1.Role of NR2B in estrogen increasing visceral hypersensitivity of female stressed rats
Junkai SU ; Jun GAO ; Zhaoshen LI ; Duowu ZOU ; Hailian CAO
Chinese Journal of Digestion 2008;28(8):527-530
Objective As estrogen increases visceral hypersensitivity induced by water avoidance stress in female rats,further experiment was designed to determine whether the influence of estrogen involves NR2B.Methods Healthy adult female Wistar rats were bilaterally ovariectomized,and then housed individually.Implantation of cannula into lateral cerebroventricle and electrodes into the abdominal muscle had been done.After 5 days recovery,rats with abnormal behavior and ehctromyography were excluded,finally a total of 48 rats were eligible,and were equipped for abdominal muscle electromyography and submitted to water avoidance stress(WAS).Visceromotor response(VMR)to 20,40,60 and 80 mmHg colorectal distension(CRD)was recorded in rats intracerebroventricular-infused with either 17β-estradiol,normal saline,AP5(NMDA receptor-antagonist)or Ro25-6981(NR2B antagonist).NR2B mRNA in anterior cingulate cortex or dorsal root ganglia were compared by real-time PCR between the rats treated with 17β-estradioI and that with normal saline.Results Bilaterally ovarieetomized rats treated with 17β-estradiol,exhibited more visceral hypersensitivity after WAS than that with normal saline on 40,60 and 80 mmHg CRD(P=0.039,P=0.033,P=O.001).The VMR on 40 and 60 mmHg CRD in 17β-estradiol treated group was not significantly different from that in 17β-estradiol plus Ro25-6981 treated group.Whilst,significant differences of VMR were noted between 17β-estradiol treated group and 17β-estradiol plus Ro25-6981 treated group on 80 mmHg CRD,17β-estradiol treated group and 17β-estradiol plus AP5 treated group on 60,80 nmmHg CRD,respectively.17β-estradiol increased NR2BmRNA in anterior cingulate cortex(0.57±0.41 vs 0.21±0.13,P=0.048),but not in dorsal root ganglia(0.35±0.45 vs 0.38±0.31,P=0.465). Stress-induced visceral hypersensitivity in the hormonally-restored visceral hyper-responsiveness of bilaterally ovariectomized rats was antagonized by AP5 or Ro25-6981.Conclusions Estrogen may be mediated through NR2B activation to enhance visceral sensitivity in female stressed rats,that probably related with the increased expression of NR2B mRNA in anterior cingulate cortex.
2.Setup error of Orfit versus vacuum bag in radiotherapy for cervical cancer
Lijuan GAO ; Jiamin HUANG ; Jun HUANG ; Jianxin SU ; Yuqi WU ; Chengguang LIN
Chinese Journal of Radiation Oncology 2017;26(9):1080-1083
Objective To compare the setup errors of the negative pressure vacuum air cushion (vacuum bag) and the Orfit body foam fixator (Orfit frame) in radiotherapy for cervical cancer.Methods A total of 40 patients receiving three-dimensional radiotherapy for cervical cancer were enrolled in this study and equally and randomly divided into vacuum bag group and Orfit frame group.And the two groups were divided into Orfit-1 group, Orfit-2 group, vacuum-1 group, and vacuum-2 group according to the treatment course.The Orfit-1 group and vacuum-1 group were the data in the first 12 treatments, while the Orfit-2 group and vacuum-2 group were the data in the following 13 treatments.A cone-beam computed tomography scan was performed before each treatment to analyze setup error and then the body position was corrected to start the treatment.Comparison of continuous data between groups was made by paired t-test, while comparison of categorical data was made by chi-square test.Results There was a significant difference in the setup error in y-axis direction between the Orfit-1 group and the Orfit-2 group (P=0.003) and the setup error in r-axis direction between the vacuum-1 group and the vacuum-2 group (P=0.013).There were no significant differences in the setup errors in four directions (x-axis, y-axis, z-axis, and r-axis) between the Orfit-1 group and the vacuum-1 group (P>0.05).There were significant differences in the setup errors in y-axis and z-axis directions between the Orfit-2 group and the vacuum-2 group (P=0.007;P=0.001).Conclusions The Orfit frame and the vacuum bag have their own advantages and disadvantages in the fixation of body position in radiotherapy for cervical cancer.The setup error can be improved by long vacuum bags, ultrasound bladder capacity scanner, image-guided radiotherapy, or sectional radiotherapy plan.
