 min on average (50-100)min;mean intraoperative bleeding volume was 145 mL(100-180)mL ;average length of hospital stay was 13 d (9-16)d;average hospitalization costs were 7 300 Yuan;average postoperative painless walking time was 11 d (7-15)d;post-operative non-manual labor work time was 27 .3 d .The VAS score was reduced from (7 .3 ± 1 .3) before operation to (3 .1 ± 0 .9) after operation and (2 .2 ± 0 .6) at the last follow-up ,which was statistically significant(P<0 .05) .Followed up for 6-33 months , average 16 .5 months .Superior efficacy in 16 cases ,gord in 7 cases ,average in 5 cases ,bad in 2 cases ,the rate of good efficacy was 76% .Conclusion The minimally invasive surgery is one of effective methods to treat postoperative recurrence of lumbar disc pro-trusion .

Objective To investigate the effects of antisense Smad2 oligodeoxynudeotides(ODN) on fibronetin(FN) and collagen Ⅳ(ColⅣ) secretion of rat mesangial cells cultured with high glucose, explore the action of Smad2 in the glomerulosclerosis and to find a new method to retard the progress of glomerular fibrosis. Methods 20-mer antisense, sense and random ODNs were designed and synthesized that were phosphorothioate modified to increase stability. The antisense ODN encompassed the ATG of the rat Smad2 gene. ODN was tranferred transiently into rat mesangial cells through liposome. Rat cells were treated with high glucose. mRNA and protein of Smad2 were detected by RT-PCR and cytochemistry. FN and ColⅣ were examined by ELISA. Results Antisense ODN significantly decreased mRNA and protein expression of Smad2 in rat mesangial cells treated with high glucose(P

4-fold elevation in renal cortex in plasma aldosterone as compared to those of the SHAM rats. The above pathologies were markedly improved in bi-ectomised rats with significantly lower aldosterone level. Being constantly infused exogenous aldosterone, bi-ectomized rats manifested greater proteinuria, hypertension, glomerulosclerosis and increased level of TGF-?1 compared to bi-ectomised rats. Indeed, these features were similar in exogenous aldosterone rats and 5/6 nephrectomized rats. Furthermore, the expression of mineralocorticoid receptor (MR) mRNA was remarkablely enhanced in SNX group and was decreased in ADX group. However, the mRNA expression of 11 ?-hydroxysteroid dehydrogenase II (11?-HSD2) in each group was opposite to that of MRmRNA. Ccr and kidney/body weight showed no differences among four experimental groups. Conclusion Aldosterone contributes to the progression of ablative nephropathy in the rat through mechanisms other than systolic blood pressure.

Objective To assess right ventricular(RV) regional and global function in patients with repaired tetralogy of Fallot (TOF) by speckle tracking imaging (STI).Methods 36 asymptomatic patients with repaired TOF whose postoperative period is more than one year and 35 healthy controls were studied.RV longitudinal peak systolic strain (ε),strain rate (SRs) were measured in RV free wall and interventricular septum(IVS) for basal,mid and apical segments by STI from the apical 4-chamber view,RV global longitudinal peak systolic strain (GLS),strain rate (GLSRs),early diastolic strain rate (GLSRe) and late diastolic strain rate (GLSRa) were measured from the above view.The correlations between RV systolic functional parameters and the postoperative period,degree of pulmonary regurgitation (PR) and QRS duration were explored.Results ①Compared with controls,εand SRs of RV free wall and IVS for most segments,GLS,GLSRs,GLSRe,and GLSRa were significantly impaired in patients with repaired TOF (P ＜ 0.05 for all).Both GLS and GLSRs were lower in postoperative patients with transannular patch than in those with RV out tract patch.②In comparison with controls,GLS,GLSRs,GLSRe and GLSRa were decreased in postoperative patients with mild,mid and severe PR,Whereas,there was no difference in RV global function among the patients with varying degree of PR.③GLSRs of RV was correlated inversely with postoperative period and QRS duration(r1--0.431,P1 =0.009 ; r2 ==-0.469,P2-0.004),RV GLS and GLSRs had no correlation with PR.Conclusions RV regional and global function in patients with late repaired TOF are decreased,STI can early detect right ventricular dysfunction in asymptomatic patients with repaired TOF.The type of operation impacts on late prognosis,RV abnormal deformations are associated with electrical depolarization abnormalities.

