Objective To construct the luciferase reporter gene vector of cell division cycle 2 (Cdc2) gene promoter and determine its transcriptional activity. Methods Primers were designed based on human Cdc2 promoter sequence from UCSC software. Then Cdc2 promoter from human genome DNA was replicated. After pGL3-Basic vector and Cdc2 promoter were digested with restriction enzymes SacⅠand XhoⅠseparately, Cdc2 promoter was inserted into pGL3-Basic vector. The recombinant plasmid named pGL3-Cdc2-promoter was transiently co-transfected into U2OS cells with control vector pRL-SV40, and then the activity of dual luciferase was detected. Results pGL3-Cdc2-promoter was constructed successfully. The restriction analysis and sequencing proved the entirely correct sequencing results. The luciferase activity was higher in pGL3-Cdc2-promoter/pRL-SV40 group than that of pGL3-Basic/pRL-SV40 group (1.591 5±0.199 8 vs 0.049 9±0.010 4). Conclusion pGL3-Cdc2-promoter can be transcribed and activated in U2OS cells. This study provided an important basis for screening and evaluation of anticancer drugs.

Background The characteristics of anisometropic amblyopia in ocular morphology are becoming a hot topic in amblyopia field.And the interocular difference in corneal parameters of anisometropic amblyopia is to be understood here.Corneal topography is a non-invasive method for in vivo corneal examination and applied in our study.Objective This study was to investigate the interocular difference of corneal topography in anisometropic amblyopic patients Methods This was a serial cases observation.Thirty anisometropic amblyopes were selected in Center for Optometry and Visual Science,People's Hospital of Guangxi Zhuang Autonomous Region.The patients were divided into amblyopic eye group and fellow eye group based on the best corrected distance visual acuity.Corneal topography was examined with Orbscan Ⅱ z,and corneal morphological parameters such as Diff values of the anterior and posterior corneal surface,Sim K 's astigmatism,Kmax,angle kappa and central corneal thickness (CCT) were measured.The interocular differences in these parameters were evaluated by paired t test,and the correlations in these parameters between the amblyopic eyes and the fellow eyes were analyzed by Pearson linear correlation and simple regression analysis.Results The Diff values of the anterior corneal surface were (0.011±0.006)mm and (0.011±0.017)mm,and those in the posterior corneal surface were (0.031 ±0.012)mm and (0.026 ±0.008)mm in the amblyopic eye group and the fellow eye group,respectively.In addition,Sim K's astigmatism were (1.8± 1.1)D and (1.1 ±0.6) D,JCC were (77±80) °and (100±80) °,J0 values were (-0.17±0.72) D and (0.02±0.41) D,J45 values were (-0.16±0.79) D and (0.13 ±0.48) D,CCT values were (551 ±37) μm and (551 ±31) μm,angle kappa values were (6.4± 1.4) ° and (4.9 ± 1.2) ° in the amblyopic eye group and the fellow eye group,respectively.Significant differences were found in the Diff values of the anterior and posterior corneal surface,Sim K's astigmatism and angle kappa between the two groups (t=-3.041,P=0.005 ;t=-4.317,P=0.000 ;t=-4.571,P=0.000).Pearson's linear correlation test demonstrated significant interocular positive correlations in parameters such as the anterior corneal surface Diff values (r =0.444),J0 (r =0.383),posterior Diff values (r =0.600),and Sim K 's astigmatism (r =0.479),and CCT (r =0.948,P＜0.05).The linear regression equation between the two groups was Y =-0.005 +1.392X (R2 =0.197,F=6.858,P=0.014) in the Diff values in the anterior corneal surface,Y =-0.013+0.421X (R2=0.360,F=15.761,P=0.000) in the Diff values in the posterior corneal surface,Y =0.616+0.27X (R2=0.230,F=8.348,P=0.007) in the Sim K's astigmatism,Y=0.060+0.219X (R2 =0.147,F=4.814,P=0.037) in theJo and Y=i08.289+0.804X (R2 =0.899,F=250.293,P=0.000) in CCT.Conclusions Corneal morphological interocular differences exist significantly in anisometropic amblyopic patients.