3.Construction and identification of recombinant adenovirus containing heat shock protein70 gene driven by carcinoembryonic antigen promoter
Yunliang SUN ; Can XU ; Changqing SU ; Jun GAO ; Jing JIN ; Hongyu WU ; Zhaoshen LI
Chinese Journal of Pancreatology 2011;11(4):251-254
Objective To construct the recombinant adenoviros containing heat shock protein70 (Hsp70) gene driven by carcinoembryonic antigen (CEA) promoter. Methods Hsp70 gene and CEA promoter were amplified by RT-PCR and PCR, and then subcloned into the shuttle vector pDC316 to construct the recombinant vector PDC316-pCEA-Hsp70. The recombinant vector was co-transfected with adenoviral backbone plasmid into HEK293 cells to generate the recombinant adenovirus Ad5-pCEA-Hsp70. The recombinant adenovirus was purified by CsCl banding and titrated by 50% tissue culture infective dose (TCID50) assay. After transfection of the recombinant adenovirus into human pancreatic cell lines SW1990 and BxPC3, the expression of mRNA and protein level of Hsp70 were determined by RT-PCR and ELISA,respectively. Results Digestion and DNA sequencing certified that the Hsp70 gene and CEA promoter was successfully inserted into pDC316 plasmid. Virus acquired through co-transfection with backbone plasmid was confirmed to be constructed successfully by PCR amplification. The particles finally expressed was 2.2 ×1011vp/ml, and the titer was 1.5 x 1010 PFU/ml. BxPC3 cancer cells with positive CEA expression showed increased expression of Hsp70 mRNA and protein after infected by recombinant adenovirus; while SW1990 cancer cells with negative CEA expression showed no change of expression of Hsp70 mRNA and protein after infected by recombinant adenovirus. Conclusions The recombinant adenovirus Ad5-pCEA-Hsp70 which can express Hsp70 gene in CEA positive cancer cells is constructed successfully.
4.Clinical analysis of reoperations for 67 bleeding patients after coronary artery bypass
Jie GAO ; Yan LIU ; Peixiong SU ; Xitao ZHANG ; Jun YAN ; Song GU
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;28(6):356-358
Objective To investigate the common sites and risk factom of reoperation for bleeding after coronary artery bypass grafts(CABG).Methods During Jan 2000 to July 2011,2765 CABG procedures hed been done in our hospital,including 874 cases with CABG by cardiopulmonary bypass,1891 cases with off-pump coronary artery bypass grafts (OPCAB),105 cases combined with other procedures (cardiac valve operation,resection of ventricular aneurysm,et al),216 cases applied intra-aortic balloon pump (LABP)).67 patients needed reoperation for bleeding,the morbidity is 2.4%.Factors that might affect the result of reoperation were analyzed by single-factor analysis.Results The following factors were related to high rates of reoperation (P < 0.05 ):the internal mammary artery (IMA) used,anfithrombotic medication discontinued less 5 days before elective surgery,concurrent hypertension,combined with other surgery procedures and ages >70 years.It was not signifieantly related to CABG or OPCAB,ff use of IABP,ff concurrent diabetes,number of grafts inserted.Major sources of bleeding found in 48 patients were the IMA and its vessels bed,vein grafts,sternum,aortic faucet,midriff,thymus glrard bed vessels.Conclusion Prepared satisfactorily before operation,avoid risk factors can reduce reoperation for bleeding,but the most important is the operation carefully during operation of hemostasis.
