1.CT diagnosis and differential diagnosis of retinoblastoma
Xingwei DU ; Weiguo ZHANG ; Yi WANG
Journal of Third Military Medical University 2003;0(19):-
Objective To explore the manifestation and diagnostic value of CT in retinoblastoma. Methods Twentythree cases of retinoblastomas proven by operation and pathologic examination were collected. Their CT images, clinical symptoms and pathologic results were retrospectively analyzed. Results Twentythree patients of retinoblastomas aged from 4 months to 9 years (mean 2.4 years). CT images showed soft tissue mass (shaped as node, crescentshaped or occupied the entire space of the eyeball) was in the globe of all cases. There was calcification in 19 cases (82.6%), including “small pinch” in 12 cases or “patches” in 7 cases. Five cases were found the optic nerve thickened (21.7%), and 1 case of intracranial invasion through the optic nerve canal (1/23), and 6 cases of retina detachment (6/23). The lesion showed mild or moderate enhancement after intravenous contrast medium were performed. Conclusion CT has an advantage on showing the size, shapes, intrastructures and invasive degree of the lesion, and is meaningful to the diagnosis and differential diagnosis of retinoblastoma.
2.THE INTERFERING EFFECT AND PROBABLE MECHANISM OF NEUREGULIN-1? ON CEREBRAL ISCHEMIA REPERFUSION INJURY IN MICE
Tao WANG ; Fang DU ; Yunliang GUO
Acta Anatomica Sinica 1953;0(01):-
Objective To study the neuroprotective effects of neuregulin-1?(NRG-1?) on the nervous behavioral function,cerebral infarction volume,brain water content(BWC),neuronal apoptosis and aquaporin-4(AQP-4) expression in astrocytes after cerebral ischemic reperfusion and the related mechanism in mice.Methods Intraluminal thread methods were applied to establish the middle cerebral artery occlusion reperfusion models in the mice.Neuregulin-1?(2?g / kg) was injected into the internal carotid artery for treatment.The nervous behavioral function was evaluated with Bederson's test.The cerebral infarction volume was observed with tetrazolium chloride staining.The BWC was measured by dry-wet weight comparing.The apoptosis positive cells were counted by immunofluorescence assay.The expression of AQP-4 was determined by immunohistochemical assay.Results Nervous behavioral malfunction appeared in all the mice with left middle cerebral artery occlusion and/or reperfusion.The infarction focus showed in the ischemic hemisphere after the injury.The BWC,the number of neuronal apoptosis cells and AQP-4 expression in astrocytes were higher than those in the sham group. In the NRG-1? treatment group,the nervous behavioral function was improved 24 hours after ischemia,the number of apoptosis positive cells reduced and the infarction volume decreased significantly compared with the control group(P0.05).In the groups of reperfusion for 22,46 and 70 hours,the five indexes mentioned above were significantly different from those in the corresponding control groups(P
3.Construction of pIRES2-hBSP-EGFP vector and stably expression in MDA-MB-231BO breast cancer cells
Hongyan DU ; Jie WANG ; Yong GUO
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To constitute a higher expression system and to obtain the breast cancer cell strains in which BSP is stably expressed. Methods hBSP gene was subcloned from pB-hBSP vector by PCR. The PCR fragment was inserted into the eukaryon expression vector, pIRES2-EGFP, which allow exogenous protein and EGFP to express respectively. The recombinant vector, pIRES2-hBSP-EGFP, was transfected into human breast cancer cells with Lipofectamine TM 2000. The expression of symbol protein EGFP, could be conveniently observed with fluorescent microscope. Results The recombinant pIRES2-hBSP-EGFP plasmid was constituted and successfully transfected into breast cancer cells. In the breast cancer cell strain hBSP and EGFP were expressed. Conclusion The successful constitution and transfection of hBSP and EGFP nonfusion expression vector laid a foundation for the further study on the effect of BSP on breast cancer metastasizing to bone in vivo or in vitro.
