1.Clinic research between vertebral endplate morphology and lumbar disc herniation
Hongsheng GU ; Wenyu ZHOU ; Zhenyu LI
Chinese Journal of Tissue Engineering Research 2010;14(50):9497-9500
BACKGROUND: Studies demonstrated that vertebral endplate can be remodeled during intervertebral disc degeneration.However,the relationship between morphology of the vertebral endplate and intervertebral disc degeneration remains poorly understood.OBJECTIVE: To explore the relationship between the morphology of the vertebral endplate and the lumbar disc herniation.METHODS: Forty cases without previous spine disorder and sixty-two cases symptomatic lumbar disc herinations lumbar vertebral were scanned by using spiral CT.Scan range was from superior L4 to superior S2.Scan protocols as below: 140 kV,345 mAs,FOV 160 mm,layer of thick 1 mm,pitch 1.0.Original images were carried through three-dimensional reconstructions using O2 image work station,W300,C80,ZOOM=1.5,Expand 1.Intervertebral disc maximum anteroposterior,transverse diameter,circumference,area and shape were measured based on curve planar reconstruction.RESULTS AND CONCLUSION: Endplate shape was strongly related to disc herniation(all the P value < 0.01).Endplate area was a less significant factor L4/5 in men and L5/S1 in men and females(P < 0.05).The shape of the intervertebral body endplate margin is an important factor contributing to the development of disc herniation at L4/5 and L5/S1.
2.Progenitor cells in mammalian maxillofacial and mandibular processes Source and differentiation phenotype
Zeyuan ZHOU ; Yan JIN ; Shuxiang YU ; Junnan SHI ; Jianping ZHANG ; Xiaoyan DUAN
Chinese Journal of Tissue Engineering Research 2010;14(50):9493-9496
BACKGROUND: It remains unclear whether ectomesenchymal cells also derived from neural crest stem cell in mammals.OBJECTIVE: To understand the specific markers and differentiation directions of maxillofacial and mandibular processes progenitor cells,and to explore the source and differentiation phenotype of ectomesenchymal stem cells.METHODS: The expression and changes of expression profiles of rat ectomesenchymal cells at E9.5,E10.5,E11.5,and E12.5days were observed by immunohistochemistry and flow cytometry.RESULTS AND CONCLUSION: The progenitors expressed multi-lineage markers,including neural system and several rnesenchymal tissue types,importantly the facts that molecule profiles were changed with time prolonged,suggesting these progenitors were in active differentiating stage,so they were stem like cells or contain stem like cells.Moreover,small populations(2%-3%)of CD57 and P75 phenotypes were detected by flow cytornetry,suggesting that ectomesenchymal stem cells were derived from neural crest,which maintained a quantitative stabilization though it is gradually differentiate after localization.
3.Construction and expression of pDsRed-human apelin receptor recombinant plasmid in human embryo kidney 293 cells
Hui DU ; Bo BAI ; Jing CHEN ; Haiqing LIU ; Yalin LI
Chinese Journal of Tissue Engineering Research 2010;14(50):9489-9492
BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.
4.Localization,cultivation and purification of sinoatrial nodes isolated from newborn rabbits
Qingzhi WANG ; Li ZHOU ; Yuqiao CHANG
Chinese Journal of Tissue Engineering Research 2010;14(50):9485-9488
BACKGROUND: The method of culture purified sinoatrial node cell is important in investigating its ultrastructural characteristics and autorhythmic mechanisms.However,the corresponding method has not been standardized.OBJECTIVE: To summarize the localization,cultivation and purification of sinoatrial nodes isolated from newborn rabbits,and to study the morphological characters of primary cultured pacemaker cells.METHODS: Hearts of the newborn rabbits(within 24 hours)were embedded in paraffin for hematoxylin-eosin staining,The location of sinoatrial nodes was observed under an optical microscope,the morphology of sinoatrial nodes cells were observed by light microscope and electron microscope.RESULTS AND CONCLUSION: Sinoatrial nodes localized in the anterior wall of the superior vena cava and the posterior laterial wall of right atrium.There was about 0.32 mm between its lowest point and sulcus terminalis.Three distinct types of cells were observed among the cultured cells of sinoatrial nodes: spindle,spider and polygon.The spindle cells occupied the greatest proportion of the cultured cells(59.6±7.3)%.The spontaneous contraction frequency of spindle cells was the highest among the constracting cells(145±9)times per minute.The ultrastructure observation showed that myofibrils and other organelles in spindle cells were sparse and significantly decreased in number compared with triangle cells.There was no significant difference between triangle cells isolated from sinoatrial nodes and from atrial muscle.Sinoatrial nodes could be harvested along the anterior root of the superior vena cava down to the posterolateral sulcus.Among the cultured cells from neonatal rabbit sinoatrial nodes,the spindle cells with small body and fast pulse frequency are pacemaker cells.
