1.Intranasal administration of the conditioned medium of human umbilical cord-derived mesenchymal stem cells for treatment of cerebral ischemia-reperfusion injury
Liping SHEN ; Shuaishuai WANG ; Liguo DONG ; Xia SHEN ; Fang HUA ; Xinchun YE ; Guiyun CUI
Chinese Journal of Tissue Engineering Research 2013;(45):7891-7897
BACKGROUND:Cytokines and neurotrophic factors secreted from human umbilical cord blood-derived mesenchymal stem cells secrete have neuroprotective effects on cerebral ischemia-reperfusion injury, but there are few reports about intranasal administration of human umbilical cord blood-derived mesenchymal stem cellconditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the protective effects of intranasal administration of human umbilical cord-derived mesenchymal stem cells-conditioned medium on neurologic function of rats with cerebral ischemia-reperfusion injury.
METHODS:Adult rats were subjected to 2 hours of right middle cerebral artery occlusion and the human umbilical cord-derived mesenchymal stem cells were isolated from the postpartum human cord. We made the conditioned medium of human umbilical cord-derived mesenchymal stem cells. Ischemic rats were randomized and assigned to three groups and were treated by intranasal routine starting 24 hours after middle cerebral artery occlusion with:(1) saline for control group;(2) Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium for medium control group;(3) conditioned medium treatment group (10mL/kg) daily for 14 days. Behavioral tests (foot fault test, and modified Neurological Severity Score) were performed before and at 1, 7, 14 days after middle cerebral artery occlusion.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperatively 1 day (P>0.05). Compared to the control and medium control group rats, respectively, rats in the conditioned medium group significantly improved functional outcome after stroke in days 7 and 14 (P<0.05). There was also no significant difference in functional tests between the control group and medium control group in days 7 and 14 (P>0.05). These results suggest that human umbilical cord-derived mesenchymal stem cells-conditioned medium via intranasal administration can significantly improve neurologic functional outcome after cerebral ischemia-reperfusion injury.
2.Diagnosis of anterior bundle injury of medial collateral ligament after elbow dislocation with 3 .0T MRI
Jingwu YU ; Guoxin SHEN ; Jie NG TA ; Yongqiang YE ; Jinlan NG HUA ; Yu SHEN ; Xiaohui NG WA
Journal of Practical Radiology 2016;32(5):761-763
Objective To apply 3 .0T MRI in diagnosing injuries of anterior bundle of medial collateral ligament after elbow dislo‐cation .Methods The MRI features of the injuries of medial collateral ligament anterior bundle were analyzed retrospectively in 20 patients with elbow dislocation .The coronal ,sagittal ,axial and lamina oblique coronal were scanned routinely with SE T1WI ,T2WI‐FS sequences .Results Varying degrees of anterior bundle injuries of medial collateral ligament were observed in all the 20 patients ,in‐cluding the mild injury(n=8) ,part avnlsion(n=5) ,completely rupture(n=7) .Furthermore ,concomitant injuries including lateral collateral ligament(n=11) ,ringlike ligament(n=5) ,flex/stretch muscle tendon(n=9) ,and the fracture(n=7) were also observed . Conclusion The injuries of medial collateral ligament anterior bundle after elbow dislocation could be diagnosed accurately with 3 .0T MRI and the degree of injuries could also be defined on image .The 3 .0T MRI could be recommended as regular examination to pa‐tients with elbow dislocation .