4.Expression of p16 and p15 Proteins in Acute Lymphoblastic Leukemia of Children
Wanru HU ; Ruiying ZHANG ; Chengji GUO ; Li'E WANG ; Shuyan DU ;
Journal of China Medical University 2001;30(1):64-65,68
Objective:Our aim was to To clarify the roles of p16 and p15 proteins in the genesis of acute lymphoblastic leukemia(ALL).Methods:Twenty-three samples of ALL were studied by the method of indirect immunofluorescence.Flow cytometer was used to estimate the cellular fluorescent intensity to determine the levels of p16 and p15 proteins.Results:Negative expression for p16 protein was found in 10 of 23 samples,and 8 of 23 were p15 negative expression.Both kinds of proteins were abscent in 6 samples.2 of 3 cases of T-ALL were negative expression of p16,p15 protein.In non T-ALL,6 of 13 were negative expression for p16 protein,5 of 13 were p15 protein deficient.The expression rates of p16,p15 protein in high leukocyte group were lower than those of non-high leukocyte group(P<0.05).The expression rates of p16,p15 proteins in HR-ALL were lower than those of SR-ALL(P<0.05).Conclusion:The p16 and p15 proteins take part in the genesis of ALL.Negative expression of p16,p15 proteins might imply the poor clinical outcome.
5.Research of Sequential Extraction Procedure for Heavy Metals in Sediments from Mariculture Area
Pengran GUO ; Dehai MOU ; Chang WANG ; Rongliang QIU ; Hong DU
Chinese Journal of Analytical Chemistry 2009;37(11):1645-1650
A sequential extraction procedure has been proposed for the evaluation of the speciation of heavy metals including Cd,Cu,Pb and Zn in sediments from mariculture area,and the speciation of heavy metals was separated and defined as acid soluble fraction,reducible fraction,fraction bound organic matter,fraction bound sulfides and residual fraction. Matrix effects of high salinity on the determination of heavy metals in sediments were eliminated by matrix matching and internal standard methods when inductively couple plasma optical emission spectroscopy (ICP-OES) and mass spectroscopy (ICP-MS) were used,respectively. The results showed that the measured values of marine sediment reference materials were consistent with the standard values when the digestion solutions were determined after dilution. The extraction results of the prepositional extraction procedure and European Community Bureau of Reference Program (BCR) procedure were compared and the selectivity of extractants was investigated. The preliminary studies indicated that this sequential extraction procedure was applicable for evaluating the speciation of heavy metals in sediment with organic substances pollution and eutrophication,especially for fraction bound organic matter and fraction bound sulfides.
6.Effects of 3-AB on PARP expression of Hela cells and apoptosis and cell cycle progression of Hela cells after X-rays irradiation
Xiang DU ; Hongguang ZHAO ; Wen WANG ; Wei GUO ; Shouliang GONG
Journal of Jilin University(Medicine Edition) 2006;0(03):-
Objective To study the changes of apoptosis and cell cycle progression of Hela cells after the poly(ADP-ribose) polymerase(PARP) was inhibited by its inhibitor 3-aminobenzamid(3-AB) and the mechanisms of PARP interaction with Hela cells damaged by irradiation.Methods Hela cell line was used.Flow cytometry(FCM) was used to examine the PARP expression of control and 3-AB groups at 0,2,4,8,12 h after administration with 5 mmol?L-1 3-AB.The percentage of apoptotic cells and cell cycle progression of control,irradiation,3-AB plus irradiation groups were measured with FCM at 2,8,12,24 h after exposure to 2 Gy irradiation following administration with 5 mmol?L-1 3-AB.Results The percentage of Hela cells with positive expression of PARP protein decreased after administration with 3-AB and there was significant difference between 3-AB plus irradiation group and control group(P
