1.Detect of P53,bax and caspase 3 genes expression after optic nerve injury in rats with SYBR green I fluorescence quantitative PCR
Yingjuan LÜ ; Xiulan, ZHAO ; Jie, YANG ; Jinguo, YU ; Hua, YAN
Chinese Ophthalmic Research 2009;27(11):973-977
Objective Previous study showed that the histopathological basis of visual function damage caused by optical nerve injury is apoptosis of retinal ganglion cells(RGCs). This procedure is regulated by P53, bax and caspase 3 genes. Present study aimed to observe the expression of bax, P53 and caspase 3 mRNA in RGCs after traumatic optic nerve damage in the rats by SYBR green I fluorescence quantitative PCR method. Methods The animal model of optic nerve injury was established in the right eyes of 56 adult Wistar rats by a fluid percussion brain injury device (FPI) . Animal were killed on days 1, 3, 5, 7, 9, 14, 28 days separately after injury. Other 16 Wistar rats were divided into normal control group and sham operation group. The total RNA was isolated from rat fresh retina tissue by Trizol method and was treated by reverse transcription to cDNA using 01igo(dt) 18 as primer and then amplified. The target fragments of bax, P53 and caspase 3 cDNA were linked with carrier pTZ57 R/T to construct recombined plasmids which were transformated to E. Coli DH5α by T/A clone method. Recombined plasmids were extracted with alkaline lysis method and the plasmids were selected in white colonies by ampicillin screening, EcoR I restrictive enzyme analysis, and their specificity was evaluated using DNA sequencing. The standard curves were created by plasmid DNA and the precise expression level of target genes in samples were determined using software. The results were expressed as the ratios of target gene mRNA to GAPDH mRNA. Results The standard curve drawn by pTZ57R/T and target gene presented a good linear tendency with the higher sensitivity and specificity. The expression of P53 and bax mRNA began to increase on the third day after the injury of optic nerve and peaked on the fifth day and started to decline on the seventh day. The expression of caspase 3 mRNA increased from the fifth day through the ninth days after injury and declined on the fourteenth day. The significant differences were found in the expression of P53, bax and caspase 3 between model group and control group (P < 0. 05) . Conclusion The pro-apoptotic protein P53, bax and caspase 3 play an important role in RGCs apoptosis.
2.Establishment of animal model of traumatic optic neuropathy and its function evaluation by F-VEP
Rongguo, YU ; Jinguo, YU ; Zhansheng SHEN ; Xing, WANG ; Yingjuan LÜ ; Wei, ZHANG ; Hua, YAN
Chinese Ophthalmic Research 2010;28(1):39-43
Background Visual electrophysiology is a sensitive index for the evaluation of visual function.It has an important value in the assessment of traumatic optic neuropathy.Rabbit is an ideal animal model of traumatic optic neuropathy,and it is simple for the record of flash visual evoked potential(F-VEP)in rabbits.ObjectiveThe present study is to establish the animal model of traumatic optic neuropathy with or without lens injury and observe the repairing procedure using F-VEP. MethodsModels of traumatic optic neuropathy associated with lens injury were established in the right eyes and only traumatic optic neuropathy were created in the left eyes of 64 healthy SPF Chinese white rabbits using fluid percussion brain injury device(FPI).F-VEP was recorded based on the Proposal of International Visual Electrophysiology on 1,2,4,7,10,14,21,28 days after injury of optic nerves.Experimental animals were sacrificed in above time points for the histopathological examination.Macrophages were labeled by ED-1 antibody and survival retinal ganglion cells (RGCs)were stained by Nissl method.Results At the first day after injury,the latencies of P_(100) in both group were longer,and the amplitudes of P_(100) in both group were lower than before injury,showing statistically significant differences among different time points(P<0.05),but no significant difference was seen between the two groups(P>0.05).The duration of latency in traumatic optic neuropathy associated with lens injury group was shorter than that in only traumatic optic neuropathy group(P<0.05).The restore of latency in traumatic optic neuropathy associated with lens injury group was much faster than that in only traumatic optic neuropathy group(P<0.05).The numbers of macrophages were significantly increased and numbers of survival RGCs were considerably decreased with lapse of injury time (P<0.05).The abnormalities of VEP P_(100) and RGCs were obviously improved in 28 days after injury in both groups. ConclusionThis animal model can be established successfully by FPI.The result of retinal histopathological examination confirms F-VEP findings in this model.
