1.Proteomic Analysis of Hypertrophied Left Ventricle in Spontaneously Hypertensive Rats
Xian JIN ; Li XIA ; Jun-Zhi SHI ; Ning-Yuan FANG ;
Chinese Journal of Hypertension 2007;0(03):-
Objective To investigate the molecular mechanisms for the development of cardiac hypertrophy in hypertension,the present study provided the differential protein expression analysis of hypertrophied heart at differ- ent stages in spontaneously hypertensive rats (SHR).Methods The profiles of protein expression of left ventricu lar myocardium in SHR and its normotensive control Wistar-Kyoto (WKY) rats at the age of 4 and 20 weeks were analyzed with two-dimensional gel electrophoresis (2-DE) in combination with matrix assisted laser desorption ioniza- tion-time of flight (MALDI-TOF-TOF) mass spectrometry.Results Although the blood pressure of SHR was normal at 4 weeks age,hypertrophy of the left ventricle had already developed.The expression pattern in the hy- pertrophic myocardium was found 27 modulated proteins,20 of which were identified.These proteins are involved in reactions of energy metabolism,mitoehondrial oxidative phosphorylation and oxidative stress,etc.The expres- sion of 13 proteins was significantly changed in SHR rats at early stage prior to the development of sustained hyper- tension,while the expression changes of other 7 proteins occurred only at late stage in SHR rats when the blood pressure was significantly elevated.Conclusions lncrease in glycolysis and decrease in oxidation of fatty acid and glucose was shown in the hypertrophied myocardium from early stage in SHR prior to the development of hyperten sion.The significant changes in protein expression of the mitochondrial electron transport chain and antioxidative molecules support the hypothesis that oxidative stress promotes and accelerates the development of hypertensive car- diac hypertrophy.
2.Expression of let-7a2 in lung adenocarcinoma
Xiaoming XIA ; Weiyun JIN ; Renzhong SHI ; Yafeng ZHANG ; Jun CHEN
Cancer Research and Clinic 2010;22(1):49-51
Objective To explore the expression and significance of let-7a2 in lung adenocarcinoma.Methods To adapt technology of realtime PCR with reference of U6, the expression of let-7a2 in tumor tissues and normal tissues was detected in 15 cases of resected lung adenocarcinoma. The laboratorial data were analysed by measure of 2~(-△△CT) method. The analysed data were managed by statistics. Results The expression of let-7a2 in lung adenocarcinoma tissue was significantly lower than that in lung normal tissue (0.8733±0.4821 vs 2.4527±1.0111). This difference possess markedness significance(t=6.816, P<0.001).Conclusion The expression of let-7a2 in lung adenocarcinoma is obviously lower and is concerned with the course of lung cancer evolution.
3.Surgical treatment for chronic pancreatitis characterized by a pancreatic mass
Jun SHI ; Feng XIA ; Guanqun LI ; Qingyu ZHANG
Chinese Journal of General Surgery 2010;25(8):649-651
Objective To summarize the diagnostic and therapeutic experience on chronic pancreatitis characterized by a pancreatic mass. Methods The clinical data of 28 cases of chronic pancreatitis with mass undergoing surgical operations were retrospectively analyzed in our hospital from June1999 to June 2009. Results Among the 28 cases, 19 were diagnosed as carcinoma, 9 cases were diagnosed as chronic pancreatitis respectively before operation. Needle aspiration biopsy and/or postoperative pathology identified chronic pancreatitis in all cases. The symptom included abdominal pain (22 cases),jaundice (15 cases), and obstruction of duodenum (4 cases). Pancreaticoduodenectomy was performed in 17 cases, choledochojejunostomy performed in 3 cases, pancreatojejunostomy performed in 1 case.Duodenum-preserving resection was performed in 4 cases, and resection of body and tail of the pancreas were performed in 3 cases. There was no operative death. Postoperative complications included pancreatic leakage (2 cases), severe gastroplegia (2 cases) and stress peptic ulcer with massive bleeding ( 1 case). All patients got follow-up ranging from 6 months to 5 years. Recurrence of abdominal pain developed in 7 cases after 2 years. Canceration of pancreatic mass was found respectively in 8 months, 1 year after operation in one each cases. Conclusion Preoperative differential diagnosis of chronic pancreas and pancreatic tumor was difficult. Although needle aspiration biopsy is the effective method for diagnosis, there may be still a possibility of missed diagnosis/misdiagnosis.
