1.Isolation,characterization and application of umbilical cord mesenchymal stem cells in the treatment of liver diseases
International Journal of Biomedical Engineering 2009;32(3):190-193
Human umbilical cord (UC) has been attracting increasing research effort in recent years.Since UC is a postnatal organ discarded after birth,the collection of its cells is non-invasive and raises no ethical con-cerns.Human umbilical cord provides large amount of mesenchymal stem cells(MSCs) which hold unlimited ca-pability of proliferation and great potential of differentiation and. MSCs have stable biological properties and low-er immunogenicity and can function well after amplifications.Studies have demonstrated that MSCs have the po-tential of inhibiting inflammatory response, generation of fibrosis, apoptosis and stimulating regeneration of hepa-tocytes.Therefore, UCMSCs could be an ideal cell source for the therapy of liver diseases.
2.Progress of tumor immunotherapy research
International Journal of Biomedical Engineering 2013;36(3):180-183
Through a series of immune evasion mechanisms,tumor cells can evade immune attack and breed wantonly in human body.In recent years,with the rapid development of oncology,immunology and molecular biology and other related disciplines,immunology treatment method which can effectively kill tumor cells and have fewer side effects draws more and more attention,while the immunotherapy method is different in therapeutic evaluation from the general treatment of solid tumors due to its special feature.This article briefly introduces the latest progress of tumor immunotherapy.
3.Advances in human placenta-derived stem cells for liver regeneration
Jian WANG ; Zhengyan ZHU ; Zhi DU
Chinese Journal of Hepatobiliary Surgery 2013;(3):236-240
Human placenta-derived stem cells (hPD-SCs) are a mixed group of stem cells.Stem cell medicine has applications for organ damage or failure through regenerative,anti-apoptotic,anti-inflammatory and anti-tumor properties in addition to cell function recovery.Presently,human placenta mesenchymal stem cells (hPMSCs) have similar characteristics to the differentiation of hepatocyte-like cells by promoting hepatocyte regeneration,anti-hepatocyte apoptosis and anti-liver fibrosis,in vitro or in animal models.To further our investigation,a summary of the origin,sorting and biological properties of hPDSCs along with a narration of hPDSCs for liver disease therapy was written.This leads to a discussion for new ideas to further explore cell treatment for liver disease.
4.The effect of intravenous thrombolysis on TIMI flow of patients with acute myocardial infarction
Zhengyan ZHU ; Dayi HU ; Aihua ZHANG
Chinese Journal of Practical Internal Medicine 2006;0(16):-
Objective To observe the effect of intravenous thrombolysis on TIMI flow of acute myocardial infarction(AMI).Methods A total of 229 patients with AMI analysed retrospectively were divided into two groups:intravenous thrombolysis(IVT)group(n=131)and primary percutaneous coronary intervention(PCI)group(n=98).The treatment time and acute-phase outcomes were compared between the IVT group and the PCI group;104 patients in the IVT group received rescued or delayed PCI(IVT+PCI);TIMI flow was analysed between the IVT+PCI group and the PCI group.Results The time from the emergency room door to initiation of treatment was shorter in the IVT group than in the PCI group(67.79 min vs 134.54 min,P=0.000).At the initial coronary angiography,TIMI 3 flow and TIMI 2+3 flow were higher in the IVT+PCI group than in the PCI group(40.4% vs 24.5%,P=0.016;63.5% vs 36.7%,P=0.000 respectively).There was no significant difference between the IVT group and the PCI group in terms of major adverse cardiac events(MACE).Conclusion IVT as an initial treatment for AMI might achieve earlier reperfusion at TIMI≥2 flow,and it should be popularized in the primary hospitals.
5.Coronary angiographic data analysis in young male patients with type 2 diabetes mellitus complicated ;coronary heart disease
Aihua ZHANG ; Zhengyan ZHU ; Hongwei ZHANG ; Xiaojuan LIU
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(2):194-196
Objective:To explore the coronary angiographic (CAG)characteristics of young male patients With type 2 diabetes mellitus (T2DM)and coronary heart disease (CHD).Methods:Clinical data of 233 young male CHD inpa-tients,Who Were confirmed by CAG and admitted in Beijing Shunyi Hospital from Jan 2011 to Jan 2013,Were retro-spectively analyzed.According to suffering from T2DM or not,they Were divided into T2DM + CHD group (n=71)and pure CHD group (n=162),baseline data and CAG results Were compared betWeen tWo groups.Results:Compared With pure CHD patients,there Were significant rise in levels of total cholesterol [TC,(4.11 ± 0.26) mmol/L vs.(5.79±0.37)mmol/L],loW density lipoprotein cholesterol [LDL-C,(2.31±0.32)mmol/L vs.(3.91 ±0.45)mmol/L],triglyceride [TG,(1.60±0.25)mmol/L vs.(3.28±0.56)mmol/L],P<0.01 all;CAG indi-cated that there Were significant increase in percentages of multi-vessel coronary disease (15.4% vs.35.2%),seg-mental lesion (13.6% vs.35.2%)and diffused lesion (31.5% vs.57.7%),and significant reduction in percentage of collateral circulation (50.6% vs.29.6%)in T2DM + CHD patients,P<0.01 all.Conclusion:There are more multi-vessel lesions,diffused lesion and less coronary collateral circulation in young male patients With type 2 diabe-tes mellitus complicated coronary heart disease.