6.Analysis of influencing factors of severe preeclampsia with exudative retinal detachment and study of pregnancy outcome
Dan WEI ; Jiumei SU ; Jun GAO ; Xiyan AN ; Zongwen TAN ; Wenjun XU
Chinese Journal of Ocular Fundus Diseases 2021;37(5):371-376
Objective:To observe and preliminary investigate the influencing factors of exudative retinal detachment in patients with severe preeclampsia (S-PE) and its influence on pregnancy outcome.Methods:A total of 89 patients of S-PE patients diagnosed in Department of Obstetrics in The Third Central Hospital of Tianjin from July 2018 to June 2020 were included in this study. According to whether the patients were complicated with exudative retinal detachment, they were divided into retinal detachment group (43 cases) and non retinal detachment group (46 cases). All patients underwent direct ophthalmoscopy, and those with suspected or definite retinal detachment underwent optical coherence tomography or B-mode ultrasonography. The age, course of disease, body mass index, systolic blood pressure, diastolic blood pressure, laboratory serum albumin level, 24 h urinary protein, platelet count, mean platelet volume, platelet distribution width and serum uric acid content were collected. The average age ( t=-1.01), body mass index ( t=-1.799), course of disease ( t=-1.657) and constituent ratio of primipara and multipara in the two groups were compared ( χ2=1.411), the difference was not statistically significant ( P>0.05). The end of pregnancy time, systemic complications, fetal outcomes and neonatal Apgar scores were analyzed, retrospectively. Independent sample t test was used to compare the quantitative data between the two groups; Chi square test was used to compare qualitative data. Binary logistic regression analysis to screen the influencing factors of exudative retinal detachment in S-PE. Results:Systolic blood pressure and diastolic blood pressure were higher in the retinal detachment group than in the non retinal detachment group, and the difference was statistically significant ( t=2.087, 2.810; P=0.040, 0.006). Serum albumin ( t=-10.404), 24-hour urinary protein quantification ( t=6.401), blood uric acid ( t=3.910), platelet count ( t=-6.124) and platelet distribution width ( t=2.992) were compared between the two groups, and the differences were statistically significant ( P<0.05). There were 30 (69.77%, 30/43) cases in retinal detachment group and 14 (30.43%, 14/46) cases in non retinal detachment group with severe systemic complications; The average time of ending pregnancy was 34.66±3.29 and 38.25±1.62 weeks; The average Apgar scores at birth were 7.28±3.40 and 9.46±1.59. The incidence of severe systemic complications in two groups ( χ2=13.755), the end of pregnancy time ( t=-6.448), Apgar score at birth ( t=-3.824), the differences were statistically significant ( P=0.000、0.000、0.000). Fetal distress and death were 8 (18.60%, 8/43), 7 (16.28%, 7/43) and 7 (15.22%, 7/46), 1 (2.17%, 1/46) in retinal detachment group and non retinal detachment group respectively. There was no significant difference in the incidence of fetal distress between the two groups ( χ2=0.182, P=0.670). The difference of fetal mortality was statistically significant ( P= 0.027). Logistic regression analysis showed that serum albumin and uric acid were associated with exudative retinal detachment in patients with S-PE ( P<0.05). Conclusions:Serum albumin and uric acid are the influencing factors of exudative retinal detachment in patients with S-PE. Patients with exudative retinal detachment are prone to adverse pregnancy outcomes.
7.Differential expression genes in various phases of chronic myeloid leukemia by cDNA microarray.
Su-Jun GAO ; Xun ZHU ; Cheng YAO ; Wei LI ; Guan-Jun WANG
Journal of Experimental Hematology 2010;18(3):575-578
The aim of this study was to explore the mechanism underlying the blast crisis of chronic myeloid leukemia. Analysis of gene expression profiles of chronic myeloid leukemia patients in chronic phase and blast crisis were analyzed by using cDNA microarray representing 4096 genes for finding the differential expression genes. The results indicated that 74 differential expression genes were identified in at least 3 gene chips in blast crisis compared with chronic phase, among them 52 genes were down-regulated and 22 genes were up-regulated in blast crisis. The differential expression genes were involved in these genes including genes related to cell structure/mobility, signal transduction, transcription factor, related immunity, metabolism, cell cycle, oncogene/anti-oncogene, cell receptor, protein translation/synthesis and some unknown functions. It is concluded that the blast crisis of CML is resulted from abnormality and interaction of multigene, among them functional abnormal genes related to signal transduction, cell cycle, cell differentiation and immunity may be the critical genes for chronic myeloid leukemia blast crisis.
Adult
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Aged
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Female
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Gene Expression Profiling
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Gene Expression Regulation, Leukemic
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Genes, Neoplasm
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Humans
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Leukemia, Myelogenous, Chronic, BCR-ABL Positive
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genetics
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metabolism
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Male
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Middle Aged
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Oligonucleotide Array Sequence Analysis
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Signal Transduction
8.Diagnostic Significance of BAT in Anaphylaxis to Non-ionic Contrast Media.
Hao-yue ZHANG ; Su-jun XU ; Xiao-xian TANG ; Ji-jun NIU ; Xiang-jie GUO ; Cai-rong GAO
Journal of Forensic Medicine 2015;31(3):188-190
OBJECTIVE:
To investigate the diagnostic significance of basophil activation test (BAT) in anaphylaxis to non-ionic contrast media through testing the content of CD63, mast cell-carboxypeptidase A3 (MC-CPA3), and terminal complement complex SC5b-9 of the individuals by testing their levels in the normal immune group and the anaphylaxis groups to β-lactam drugs and non -ionic contrast media.
METHODS:
The CD63 expression of basophilic granulocyte in blood was detected by flow cytometry. The levels of MC-CPA3 in blood serum and SC5b-9 in blood plasma were detected by ELISA.