3.Clinical investigation of RF transparent heating combined with TACE in the treatment of advanced primary hepatic carcinoma
Hua BAI ; Weixi SHEN ; Yongchong DOU ; Yanfang ZHANG ; Jin ZHENG ; Tao ZHENG ; Jianzeng YE ; Min XU
Chinese Journal of Postgraduates of Medicine 2008;31(18):4-6
Objective To observe the efficacy and toxicities of RF transparent heating (RTH) combined with transcatheter hepatic arterial chemoembolization (TACE) in the treatment of advanced primary hepatic carcinoma. Methods In a randomized manner, 69 patients with advanced primary hepatic carcinoma were divided into two groups: study group (TACE+RTH) 34 cases and control group (TACE alone) 35 cases, the control group were treated with DDP 80mg, FU 1000mg and E-ADM 60mg, E-ADM was used with iodized oil embolism 10ml. Results The total effective rate in the near future were 70.59% and 45.71%, the overall survival rates of 0.5, 1.0, 1.5, 2.0 years were 82.35%, 73.53%, 58.82%, 38.24% in study group and 74.29%, 75.14%, 45.71%, 22.86% in control group, respectively. Toxicities were similar comparing with the two groups. Conclusions RTH combined with TACE in the treatment of advanced primary hepatic carcinoma is better than TACE alone, at the same time TACE +RTH method is no increasing toxicity and is an effective safe combined one.
4.Clinical study with the method of purging liver and the heart fire,nourishing yin and moistering arid on patients with type 2 diabetes
Qing XUE ; Xingkuan WANG ; Hua ZHANG ; Yanan WANG ; Liao SUN ; Jin SHEN ; Lihong YE ; Hongyun LU
Chinese Traditional Patent Medicine 1992;0(11):-
0.05). group 1 and group 4 could significantly reduce the level of lipid、FINS, increase the level of IAI (P0.05). After treatment, the clinical symptom score dropped in group 1、2、3 (P0.05); serum levels of resistin decreased and adiponectin increased in group 1、4 (P0.05). More significant difference was observed in group 1、4 between the four groups about the serum level of resistin、adiponectin、Leptin、TNF-?、CRP、IL-6 (P
5.Transplantation of autologous bone marrow mesenchymal stem cells modified with AKT1 for treating pig ischemic cardiomyopathy
Yunsheng YU ; Shiqiang GUO ; Guiping YU ; Wenxue YE ; Haoyue HUANG ; Yihuan CHEN ; Fei HUA ; Yongquan GU ; Zhenya SHEN
Chinese Journal of Tissue Engineering Research 2009;13(49):9616-9624
BACKGROUND: A great quantity of cell loss in early stage following stem cell transplantation can significantly affect transplantation effect. Presently, it is confirmed that overexpression of AKT1 gene significantly inhibit cell apoptosis. OBJECTIVE: To explore whether AKT1 gene overexpression can block stem cell apoptosis under hypoxic condition following pig autologous bone marrow mesenchymal stem cell (BMSC) transplantation, and the effect of repairing damaged myocardium. DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Soochow University from August 2005 to February 2007.MATERIALS: A total of 24 healthy male Meishan pigs were supplied by the Animal Experimental Center of Soochow University. METHODS: The CDS (regulation domin of AKT1) AKT1-cDNA fragment was amplified. Lentivector Packaging Kit was used to transfect BMSCs after synthesized with pCDH1-AKT1 shuttling plasmid. Following BrdU labeling, models of myocardial infarction were constructed by occluding the distal left anterior descending coronary artery in pigs with gelatin sponge. 4 weeks later, pigs were randomly divided into four groups: the model control group, the DMEM group, the BMSCs group, and the AKT-transfected group. In model control group, there was no other injection after occluding the left anterior descending coronary artery. In the DMEM group, 5 mL DMEM was injected into the coronary artery. 5 mL BMSCs (1×10~7 cells) were infused into the coronary artery in the BMSCs group. 5 mL BMSCs transfected with the AKT1 gene were injected in the AKT-transfected group MAIN OUTCOME MEASURES: Western blot analysis and real time RT-PCR were used to test the plasmid. The cardiac function was evaluated by magnetic resonance image. Histological characteristics of the myocardium were observed using immunohistochemistry. Serum vascular endothelial growth factor and transforming growth factor β1 levels were determined by ELISA. RESULTS: AKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed in comparison with the published one. AKT mRNA expression could be detected distinctly 24 and 48 hours after transfecting cells. The expression of AKT1 intensity in MSCs remained strong 2 weeks later with detected by real time RT-PCR and Western blot analysis. AKT1-mRNA transcriptional levels were 120 times of primary cells. Before the cell implantation, the left ventricular end-diastolic dimension increased and the stroke volume decreased in the myocardial infarction hearts. The cardiac function was significantly improved after cell implantation, and the implanted MSCs prevented the infarct region from thinning and expanding, improved contraction and increased perfusion in all groups relative to the control hearts. The left ventricular chamber size was smaller in the hearts with being