1.Abnormal myocardial contractile responses in patients with idiopathic dilated cardiomyopathy
Chinese Journal of Ultrasonography 2003;0(08):-
Objective To investigate abnormal myocardial contractile responses induced by dobutamine in patients with idiopathic dilated cardiomyopathy (DCM).Methods Eighteen DCM patients underwent low dosage dobutamine stress echocardiography (5,10,20 ?g?kg~ -1?min~ -1).Transthoracic echocardiogram was recorded with the use of a HP Sonos 5500 color echocardiographic diagnostic system.Wall motions in 16 myocardial segments were assessed using a four-point scale recommended by the American Society of Echocardiography.During dobutamine infusion,abnormally contracting segments were classified into four different patterns of contractile response: improved,unchanged,worsened and biphasic.Unchanged,worsened and biphasic segments were defined as abnormal contractile responses.Worsened and biphasic segments were judged to be ischemia-like responses.Results All patients showed abnormal myocardial contractile responses of wall motions (100%),and thirteen were ischemia-like responses ( 72.2%).In total 225 segments,126 segments showed abnormal contractile responses ( 56.0%).Among them,97 segments were unchanged segments ( 43.1%),29 segments were ischemia-like responses ( 12.9%),in which 16 segments were worsened ( 7.1%),and 13 segments were biphasic ( 5.8%).The statistical analysis showed that incidence of improved segments was highest in two point segments( 62.1%) compared with three point and four point segments; incidence of ischemia-like responses was highest in three point segments( 24.6%); incidence of unchanged segments was highest in four point segments( 68.4%).Conclusions DCM has the abnormal myocardial contractile responses induced by dobutamine stress echocardiography.It indicates there is the ischemia-like energy mismatch between demand and supply in DCM,which would be related to etiology and progress of DCM.
2.Study of dobutamine stress with Doppler tissue imaging echocardiography on synchronism of electromechanical activity in idiopathic dilated cardiomyopathy
Chinese Journal of Ultrasonography 1993;0(04):-
Objective To investigate whether dobutamine could induce or worsen electromechanical activity synchronism in the patients with idiopathic dilated cardiomyopathy(DCM). Methods Eighteen patients with DCM[mean age ( 45.8 ? 13.9 ) years, NYHA class Ⅱ-Ⅲ, left ventricular ejection fraction(LVEF)
3.Strategy and clinical translational advances of targeting the costimulatory molecule CD_(40)/CD_(40L) for neoplasm therapy
Songbing HE ; Liang WANG ; Dechun LI
Cancer Research and Clinic 2009;21(12):860-862
Costimulatory molecule CD_(40) is extensively expressed by immune, hematopoietic, epithelial, and a wide range of tumor cells. As a potential target for novel cancer therapy, CD_(40)/CD_(40L) may mediate tumor regression through both an indirect effect of immune activation and a direct cytotoxic effect on the tumor. Several drug formulations that target the CD_(40)/CD_(40L) pathway have undergone phase I clinical evaluation in advanced-stage cancer patients, and initial findings show objective clinical responses immune modulation function and have not serious toxicity.
4.Trypsin plus type II collagenase digestion for isolation of nucleus pulposus cells:the optimal glucose concentration in complete medium
Cunxin ZHANG ; Jinfeng MA ; Dechun WANG
Chinese Journal of Tissue Engineering Research 2016;20(20):2899-2906
BACKGROUND:Intervertebral disc degeneration is the pathological basis of degenerative spinal diseases. Studies on the influentialfactors of intervertebral disc degeneration contribute to the prevention and treatment of degenerative spinal disease.
OBJECTIVE:To observe the growth and proliferation of nucleus pulposus cels isolated by trypsin plus type II colagenase digestion in complete medium with different glucose concentrations, exploring the optimal glucose concentration for growth of nucleus pulposus cels.
METHODS:Nucleus pulposus cels isolated and cultured by trypsin plus type II colagenase digestion method were observed under an inverted microscope, and thecelnumber was counted. Morphology of nucleus pulposus cels was observed afterhematoxylin-eosinstaining and toluidine blue staining. Colagen type II immunoreactivity was detected by immunohistochemical staining combined with immunofluorescent staining.Nucleus pulposuscels were incubated in complete medium containing various glucose concentrations (0, 6.25, 12.5, 17.5, and 25 mmol/L) for 24 hours, and then cel proliferation and apoptosis were determined.
