1.In vitro magnetic resonance imaging of porcine bone marrow mesenchymal stem cells labeled by various kinds of superparamagnetic iron oxide nanoparticles
Yonghao LIU ; Liang GUO ; Jianhua CHEN
Chinese Journal of Tissue Engineering Research 2009;13(36):7113-7118
BACKGROUND: Magnetic labeling of stem cells is a recently developed stem cell in vitro labeling technique. Through in conjunction with magnetic resonance imaging (MRI), it can monitor transplanted stem cells in vivo. OBJECTIVE: To identify the method of superparamagnetic iron oxide (SPIO) labeling pig bone marrow mesenchymal stem cells (BMSCs), to investigate the characteristics of stem cells labeled by various SPIO following MRI, and to determine the minimum amount of labeled cells fer MRI. DESIGN, TIME AND SETTING: A control observation was performed at the laboratories of Department of Cardiovascular Surgery, Medical College of Soochow University, and Medical Imaging Centre, First Affiliated Hospital, Soochow University between September 2006 and March 2007. MATERIALS: Fresh porcine iliac bone marrow was collected from Taihu Meishan pigs. SPIO nanometer particles were purchased from Schedng, Germany. Ultramicro SPIO (USPIO) nanoparticles were provided by School of Chemistry and Chemical Engineering, Soochow University. For such particles, crystal nucleus surface was coated with dextran, and following coating, they were named 1#, 2#, 3# for short according to particle size. METHODS: Following isolation, purification, and culture, BMSCs were in vitro labeled by various kinds of SPIO nanoparticles. The labeled cells were subjected to Prussian blue staining and fluorescence microscope observation. The cell growth was observed using MTT method and the growth curve was plotted. For Feridex-labeled cells, 1×106), 5×105, and 1×105 three cell amount groups were set, for unlabeled cells, a 5×105 cell amount group was included, and for 1#, 2#, and 3# SPIO-labeled cells, only 5×105 cell amount group was used. MRI was conduced for measurement of signal intensity of cells labeled by different scanning sequences, followed by statistical analysis. MAIN OUTCOME MEASURES: Detection of SPIO nandparticles-labeled cells by Prussian blue staining; Growth curves of SPIO nanoparticles-labeled cells; Detection of cellular apoptosis by double staining; Determination of signal intensity of cell masses from different Ependoff tubes using MRI with T1WI, T2WI, and fast field echo (FFE) sequences. RESULTS: BMSCs could be labeled with SPIO and the labeling efficiency was 100%. Different amounts of blue-stained Fe particles could be observed in the cytoplasm by Prussian blue staining. SPIO labeling caused a significantly lower signal attenuation effect in T2WI and FFE (T2*WI) images than in T1WI images. In a labeling concentration of 25 mg/L Fe solution, the minimum cell amount for MRI was 1 x 105. The signal intensity exhibited significant difference in 2#, 3#USPIO- and Feridex-labeled cells in no matter T2WI or T2*Wl images(P < 0.01). But no significant difference was found between 1#USPIO- and Feridex-labeled cellss in no mater T2WI or T2*WI images(P > 0.05). There was significant difference in signal intensity of Feridex-labeled BMSCs between T2WI, T2*WI and T1Wl images (P < 0.01). CONCLUSION: BMSCs can be easily and efficiently labeled by SPIO nanoparticles without interference, at proper concentrations on cell viability and proliferation. MRI visualization of SPIO-labeled BMSCs is feasible in both T2WI and T2*WI images.
2.Radiological features of extraskeletai mesenchymal chondrosarcoma
Yinyin CHEN ; Ximing WANG ; Lingchuan GUO ; Yonggang LI ; Yonghao LIU ; Liang GUO
Chinese Journal of Radiology 2012;46(3):248-251
Objective To explore CT,MRI findings of extraskeletal mesenchymal chondrosarcoma (EMC).Methods Imaging information of all 8 cases of EMC verified by pathology were retrospectively analyzed.Results The location of lesions included lower extremity in 4 cases,forearm in 1 case,trunk in 2 cases and right lung in 1 case.The CT examination was performed in 7 cases,and 5 cases contained different patterns of ring-and-arc,granular,clump or irregular streaky mineralization.Dense calcification was detected in 3 cases,and focal in 2 cases.The nonmineralized component had slightly lower attenuation on CT scans than adjacent muscle.Four cases of peripheral located EMC demonstrated isointense on T1 WI,and mixed signal intensity on T2WI.For the cases of fine and dense calcification in 2 cases,numbers of dot-like low-intensity signals were detected resembling “pepper sign”; while for the cases of focal mineralization in 1 case,the low intensity area was located centrally in the high intensity area.Heterogeneous enhancement was found both in the calcified and uncalcified areas.One case of central located tumor exhibited low and high intensity on T1 and T2 weighted images,and nodual enhancement was observed.Conclusion EMC has several characteristic imaging features,including various mineralization pattern,enhancement of calcified area and signal intensity,which might have diagnostic value for this rare tumor.
