OBJECTIVE:To establish the method to determine the concentration of human plasma felodipine by HPLC-MS.METHODS:The HPLC separating system was adopted and the mobile phase was a mixture of methanol-water-acetic acid with a flow rate at1ml/min and split ratio at2∶1,the fixed phase was Hypersil C 18 column;The mass spectrometric de-tection system and AP-ESI positive ion model were adopted with the atmosperic pressure at0.054Pa(400psi),the protecting gas at N 2 10L/min,capillary voltage at4000V,the ion source temperature at100℃and the fragment voltage at220V,the ions collected by SIM were felodipine m/z411(M+Na),Nimodipine m/z441(M+Na).RESULTS:The linear concentration range for felodipine was0.2～50ng/ml with a minimum detection limit at0.15ng/ml,the recovery was above90.93%with both the intra-day RSD and the inter-day RSD at less than9.8%(RSD

Objective:To transfect the tongue squamous cell carcinoma Tca8113 cells with NICD, and to investigate the effects of Notch 1 on Tca8113 cells and the expression of epidermal growth factor receptor (EGFR) in tongue squamous cell carcinoma.Methods: The Notch plasmid pRAMIC-IRES2-EGFP encoding exogenous and control plasmid pIRES2-EGFP were transiently transfected into the Tca8113 cells by LipofectamineTM 2000. In the experiment, target plasmid pRAMIC-IRES2-EGFP transfection group, control plasmid pIRES2-EGFP transfection group and non-transfection group were set up.RT-PCR, Western blotting and immunocytochemical method were used to observe the mRNA and protein expression levels of Notch 1 and EGFR.The proliferation activities of Tca8113 cells were detected by MTT assay,and the survival rates were calculated.Results:Compared with non-transfection group and control plasmid pIRES2-EGFP transfection group, the mRNA and protein expression levels of Notch 1 in Tca8113 cells in pRAMIC-IRES2-EGFP group was significantly increased(P<0.05), the mRNA and protein expression levels of EGFR in Tca8113 cells were significantly decreased (P<0.05),and the proliferation rate of Tca8113 cells was significantly decreased(P<0.05) after transfection.There were no significant differences in the expression levels of Notch 1 and EGFR mRNA and protein and the proliferation rates in Tca8113 cells between non-transfection group and pRAMIC-IRES2-EGFP group (P>0.05).Conclusion:The effect of Notch 1 on the proliferation of Tca8113 cells may be related to down-ragulatoin of expression level of EGFR.

Objective: To study the association of dopamine ?-hydroxylase gene 5′ flanking region polymorphism -1021 C→T with attention deficit hyperactivity disorder (ADHD) in Han children. Methods: ADHD was diagnosed according to the DSM-Ⅳ criteria . DNA was extracted and PCR was performed to examine the DBH-1021C→T polymorphism. HHRR was used to test the association of dopamine ?-hydroxylase gene with different subtypes of ADHD. Results: HHRR results showed this polymorphism had a tendency to be associated with the inattention subtype (P=0.067) and the combined subtype (P=0.076). The T allele was the protective factor of the inattentive subtype (P= 0.07), and the risk factor of the combined subtype (P=0.08). After dividing the 292 nuclear families according to sex, DBH-1021C→T polymorphism was found to be associated with the combined subtype (P=0.04) with the T allele as the risk factor (P=0.02). There were no positive findings among the girl nuclear families of ADHD. Conclusion: DBH gene is related to ADHD combined subtype and inattention subtype. The genetic basis of ADHD combined subtype and is inattention is different, and is influenced by sex factor.

Objective To establish a way to measure the injury severity of articular surface due to posterior malleolus fracture and investigate its clinical significance based on three-dimensional reconstruction technique.Methods Between May 2009 and March 2014,138 cases of peri-ankle injury combined with posterior malleolus fracture were treated.Lateral radiographs were reviewed to measure the ratio of posterior fragment area to distal tibial plafond area using the common method.Three-dimensional CT images were examined to measure ratio of the area of injury using the three-dimensional measuring method.Statistical comparison was made using the t-test and intraclass correlation coefficient (ICC).Results Measurement failed on 33 lateral radiographs and 13 three-dimensional CT images.Six cases cannot be measured by both imaging technology,so results of 98 cases were compared.Ratio of posterior fragment area to tibial plafond area was (16.2 ± 7.2) ％ measured by radiographs and was (29.6 ±10.1) ％ by three-dimensional CT scans (P ＜ 0.05).Three-dimensional CT measurements showed higher intra and inter-observer agreement (ICC of 0.973 to 0.942) than that in radiographic measurements (ICC of 0.875 to 0.860).Conclusion Three-dimension CT assessment of the extent of injury to articular surface because of posterior malleolus fracture is reliable and reproducible,but radiographic assessment may underestimate the injury and influence the choice of surgical indications.

