4.Significance of siRNA-mediated TGFBR2 gene silencing on HepG2 cell proliferation
Lin CHENG ; Wujian DENG ; Xiaofeng JIANG ; Kun YU ; De CHEN
The Journal of Practical Medicine 2014;(14):2200-2203
Objective To value the significance of TGFBR2 gene in mediating HepG2 cell proliferation by RNA interference technology. Methods Three kinds of siRNAs targeting TGFBR2 gene were designed, synthesized and transfected into HepG2 cells via lipofetamine2000. Among three kinds of siRNAs, only the one with the most interference efficacy was selected and the correspondent DNA sequence was inserted into plasmid pEGFP-N3. Then the recombinant plasmid of siRNA-pEGFP-N3 was transfected into HepG2 cell and western blot was used to detect the protein level of TGFBR2. Then, TGF-β1 was used to stimulate HepG2 cells with or without siRNA interference and proliferation of HepG2 cells was observed. Results Among these three siRNAs, siRN-1 appeared to be the most effective. After stimulated by 5ng/mL TGF-β1, proliferation of HepG2 cells showed a marked increase in siRNA-1 group compared with blank and siRNA-NC groups (P<0.05). For all that, the proliferation rate was still lower than that in normal HepG2 cell group without TGF-β1 stimulation. Conclusion By silencing TGFBR2 gene, inhibition of TGF-β1 signaling pathway to HepG2 cells could be decreased, thereby enhancing the cell proliferation.
5.Co-transfection of tPA gene and c-myc antisense oligodeoxynucleotides for the prevention of intimal hyperplasia of rabbit auto-transplantion artery
Zhongjun WU ; Weiwei WU ; Lin YU ; De SHI ; Shusen ZHENG
Chinese Journal of General Surgery 2001;0(07):-
Objective To observe the effects of co-transfection of proto-oncogene c-myc antisense oligodeoxynucleotides(c-myc-AODN) and tissue-type plasminogen activator(tPA) gene on intimal hyperplasia of auto-transplantion artery in rabbit. Methods The left and right external iliac artery(length 1.0 cm) of rabbits were cross transplanted. The artery grafts and sutures were respectively soaked in Lipofection, c-myc-AODN, pBudCE4.1/tPA, c-myc-AODN and pBudCE4.1/tPA solution for 15 minutes. Each group were divided into five subgroups(n=5, in each subgroup) according to the sacrifice times(3, 7, 14, 28 and 56 d after operation). Specimens were harvested for pathology, chromogenic substrate test, 3H-TdR incorporation test and immunochemisty coloration study. Result The intimal area, stenosis ratio, 3H-TdR incorporation, PCNA positive cell in c-myc-AODN adding tPA co-transfection group were significantly lower than that of control group(P0.01), and that were lower than c-myc-AODN transfection group and tPA gene transfection group(P0.05). Conclusion Vascular local co-transfection of tPA gene and c-myc-AODN effectively inhibits the proliferation of vascular smooth muscle cell(VSMC) and hyperplasia of intima of the transplanted artery.