4.Relationship of detection of serological endothelin and coronary artery lesion in children with kawasaki disease
shi-mei, JI ; yi-chang, LIANG ; shu-jun, XIA
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To investigate the alterations of serological endothelin(ET) in patients with kawasaki disease(KD) and its relation with coronary dilatation(CAD).Methods Serological ET were measured in 50 cases of patients with KD in acute phase as well as subacute phase; 30 cases of patients with acute febrile infection(IC) in infective phases and 30 healthy children(HC).Results 1.ET in subacute phase significantly increased than that in acute phase.2.In both acute and subacute phases of KD, ET significantly elevated higher than that in HC; and ET in subacute phase significantly elevated than that of IC. 3. Fifty-four percentage patients with KD were complicated with CAD. For CAD subgroup, ET had no difference with CAD subgroup in subacute phase.Conclusion ET has still increases in subacute of KD,which indicates the relation with coronary artery lesion.
6.A pharmacodynamic study of intervals of Mitomycin C treatment on human bladder cancer cell line in vitro
Yun-Xia WAN ; Jun TIAN ; Jian-Hui MA ; Han-Zhong LI ; Yan-Fen LI ; Wei SHI ;
Cancer Research and Clinic 2006;0(11):-
Objective To study the relationship between the intervals of Mitomycin C treatment and cytotoxicity, apoptosis and drug resistance for bladder cancer cells.Methods The bladder transitional cell cancer line BIU-87 was treated for two hours every time for five times with intervals of 24, 48, 72 and 96 hours respectively.Cytotoxicity was measured by MTT.p53,bcl-2,Bax and p170 expression were analyzed by Western blot.Results The IC_(50)(?g/ml)were 4.41,0.71,2.83,4.51and 6.16 with treatment intervals of 24, 48, 72 and 96 hours respectively, p53 and bcl-2 were significantly down-regulated and bcl-2/Bax was re- duced at 24 hour treatment interval but not changed at 48,72 and 96 hour intervals,p170 was not detected at 24 hour treatment interval but increasingly expressed at 48,72 and 96 hours intervals.Conclusion The in- terval of Mitomycin C treatment is closely related with cytotoxieity and apoptosis and drug resistance of blad- der cancer cells.The intervals of intravesical instillations may play an important role in the effect of chemotherapy.
7.The Effect of Fructose-1,6-diphosphate and HTK Solution on Protecting Primary Cardiac Muscle Cells of Rat with Cold Preservation
Xiaofeng SHI ; Jun CHENG ; Suisheng XIA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(3):292-293,302
Summary: In this study we tried to investigate the effect of fructose-1,6-diphosphate and HTK solution on protecting primary cardiac muscle cells of rat with cold preservation. The primary cardiac muscle cells of rat were cultured in vitro with four preservation solutions respectively: 0.9 % sodium chloride solution (group A), FDP (group B), HTK solution (group C) and a mixture of FDP and HTK solution (group D). The cells were preserved for 6, 8 and 10 h at 0-4 ℃. The values of AST and LDH-L and the Na+-K+ ATPase activity in cardiac muscle cells were detected, and the survival rate of cardiac muscle cells was detected with trypan blue staining. The values of AST and LDH-L in group C and group D were remarkable lower those in group A and group B (P<0.001), while the Na+-K+ ATPase activity and the survival rate of cells in group C and group D were much higher than those in group A and group B (P<0.001). The values of AST and LDH-L after 6 hours in group D decreased much more than those in group C (P<0.01), while the Na+-K+ ATPase activity and the survival rate of cells in group D improved more than those in group C (P<0.01). Both of the HTK solution and the mixture of HTK and FDP solution have an evident effect on protecting the primary cardiac muscle cells of rat in vitro with cold preservation, Compared with the HTK solution, the mixture solution has a better short-term protective effect.
8.Construction of bicistronic green fluorescent protein labeled pSELECT GFPzeo human bone morphogenetic protein 2 eukaryotic expression vector
Hongchao HUANG ; Jun MIAO ; Chunrong LIU ; Yongcheng HU ; Qun XIA ; Kesong SHI
Chinese Journal of Orthopaedics 2010;30(12):1228-1234
Objective To construct green fluorescent protein (GFP)-labeled pSELECT-GFP zeohBMP2 eukaryotic expression vector. Methods The encoding fragment of hBMP2 gene was obtained from a recombinant plasmid pcDNA3.1/CT-hBMP2 by using polymerase chain reaction (PCR). hBMP2 gene was inserted into pTA2-T-easy and pSELECT-GFPzeo-MCS eukaryotic expression vector, and then transferred into competence DHSα cells. After screening, pSELEC-GFPzeo-hBMP2 was obtained and identified by sequence analysis. The recombinant vector pSELECT-GFP zeo-rhBMP2 was transfected into CHO cells. The successful trasfection was verified by fluorescence microscope in 48-72 hours. The RT-PCR and immunofluorescence was used to confirm the hBMP2 expression. Western Blotting was used to detect the secretion of hBMP2.Results A 1216 bp fragment was obtained by PCR, the same as expectant fragment. The recombined pSE-LECT-GFPzeo-hBMP2 eukaryotic expression vector was identified by restriction mapping and sequence analysis. The results were identical with that of reported hBMP2 sequence (Genebank NM-001200). The successful transfection was verified by fluorescence microscope in 48-72 hours. The stable expression in eukaryotic cells was confirmed by immunofluorescence and RT-PCR which showed an obvious band between 1000-2000 bp. Western Blotting identified the immunogenicity of recombinant human BMP2 with the molecular weight of about 17×103. Conclusion The pSELECT-GFPzeo-hBMP2 eukaryotic expression vector was constructed successfully.