Objective To investigate the effect of sustentaculum tali screw placement on outcomes following open reduction and internal fixation (ORIF) of intraarticular calcaneal fractures.Methods A retrospective analysis was made on 139 patients with intraarticular calcaneal fractures treated by ORIF from April 2008 to January 2012.According to Sanders classification,there were 9 patients with type Ⅱ fractures,87 with type Ⅲ fractures and 43 with type Ⅳ fractures.Fracture reduction followed by placement of calcaneal anatomic plate or anatomic locking plate was performed via calcaneal lateral L-shape incision.All the patients received X-rays and CT scans within postoperative 2 weeks.Placement of screws in sustentaculum tail was detected by CT volume rendering to visualize implants combined with multiplanar reconstruction imaging (MPR).Patients were divided into accurate fixation group (Group A,n =28),marginal fixation group (Group B,n =58),and non-fixation group (Group C,n =53) based on radiological results.Intra-and peri-operative parameters,time to partial weight-bearing,time to full weight-bearing,fracture healing time,and clinical outcome were compared among groups.Functional outcome was assessed using American Orthopedic Foot and Ankle Society (AOFAS) Ankle Hindfoot Scale.Results Postoperative X-rays indicated accurate fixation of sustentaculum tail in 64 patients,marginal fixation in 22,and non-fixation in 53,but CT images manifested accurate fixation in 28 patients,marginal fixation in 58,and non-fixation in 53.Group A had operation duration of (93 ±11) min and intraoperative blood loss of (252 ± 27) ml,longer or higher than those in Group B [(85 ±8) min,(194±16) ml] and Group C [(57±6) min,(136 ±13) ml] (P＜O.05).There was no significant difference in mean hospital stay among the three groups (P ＞ 0.05).Ninety-two patients were followed-up for 12-38 months (mean 18.5 months) and 85 patients completed foot and ankle exercise as planned.Moreover,no significant differences were observed among groups concerning time to partial and full weight-bearing,fracture healing time and AOFAS score at the final follow-up (P ＞ 0.05).Conclusion Sustentaculum tali screw placement has no apparent effect on the short-term outcome of surgical treatment for calcaneal fractures.

Objective To investigate the protective effect and potential mechanisms of hypertonic sodium chloride hydroxyethyl starch solution (HSH) against the cerebral vasospasm (CVS) following subarachnoid hemorrhage (SAH).Methods Twenty-four male Sprague-Dawley (SD) rats were randomly assigned to four groups according to the random number table,with 6 rats in each group.The SAH-CVS model was reproduced by injection of the blood twice through the cisterna magna.Rats in both model and HSH treatment groups received 8 mL/kg normal saline (NS) or HSH treatment everyday via caudal vein.Rats in sham group were injected with 1.5 mL/kg NS into cisterna magna followed by 8 mL/kg NS treatment.Rats in normal group received no treatment.Rats were sacrificed to harvest basilar artery after 7 days.The thickness of vessel wall and lumen area were measured using hematoxylin-eosin (HE) staining.The rate of apoptosis of vascular smooth muscle cell (VSMC) was assessed using flow cytometry.Caspase-3 activity was measured by a fluorometric assay.The expressions of Bax and Bcl-2 were determined by Western Blot.Intracellular reactive oxygen species (ROS) was detected by H2DCFDA.Results Compared with normal group,increased thickness of vessel wall (μm:27.72 ± 1.94 vs.18.30 ± 1.10,P＜0.05),decreased lumen area (μm2:26 115 ± 1 991 vs.55 080 ± 2 091,P＜0.05),and elevation of rate of apoptosis of VSMCs [(35.05 ± 5.54) ％ vs.(5.93 ± 1.53) ％,P＜ 0.05] were found in model group.Compared with model group,decreased thickness of vessel wall (μm:22.55 ± 1.50 vs.27.72 ± 1.94,P＜0.05),increase of lumen area (μm2:48 115 ±2 460 vs.26 115 ± 1 991,P＜0.05),and depressed rate of apoptosis of VSMCs [(16.54 ± 5.94) ％ vs.(35.05 ± 5.54) ％,P＜ 0.05] were found in HSH treatment group.Caspase-3 activity,intracellular ROS level,Bax and Bcl-2 expressions in model group were (188.40 ± 19.35)％,(163.50 ± 17.02)％,(208.71 ± 26.04)％ and (44.52 ± 9.61) ％ of those of normal group,and the differences of these parameters between model and normal groups were statistically significant (all P＜0.05).Caspase-3 activity,intracellular ROS level,Bax and Bcl-2 expressions in HSH treatment group were (135.05 ± 19.52)％,(119.44 ± 11.50)％,(139.20 ± 18.04)％ and (85.35 ± 13.12)％ of those of normal group,respectively,and the differences of these parameters between HSH treatment and model groups were statistically significant (all P＜0.05).The differences of all measurements between sham and normal groups were not statistically significant.Conclusion The current results demonstrate that HSH attenuates the SAH-induced CVS,alleviates thickness of vessel wall,and increases lumen area via inhibition of VSMCs apoptosis.

