1.Application of induced pluripotent stem cells
Rui WANG ; Zhengjie HUANG ; Qi LUO
International Journal of Surgery 2010;37(6):414-416
The research of induced phripotent stem(ips)cells is a hotspot in the area of life sciences at present.Currently,the research of ips cells has focused on cells'induction methods,source and differentiation.At the same time,the research of ips cells'application has also made some achievements.With the extensive research of application,ips cells will have an inestimable effect on the future of biomedical development.This article will give an reviews the latest application and future research trends of ips cells.
2.Nitric oxide production, NOS activity and expression in pulmonary arterioles of rats with chronic hypoxic hepercapnic pulmonary hypertension
Zhengjie XU ; Liangxing WANG ; Qunji WANG ; Hongqi ZHANG
Chinese Journal of Pathophysiology 2000;0(12):-
AIM: To clarify the role of nitric oxide (NO) system in development of chronic hypoxic hypercapnic pulmonary hepertension. METHODS: Male Sprague-Dawley rats were randomly divided into control group and hypoxic hypercapnic group. NO content of plasma was determined, constitutive nitric oxide synthase (cNOS) and inducible nitric oxide synthase (iNOS) were examined using the technique of immunohistochemistry, expression of cNOS mRNA and iNOS mRNA of arteriole were detected by in situ hybridization. RESULTS: Plasma NO concentration, cNOS activity and cNOS mRNA expression in arteriole of chronic hypoxic hypecapnic group were significantly lower than that of control group ( P
3.Influence of injection carthamus tinctorius D on the expression of ICAM-1 in lung injury induced by ischemia- reperfusion in rabbits
Xiaolong ZHANG ; Fangyan WANG ; Zhengjie XU ; Wantie WANG ; Maolin HAO
Chinese Journal of Emergency Medicine 2008;17(6):610-613
Objective To investigate the influence of injection carthamus tinctorius D. (1C) on the expression of intercellular adhesion molecule-1(ICAM-1) during the ischemia-reperfusion injury of lung (LIRI) in rabbits and its potential mechanism. Method Single lung ischemia-reperfusion animal model was induced in rabbits. A total of 30 Japanese white rabbits were randomly divided into sham-operation group (S group, n =10), ischemia-reperfusion group (I/R group, re = 10) and ischemia-reperfusion plus 1C group (1C group, n = 10) .The rabbits of 1C group received 1C 2.0 ml/kg injected intravenously just at 20 min before ligation of artery involved and the same dose of 1C instantly at the initiation of reperfusion. Malondialdehyde (MDA) , superoxide dismutase (SOD) and xanthine oxidase(XO) in serum were measured. The lung tissue was sampled and assayed wet/dry weight ratio (W/D), contents of myeloperoxidase (MPO) at the end of the experiment, and ultrastructure changes were observed under electron microscope. The expression of ICAM-1 was measured by using immunohistochemistry(IHC) . snd one-way ANOVA was used for statistical analysis. Results In I/R group, serum XO and MDA increased and SOD decreased, whereas the same pattern of changes but less magnitude happened in 1C group ( P < 0.01). The values of W/D and MPO were much higher in I/R group, but lower in 1C group. Under electron microscope, the ultrastructure of lung tissue showed pathological changes in the rabbits of I/R group,and these changes were greatly attenuated in the rabbits of 1C group . The IHC showed that ICAM - 1 in lung tissue of I / R group was (2.94±0.48) which was significantly higher than that of 1C group(1.75 (P < 0.01). Conclusions Injection Carthamus tinctorius D. may meliorate the ischemia-reperfusion injury of lung by way of suppressing the expression of ICAM-1, inhibiting neutrophil aggregation, lowering oxygen free radical level and decreasing lipid peroxidation.