3.MRI study of transplanting autologous bone marrow mesenchymal stem cells at different time points on cardiac function amelioration following myocardial infarction
Yonghao LIU ; Liang GUO ; Jianhua CHEN ; Shiqiang GUO ; Guiping YU ; Zhenya SHEN
Chinese Journal of Tissue Engineering Research 2009;13(49):9625-9630
OBJECTIVE: To study the effect of transplanting autologous bone marrow mesenchymal stem cells (BMSCs) at different time point after myocardial infarction on cardiac function, and to approach its mechanism. METHODS: Thirty healthy Taihu Meishan swine were prepared for myocardial infarction models, and divided into 6 experimental groups, with 5 animals in each group. BMSCs were transplanted into 3 groups through coronary artery at 3 hours, 2 weeks and 4 weeks after myocardial infarction, named G1, G2 and G4, respectively. Meantime, DMEM culture medium was injected in the control group at correspond periods. Each swine was examined by MRI and Doppler before infarction, before transplantation, and at 8 weeks after infarction, respectively, to observe the change of cardiac function. The VEGF values of blood serum in different periods after transplantation were detected. All swine hearts were harvested after 8 weeks (the experimental terminus), and the planting and differentiation of transplanted cells in cardiac muscle were detected by the method of immunity histochemistry. The density of blood vessels in cardiac muscle was acquired simultaneously. RESULTS: There was no statistic difference of cardiac function between G1 and its control groups. The groups of G2 and G4 could improve cardiac function compared to the control groups, and G4 was superior to G2 (P < 0.05). There was no statistics difference of the decreased absolute value of myocardial infarcted area between G1 and the control groups. The myocardial infarcted area of G4 was greater than G2 (P < 0.05). The value of blood serum VEGF rose obviously in the G2 and G4, while G1 and all control groups did not present any marked changes, the rising amplitude of G4 was larger than G2 (P < 0.05). There were not any planting and differentiation of transplanted stem cells in G1 and all control groups at 8 weeks after infarction, but G2 and G4 could display, especially in G4 group (P < 0.05). There was no statistic difference of the density of blood vessels in cardiac muscle between G1 and all control groups at 8 weeks after infarction, but the differences were significant in all experimental groups, which was superior in G4 group to G1 and G2 groups (P < 0.05).CONCLUSION: There is disparity of transplanting BMSCs at different time point after myocardial infarction on cardiac function. Transplantation in acute period of myocardial infarction has no significant effect, but transplantation in non-acute period can ameliorate cardiac function. The therapeutic effect of transplanted at 4 weeks is superior to other time point. The MRI can display the location and compass of infarct cardiac muscle, and reflect the variation of cardiac function.
4.Synthesis and activity of some new histone deacetylases inhibitors
Yonghao CHENG ; Yanshen GUO ; Haizhu HAN ; Nan WANG ; Guohong ZHANG ; Zongru GUO ; Song WU
Acta Pharmaceutica Sinica 2010;45(6):735-41
To explore novel histone deacetylase (HDAC) inhibitors with anti-tumor activity, twelve target compounds were synthesized, and their structures were confirmed by 1H NMR, MS and elemental analyses. Evaluation results in vitro showed that compound Ia exhibited potent inhibition against HDAC and is worth for further investigation. And compounds IIa, IIb, IIIa-IIIi possessed moderate HDAC inhibitory activity.