RESULTS AND CONCLUSION:The stained nucleus pulposus cels showed polygonal and short spindle, with one or two nuclei. Celularpseudopod appeared gradualy and then became slim with increased passage numbers. The isolated and cultured nucleus pulposus cels positively expressed colagen type II and aggrecanProliferative activity of nucleus pulposus cels cultured in medium with 17.5 mmol/L glucose was significantly higher than that in medium with 0 and 25 mmol/L glucose (P< 0.05 orP< 0.01). There wasno significant differencein cel apoptosis between these groups except for 0 mmol/L glucose (P<0.05). These results confirm that a large number of nucleus pulposus celscan beharvested by trypsin plustype II colagenase digestion and the optimal glucose concentration is 17.5 mmol/L.
5.Memory Consolidating During Sleep
Yang LI ; Dechun WANG ; Zhian HU
Progress in Biochemistry and Biophysics 2006;0(11):-
Sleep and memory are the basic function of the brain. A large number of studies from both humans and animals experiments have offered a substantive body of evidence supporting that sleep contributes crucially to memory consolidation. The processes of memory consolidation in hippocampus and cortex during sleep was reviewed and the primary cellular and molecular mechanism were briefly introduced.
6.Effects of oxidative stress on the apoptosis of nucleus pulposus cells in rats
Hongtao LI ; Dechun WANG ; Jianwei WEI
Chinese Journal of Tissue Engineering Research 2007;0(20):-
BACKGROUND:Apoptosis,which result in various reasons,plays an important role in intervertebral disc degeneration. One of important reasons for apoptosis is oxidative stress. OBJECTIVE:To investigate effects and mechanism of hydrogen-peroxide(H2O2) on nucleus pulposus cell injury induced by oxidative stress at intracellular signal transduction level in rats. DESIGN,TIME AND SETTING:A single sample observation was performed at the institute of Traumatology and Orthopaedics of Shandong Province from March to October in 2008. MATERIALS:The specific p38 mitogen-activated protein kinase(p38 MAPK) inhibitor SB203580,the specific JNK inhibitor SP600125 were purchased from Beyotime Institute of Biotechnology;Two 24-hour-old SD rats of SPF degree were purchased from Qingdao Institute for Drug Control. METHODS:The primary cultured nucleus pulposus cells were divided into four groups:the H2O2 group,stimulated by H2O2 with concentrations of 0,50,100,200,400,and 800 ?mol/L;Control group;SB203580+H2O2 group:cells were preincubated with SB203580,and then were treated by stimulated by H2O2;SP600125+H2O2 group,cells were preincubated with SP600125,followed by stimulated by H2O2. MAIN OUTCOME MEASURES:The expression and cellular localization of P-p38MAPK and P-JNK was detected by immunohistochemistry,and the expression of total and phosphorylated SAPK/JNK,p38MAPK were measured by Western blotting. RESULTS:H2O2 could activate the activity of P38 and JNK. The expression of P38MAPK was significantly inhibited with the pretreatment of SB203580;however,the SP600125 could inhibit the expression of JNK. Immunofluorescence analysis demonstrated that P-p38MAPK and P-JNK were expressed and distributed mainly in cytoplasmic and nuclear exception of the control group. CONCLUSIONS:Apoptosis of rat nucleus pulposus cells are induced by oxidative stress via p38MAPK and JNK signal pathway.
7.Expression,purification and bioactivity identification of recombinant SCR15-18 domain of human soluble complement receptor type 1
Bing TAN ; Dechun ZHANG ; Zhengqing WANG
Journal of Third Military Medical University 2003;0(10):-
Objective To prepare highly expressed, purified and refolded SCR15-18 of human soluble complement receptor type 1 (sCR1-SCR15-18) protein. Methods The expression of recombinant pET32-sCR1-SCR15-18 in E.coli.. BL21 was induced by IPTG of different concentrations for different time period under different temperatures and the bacteria were split by sonication. The sCR1-SCR15-18 protein was purified by Ni2+-NTA resin affinity chromatography. The purified protein was refolded under different conditions. Then the bioactivity of the protein was analyzed. Results The sCR1-SCR15-18 protein of high expression, purity and bioactivity was attained. Conclusion The parameters of expression, purification and refolding of sCR1-SCR15-18 protein were optimized, which may pave a way for further studies.