transplanted cells than that in the control hearts. Moreover, the improvement was even markedly greater in AKT-transfected group (P < 0.05). Hematoxylin-eosin staining results showed that fibering was significant in the model control group and DMEM group. Island-like myocardium was observed in the infarct zone of the BMSCs group and AKT-transfected group, and plenty of small vessels-shape structure was detected in the AKT-transfected group. Immunohistochemistry demonstrated that Von Willebrand Factor (vWF) and Cx-43 expression was determined in the myocardium in the BMSCs group and AKT-transfected group, and the proportion of BrdU and Cx-43-positive cells to BrdU-positive cells was significantly greater in the AKT-transfected group compared with the BMSCs group 4 weeks following transplantation (P < 0.05). Following cell transplantation, vascular endothelial growth factor levels were gradually increased, peaked at 1 week, gradually decreased, and reached a normal level at 4 weeks. Transforming growth factor p1 levels were gradually reduced, and significantly less than the model control group, DMEM group 4 weeks later (P < 0.05), and significantly lower than that pretransplantation (P < 0.05).CONCLUSION: Using lentiviral vector to construct with AKT1 gene could stably make BMSCs overexpress AKT1. The BMSCs engraftment in host myocardium might improve the left ventricle function by attenuating the contractile dysfunction and pathologic thinning in this model of left ventricular wall infarction. AKT1 overexpression can significantly improve cardiac function following infarction.
6.Fluoroscopic image capturing and DICOM storage application in the cardio-catheter room.
Ming YAO ; Bao-Hua WANG ; Zhong-Bing GONG ; Hai-Dong SHEN ; You-Li YE
Chinese Journal of Medical Instrumentation 2005;29(2):104-108
This paper analyses the data structure of DICOM standard by the applications in the Non-DICOM format fluoroscopic images converted into the DICOM format images. It puts forward a solution to integrate the Multi-Channel Electrophysiology Recorder System with the X -ray system in the cardio-catheter room.
Computer Communication Networks
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Diagnostic Imaging
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instrumentation
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methods
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Electrophysiologic Techniques, Cardiac
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instrumentation
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Fluoroscopy
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instrumentation
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Humans
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Image Processing, Computer-Assisted
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instrumentation
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methods
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standards
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Information Storage and Retrieval
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Radiology Information Systems
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standards
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Software Design
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Video Recording
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instrumentation
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methods
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standards
7.Conditioned medium of bone marrow mesenchymal stem cells via intravenous injection to treat cerebral ischemia-reperfusion injury
Feng CHEN ; Bin LIU ; Yuanyuan MING ; Suqin ZHOU ; Xia SHEN ; Fang HUA ; Guiyun CUI ; Xuanye YUE ; Kun ZAN ; Xinchun YE
Chinese Journal of Tissue Engineering Research 2015;(28):4544-4548
BACKGROUND:Large numbers of experimental data have confirmed that bone marrow mesenchymal stem cel s have a positive therapeutic effect on cerebral ischemia-reperfusion injury, but there are few reports about intravenous administration of bone marrow mesenchymal stem cel conditioned medium in the treatment of stroke.
OBJECTIVE:To investigate the effects of the conditioned medium of rat bone marrow mesenchymal stem cel s on the recovery of neurological function in rats after cerebral ischemia-reperfusion injury.
METHODS:The bone marrow mesenchymal stem cel s were isolated from rat bone marrow. When cel s at passage 2 or 3 reached 90%confluence, the original culture medium was removed. Then the cel s were cultured in serum-free DMEM for 18 hours. After that, the culture solution was col ected as the conditioned medium of rat bone marrow mesenchymal stem cel s. Adult rats were subjected to 2 hours of right middle cerebral artery occlusion. Ischemia-reperfusion injury rats were randomly assigned to three groups:control group, simple culture medium group and conditioned medium group, and respectively given injection of normal saline, DMEM, conditioned medium (10 mL/kg) via the tail vein at 2, 24, 48 hours after operation.