7.Establishment and evaluation of the model of middle cerebral artery occlusion reperfusion in mice
Fang DU ; Tao WANG ; Xuewei YANG ; Yonggang LI ; Yunliang GUO
Chinese Journal of Tissue Engineering Research 2006;10(46):201-204
BACKGROUND: Most animal experiments of transgene are derived from mice; therefore, it is necessary to establish a focal cerebral ischemiareperfusion model and significant to prevent and cure ischemic cerebrovascular diseases.OBJECTIVE: To establish a convenient and reliable model with middle cerebral artery occlusion reperfusion (MCAO/R) in mice.DESIGN: Randomized controlled animal study.SETTING: Institute of Cerebrovascular disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: Twenty healthy BALB/c mice, of both genders, weighing 25-30 g, of SPF grade, were divided into sham operation group (n=5), ischemia group (n=10) and 22-hour reperfusion group (n=5) on the basis of digital table. In addition, according to digital table, 130 healthy male Kunming mice were divided into sham operation group (n=10), 24-hour ischemia group (n=30), 2-hour ischemia/22-hour, 46-hour and 70-hour reperfusion groups with 30 in each group; meanwhile, 30 female mice were divided into sham operation group, 24-hour ischemia group and 2-hour ischemia/22-hour reperfusion group with 10 in each group. All Kunming mice were weighing 25-30 g and of SPF grade.METHODS: The experiment was carried out in the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from July 2005 to March 2006. The 6-0 suture with silica gel covered at an end was sent from the left external carotid artery (ECA) into internal carotid artery (ICA) till arriving at the initiation of middle cerebral artery (MCA) to block the blood stream in it, then drawing the suture from ICA 2 hours after occlusion to accomplish reperfusion. Mice were cut off their heads in sham operation group at 24 hours after operation, in ischemia group at 24 hours after blocking blood stream and in reperfusion group at 24, 48 and 72 hours after operation. Reliability of models was evaluated with neurology score and tetrazolium chloride stain. Longa standard scores: neurology score ≥ 1 point was regarded as successful models;coronal sections of brain tissue were stained with tetrazolium chloride, and the white region was regarded as infarcted volume.MAIN OUTCOME MEASURES: Neurology score and infarcted volume after staining of triphenyltetrazolium chloride in brain tissue.RESULTS: All mice were involved in the final analysis. ① Successful rate was 20% of BALB/c mice, 66.7%-73.3% of male Kunming mice and 40%-50% of female Kunming mice. ② Brain sections of BALB/c mice in sham operation group were orange at both sides of cortex and infarction focus was not observed. A big infarcted volume was observed on brain sections of mice in ischemia group, and infarcted volume counted for 50%-70% as homonymy hemisphere on optochiasmatic coronal sections. The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%. In addition, condition in ischemiareperfusion group was similar to that in ischemia group. A big infarcted volume was observed on brain sections, and infarcted volume counted for 50%-75% as homonymy hemisphere on optochiasmatic coronal sections.The condition of Kunming mice was similar to that of BALB/c mice, but infarcted volume counted for 40%-65%.CONCLUSION: The model with MCAO/R in mice characterizes by relatively smaller trauma, and the ischemic region is stable; therefore, it can be used to accurate timing control of ischemia/reperfusion. This model is an ideal one for researching pathophysiological changes, prognosis and therapy in cerebrovascular disease.