6.Effects of levodopa On visual evoked potential and visual cortex neuron in monocular deprivation rat
Xiao-ying, WU ; Yu-lin, LUO ; De-lin, LIU ; Shuang-zhen LIU
Chinese Journal of Experimental Ophthalmology 2011;29(3):220-225
Background Nearly over 40 years have elapsed since the original findings of visual cortical plasticity,but none of drug has been found for curing amblyopia effectively. Objective The goal of this study was to investigate the effects of different dose of levodopa on flash visual evoked potential(F-VEP)and morphology of visual cortex cells in monocular deprivation rat and explore the possible mechanism of curing amblyopia.Methods Monocular deprivation model were established by suturing eyelids of 30 2-week-old Sprague Dawley(SD)rats for 4 weeks.The 30 SD rats were then divided into 3 groups randomly and 10 rats for each group.Normal saline.20 ms/kg levodopa,80 ms/kg levodopa were intragastrically administered once per day after modeling respectively for 4 weeks.F-VEP was recorded after establishment of model and administration of drug respectively.The rats were sacrificed and the visual cogex was obtained for histological examination,and TUNEL technique was used to assess the structural change of visual cortex.Results The latency of P1 wave was significantly longer in the deprived eye than the normal eyes(P<0.05).After administration of levodopa,the latent periods of Pl wave in the deprived eye were obviously shortened in comparison with before administration of levodopa in 20 ms/kg and 80 mg/kg levodopa group (P<0.05).The difference values of latent period of P1 wave between before and after administration of drug showed statistically significant change in three groups(P<0.05).No evidently alterations were found in the amplitude differences of N1 P1 and P1 N2 waves among three groups(P>0.05).The number and structure of neurons in contralateral visual cortex of non-deprived eye were normal.However,the numbers of neurons in deprived eye were significantly less and presented the signs of para-apoptosis in normal saline group.In 20 mg/kg levodopa groups,the alterations of number and morphology in neurons of rat visual eogex were slight.TUNEL assay revealed that the numbers of positive neurons in contralateral visual codex of non-deprived eye were 2.20±1.23.while those in deprived eye were 53.7±9.36,27.20 4±5.96 and 10.70±3.23 in normal saline group,20 mg/kg and 80 mg/kg levodopa group respectively,showing a significant difference among them(P>0.05).After usage of levodopa,the numbers of positive neurons was negatively correlated with the difference value of P,latent period of VEP(r=-0.815,P=0.000).Conclusion Levodopa has a therapeutic effect on rat deprived eye,and its possible mechanism is inhibiting the para-apoptosis of neurons and participating in the development and plasticity of visual system.
7.Splitted fractions and unoverlapping analysis of chemical constituents of Poria cocos.
Zhe LIN ; Yu-Bin XU ; Xiao-Ku RAN ; De-Qiang DOU
China Journal of Chinese Materia Medica 2013;38(24):4340-4346
With the combined applications of steam distillation, water extraction and alcohol precipitation, liquid-liquid extraction and column chromatography over macroporous resin, a splitted-fractions method of the chemical constituents of Poria cocos was established. The unoverlapping property of the fractions of P. cocos was qualitatively analysed by using principal component analysis and cluster analysis. With angle cosine, squared euclidean distance and the overlapping analysis of peak area of crude herbs, the unoverlapping property of the fractions of P. cocos was half-quantitatively analysed. The chemical components of P. cocos was divided into the fractions of polysaccharide, petroleum ether, ethyl acetate, alcohol eluate from macroporous resin and water eluate from macroporous resin. Non similarity degree among each chemical fraction was above 80% and main chemical components were identified. The established method for splitting fractions of P. cocos has good stability and repeatability and all chemical components in P. cocos could be completely divided into six fractions. It is the first time that the author half-quantitatively analyse the unoverlapping property of the chemical fractions of P. cocos.
Chromatography, High Pressure Liquid
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Cluster Analysis
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Medicine, Chinese Traditional
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Poria
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chemistry
8.Application of intra- and post-operative ultrasound in traumatic brain injury
De-lin, YU ; Ping, MA ; Zeng-hui, LI ; Kai, XUE ; Gui-lan, QU
Chinese Journal of Medical Ultrasound (Electronic Edition) 2008;5(2):248-254
Objective To explore the application of B-mode intra- and post-operative ultrasound in patients with traumatic brain injuries. Methods B-mode intra- and post-operative ultrasound were performed 35 times in 24 patients, including 7 times of intra-operation in 7 patients and 28 times of post-operation in 17 patients. Results During operation epidural hematomas in 3 cases as well as intracranial hematomas in 2 cases, subdural hematomas in 1 case, diffused brain swelling in 1 case were found. After operation epidural hematomas were examined in 4 cases as well as subdural hematomas in 1 case, delayed intracranial hematomas in 6 cases, scattered contusion and laceration of the brain in 12 cases, acute obstructive hydrocephalus in 1 case, and subdural hydroma in 4 cases. Six cases underwent operations again because of delayed hematomas, contusion and laceration of the brain. B-mode ultrasound was compared with CT scan and the total coincidence rate was 85.4%.Conclusions B-mode ultrasound is helpful to examine delayed hematomas and other emergent complications. It may save time and may improve rescue rate and has a good coincidence with CT scan. B-mode ultrasound may be widely used as a routine way in neurotrauma ICU.