Objective To compare the clinical results of percutaneous compression plating (PCCP)and Trigen short reconstruction intramedullary nail for intertrochanteric hip fractures. Methods During 2005 and 2008, the patients suffered with AO/OTA Al and A2 intertrochanteric hip fractures were divided into two groups; 36 fractures were treated with PCCP and 48 fractures with Trigen short reconstruction nail.During an average of (16.3±3.2) months follow-up, clinical evaluation involved visual analogue scale(VAS)score for pain in the 1st week, the 1st month, the 3rd month, the 6th month and the 12th month, and a Harris hip score one year post operation. Radiographs were examined for fracture healing-time, displacement scale of the neck screws and fracture impaction scale. All the complication in both groups was recorded. Results There were no difference in blood loss and operation time in both groups. The postoperative pain was significantly lower in the PCCP group in the initial three months after the surgery. Larger scale of fracture impaction and screw telescoping were seen in PCCP group. Also shorter healing time, higher Harris score results were achieved in PCCP group than those of Trigen short reconstruction nail group. Four peri-implant fractures occurred in Trigen short reconstruction nail group, which included one in the greater trochanter and three in the femoral shaft, but only one case need revision for bone displacement. In PCCP group, it was found that the superior neck screw was slightly displaced for tendency to cut-out in one patient. Conclusion Both PCCP and Trigen short reconstruction intramedullary nail can be successfully used to treat Al and A2 intertrochanteric hip fractures with minimal invasive technique. And the PCCP showed more rapid pain relief and bone healing, easily bony reduction and fewer complications.

G. The current results indicate that ADHD with DBD has more heritable backgrounds when compared with ADHD without DBD.

T were genotyped by restriction fragment length polymorphism analysis.Transmit/disequilibrium test and haplotype analysis were used to test the association of the three polymorphisms with ADHD comorbid or not comorbid DBD separately.Results:Haplotype T/G/T showed tendency of overtransmission(?2=3.470,P=0.062) to probands of ADHD with DBD, while haplotype C/G/T(?2=4.568,P=0.032) and C/G/C(?2=5.333,P=0.021) were undertransmitted to probands of ADHD without DBD,No biased transmissions of any allele were found in families with probands of ADHD with and without DBD.Conclusion:whether ADHD comorbid DBD or not comorbid DBD makes difference at the level of HTR4 gene polymrohisms.

Objective To investigate the incidence and the related factors of the depression in Parkinson's disease. Methods 52 cases with idiopathic Parkinson's disease were included in this study. Such scales as Hamilton depression scale-24 item (HAMD-24), Non Motor Symptoms Scale for Parkinson's disease (NMSS), 39-item Parkinson's Disease Questionnaire (PDQ-39), unified Parkinson's disease rating scale (UPDRS), revised Hoehn-Yahr rating scale (H-Y), revised Schwab and English daily life event scale (S-E), Minimum Mental State Examination (MMSE), etc. were applied and underwent Spearman correlation analysis. Results Thirty-six patients with Parkinson's disease (69.23%) met the criteria for depression. The spearman rank correlation analysis showed that MMSE (r=-0.294, P=0.036) and S-E (r=-0.506, P<0.001) had negative correlation with depression in Parkinson's disease, and the course of the disease (r=0.274, P=0.049), UPDRS (r=0.485, P<0.001), H-Y (r = 0.459, P = 0.001), NMSE (r = 0.729, P < 0.001) and PDQ-39 (r = 0.619, P < 0.001) had positive correlation with depression in Parkinson's disease. Conclusions The patients with Parkinson's disease had high incidence of depression. The depression in Parkinson's disease is associated with many factors and affected the quality of life of the patients.