8.Construction of pIRES2-HBSP-EGFP vector and its expression in breast cancer cells
Hongyan DU ; Jie WANG ; Yong GUO ; Lin ZHENG ; Jing YANG
Chinese Journal of Tissue Engineering Research 2005;9(38):152-154
BACKGROUND: Bone sialoprotein (BSP) gene is expressed in human breast cancer cells, in which bone metastasis occurs easily outside the mineralized tissue. Clinical observation shows that the expression level of BSP of breast cancer cells at bone metastasis is higher that at the primary site;therefore, BSP may be closely related to tumor specific bone metastasis.The study on breast cancer bone metastasis can provide new drug target for clinical prevention and treatment.OBJECTIVE: To establish breast cancer cell strains of BSP with stable expression and observe the effect of BSP in the whole process of breast cancer bone metastasis.DESIGN: Controlled experiment.SETTING: College of Biological Sciences and Engineering, South China University of Science and Technology; Medical Experiment Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.MATERIALS: This experiment was conducted in the Medical Experimental Center,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA,betweer November 2003 and March 2004..pIRES2-EGFP vector (5.3 kb) was purchased from BD Biosciences Clontech Inc.; E.Coli.Top10, pB-hBSP plasmid containing the coding region of hbsp, and human breast carcinoma cells, MDA-MB-231BR that was specifically transferred to brain and MDA -MB-231BO that was specifically transferred to bone.METHODS: hbsp gene was subcloned from pB-hBSP vector by PCR. Bg1Ⅱ and Pst Ⅰ restriction enzyme sites were inserted at 5' and 3' ends, orientation cloned to eukaryon expression vector pIRES2-EGFP, and constructed recombinant vector pIRES2-EGFP. The constructed recombinant vector was transfected into MDA-MB-231BR that was specifically transferred to brain and MDA-MB-231BO that was specifically transferred to bone.MAIN OUTCOME MEASURES: Construction of pIRES2-hBSP-EGFP recombinant expression vector; recombinant expression vector pIRES2-hBSP-EGFP transfecting breast cancer cells.Breast cancer strains specific in bone metastasis and brain metastasis were successfully transfected. The fluorescence labeling could be observed under the fluorescence microscope, and BSP had corresponding expression.CONCLUSION: The successful construction and transfection of pIRES2hBSP-EGFP of eukaryon expression vector would lay foundation for further study on the role of BSP in breast cancer metastasizing to bone in vivo or in vitro.
9.Value of ultrasound in diagnosing ductal carcinoma in situ
Rongrong GUO ; Fang CHANG ; Peixia WANG ; Chenxiang DU ; Li ZHANG
Cancer Research and Clinic 2009;21(11):739-741
Objective To evaluate the value of ultrasound in diagnosing ductal carcinoma in situ (DCIS). Methods The sonographic characteristics of 12 DCIS which were confirmed by pathology were analyzed retrospectively. Results The ultrasound image of DCIS could be divided into four types;the solid mass nodule, mammary dysplasia, mix mass nodule, the dilated duct type. Micro calcification had high incidence rate. Ultrasonic diagnosis accordance rate was 50.0 %. On molybdenum target mammograms, the tumor appeared as a cluster of calcified spots in 8 cases, and the accuracy rate of diagnosis of was 66.7 %.Conclusion There are no typical characters of DCIS in ultrasound image. However, some characteristics are suggestive and can help to differentiate them from the benign tumors, such as small nodule, irregular shape,obscure boundary, and microcalcification. When sonography combine with molybdenum target mammography,the accuracy rate of diagnosis will be improved.
10.High-throughput screening of human soluble epoxide hydrolase inhibitors.
Shoubao WANG ; Jing GUO ; Xiaoming YU ; Guanhua DU
Acta Pharmaceutica Sinica 2010;45(11):1367-72
To screen potential human soluble epoxide hydrolase (hsEH) inhibitors, a high-throughput screening model in 384-well microplate with total volume of 50 microL was established. Recombinant hsEH was cloned and expressed in E. coli. and its specific substrate PHOME was synthesized. The HTS model was based on fluorescence analysis with enhanced sensitivity and specificity (Z' = 0.65). A total of 47 360 samples (including 25 040 compounds and 22 320 natural products) were screened, of which 950 samples with inhibition greater than 80% were selected for further rescreening. Finally, two compounds with high inhibitory activity were identified, whose IC50 value were 8.56 and 4.31 micromol x L(-1), separately. The results indicated that the method was stable, sensitive, reproducible and also suitable for high-throughput screening.