6.Growth activity of fetal liver stem cells in polyanhydride-three-dimensional vector-glucan
Meili YU ; Zhi DU ; Zhengyan ZHU ; Yingtang GAO ; Qiang GAO
Chinese Journal of Tissue Engineering Research 2009;13(47):9302-9304
OBJECTIVE: To observe effects of polyanhydride-three-dimensional vector-glucan material on the fetal liver stem cell adhesion and proliferation.METHODS: The two-step collagenase perfusion digestion and bliquid percoll discontinuous density gradient centrifugation was used to isolate fetal liver stem cells. Fetal liver stem cells at the third passage were incubated on the polyanhydride-three-dimensional vector-glucan material. Inverted microscope was utilized to observe cell adhesion and growth status. Cell adherent rate, proliferation activity were calculated, and cell number was counted. Cell-vector was obtained for tissue section. Using hematoxylin-eosin staining, cell growth in the vector was observed under the optical microscope. At 7 days,immunofluorescence staining and flow cytometry were used to determine marker expression.RESULTS: Polyanhydride-three-dimensional vector-glucan promoted grow and adhesion of liver stem cells. There was the active function of the liver stem cells within carrier materials. In the three-dimensional surface and the internal culture, liver stem cell proliferation was sustained. After 10 days, the polyanhydride common culture-three-dimensional vector-glucan on stem cells was non-toxic, and human fetal liver stem cells could be attached to the polyanhydride-three-dimensional vector-glucan stent. The cell proliferation was better and dynamic sustained expression of markers. 7-days training received 19.7 percent increase in the number of cells.CONCLUSION: Polyanhydride-three-dimensional vector-glucan promotes the proliferation of liver stem cells, and liver stem cells can be used as the vector in liver tissue engineering.
7.Isolation, culture, induction and differentiation of fetal bone marrow-derived postembryonic pluripotent stem cells
Zhengyan ZHU ; Jinjuan ZHANG ; Tao LI ; Zhi DU
Chinese Journal of Tissue Engineering Research 2009;13(1):192-196
BACKGROUND: At present, studies show that a kind of stem cell community which in many kinds of organizations can differentiate into tissue cells of different embryonic layers; but those are different from embryonic stem cells, embryonic stem cell will lose the part differentiation potential gradually during the development of pregnancy, and will present some special phenotypes or the molecular markers, as CD105 and so on, will cell it postembryonic pluripotent stem cells.OBJECTIVE: To study the isolation of postembryonic pluripotent stem cells from fetal bone marrow, proliferative culture in vitro, induction and differentiation; transplantation to the liver of SCID mice with hepatic failure, and detect therapy effects.DESIGN, TIME AND SETTING: Cell observation and animal randomization experiment which was completed in the Ministry of Health of Cell Engineering Technology Research Center, Tianjin Third Central Hospital from March 2003 to March 2005.MATERIALS: The postembryonic pluripotent stem cells were extracted from thighbone and shinbone of 22-week old fetuses under sterile circumstance. Adult female SCID mice ware regarded as the recipients. CD105 immunornagnetic beads were provided by Miltenyi Biotec, Germany; mouse-anti-human albumin by Sigma, USA; basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) by PEPROTECH, UK.METHODS: Postembryonic pluripotant stem cells obtained from fetal bone marrow were isolated using density gradient centrifugation and micromagnetic beads technique. The hepatocyte-like cells were induced and differentiated with culture media containing HGF (30 ng/mL) and bFGF (20 ng/mL). Twenty-four SCID mice were randomly divided into experimental group and control group with 12 mice in each group. Hepatic injury models were established with intraperitoneal injection of D-galactosamine On the next day, about 106 CD105(+) cells were perfused into liver in situ in the experimental group, and about 106 CD105(-) cells or isovolumic culture medium were perfused in the control group.MAIN OUTCOME MEASURES: Two, seven days, one and three months after the transplantation of cells, human albumin expression in the liver tissue was detected by immunohistochemistry.RESULTS: The immunocytochemical assay of the cells after micromagnetic beads selection showed that the CD105 expression was slightly positive; the doubling time of the cells in the logarithmic growth period was around 30 hours; after being expanded for 10 population doublings, the cells entered decline period. The cells were transplanted into SCID mice's liver; 3 months later, the human serum albumin in the mouse liver was assessed by using monoclonal antibody of mouse-anti-human serum albumin, dotted or small focal expression of the protein could be detected. However, any expression was not observed in the control group.CONCLUSION: The bone marrow-derived pluripotent stem cells are able to transform to hepatocytas in the hepatic microenvironment.