4.Study of MK promoting breast cancer cell MDA-MB-231 angiogenesis through EPCR/PAR1 pathway in vitro
Qingling WANG ; Dongmei LIU ; Zhengjie HAN ; Yongping WU
Chinese Journal of Clinical and Experimental Pathology 2015;(9):961-965
Purpose To observe the effects of midkine ( MK) on human breast cancer cell line MDA-MB-231 angiogenesis in vitro, and to explore its mechanism. Method shRNA interference was performed to silence the expression of MK in MDA-MB-231 cells, and Western blot was used to identify the expression of MK and EPCR. After MK and EPCR knockdown, or treated with anti protease-activated receptor 1 (PAR1) antibody, the culture medium of MDA-MB-231 cells were collected and the conditioned medium were pre-pared. Human umbilical vein endothelial cells ( HUVECs) were cultured with conditioned medium, and the endothelial cells prolifera-tion was detected by CCK-8 assay, cell migration was detected by transwell method, vasculogenic activity was assessed by Matrigel-based tube formation assay. Results After knockdown of MK, the protein level of EPCR was decreased in MDA-MB-231 cells. Com-pared with control, knockdown of MK and EPCR decreased the proliferation, migration and angiogenesis ability of HUVECs significant-ly (P<0. 05), and the effect of EPCR knockdown group was stronger than MK knockdown group (P<0. 05). After treated with anti-PAR1 antibody, the proliferation, migration and angiogenesis ability of HUVECs were decreased compared with control and EPCR knockdown group (P<0. 05). Conclusion MK promotes human breast cancer cell line MDA-MB-231 angiogenesis through EPCR /PAR1 signaling pathway in vitro.
5.Experimental study of (RGD)_3-tTF fusion proteins binding specifically to tumor vasculature in the colonic carcinoma nude mice model
Zhengjie HUANG ; Qi LUO ; Jianghua YAN ; Shengyu WANG
China Oncology 2009;19(10):735-741
Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.
6.Clinical application of a new skin-stretching device
Zhengjie DANG ; Baoquan ZHANG ; Shoufeng WANG ; Xingang YANG ; Wei GAO
Chinese Journal of Tissue Engineering Research 2015;(43):7047-7052
BACKGROUND:Previously deep burn wound or skin defects are generaly repaired with skin grafting or flap of skin grafting. Obvious scar hyperplasia usualy appears after operation, which requires multiple surgeries. Meanwhile, patients have to suffer from great pain and bear high cost. OBJECTIVE: To observe the clinical effects on deep wounds by continuous traction of self-designed skin-stretching device (patent No. ZL 2012 2 0022443.7). METHODS: Thirty patients with deep burn wound, skin defect or funicular scar were enroled, including 22 males and 8 females, aged 18-49 years, and randomly divided into two groups. Skin-stretching device was adopted for skin traction treatment. Twenty cases underwent skin traction from 1 kg puling force to 5 kg, with an increase of 1 kg per 2 days, 6 hours a day for 10 days. Blood flow at the beginning, 1, 5, 10, 15, 20, 30, 60 minutes of the skin traction, and the changes of wound edge skin as wel as histological changes of the skin were observed. Of the remaining 10 cases, 2, 6, and 2 cases underwent skin traction of 2, 4, 7 kg, respectively. Blood flow and skin changes were also observed to find out the most suitable and safe force. RESULTS AND CONLUSION:Al the 30 cases achieved primary healing without necrosis of skin, infection or peripheral circulatory disorders, and the appearance and function recovered wel. The healing time was 8-24 days. The skin-stretching device was most safe under 4 kg puling force, by which, there was neither blood circulation obstacle nor tear of skin. After traction, the skin blood flow and the number of cels increased, especialy the epithelial basal cels. The colagen fibers became thicker and denser, and the elastic fibers regenerated significantly; the fibroblasts and capillary density increased. It has been proved that we can better close the wound and reduce scar formation effectively with the self-designed skin-stretching device for skin traction.