Objective To evaluate the value of diffusion registration in diffusion tensor imaging (DTI) through comparing the diffusion tensor data to conduct the two kinds of post-processing methods for obtaining the ADC ,eADC and FA values and the vis-ual neural fiber length .Methods 20 cases of DTI data were analyzed retrospectively .The original data were adopted to directly process for obtaining the ADC ,eADC and FA values respectively ,after exerting diffusion registration the ADC ,eADC and FA val-ues were measured again ,at the same time the two different processing modes were performed the fiber trace imaging ,.The changes of ADC ,eADC and FA values in each case were compared before and after exerting the diffusion registration ,and at the same time the differences of fiber length in the same interest area in the fiber trace image were compared before and after exerting the diffusion registration .Results The difference of the ADC and eADC values obtained before and after exerting the diffusion registration in the normal group showed no statistical significance (P=0 .695 3 ,P=0 .632 1) and the FA value difference between before and after ex-erting the diffusion registration had statistical significance (P=0 .032 1);the difference of the ADC and eADC values obtained be-fore and after exerting the diffusion registration in the clinical patients group had no statistical significance ( P= 0 .203 9 ,P=0 .075 4) ,the FA value difference had statistical significance (P=0 .011 4) ,the visual neural fiber length was elongated after exer-ting the diffusion registration .Conclusion The diffusion registration processing can obtain better quality of ADC ,eADC ,FA images and the fiber trace image ,more reliable ADC ,eADC and FA values ,the visual fiber length is elongated ,which has large application value in MR DTI .

Renal function was studied in 55 prematures (PTS) without complication, 28 male and 27 female. All PTS were divided into three groups according to age of days(AD), gestational age(GA), and body weight(BW)。BUN and SCr were significantly correlated conversely with AD, GA and BW (P

OBJECTIVE: To develop an analytical method for determination of unknown residual solvents in solid drug products by GC-MS in order to meet the need of supervision. METHODS: Fifty residual solvents including benzene, carbontetrachloride, 1, 2-di-chloroethane, 1, 1, 1-trichloroethane, acelonitrile, chlorobenzene, acetone, etc, were determined by GC/MS using selected ion monitoring (SIM) mode. RESULTS: Good linearity was obtained with correlation coefficients of more than 0.9975 for all the solvents. The average recoveries of the quality control samples at low, middle and high concentrations were from 88.8% to 109.8% and the relative standard deviations (RSDs) were lower than 11.6%. The limits of detection (TODs) were in the range of 0.00003-0.3μg·mL-1, while the limits of quantitation (LOQs) were in the range of 0.00009-1 μg ·mL-1. CONCLUSION: The method is simple, rapid, sensitive and accurate, and can be used for the simultaneous determination of 50 residual solvents in solid preparations such as tablets and capsules.

Objective To evaluate the effect of anisodamine on endoplasmic reticulum stress during acute kidney injury (AKI) in rats.Methods Forty-two nale Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups:control group (group C,n =6),AKI group (n =18) and anisodamine group (group AD,n =18).AKI was induced by intramuscular injection of 50％ (v/v) glycerol 10 ml/kg into bilateral hind limbs in groups AKI and AD.In addition,anisodamine 10 mg/kg was injected intraperitoneally 20 min before intramuscular injection of glycerol in group AD.In group C the rats received intramuscular injection of normal saline 10 ml/kg into bilateral hind limbs.Six rats were chosen immediately after injection of normal saline in group C or at 1,6 and 24 h after glycerol injection in groups AKI and AD and then anethetized with intraperitoneal pentobarbital.The animals were sacrificed and kidney specimens were obtained and cut into sections which were stained with hematoxylin and eosin for pathological examination.The pathological changes of the renal tubules were scored.The expression of glucose-regulated protein 78 (GRP78) and oxygen-regulated protein 150 (ORP150) in renal tissues was determined by immuno-histochemistry and Western blot.Results Compared with group C,the pathological scores were significantly increased and the expression of GRP78 and ORP150 was up-regulated at all time points in groups AKI and AD (P ＜ 0.01).Compared with group AKI,the pathological scores were significantly decreased and the expression of GRP78 and ORP150 was down-regulated at all time points in group AD (P ＜ 0.05 or 0.01).Conclusion Anisodamine can ameliorate AKI through inhibiting endoplasmic reticulum stress in renal tubular epithelial cells and decreasing endoplasmic reticulum stress-induced cell apoptosis in rats.