3.The preliminary study of the value of MMP9, MPO and sCD40L in detection of the characteristics of coronary artery plaque
Na JING ; Bin Lü ; Jinsuo KANG ; Chunling ZHANG ; Jinguo LU ; Li ZHANG ; Xiongbiao CHEN ; Zhihui HOU ; Xi CHEN ; Xiangfeng CONG
Chinese Journal of Laboratory Medicine 2011;34(10):889-892
Objective To evaluate the utility of MMP9,MPO and sCD40L in detection of the character of coronary artery plaque.Methods From April 2008 to January 2010,118 patients from outpatient of Fu Wai Hospital with chest pain were enrolled.All of them underwent 64 Multiple-detector row spiral computer tomography (64-MDCT),the CT value < 130 Hu patients were enrolled in non-calcified plaque group (71 cases),CT value ≥ 130 Hu patients were enrolled in the calcified plaque group (47 cases).Ninty healthy volunteers were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum markers,including MMP9,MPO and sCD40L.Levels of MMP9,MPO and sCD40L of each group were compared.ROC curve was used to evaluate the sensitivity and specificity of the markers in diagnosis of non-calcified plaque.Results MMP9,MPO and sCD40L levels of non-calcified were ( 762.25 ± 368.71 ),[ 844.10 (582.00 - 1220.70) ],(9.37 ± 3.15) μg/L,higher than the healthy control group (342.70 ± 178.53),[426.35 ( 283.20 - 592.00) ],(6.55 ± 2.96) μg/L and calcified plaque group ( 483.12 ± 219.09 ),[ 469.00 ( 302.45 - 723.55) ],( 7.24 ± 2.86) μg/L The difference was statistically significant ( F =42.47,H =50.28,F =17.94,all P < 0.01 ). Areas of MMP9,MPO and sCD40L under the ROC curve to predict non-calcified plaque were 0.854,0.792,0.751 respectively,when the identification threshold for non-calcified plaque were 510.13,537.82,7.05 μg/L respectively,the diagnostic sensitivity was 80%,80%,80% respectively,and specificity was 80%,67% and 55% respectively.Conclusion The serum MMP9,MPO and sCD40L levels can help to determine the character of coronary plaque.
4.Study of Morphological Characteristics of Calcified Plaques and Luminal Stenosis of Coronary Artery at 64-slice Computed Tomographic Coronary Angiography
Jinguo Lü ; Bin LU ; Xiang TANG ; Xiongbiao CHEN ; Zhihui HOU ; Shihang JIANG ; Ruping DAI ; Yongjian WU ; Shubin QIAO ; Yuejin YANG
Journal of Practical Radiology 2010;26(4):485-491
Objective To study the correlation between morphological characteristics of coronary artery calcified plaques and luminal stenosis of local coronary artery segments with 64-slice computed tomography.Methods One hundred and eleven patients who had undergone 64-slice computed tomographic coronary angiography(CTCA)and conventional coronary angiography(CAG)were retrospectively analyzed.The calcified plaques were classified as punctate,nodular,strip-like and nubbly in long-axis view of coronary artery lumen,and were classified as crescent,semilunar,round moon and circinate in short-axis view.The morphologic characteristics of these calcified plaques on CTCA were retrospectively analyzed and compared with luminal stenosis of CAG results.Results Among 528 calcified coronary segments which were analyzed in 111 patients,there were 383(72.5%)punctate calcified plaque segments and 145(27.5%)of non-punctate plaques.There were 34(23.4%,34/145)non-punctate calcified plaques which caused severe stenosis(≥75%),including 4(11.8%)nodular,8(23.5%)stripe-like and 22(64.7%)nubbly calcified plaques on the long-axis view,and 0(0.0%)cresent,8(23.5%)semilunar,18(52.9%)round moon and 8(23.5%)circinate calcified plaques on the short-axis view.The ratios of different morphological coronary artery calcifications which caused severe stenoses were significantly different with each other(all P<0.01).Conclusion Different figures of coronary artery calcified plaques demonstrate different degrees of stenoses of local coronary artery lumen.Severe stenoses were mostly caused by nubbly calcified plaque on long-axis view,round moon and circinate calcified plaque on short-axis view.