9.Constructing bicistronic eukaryotic expression plasmid containing human bone morphogenetic protein-2 labeled with green fluorescent protein
Jun MIAO ; Chunrong LIU ; Hongchao HUANG ; Qun XIA ; Kesong SHI ; Guosheng CUI
Chinese Journal of Tissue Engineering Research 2008;12(20):3984-3987
BACKGROUND: Bone morphogenetic protein-2(BMP-2) production of targeted cells is promoted by transfection of adenoviral vectors containing gene, but there are some immune responses. Transfection with plasmid as vector holds promise.OBJECTIVE: To explore the feasibility to construct human bone morphogenetic protein-2 eukaryotic expression vector labeled with green fluorescent protein (GFP).DESIGN: Single sample observation.SETTING: Tianjin Hospital.MATERIALS: The experiment was performed at the Key Laboratory of Hormone and Development, Ministry of Health, Tianjin Medical University from March 2006 to March 2007. pcDNA3.1/CT-hBMP2 plasmid containing full-length hBMP2 gene fragment was provided by Dr. Li; bicistronic eukaryotic expression vector pSELECT-GFPzeo-MCS and Zeo was provided by Invivogen; pTA2(R)-T Easy by Dingguo, China; restriction enzymes BamHI and NheI, T4 DNA ligase by Jingmei Biotech; PCR upstream and downstream primer synthesis and sequencing by Augct, Beijing.METHODS: With pcDNA3.1/CT-hBMP2 as template, hBMP2 target fragment was subcloned by PCR binding with designed specific primers. The fragment was bound with pTA2-T-easy and pSELECT-GFPzeo-MCS, separately, and transfected into DH5 α cells. pSELECT-GFPzeo-hBMP2 containing GFP was obtained after screening.MAIN OUTCOME MEASURES: hBMP2 sequence was identified by PCR; whether hBMP2 was cloned into pTA2-hBMP2 and pSELECT-GFPzeo-MCS was identified by digestion and sequencing.RESULTS: A target fragment of 1 216 bp was obtained by PCR amplification, and cloned into pTA2-T-easy and pSELECT-GFPZeo-MCS. The screening and sequencing results showed that the target fragment was 100% matched with BMP2cDNA sequence (NM-001200) from GenBank.CONCLUSION: hBMP2 eukaryotic expression vector labeled with green fluorescent protein is successfully constructed.
10.Research on the correlation between the rescue time and the spot survival rate during Wenchuan earth-quake
Fuxiang LI ; Zhaoxia XU ; Guomin CAO ; Hongmei WANG ; Kai ZHOU ; Li SHI ; Jun LI ; Xun XIA
Chinese Journal of Emergency Medicine 2009;18(4):420-422
Objective To study the correlation between the rescue time and the spot survival rate.Method The data of spot-rescued victims in a large public place of Dujiangyan City from 14:35 on May 12,2008 to 11:40 on May 15,2008 were analyzed.The searched-out victims included the spot death and spot survival,and they were statistically analyzed with Chi-Square test and Partitions of X2 method in order to find out correlation between rescue time and survival rate.Results Out of the 366 spot-rescued victims from the ruins,87 ones survived and the spot survival rate was 23.77%.The spot survival rate in the first 24 hours was much higher than that in the second 24 hours(X2=22.62,P<0.0125)and that in the third 24 hours(X2=37.84,P<0.0125),and no obvious difference in the spot survival rate between the second and the third 24 hours was found(X2=1.92,P>0.0125).The first 24 hours was further divided into 3 periods in equal length of time in order to find more subtle differences in early rescue.The spot survival rates in the first and the sccond 8 hours were much higher than that in the third 8 hours(x2=19.33 and 7.11,respectively,P<0.012 5)while there was no statistical difference in the spot survival rate between the first 8 hours and the second 8 hours(X2=1.75,P>0.012 5).Conclusions The"golden time"for spot rescuing the victims is the first 24 hours after seismic disaster,the chances to find the survivals is decreasing as the time elapsing.The earlier spot rescue starts in the first golden24 hours,the higher spot survival rate of the seismic victims will be.