8.Predictive value of serum cTnI and NT-proBNP for patients with chronic stable heart failure
Yong ZHAO ; Qingqing CHEN ; Feng GUO ; Zhengyan ZHU
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(4):401-404
Objective:To explore the correlation among serum cardiac troponin I (cTnI),N terminal pro brain natri-uretic peptide (NT-proBNP)and main endpoint events of heart in patients with chronic stable heart failure.Meth-ods:The present study enrolled 95 patients with NYHA cardiac function class III~IV from Feb 2010 to Feb 2011.According to levels of cTnI and NT-proBNP,the patients were divided into cTnI negative group (n=60)and cTnI positive group (n=35);NT-proBNP negative group (n=40)and NT-proBNP positive group (n=55),all patients were followed up for two years,and the main endpoint events were cardiogenic sudden death and rehospitalization caused by acute aggravation of heart failure.Results:Compared with negative group,the hazard ratio (HR)of end-point events was 2.69 and confidence interval (CI)was 1.54~ 4.72,P = 0.002 in cTnI positive group;HR was 2.54 and CI was 1.35~4.78,P =0.003 in NT-proBNP positive group;further interclass crossover analysis found that,when patients'cTnI and NT-proBNP were both positive,the hazard ratio of cardiac endpoint events was the highest (HR=6.34,CI 2.26~17.9,P <0.001).Conclusion:In patients with chronic stable heart failure,serum elevated levels of cardiac troponin I and N terminal pro brain natriuretic peptide are important predictors reflecting prognosis of patients with heart failure.
9.Effect of human umbilical cord mesenchymal stem cell-secretion on proliferation and apoptosis in hepatocytes
Jiao LI ; Zhengyan ZHU ; Zhi DU ; Ying LUO ; Peng WANG ; Yingtang GAO
Chinese Journal of Hepatobiliary Surgery 2010;16(6):460-464
Objective To investigate the effect of human umbilical cord mesenchymal stem cell paracrine substance on proliferation and apoptosis of liver cells in vitro. Methods Mesenchymal stem cells (MSC)were separated from human umbilical cord with type Ⅳ collagenase and trypsogen digestion method and cultured in vitro. The human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) which contain paracrine substance of human umbilical cord mesenchymal stem cells (HUCMSC) was prepared. Hepatocytes were isolated from SD rats by low concentration collagenase perfusion procedure. There were three groups in the experiment, control group, 2% MSC-CM group and 8% MSC-CM group. The proliferation of normal hepatocytes were assayed with MTT method. We detected the urea and albumin level in culture supernatant to assay the hepatocyte function under different concentration MSC-CM. Hepatocytes were induced for apoptosis by Actinomycin D and tumor necrosis factor alpha (TNF-α),and the apoptosis effect of different concentration MSC-CM was assayed with LIVE/DEAD Viability/Cytotoxicity Kit. Results The MTT assay showed that the absorbance of 2% MSC-CM group was significantly increased (P<0. 01), and the urea and albumin levels of 2 % MSC-CM group were also significantly increased when compared with control group(P<0. 01).LIVE/DEAD Viability/Cytotoxicity Kit revealed that hepatocyte survival rate of 2 % MSC-CM group was increased when compared with control group(P<0. 05), there were no significant differences in above-mentioned experiments when 8% MSC-CM group compared with control group. Conclusion The low concentration MSC-CM could stimulate normal hepatocyte proliferation, inhibit impaired hepatocyte apoptosis and improve hepatocyte function.
10.Dysfunction of branded-chain amino acids catabolism in rat cardiac allograft
Qingchun ZHANG ; Haihui YIN ; Zhongya YAN ; Yueheng WU ; Zhengyan ZHU ; Hong LEI ; Zhong LU
Chinese Journal of Organ Transplantation 2011;32(8):492-496
Objective Allograft vasculopathy (AV), feature of chronic rejection, is a major serious long-term post-operation complication in organ transplantation. The accurate mechanisms for AV have not been definitively established, but extensive basic and clinical studies demonstrate AV is triggered by immune reaction and nonimmunologic factors, and also possibly attributed to the metabolism of branched-chain amino acids (BCAA). Methods The transplanted hearts from Lewis to Sprague-Dawely rats served as allografts and those from Lewis to Lewis rats as isografts based on Ono 's model. The differential proteins in transplanted hearts were separated by comparative proteomic technique, and some enzymes which regulated the metabolism of BCAA were identified and validated.Results All transplanted hearts at second week postoperation were characterized by lumen loss (total area-luminal area/total area) in coronary artery, but more predominant at 8th week. All samples from the left ventricles were analyzed by proteomic techniques and the subunits E1 a, E1β and E3 of branched-chain α-ketoacid dehydrogenase (BCKDH) complex were decreased in the heart allografts.Immunohistological detection also showed the expression of BCKDH was reduced not only in the cardiac muscle but also more significantly in blool vessels with cardiac allograft vasculopathy (CAV).BCAA concentrations were increased in the cardiac allografts, but there was no difference in the serum. Conclusion These findings suggest that the catabolic pathways of the BCAA may be inhibited owing to the reduced expression of BCKDH complex, and elevated intracellular concentrations of leucine. The vascular smooth muscle cell and cardiac muscle cell proliferation is stimulated via mTOR-dependent and mTOR-independent pathways, which is associated with the formation of myocardial hypertrophy and AV in the heart allografts.