Objective To evaluate the efficacy and safety of argatroban in the treatment of progressive cerebral infarction.Methods Three hundred hospitalized patients with progressive cerebral infarction from October 2006 to September 2011 were collected,among which 150 cases who agreed to the use of argatroban as experimental group,and 150 cases who didn' t agree to the use of argatroban for economic reasons as control group.Experimental group was treated with venous argatroban.It was 60 mg/d within the first 2 days and 24 h continuous intravenous drip.From the beginning of the 3rd day,the dosage was changed to 10 mg each time,twice a day for 5 days.Additionally,75 mg clopidogrel was given once a day for 12 days.Control group was only treated with 75 mg clopidogrel once a day for 14 days.The degree of neurological deficit was compared using the National Institutes of Health Stroke Scale (NIHSS) scoring and the rehabilitation condition was evaluated by Barthel index (BI) and modified Rankin Scale (mRS) scoring between 2 groups before treatment and 3 d,14 d,30 d and 90 d after treatment respectively,and further compared to patients with large artery stenosis and anterior or posterior circulation infarction.The indexes of coagulation function of 2 groups were monitored meanwhile.Results The degree of neurological deficit was significantly improved in the short term (before treatment:NIHSS 15.19 ± 2.70,after treatment of 3 d 10.75 ±2.09,t =2.l14,P =0.037 ;14 d 8.77 ± 1.50,t =2.092,P =0.039;30 d 6.89 ± 0.79,t =2.520,P =0.013 ;90 d 4.85 ± 0.38,t =2.723,P =0.008),and the activities of daily living were significant enhanced in the experimental group (after treatment of 30 d BI 70.89 ± 12.69,90 d 88.16 ± 11.96,90 d mRS 1.57 + 0.39) when compared with the control group (after treatment of 30 d BI 60.26 ± 11.85,t =2.292,P=0.023;90 d 69.90 ± 12.63,t =2.790,P =0.006;90 d mRS 2.14 ±0.52,t =2.124,P =0.035).For large artery stenosis or posterior circulation infarction,the efficacy of argatroban was better.The indexes of coagulation function were in the normal range both before and after medication.No serious adverse reaction occurred during treatment.Conclusion Argatroban which has a higher security probably improves the prognosis and reduces the disability of patients with progressive cerebral infarction.

Objective To evaluate therapeutic effects of endovascular therapy for in-stent restenosis of superficial femoral artery,and the risk factors of restenosis of re-intervention therapy.Methods We retrospectively summarized the clinical data of 35 patients(38limbs)who suffered from in-stent restenosis of superficial femoral artery(SFA)stent from 2010 to 2012.Results 35 patients (38 limbs)were enrolled in this study,there were 24 males and 11 females.Mean age was (68 ±8)years.The success rate of endovascular therapy was 89.5％ (34/38),including in-stent angioplasty in 14 limbs,in-stent and beyond-stent angioplasty in 5 limbs,in-stent angioplasty and beyond-stent re-stenting in 15 cases.The primary patency and sencondary patency rate for 6,12 and 24 months was 65.2％,46.5％,46.5％,and 87.5％,80.2％,55.8％ respectively.The independent risk factors for re-obliteration were age ＞ 70 years (P ＜ 0.05) and diabetes mellitus (P ＜ 0.05).Conclusions Endovascular therapy is effective and safe in treatment in-stent restenosis of superficial femoral artery.The long-term in-stent patency is not satisfactory,with age ＞ 70 years and diabetes mellitus as independent risk factors.