7.Effects of panax notoginseng saponins on pneumocyte apoptosis and Fas/FasL expression in rabbits with lung ischemia/reperfusion injury
Zhengjie XU ; Shirong NI ; Wantie WANG ; Weibin ZHOU ; Xiwen CHEN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM: AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and observe effects of panax notoginseng saponins (PNS) on apoptosis in lung ischemia/reperfusion injury. METHODS: Single lung in situ ischemia/reperfusion animal model was used. Eighty four Japanese white rabbits were randomly divided into control group (control), ischemia/reperfusion 1 h group (IR1h), IR3h, IR5h, Panax Notoginseng Saponins 1 h group (PNS1h), PNS3h and PNS5h. TUNEL, immunocytochemistry and in situ hybridization techniques were used to observe apoptosis and Fas/FasL expression in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues were significantly high, Fas/FasL mRNA and its protein were up-regulated in lung tissues of lung ischemia/reperfusion injury compared with control (all of P
8.Effects of safflor injection on expression of cyclooxygenase-2 mRNA during lung ischemia/reperfusion injury in rabbits
Maoping CHU ; Xiaolong ZHANG ; Wantie WANG ; Zhengjie XU
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To investigate the effects of safflor injection(SI) on expression of cyclooxygenase-2 mRNA during lung ischemia/reperfusion injury(PIRI) in rabbits.METHODS: Rabbit lung model of ischemia/reperfusion injury was constructed in vivo.The rabbits were randomly divided into three groups: sham-operation group(group S),ischemia-reperfusion group(group I/R) and ischemia/reperfusion plus safflor injection group(group SI).The lung tissue sampled at the end of the experiment was assayed for wet/dry weight ratio(W/D),injured alveoli rate(IAR) and observed ultrastructure changes under electron microscope.The expressions of COX-1 and COX-2 were measured by immunohistochemistry(IHC).The expression of COX-1 mRNA and COX-2 mRNA were observed by in situ hybridization(ISH).RESULTS: The value of W/D and IAR was much higher in I/R group,but decreased in SI group.Electron microscope showed obvious ultrastructure injury brought by PIRI in I/R group,which was greatly attenuated in SI group.The IHC and ISH demonstrated that COX-2 and COX-2 expressions in pulmonary tissue of I/R group were significantly higher than those in S group(P
9.Change of caspase-3 in rabbits after lung ischemia-reperfusion and the effect of puerarin(葛根素)
Yixiao XU ; Wantie WANG ; Zhengjie XU ; Xiaofeng JIN ; Maolin HAO
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(02):-
Objective To investigate the change of caspase-3 in rabbits after lung ischemia-reperfusion injury(LIRI) and the effect of puerarin(葛根素).Methods Thirty healthy rabbits used for unilateral lung ischemia-reperfusion model were randomly divided into 3 groups(each n=10): control group(C group),lung ischemia-reperfusion group(I/R group) and puerarin group.The activity of serum superoxide dismutase(SOD),the contents of serum malondialdehyde(MDA) and nitric oxide(NO),the wet to dry weight(W/D) ratio of lung tissue and the index of quantitative assessment of histological lung injury(IQA) were measured respectively in different groups;the pneumocyte apoptosis index(AI) was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL);caspase-3 protein and mRNA expression were studied by using in situ hybridization(ISH) and immunocytochemistry(IHC) techniques in the groups mentioned above.Results The activity of SOD and content of NO were significantly lower in I/R group than those in C group(both P
10.Polydatin downregulates TLR4 expression in lung ischemia reperfusion injury in rabbits
Xiaofeng JIN ; Zhengjie XU ; Wantie WANG ; Yixiao XU
Chinese Journal of Pathophysiology 1986;0(04):-
0.05) was observed. The protein expressions of TLR-4, NF-?B p65, HSP70 and ICAM-1mRNA in IR group were significantly increased as compared to C group and PD group, while those expressions in PD group were evidently higher than those in C group (all P