1.Construction and ldentification of Cell Division Cycle 2 Promoter Reporter Gene Vector
Tianjin Medical Journal 2014;(2):101-104
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.
2.Application value of diffusion registration in diffusion tensor imaging
Wei WU ; Jiannong ZHAO ; Weibo XIE
Chongqing Medicine 2014;(27):3602-3604,3607
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 .
3.Difference analysis of Orbscan Ⅱz corneal topography in anisometropic amblyopes:a contralateral eye study
Chinese Journal of Experimental Ophthalmology 2013;(6):582-586
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.
4.RENAL FUNCTION IN THE PREMATURES
Yubin WU ; Kelun WEI ; Yazhen ZHAO
Chinese Journal of Nephrology 1994;0(04):-
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
5. Simultaneous determination of 50 residual solvents in solid drug preparations by GC-MS
Chinese Pharmaceutical Journal 2014;49(9):764-768
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.
6.Effect of anisodamine on endoplasmic reticulum stress during acute kidney injury in rats
Wei ZHAO ; Guangli WU ; Bin ZHAO ; Zhengjia HE ; Junfang RONG
Chinese Journal of Anesthesiology 2013;(3):366-370
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.
7.Clinical study on the efficacy and safety of argatroban in patients with progressive cerebral infarction
Li REN ; Wei WU ; Wei ZHAO ; Rong XUE
Chinese Journal of Neurology 2013;46(8):551-554
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.
8.Effects of 5-L0 inhibitor AA-861 on the development and progression of liver fibrosis
Zhiqi ZHANG ; Jiangfeng QIU ; Gang ZHAO ; Wei CHEN ; Zhiyong WU
Chinese Journal of General Surgery 2008;23(8):606-609
Objective To investigate the anti-fibrosis effect of a 5-lipoxygenase (5-LO)specific inhibitor AA-861 on liver fibrosis. Methods Liver fibrosis was induced in male Sprague Dawley rats by intraperitoneal injection of carbon tetrachloride(CCI4).AA-861(0.2 mg/100 g/d)was administrated by intraperitoneal injection starting 2 days before the first dosage of CCI4 and rats were killed at weeks 2,4,and 6.Liver specimens were obtained from each animal and fixed with 4%formaldehyde for histological analysis. The Mrna expression of 5-LO,smooth muscle-alpha(α-SMA),Collagen-1,matrix metalloproteinase-2(MMP-2)and tissue inhibitors of metalloproteinase-1(TIMP-1),the protein expression of 5-LO were evaluated by real time PCR and Western blot respectively. Histological analysis was performed by microscopy observation. Fibrosis conditions in rat liver in each group were evaluated according to the semi-quantitative scoring system (SSS), Hyp in rat livers and the hepatic functional biochemistry were also detected. Results Along with the aggravation of liver fibrosis, the gene expression of 5-Lo,α-SMA,Collagen-1,MMP-2 and TIMP-1 increased gradually, and the level of ALT, TBA, Hyp and SSS score also increased gradually. With the administration of AA-861,the Mrna expression of 5-LO,α-SMA,Collagen-1,MMP-2,TIMP-1 and the protein expression of 5-LO decreased remarkably, and the reduction in TIMP-1 Mrna expression was more significant than that in MMP-2.Six weeks after AA-861 treatment, the level of ALT, TBA, Hyp and SSS score were also decreased significantly. Conclusion AA-861 ameliorates CCI4 induced rat liver fibrosis by inhibiting the 5-LO pathway and decreasing the expression of 5-LO,α-SMA,Collagen-1,MMP-2 and TIMP-1.
9.The correlation between non-motor symptoms and the quality of life about the mild Parkinson's disease patients of different gender
Chunlin YAN ; Qingwen WU ; Suchen ZHAO ; Wei SUN
Chinese Journal of Practical Nursing 2014;30(29):25-26
Objective To analyze the correlation between non-motor symptoms (NMS) and the quality of life in the mild Parkinson's disease (PD) patients of different genders.Methods 85 mild PD patients (H-Y stage 1~2) were investigated.The PD NMS Quest was used to assess NMS.The quality of life was assessed with SF-36 questionnaire.Results Stepwise linear regression analysis showed that nocturia,restless legs,depression,daytime sleepiness,decreased interest,smell taste problems were positively correlated with the score of life quality of male and pain,increased sweating,taste smell problems,sleep problems,changes in libido,decreased memory,diplopia influenced life quality score of female.The scores of SF-36 questionnaire of the male with PD were lower than that of female patients.Conclusions The nonmotor symptom (NMS) There was a gender difference about the influence that NMS has made on PD patients.The male's life quality were lower than that of female patients about mild PD patients.
10.Soybean isoflavone attenuates trans fatty acids-induced atherosclerosis in rats
Wei MI ; Qian ZHAO ; Sijie MI ; Wu LIAN ; Tala SHI
Basic & Clinical Medicine 2015;(1):7-11
Objective To observe the effects of soybean isoflavone in attenuating trans fatty acids-induced athero-sclerosis in rats .Methods The rats were randomly divided into normal diet group、trans fatty acids group、soybean isoflavone group and trans fatty acids +soybean isoflavone group .Research kit was used to test the amounts of TG , HDL,LDL and IL-6,TNF-αin rats'blood.Bovine aortic endothelial cells were cultured and divided into control group,trans fatty acids group and trans fatty acids group +soybean isoflavone group .Western blot was performed to test the NF-κB phosphorylation ,as well as to determine ICAM-1 and VCAM-1 protein contents in bovine aortic endo-thelial cells.Results Compared with normal diet group ,the amounts of TG,LDL,IL-6,TNF-αincreased evidently (P<0.05), whereas the amounts of HDL decreased evidently (P<0.05) in the rats'blood of trans fatty acids group . After the soybean isoflavone intervened ,the amounts of TG,LDL,IL-6,TNF-αall decreased (P<0.01), meanwhile the amounts of HDL increased significantly (P<0.01).Soybean isoflavone inhibited NF-κB phosphorylation induced by trans fatty acids ( P<0.01) .The high expression of ICAM-1 and VCAM-1 induced by trans fatty acids was de-creased by soybean isoflavone when comparing with those in the control group ( P<0.01 ) .Conclusions Soybean isoflavone has a role in attenuating trans fatty acids trans fatty acids-induced atherosclerosis in rats .