Objective To assess the value of combined detection of enterovirus nucleic acid and antibody in early etiological diagnosis for hand-foot-and-mouth disease ( HFMD).Methods A case-control study was conducted.A total of 1 066 cases of children clinically diagnosed with HFMD from Hangzhou Children′s Hospital were involved into the research group from January to June 2014, consisting of 401 common cases and 665 severe cases; Throat swabs and serum samples from these children underwent combined detection for EV71/CA16/EV of enterovirus nucleic acid by fluorescence quantitative RT-PCR and for EV71/CA16-IgM by ELISA.All data were analyzed with SPSS 16.0.Results The total positive rate of enterovirus nucleic acid EV71/CA16/EV by fluorescence quantitative RT-PCR in the 1 066 cases of children clinically diagnosed with HFMD was 75.52%( 805/1 066 ) ( 95%CI: 72.80%-78.05%).But the total positive rate of combined detection was 91.46%( 975/1 066 ) ( 95%CI:89%.58-93.04%).The total positive rate of combined detection is higher than that of RT-PCR test(χ2 =98.338,P=0.000).The positive rate of EV71 type of combined detection was 64.63%(689/1 066)(95%CI:61.67%-67.49%),which is 15.38%higher than that of RT-PCR test 49.25%(525/1 066)(95%CI:46.21%-52.29%)(χ2 =51.453, P=0.000).In 665 severe cases of HFMD, the total positive rate of combined detection was 96.69%(643/665)(95%CI:94.95%-97.87%), which is higher than that of RT-PCR test 79.25%(527/665)(95%CI:75.92%-82.22%)(χ2 =95.607, P =0.000).In the severe cases, the positive rate of EV71 type of combined detection was 87.52%( 582/665 ) ( 95%CI:84.71%-89.89%) , which is 18.95% higher than that of RT-PCR test 68.57%(456/665) (95%CI:64.87%-72.06%) (χ2 =69.665, P=0.000).In the fatal cases, the positive rate of EV71 type of combined detection was 95.92%(94/98) (95%CI:89.28%-98.68%).Conclusions The combined detection of enterovirus nucleic acid and specific IgM antibody can significantly increase the positive rate of HFMD, especially for severe cases.The combine detection increases both the total positive rate and EV71 positive rate.Thus it has a high potential for becoming a new guidelines for laboratory diagnosis of HFMD.

Objective To investigate the effects ofTianqi-Yizhi granules on oxidative stress in the brain tissue, and learning and memory in Alzheimer's disease model rats.Methods A total of 90 male SD rats were randomly divided into 6 groups by random number table method: sham operation group, model group, huperzine A group and groups of low-, medium- and high-doseTianqi-Yizhi granules, with 15 rats in each group. The AD rat model was prepared by the left lateral ventricle injection of amyloid-β1-42. One week after modeling, the rats in the groups of low-, medium- and high-doseTianqi-Yizhi granules received intragastric administration of 0.8, 1.6 and 3.2 g/kgTianqi-Yizhi granules, respectively; the rats in the huperzine A group received intragastric administration of 0.02 mg/kg huperzine A solution; and the rats in the sham operation and model groups received intragastric administration of equivalent volume of normal saline for 30 days. Learning and memory were evaluated using the dark avoidance test. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and malonaldehyde (MDA) level in the brain tissue were determined. ResultsIn comparison with the model group, the latencies to step into the dark chamber in the high- and medium-dose Tianqi-Yizhi granules groups were significantly longer (239.05 ± 48.42 s, 214.35 ± 74.52 svs. 97.39 ± 30.69 s; allP<0.01), the numbers of errors significantly decreased (1.93 ± 3.25, 2.27 ± 3.09vs. 6.62 ± 3.45; allP<0.05), the activities of SOD (177.27 ± 63.10 U/mg, 164.53 ± 72.58 U/mgvs.72.56±21.04 U/mg; all P<0.01) and GSH-Px (2 899.36 ± 362.27 U/g, 2 407.68 ± 472.14 U/gvs. 1 397.64 ± 442.17 U/g; allP<0.01), and MDA level (24.75 ± 9.94 nmol/mg, 27.74 ± 5.82 nmol/mgvs. 37.56 ± 17.23 nmol/mg; allP<0.01) in the brain tissue significantly increased. ConclusionTianqi-Yizhi granules could attenuate oxidative stress in the brain tissue, and improve learning and memory in AD rats.