Objective To observe and compare the curative effects of zoledronic acid (ZA) combined with radiotherapy and ZA combined with chemotherapy in the treatment of bone metastasis of non-small cell lung cancer (NSCLC). Methods Seventy-eight patients with NSCLC bone metastasis treated by radiotherapy or chemotherapy were taken in our hospital from January 2010 to June 2014, including 39 cases of ZA combined conventional fractionation radiotherapy (radiotherapy combined group), and 39 cases of ZA combined with chemotherapy (paclitaxel liposome + cisplatin) (chemotherapy combined group). Each group had 39 cases. WHO objective evaluation standard, efficacy evalulation of solid tumor metastasis and curative effect standard grading of pain, anticancer agent toxicity classification standard, Karnofsky standard were used for evaluating and analyzing the patients with primary lung tumor, bone metastasis, degree of pain, adverse reactions and functional status (once before and after the treatment). Results The efficiency rate of primary lung tumor, the efficiency rate of bone metastasis, the total effective rate of pain relief and the improvement rate of functional status (Karnofsky score increased by 10 points or more)in the radiotherapy combined group and chemotherapy combined group were 82.05 % (32/39) vs. 79.49 % (31/39), 48.72 % (19/39) vs. 51.28 %(20/39), 82.05 % (32/39) vs. 84.62 % (33/39), 66.67 % (26/39) vs. 71.79 % (28/39) respectively, and the differences were not statistically significant (the values of x2 were 0.224, 0.237, 0.195, 0.259 respectively, all P> 0.05); Although the two groups showed low-grade fever, bone marrow suppression, esophagitis, liver and kidney damage, gastrointestinal reactions and other adverse reactions, the adverse reactions of two groups were close to [28.21 % (11/39) vs. 30.77 % (12/39)] (x2 = 0.314, P> 0.05). Fortunately, these reactions were controlled well after symptomatic treatment. Conclusion ZA combined with radiotherapy or chemotherapy is a safe and effective way for bone metastasis of NSCLC, which should be taken based on the individual condition of the patients.

Objective To examine the expression of protein kinase signaling cascade protein kinase B (Akt)-which plays an im-portant role in a number of key biological functions in cellular processes after spinal cord injury (SCI) in rats ,and its effects on SCI induced motor function defects ,and to provide the molecular mechanism for repairing SCI .Methods SCI was produced by extra-dural contusion using modified Allen′s stall with damage energy .The rats were divided into three groups :sham-operated group (Sham) ,interference group (Int) ,and control group (Con) .Using immunostaining studies ,Western blot analyses and real-time qualitative RT-PCR analyses ,we detected the changes of Akt expression at the protein and mRNA level in spinal cord tissues at 1 , 7 ,and 14 d after surgery ,and evaluated the presence and extent of neurological impairment after SCI by the BBB locomotor rating scale .Results The sham operated groups exhibited low expression of the Akt signaling at the protein and mRNA level ,and the ex-pressions increased following SCI .Compared to the animals in the sham operated groups ,prominently elevated level of Akt signaling was detected in the injured spinal cords of SCI group .LY294002 ,an inhibitor of phosphatidylinositol 3-kinase (PI3K) that initiates Akt phosphorylation ,prominently inhibited the expression of Akt produced by SCI and the spontaneous recovery of motor function after SCI .Conclusion Activated protein kinase signaling cascade Akt might be the important intervention aspects of involving in neuroprotective and repair process after SCI .

Objective To investigate the effects of trimebutine maleate (TM) on the expression of large conductance calcium-activated potassium channel (BKCa) and ryanodine receptors (RyR)channels at mRNA and protein level in colonic smooth muscle cell of cold restraint stress(CRS)induced rats.Methods A total of 24 Wistar rats were divided into CRS group,CRS with TM group and control group equally.The rats of CRS group were gavaged with 0.9％NaCl (6 ml/kg) daily; the rats of CRS with TM group were gavaged with 15 g/L TM (6 ml/kg) daily and activity was restricted in wire cage at 4 ℃ for two hours,continuously for five days.The rats of control group were gavaged with 0.9 ％ NaCl (6 ml/kg) once without CRS.The amount and characteristics of stool of rats in each group were observed.The colonic smooth muscle was isolated to detect the expression of BKCa and RyR at mRNA and protein level by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results The median of rats defecation particles of CRS group was six,control group was one and CRS with TM group was five.Compared with control group,the defecation appearance of CRS group and CRS with TM group was looser and wetter observed by naked eyes.Compared with control group,there was no obvious pathological changes in CRS and CRS with TM group.There was no significant difference in the mRNA expression of BKCa and RyR channels between control group and CRS group.Compared with control group,the BKCa expression at mRNA level of CRS with TM group increased 1.45 fold.Compared with control group,the RyR2 expression at mRNA level of CRS with TM group increased 1.32 fold.Compared with control group,the BKCa expression at protein level of CRS with TM group increased 1.39 fold,and there was no RyR2 expression band at protein level.Conclusion TM might affect colonic smooth muscle contraction through the upregulation of BKCa expression at mRNA and protein level and RyR expression at mRNA level.