9.The effect of histotype and histodifferentiation on the standardized uptake value of no-small cell lung cancer ~(18)F-FDG PET/CT imaging
De-jun, LIU ; Yan-lin, FENG ; Feng-wen, YU ; Xiao-hong, HE
Chinese Journal of Nuclear Medicine 2010;30(1):5-9
Objective To determine the effect of histotype and histodifferentiation on the maximum standardized uptake value (SUV_(max)) of non-small cell lung cancer (NSCLC) ~(18)F-fluorodeoxyglucose (FDG) PET/CT imaging.Methods Two hundred and sixty patients with NSCLc underwent ~(18)F-FDG PET/CT imaging.They were classified according to (1) histotype:as adenocarcinoma (AC),squamous cell carcinoma(SQC),adenosquamous carcinoma (ASC) and other type carcinoma (OTC),and (2) histodifferentiation:as grade Ⅰ (well-differentiated),grade Ⅱ (moderate-differentiated) and grade Ⅲ (poor-differentiated).The SUV_(max) and size(long diameter)of the primary lesions were measured.Multivariate regression analysis was used to analyze the relationship between the SUV_(max) and variable factors including histotype,histodifferentiation,lesion size,age,sex,body height,body weight,body mass index (BMI),blood glucose level,dose,and rate of dose.Results Two hundred and sixty patients had 260 primary NSCLC tumors.There were 161 AC(15 grade Ⅰ,88 grade Ⅱ,58 grade Ⅲ),74 SQC(6 grade Ⅰ,39 grade Ⅱ,29 grade Ⅲ),15 ASC(7 grade Ⅱ,8 gradeⅢ)and OTC(8 large cell,2 carcinosarcoma).Only lesion size (F=87.046.P<0.001),histodifferentiation (F=87.604,P<0.001) and histotype (F=66.663,P<0.001) were included for multivariate regression analysis with SUV_(max).After adjustment for lesion size,the SUV_(max)(mean and 95%confidence interval) in ascending order was AC Ⅰ:3.3(2.1-4.5),ACⅡ:6.0(5.5-6.6),SQCⅠ:6.1(4.2-8,0),ASC Ⅱ:6.6(4.8-8.4),SQCⅡ.7.8(7.0-8.6),OTC:8.1(6.6-9.6),AC Ⅲ:8.3(7.6-8.9),ASC Ⅲ:8.7(7.0-10.4),and SQC Ⅲ:8.9(8.0-9.8).11he SUV_(max) of AC Ⅰ was significantly lower than that of SQC Ⅰ(q=-2.786,P=0.017),same for AC Ⅱ and SQC Ⅱ(q=-1.776,P<0.001),but no statistically significant differences were found among AC Ⅲ,ASC Ⅲ and SQC Ⅲ(q=-0.593,-0.422,0.171,P=0.288,0.642,0.860,respectively).For the same histotype lesions,the difference of SUV_(max) among AC Ⅰ,Ⅱ and Ⅲ was statistically significant(q=-2.720,-4.943,-2.223,all P<0.001),as also for SQC Ⅰ and Ⅲ(q=-2.751,P=0.012).Conclusion Histotype and histodifferentiation are significant correlative factors for ~(18)F-FDG uptake of NSCLC,with histodifferentiation being the factor with greater impact.