5.Different contrast injection protocols for 64-slice spiral CT coronary angiography
Jinguo LU ; Bin Lü ; Jinhai QIU ; Hua BAI ; Xiang TANG ; Xinling YANG ; Shiliang JIANG ; Ruping DAI ; Tao CHEN
Chinese Journal of Radiology 2008;42(6):586-591
0bjective To determine the optimal contrast protocols for 64.slice spiral CT coronary angiography in order to reduce the volume of contrast injection.Methods One hundred fifty pailents scheduled to undergo 64-slice spiral CT coronary angiography were prospectively randomized into the following five groups with different injection protocols:group l:uniphasic injection without a flush;group 2:biphasic injection with a flush;group 3,group 4 and group 5:triphasic injection with a diluted contrast material with 3:7.5:5.7:3 contrast/saline ratio respectively.Attenuation was measured in the right atrium,right ventricle,left atrium,left ventricle,ascending aorta,fight coronary artery and left coronary artery and analyzed with single factor variance test(ANOVA).The quality the coronary artery images was evaluated and compared using person Chi-Square.Results The total contrast material vohme were (67.0±5.3)ml,(59.9±4.9)ml,(62.9±3.2)ml,(69.2 4±5.7)ml and(70.9 4-4.6)ml in five groups respectively(F=27.43,P<0.01).Image quali~scores of coronary arteries were significant different among five groups(X2=18.81,P<0.05).There were signiflcandy differences in artifacts of the superior vena cava among five groups(X2=31.44,P<0.01).The artifacts in the superior vena cava in group 1 was the most,and in group 2 was the least.The mean enhancement values of right and left coronary arteries in group 2 were significantly greater than those in other groups(F=2.47 and 4.10,P<0.05).The visualization of both left ventricle and right ventricle cavities W88 the best in group 3.Conclusion Biphasic injection and triphasic injection are better than uniphasic injection for 64-slice spiral CT coronary angiography and triphasic injecfion is better than biphasic injection for the visualization of both left ventricle and right ventricle cavities.
6.Integrated multi-model imaging in study of acute myocardial ischemia in Chinese mini-swine model
Xiang TANG ; Bin Lü ; Wei FANG ; Minjie LU ; Jinguo LU ; Xiongbiao CHEN ; Zhihui HOU ; Shihua ZHAO ; Zuoxiang HE ; Yue TANG ; Fengying Lü ; Shiliang JIANG
Chinese Journal of Radiology 2010;44(10):1094-1099
Objective To evaluate the value of MSCT, MRI and SPECT in detecting acute myocardial ischemia in Chinese mini-swine model. Methods A total of six male mini-pigs were recruited with a mean body weight of (21.6 ± 1.2) kg. All pigs were scanned on MSCT before the ligation of distal segment of Left anterior descending artery. Then, MSCT was rescanned every 2 h from ligation till 8 h latter.MRI, SPECT and the last MSCT scan were performed within 24 h one by one. Finally pathological examination was carried out right after the pig killed. Results One pig died during operation, the other 5 finished all the examinations. The pathological staining showed the same areas of myocardial infarction in the left ventricular anterior wall with the all the imaging findings, including low perfusion region of MSCT arterial phase at 2-24 h, low perfusion region of SPECT at 24 h and low perfusion region of MRI first pass phase at 24 h. Three of 5 pigs showed enhanced edge of low perfusion region on MSCT delayed scan at 4-8 h. The mean CT values in the region with reduced first-pass perfusion were 75.9,36.4, 35. 2,37. 8,37.4,33.3 HU on MDCT image at baseline, 2-8 h after operation and within 24 h after operation, respectively,and there were statistically significant difference of CT values ( F = 12. 341, P <0. 01 ) between preoperative and all postoperative MSCT scan. There were no statistically significant difference (F = 2. 278, P = 0. 