RESULTS:
The CD63 expression of basophilic granulocyte in blood, the levels of MC-CPA3 and SC5b-9 of anaphylaxis to non-ionic contrast media and β-lactam drugs were significantly higher than that in normal immune group (P < 0.05).
CONCLUSION
There is activation of basophilic granulocytes, mast cells and complement system in anaphylaxis to non-ionic contrast media. BAT can be used to diagnose the anaphylaxis to non-ionic contrast media.
Anaphylaxis/diagnosis*
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Basophils/cytology*
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Carboxypeptidases A/metabolism*
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Complement Membrane Attack Complex/metabolism*
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Contrast Media
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Flow Cytometry
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Granulocytes/cytology*
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Humans
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Mast Cells/cytology*
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Tetraspanin 30/metabolism*
9.Expressing the specific vessels signs in lung cancer by 64-slice spiral CT in first phase perfusion imaging *
Guanqin SU ; Xiaoqing BO ; Shu YANG ; Guopeng SUN ; Jun CHAI ; Ke ZHOU ; Amei GAO ; Hubing DUAN ; Xiuping DONG
Chongqing Medicine 2013;(24):2884-2886,2909
Objective To investigate the specific vessels signs in lung cancer by 64-slice spiral CT in first phase perfusion ima-ging so as to get more reasonable time about CT angiography .Methods Among the 47 cases ,there were 38 cases of central and 9 cases of peripheral primary lung cancer underwent 64-slice spiral CT in first phase perfusion imaging :one scan was obtained every 1 seconds during 8-38 seconds with 8 section × I without scanning interval after injection .Precontrast and postcontrast attenuation on every leisions was recorded ,to calculate the peak height(PH) and peak height time by time density curves TDC of pulmonary le-sions to aorta ,those reflect the the tumor feeding artery Imaging about lesions of lung cancer and inflammation .observed and calcu-lated the display rate and display time of lesions of lung cancer and inflammation .means of t test was used for statistics .Results a-bout the peak heights of in pulmonary artery and aorta phese ,statistically significant differences were found between inflammation and lung cancer(P<0 .05) .no statistically significant differences were found between central and peripheral lung cancer (P>0 .05) . about the peak heights time in aorta phase ,no statistically significant differences were found among three groups (P>0 .05) .The tumor vascularity were discoved in lesions in patients with lung cancer (44/47 cases ,93 .62% );The erosion narrow pulmonary ar-tery were discoved in central lung cancer (37/38 cases ,97 .37% ) ,in peripheral lung cancer(6/9 cases 66 .67% );no abnormal pulmo-nary artery were discoved only in 4 patients with lung cancer .Both tumor vascularity and abnormal pulmonary artery were most dis-plaied in 18-31s in CT angiography .Conclusion 64-slice spiral CT in first phase dynamic perfusion imaging can order completely show the specific vessels signs in lung cancer and reflect the tumor feeding artery Imaging of lung cancer .Analysising those benefi-ted to select reasonably the time of CT angiography and Improve the rate of lung cancer diagnosis .
10.Cloning and expression analysis of 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase gene in Tripterygium wilfordii.
Yu-ru TONG ; Ping SU ; Meng ZHANG ; Yu-jun ZHAO ; Xiu-juan WANG ; Wei GAO ; Lu-qi HUANG
China Journal of Chinese Materia Medica 2015;40(22):4378-4383
To clone the 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (TwMCT) full length cDNA from Tripterygium wilfordii, the specific primers were designed according to the transcriptome data and the LCPCR were carried out. After a series of bioinformatics analysis on the TwMCT, the MeJA induced expression content were investigated by real-time fluorescence quantification polymerase chain reaction (RT-qPCR). The result showed that the full of TwMCTcDNA was 1 318 bp nucleotides encoding 311 amino acids. The molecular weight of the deduced TwMCT protein was about 34.14 kDa and the theoretical isoelectric point was 8.65. Result of the RT-qPCR analysis indicated that the content of TwMCT mRNA expression in T. wilfordii suspension cell was rising after treating with MeJA and reached the maximum in 24 h. Cloning and analyzing TwMCT gene from T. wilfordii provided gene element for studying the function and expression regulation of secondary metabolites.
Amino Acid Sequence
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Cloning, Molecular
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Erythritol
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analogs & derivatives
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metabolism
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Gene Expression Regulation, Plant
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Molecular Sequence Data
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Nucleotidyltransferases
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chemistry
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genetics
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metabolism
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Phylogeny
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Plant Proteins
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chemistry
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genetics
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metabolism
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Protein Structure, Secondary
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Sequence Alignment
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Sugar Phosphates
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metabolism
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Tripterygium
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chemistry
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enzymology
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genetics