5.Further study of sonographic examination skills and classifications of the inferior vena cava lesions in patients with Budd-Chiari syndrome
Yonghao GAI ; Shuang MA ; Wenbin GUO ; Bo LIANG ; Tao JIA ; Suzhen ZHANG ; Jianzhong ZHAO
Chinese Journal of Ultrasonography 2012;(11):965-968
Objective To explore a compatible approach to detect and classify the lesions of inferior vena cavas (IVCs) on sonogram in patients with Budd-Chiari syndrome(BCS).Methods Ultrasonogram of the IVCs were observed detailedly in 300 patients with BCS by using trans-abdomen and trans-thorax-right atrium-inferior vena cava ingress sections.Transducers usually used for heart examination were applied in the latter.Lesions of the IVCs found in 277 out of 300 patients were classified.All lesions were confirmed by digital subtraction angiography (DSA) and among them,52 cases underwent computed tomography angiography (CTA).Results Lesions of IVCs were classified into 3 categories as follows:membranous type,segmental type,and ex-pressed type.① Membranous type (thickness ≤ 15 mm) included membranous stenosis type and membranous occlusion type.On the basis of the thickness,the membranous stenosis type was further classified into thinner membranous stenosis type (thickness ≤5mm) and thicker membranous stenosis type (5 mm<thickness≤ 15 mm).The membranous occlusion type was further classified into thinner membranous occlusion type (thickness ≤5 mm) and thicker membranous occlusion type (5 mm<thickness ≤15 mm).② Segmental type (lengtb > 15 mm) was consist of segmental stenosis type and segmental occlusion type.Based on the length of the lesion,the segmental stenosis type was further divided into longer segmental stenosis type (length > 30 mm) and shorter segmental stenosis type (15 mm<length ≤30 mm).The segmental occlusion type was further divided into longer segmental occlusion type (length > 20mm) and shorter segmental occlusion type (15 mm< length ≤20 mm).③ Ex-pressed type of IVCs was mainly caused by compression of intumescent caudate lobes.Corresponding sonographic features were demonstrated in each type.Conclusions Ultrasonogram of trans-abdomen and trans-thorax-right atrium-inferior vena cava ingress sections could accurately classify the lesions of IVCs.It is of important significance for the clinical treatment.
6.A case of cutaneous phaeohyphomycosis caused by Phialophora verrucosa
Yonghao XU ; Chunyang LI ; Jie ZHAO ; Keyu WANG ; Shulan GUO ; Ying SU ; Jian JIAO ; Qinfeng HU
Chinese Journal of Dermatology 2011;44(11):809-811
A 16-year-old male presented with a 11-year history of progressively enlarging erythema and crusting on the right cheek.Physical examination revealed an irregularly shaped,sharply marginated,dark erythematous patch sized 6 cm x 10 cm and plaques with mild verrucous proliferation.There were strip-like scar at the margin of lesions and multiple ulcers measuring 0.5 to 1 cm in diameter with firm crusts.No small jellycolored nodules were observed.Direct microscopy of multiple scrapings under the crusts showed many light brown,septate,branching and irregular hyphae.Olivaceous-black woolly colonies grew at 25 C and 35 C on Sabouraud's dextrose agar and potato dextrose agar; flask-shaped conidiogenous cells with funnel-shaped collarettes and ellipsoidal conidia arranged in flower-like shape were observed microscopically.PAS staining showed numerous septate and branching hyphae,pseudohyphae and yeast-like cells.There was a 99.73% similarity in the species-specific rDNA sequence between the isolate and phialophora verrucosa standard strain CDC-B2152.The patient was diagnosed with cutaneous phaeohyphomycosis caused by Phialophora verrucosa.The lesion subsided after treatment with amphotericin B and itraconazole,but recurred after drug withdrawal.Itraconazole and terbinafine were administered for the retreatment of this patient.
7.Effect of Toll-like receptor 7 agonist on JNK signaling pathway during liver injury in septic mice
Suqian GUO ; Yu ZHANG ; Qi ZHAO ; Lan WU ; Yonghao YU ; Guolin WANG
Chinese Journal of Anesthesiology 2016;36(5):613-615
Objective To evaluate the effect of Toll-like receptor 7 (TLR7) agonist on c-Jun Nterminal kinase (JNK) signaling pathway during liver injury in the septic mice.Methods One hundred eighty pathogen-free adult male C57BL/6 mice,aged 10-14 weeks,weighing 20-26 g,were randomly divided into 3 groups (n=60 each) using a random number table:sham operation group (group S);sepsis group (group Sep);TLR7 agonist group (group GDQ).Sepsis was induced by cecum ligation and puncture.In group GDQ,TLR7 agonist 1.5 g/kg was injected intraperitoneally at 24 h before establishment of the model.At 6,12 and 24 h after operation,10 mice in each group were sacrificed,and the livers were removed to detect the expression of interleukin-6 (IL-6) and IL-10 by Western blot.The expression of JNK was determined by immuno-histochemistry,and the histopathologic changes of livers were examined with a light microscope at 24 h after operation.The survival of mice was observed at 14 days after operation,and the 14-day survival rates were calculated.Results Compared with group S,the 14-day survival rates were significantly decreased,and the expression of IL-6,IL-10 and JNK was significantly up-regulated at 6,12 and 24 h after operation in Sep and GDQ groups (P<0.05).Compared with group Sep,the 14-day survival rates were significantly increased,and the expression of IL-6,IL-10 and JNK was significantly down-regulated at 6,12 and 24 h after operation in group GDQ (P<0.05).The pathological changes of livers were significantly attenuated in group GDQ as compared with group Sep.Conclusion TLR7 agonist can reduce the liver injury through blocking the JNK signaling pathway in the septic mice.