RESULTS AND CONCLUSION:There was no difference in the behavioral tests among the three groups at postoperative 2 hours (P>0.05). Compared with the control and simple culture medium group, neurological impairment was significantly improved in the conditioned medium group at postoperative 1, 3, 5 days (P<0.05), but there was no significant difference between the control and simple culture medium groups. At postoperative 5 days, brain edema was significantly eased in the conditioned medium group in comparison to the control and simple culture medium groups (P<0.05), and there was also no difference between the latter two groups (P>0.05). These results suggest that rat bone marrow mesenchymal stem cel s-conditioned medium via intravenous administration can significantly ease brain edema and improve the neurologic function after cerebral ischemia-reperfusion injury.
8.Utility of 64-MSCT in assessing acute non-reperfused myocardial infarct size
Xinkai QU ; Weiyi FANG ; Jianding YE ; Shaofeng GUAN ; Ruogu LI ; Yingjia XU ; Yan SHEN ; Min ZHANG ; Hua LIU
Journal of Geriatric Cardiology 2013;(3):247-252
Objective To evaluate the utility of multi-slice computed tomography (MSCT) in assessing acute non-reperfused myocardial infarct size. Methods Seven domestic pigs (mean weight 17.3 ± 1.9 kg) underwent ligation of the distal left anterior descending artery to establish a model of acute myocardial infarction (MI). MSCT and triphenyltetrazolium chloride (TTC) staining were performed two hours later. The following data were acquired and analyzed:MI volume (%), CT values of the infarcted region, left ventricular cavity and normal cardiac tissue at various scanning time-points (1, 5, 10, 15, 20 min after contrast injection). Results Using MSCT, the overall MI volume showed a time-dependent decrease, with a reduction of 28.87%after 20 min. The greatest reduction occurred at the 5 min time-point. In TTC staining, MI volume was 9.87%± 2.44%. When MI size, as determined by MSCT, was compared with that by TTC staining in Bland-Altman plots, there was a better agreement at 5, 10, and 15 min time-points at 1 and 20 min. Conclusions The study indicates that double-phase scanning examination using MSCT is a useful tool to assess MI size, and the optimal late-phase scanning time-point set within 5-15 min of contrast injection.
9.Optimizing expression of recombinant jasmonate ZIM-domain protein from Salvia miltiorrhiza.
Li-Hua ZHANG ; Wen-Yan WU ; Lu-Qi HUANG ; Ye SHEN
China Journal of Chinese Materia Medica 2012;37(24):3712-3716
OBJECTIVEAccumulation of tanshinton in Salvia miltiorrhiza are enhanced by exogenous application of jasmonates. The core JA signaling module COI1/JAZ/MYC2 play a central role on control of downstream gene expression in the JA pathway. To obtained the antibody of SmJAZ, SmJAZ recombinant protein was expressed in Escherichia coli and optimal expression was performed.
METHODThe full-length SmJAZ1 ORF was sub-cloned in a prokaryotic expression vector pET32a. The recombinant fusion protein had high expression level in BL21 (DE3) strain of E. coli, and SDS-PAGE analysis showed its molecular weight was about 24 kDa.
RESULTThe induction of E. coli [pET32-JAZ1] in different temperature, induction time, IPTG concentrations and IPTG adding time of E. coli were performed. The induction time and the induction temperature are positively related trends with SmJAZ1 protein expression, and IPTG concentration had no significant impact in protein expression, whereas IPTG adding time had significant impact on protein expression.
CONCLUSIONShaking the culture at 30 degrees C until the A600 is approximately 0.9 (2 h in LB), and add IPTG to a final concentration of 0.1 mmol x L(-1), and then the optimal expression of SmJAZ1 recombinant protein were accumulated after the induction time of 20 h.
Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Plant Proteins ; genetics ; metabolism ; Plasmids ; genetics ; metabolism ; Recombinant Proteins ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Salvia miltiorrhiza ; genetics ; metabolism ; Time Factors