792)among all postoperative MSCT scan. At baseline, 2-8 h after operation and within 24 h, the average volumes of stroke volume(SV)were 21.7,11.9,10.3,11.4,12. 3,12.6 ml, respectively, while the average volume of end-systolic volume( ESV)were 15.2,23.4,25.0,24. 4,25.3,22. 8 ml,respectively. The average volume of end-diastolic volume ( EDV ) at these time point were 37. 0,35.4,35.0,35.7,37. 6,37.5 ml,respectively and the average percentage of ejection fraction (EF) were 58.9% ,33.8% ,29. 0%, 31.9%,32.6% ,33.5% ,respectively. SV(F =22. 349, P<0.01) ,ESV (F=8. 810, P<0.01) ,EF(F =27. 240,P < 0. 01 ) were all significantly different among all postoperative MSCT scan except EDV ( F = 2. 339, P =0. 079). Infarct size, which was defined as the proportion of the area of infarction to that of the entire heart,were (39.4 ±12.6)% for MSCT,(37.2 ± 10.0)% for MRI, (35.9 ±9.6)% for TTC, respectively.There were no significant differences of infarct size between TTC and MSCT (t =0. 612, P =0. 574), TTC and MRI (t=0.820, P=0.458), MSCTand MR (t=0. 425 ,P =0. 692 ). Conclusions MSCT,MRI and SPECT were all able to be used to detect the myocardial infarction in acute myocardial ischemia model The infarct size defined on MSCT, MRI and pathology were consistent. The density of ischemic myocardium and cardiac function did not change over the time within 24 h right after infarction.
7.Analysis of dual-source CT coronary angiography of congenital heart disease in children
Xiang TANG ; Bin Lü ; Lei HAN ; Hua BAI ; Jinguo LU ; Xiongbiao CHEN ; Hongyu WANG ; Xinling YANG ; Yanling WU ; Zhihui HOU ; Ruping DAI ; Shiliang JIANG
Chinese Journal of Radiology 2010;44(1):48-52
Objective To analyze the image quality of coronary arteries by dual-source computed tomography (DSCT) in children with congenital heart disease(CHD). Methods Image quality of coronary arteries were scored by two radiologists with an ordinal grading system. Score 3 or 4 was considered to be good image quality and diagnostic. Score differences among 3 groups of various baseline heart rate levels, 3 groups of various age levels and 4 coronary branches were analyzed by using ANOVA. Multiple linear regression analysis was used to test the effects of age, heart rate, tube voltage, tube current, pitch, contrast volume, contrast injection rate on the coronary image scores. Results A total of 72 consecutive patients were enrolled. 71.2% (205/288) of coronary artery branches was good enough for diagnosis. Image quality of left main coronary artery (LM, score 3.64±0.49), left anterior descending (LAD, score 2.97±0.40), right coronary artery (RCA, score of 2.82±0.70), left circumflex (LCX, score 2.76±0.73) were in a descending order (F=29.00, P<0.01). Image scores of age groups of 0--1 years old (2.80±0.65), >1--3 years old (2.81±0.74), >3--14 years old (3.58±0.52) were in an ascending order (F= 20.16,P<0.01). Image scores of heart rate groups of<100 bpm (3.54±0.56),100--150 bpm (2.86± 0.70) and >150 bpm (2.81±0.63) were in a descending order (F=16.72, P<0.01). Multiple linearregression analysis demonstrated that multiple impact factors affected image quality of LM, LAD, LCX and RCA. Age(r=0.013 ,P<0.01)and contrast volume(r=0.292, P<0.01)had positive correlation with LM score. Age(r=0.509, P<0.01) and voltage (r=0.292, P=0.011) had positive correlation with LAD score. Heart rate(r= -0.179, P<0.05) had negative correlation with LAD score. Age (r=0.063, P< 0.01) had negative correlation with LCX score. Age (r=0.486,P<0.01) and voltage (r=0.220, P<0.05) had positive correlation with RCA score. Conclusions Dual-source CT could clearly shows coronary arteries of children with CHD over the age of 3 or with heart rate less than 100 bpm. Limitations of Dual-source CT coronary angiography for CHD children included young age and fast heart rate.