8.Application of MR 3D FIESTA Technology in Spinal Malformation
Jinfeng LI ; Yonghao WANG ; Hongmei GUO ; Zhiwei FAN ; Na YANG ; Si CHEN
Chinese Medical Equipment Journal 2003;0(10):-
Objective To study the value of MR 3D FIESTA technology in spinal malformation. Materials and Methods 70 patients with spinal malformation and GE 1.5T superconducting MR machine got involved in. The scanning sequences included FSET2WI scanning at 2D axial position, coronal position and sagital positon, and FIESTA scanning at 3D coronal position and sagital position. Results FIESTA scanning could be used in the achievement of multi-slice and multi-angle coronal, sagital and axial images. Conclusion 3D MR FIESTA can be applied to rapid, multi-angle, multi-slice and continuous display of the spinal cord.
9.Effect of dexmedetomidine on necroptosis during liver injury in septic rats
Yu ZHANG ; Yu MENG ; Suqian GUO ; Lan WU ; Qi ZHAO ; Chengcheng SONG ; Yonghao YU ; Guolin WANG
Chinese Journal of Anesthesiology 2017;37(7):888-890
Objective To evaluate the effect of dexmedetomidine on necroptosis during liver injury in septic rats.Methods Eighteen SPF adult male Sprague-Dawley rats,weighing 200-220 g,were divided into 3 groups (n=6 each) using a random number table:sham operation group (group SH),sepsis group (group SEP) and dexmedetomidine group (group DEX).Sepsis was induced by cecal ligation and puncture in chloral hydrate-anesthetized rats in SEP and DEX groups.Dexmedetomidine 5 μg/kg was injected via the caudal vein at 1 h before operation in group DEX.Blood samples were collected from the caudal vein at 6 h after operation for determination of serum aspartate amino-transferase (AST) and alanine aminotransferase (ALT) concentrations.The rats were then sacrificed and livers were removed for determination of the level of reactive oxygen species (ROS) in liver tissues (using chemiluminescence assay) and expression of receptor-interacting protein 1 (RIP1),RIP3,mixed lineage kinase domain-like (MLKL),high-mobility group box 1 protein (HMGB1) and dynamin-related protein 1 (Drpl) in liver tissues (by Western blot).Results Compared with group SH,the serum AST and ALT concentrations were significantly increased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drpl in liver tissues was up-regulated,and the level of ROS in liver tissues was increased in SEP and DEX groups (P<0.05).Compared with group SEP,the serum AST and ALT concentrations were significantly decreased,the expression of RIP1,RIP3,MLKL,HMGB1 and Drp1 in liver tissues was down-regulated,and the level of ROS in liver tissues was decreased in group DEX (P<0.05).Conclusion The mechanism by which dexmedetomidine attenuates liver injury may be related to inhibition of necroptosis in septic rats.
10.Role of necroptosis in liver injury in septic rats
Yu ZHANG ; Yu MENG ; Suqian GUO ; Lan WU ; Qi ZHAO ; Yonghao YU ; Guolin WANG
Chinese Journal of Anesthesiology 2017;37(6):748-750
Objective To evaluate the role of necroptosis in liver injury in septic rats.Methods Twenty-four SPF healthy male adult Sprague-Dawley rats,weighing 200-220 g,aged 6-8 weeks,were divided into 3 groups (n=8 each) using a random number table:sham operation group (Sh group),sepsis group (Sep group) and specific necroptosis inhibitor necrostatin-1 group (N group).Sepsis was induced by cecal ligation and puncture in anesthetized rats in N and Sep groups.Necrostatin-1 1.0 mg/kg was intravenously injected at 1 h before operation in group N,while the equal volume of dimethyl sulfoxide was given instead in group Sep.Rats were sacrificed at 6 h after operation,and livers were removed for examination of the pathological changes (with a light microscope) and for determination of the level of reactive oxygen species (ROS) in liver tissues (by using chemiluminescence assay) and expression of receptor-interacting protein kinase-1 (RIPK1),RIPK3,mixed lineage kinase domain-like (MLKL) and high-mobility group box 1 protein (HMGB1) in liver tissues (using Western blot).Results Compared with Sh group,the ROS level in liver tissues was significantly increased,and the expression of RIPK1,RIPK3,MLKL and HMGB1 in liver tissues was up-regulated in Sep and N groups (P<0.05).Compared with Sep group,the ROS level in liver tissues was significantly decreased,and the expression of RIPK1,RIPK3,MLKL and HMGB 1 in liver tissues was down-regulated in group N (P<0.05).The pathological changes of liver tissues were significantly attenuated in group N when compared with group Sep.Conclusion Neeroptosis is involved in liver injury in septic rats.