8.Diagnosis and treatment of the posterior margin separation of lumbar vertebral epiphysis
Dechun WANG ; Xiaoliang CHEN ; Xiujun ZHENG
Orthopedic Journal of China 2006;0(03):-
[Objective]To study the pathogenesis,clinical manifestation and surgical treatment of the posterior margin separation of lumbar vertebral epiphysis.[Method]Sixteen patients suffering from the posterior margin separation of lumbar vertebral epiphysis were followed up.The clinical manifestation and radiologic examination were analyzed and results of surgical treatment were evaluated.[Result]The patients were usually young and manifested with the sign and symptom of lumbar disc herniation and/or lumbar stenosis.CT was helpful for the accurate diagnosis of this disease.The different surgical measures were taken for the treatment according to the type and range of protrusion.[Conclusion]The posterior margin separation of lumbar vertebral epiphysis were divided into three types:end plate separation and moving into posterior margin,Schmorl node and avulsion fracture.The good results can be obtained with surgery.
9.Biological effects of adeno-associated virus vectors of human transforming growth factor ?_1 and ?_3 for reversion of rabbit intervertebral disc degeneration
Jiaming SAI ; Yougu HU ; Dechun WANG
Chinese Journal of Orthopaedics 1999;0(04):-
Objective To study the biological effects of pSNAV2-hTGF?1 and pSNAV2-hTGF?3 on the reversion of rabbit disc degeneration. Methods Rabbit nucleus pulpous and annulus fibrosus cells were isolated and cultured. The fluorescence labled pSNAV2 were used to detect the transfect rates of rabbit disc cells at first. Then, the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 were transfected into the degenerated rabbit disc cells respectively. The biological effects of hTGF?1 and hTGF?3 on degenerated rabbit disc cells were detected with Western-bloting and 35S detection to analyze and compare the matrix synthesis of the tranfected cells. Results pSNAV2 could transfect degenerated disc cells effectively in the early stages. Both the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 could stimulate the synthesis of collagen Ⅱ and proteoglycan of the rabbit disc cells. For the early stage of degenerated disc cells, the synthesis of collagen Ⅱ and proteoglycan were greater transfected with pSNAV2-hTGF?1 than transfected with pSNAV2-hTGF?3. The pSNAV2-hTGF?1 could promote the degenerated rabbit annulus fibrosus cells to synthesize collagen Ⅰ and pSNAV2-hTGF?3 could promote the degenerated nucleus pulpous cells of later stage to synthesize the collagen Ⅱ. Conclusion Both pSNAV2-hTGF?1 and pSNAV2-hTGF?3 can promote the degenerated rabbit disc cells of early stage to synthesize the matrix. pSNAV2-hTGF?3 can efficently promote the seriously degenerated nucleus pulpous cells to synthesize the collagen Ⅱ.
10.Construction of AAV-hVEGF_(165) and the biological effect of hVEGF_(165) and TGF?1 in degenerative annulus fibrosus cell of intervertebral disc
Yuefu DONG ; Yougu HU ; Dechun WANG
Chinese Journal of Orthopaedics 2001;0(06):-
Objective To evaluate the reversion possibility of hVEGF165 and TGF?1 to intervertebral disc degeneration by gene method. Methods The hVEGF165cDNA obtained from plasmid pcDNA3(+)-hVEGF165 was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hVEGF165 was identified by restriction enzymes analysis and sequencing analysis, and then transfered to the HEK293 cell and VEC by lipofectamine mediated gene transfer method. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry and explored the influence to the proliferation of vascular endothelial cell by MTT. Whereafter the AAV-hVEGF165 was packaged by Benyuan Zhengyang Company. AAV-hVEGF165 and AAV-TGF?1 were cotransfected into annulus fibrosus cell of intervertebral disc, then the expression of hVEGF165 and TGF?1, and the change of collagen Ⅰin annulus fibrosus cell were detected by Western blot. Results The recombinant pSNAV-hVEGF165 was completely constructed and confirmed by restriction enzymes analysis and sequencing analysis. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry in experimental group, and hVEGF165 could promote the proliferation of vascular endothelial cell. The bioactive AAV-hVEGF165 was successfully constructed. The expression of AAV-hVEGF165 and AAV-TGF?1 were manifested in degenerative annulus fibrosus cell by Western blot, and the expression of collagen Ⅰin annulus fibrosus cell cotransfected by AAV-hVEGF165 and AAV-TGF?1 was markedly more than that of the monogenic transfected cell. Conclusion hVEGF165 could cooperate with TGF?1 to promote the expression of collagen Ⅰ.