Objective To evaluate right ventricular(RV) global and regional volume and systolic function by real-time three-dimensional echocardiography (RT-3DE) in normal children with different age.Methods One hundred and ninty-two normal children were divided into five groups by age:group Ⅰ,＜1 years old,32 cases;group Ⅱ,≥1 years old-＜3 years old,46 cases;group Ⅲ,≥3 years old-＜6 years old,36 cases;group Ⅳ,≥6 years old-＜9 years old,41 cases;group Ⅴ,≥9 years old-＜14 years old,37 cases.Full volume imaging of RV was obtained at the parasternal four-chamber view near the apex by RT-3DE.RT-3DE data set were analyzed off-line by TomTec RV-Function.RV were divided into three parts:inflow,body,outflow.The measurements were including RV global and regional end-diastolic volume (EDV),end-systolic volume (ESV),ejection fraction (EF) and the ratio of regional parts EDV to RVEDV. The volume and systolic function were compared in three regional parts.RV global and regional parts EF and the ratio of regional parts EDV to RVEDV were also compared in five age groups.Correlation analysis and curve estimation were studied on RV global and regional EDV with age and physical development indexes.Results In the comparison of three regional parts:inflow EDV and EF were higher than outflow and body parts (P＜0.05).No significant different was found between outflow EDV and body EDV (P＞0.05),however,outflow EF was significant higher than body EF(P＜0.05).The comparison of RV global and regional EF in five age groups were no statistical different (P＞0.12). The ratio of regional parts EDV to RVEDV remained constant in five age groups(P＞0.58).Correlation analysis showed the global and regional RV volume were strongly correlated with age,height,weight and BSA (r＞0.77,P=0.000).The best correlation was found with BSA (r＞0.83,P=0.000).Curve estimation demonstrated that the relationship of RV global and regional EDV with age and physical development indexes could be best expressed by power model,the best matched model were found with BSA.Conclusions Among three regional parts of RV,inflow and outflow parts volume contraction were the two main contribution factors for RV function.In childhood RV volume didn't increase linearly with age and physical development indexes,but in an exponential model.

Objective To investigate the effect and mechanism of murine bone marrow mesenchymal stem cells(BM-MSCs)transplantation on B-cell activating factor(BAFF)expression and B cell activation of MRL/Ipr mice.Methods Eighteen female MRL/Ipr mice were divided into the treatment group and the control group.Five female BAL B/C mice were used as negative controls.At the age of 18 weeks,the treatment group was transplanted with 1×10~6 murine BM-MSCs through vena caudalis,the control group was treated with 0.5ml sodium chloride.Enzyme linked immunoserbent assay(ELISA)was used to measure the level of the BAFF,IFN-γ,IL-2 and IL-10 in the serum.The percentage and numbers of Marginal zone,T1 and T2 B cells in spleen were detected by flow cytometry.Results①Eight weeks after transplantation,the level of BAFF [(32±14)ng/ml]in serum of the treatment group decreased significantly than the control group[(47±13)ng/ml](P＜0.05)as well as the level of serum IL- 10,IFN-γ and IL-2 levels[(19±7)vs(40±13)pg/ml](P＜0.01)[(25±20)pg/ml vs(38±25)pg/ml][(73±10)pg/ml vs(80±15)pg/ml].② Eight weeks after trans-plantation,the mice in the treatment group had lower percentages of marginal zone B cells[(15±4)% vs (21±5)%],and the numbers of marginal zone B cells were significantly decreased in the treatment group as compared with the control group[(9±6)×10~6 vs(19±10)×10~6,P＜0.05].③ Eight weeks after transplantation,the mice in the treatment group had lower percentages of T1 and T2 B cells[(3.4±2.1)% vs(7.3±4.0)%][(2.6+1.4)%vs(4.8±2.7)%],and the numbers of T1[(2.7±1.7)×10~6 vs(5.1±2.0)×10~6,P＜0.05]and T2 B cells[(2.0±1.2)×10~6 vs(3.7±1.7)×10~6,P＜0.05]were both significantly decreased in the treatment group as compared with the control group.Conclusion BM-MSCs transplantation decreases the expression of BAFF in association with the diminished production of the pathogenic cytokines IFN-γ and IL-10.Inhibition of BAFF also results in decreased numbers of T1 and T2 B cells and MZ B cells.