5.Length changes of knee lateral collateral ligaments following anterior cruciate ligament injury
Pingyue LI ; Qingshui YIN ; Huayang HUANG ; Jianyi LI ; Hongyuan SHEN ; Zejin WANG ; Qing WANG
Chinese Journal of Tissue Engineering Research 2010;14(50):9480-9484
BACKGROUND: Lateral collateral ligaments play an important role in maintaining knee stability.Motion reduction of knee joint can be realized and the changes laws of medial and lateral collateral ligaments' length after anterior cruciate ligament(ACL)injury during weight-bearing flexion can be obtained via 2D/3D image registration technique.OBJECTIVE: To study in vivo stability of length changes of the medial and lateral collateral ligaments of ACL injury knee during weight-bearing flexion.METHODS: Eight volunteers with unilateral ACL rupture and contralateral normal knees,was captured CT images and 2orthogonal images of the knee at 0,15°,30°,60°,and 90° of weight-bearing flexion.These orthogonal images were used to recreate the in vivo knee positions at each of the targeted flexion angles by the method of 2D/3D image registration.Through the bone insertion of medial and lateral collateral ligaments,the elongation changes of medial and lateral collateral ligaments were obtained.RESULTS AND CONCLUSION: At 0°,15° and 30°,the length of medial collateral ligament of ACL injury knees was longer than normal knees,but the lateral collateral ligaments length of ACL injury knee was shorter than that of normal knees.All the differences have statistical significances(P < 0.05).The findings demonstrated that,at 0°,15° and 30°,the medial collateral ligament length of ACL injury knees was longer than normal knees,but lateral collateral ligaments length of ACL injury knees was shorter than normal knees.
6.Induction of umbilical cord blood-derived mesenchymal stem cells differentiating into pancreatic islet β-like cells in vitro
Zuohua CHI ; Yan LU ; Yuan ZHANG
Chinese Journal of Tissue Engineering Research 2010;14(49):9307-9310
BACKGROUND: At present,there are many reports regarding the differentiation of bone marrow-derived mesenchymal stem cells or pancreatic gland stem cells into pancreatic islet β-like cells.But little is about umbilical cord blood-derived mesenchymal stem cells(UCBMSCs)differentiation into pancreatic islet β-like cells in vitro.OBJECTIVE: To investigate whether UCBMSCs can differentiate into pancreatic islet β-like cells in vitro and the optimal inducing condition.METHODS: UCB samples were obtained sterilely from healthy parturients.Nucleated cells were isolated by sedimentation with hydroxyethyl starch and MSCs were obtained by adherent method.Then purified UCBMSCs were induced with epidermal growth factor,β-mercaptoethanol,high glucose,Activin A and hepatocyte growth factor(HGF).Following cell morphology observation,induced cells were identified by insulin immunofluorescence.In addition,insulin secretion and glucose-stimulated insulin release were examined by chemiluminescence immunoassay.RESULTS AND CONCLUSION: After induction,many cells exhibited a round appearance and produced islet-like cell clusters.Immunofluorescence assay showed insulin positive in the treated cells.In addition,chemiluminescence immunoassay demonstrated low expression of insulin and secretion of insulin upon glucose challenge.UCBMSCs can differentiate into pancreatic islet β-like cells in vitro.
7.Effects of natural killer cells on graft rejection,hematopoietic and immune reconstitution following allogeneic bone marrow transplantation
Zhigang YANG ; Dan XIONG ; Yaoying ZENG ; Liang LIANG ; Qinghua LI ; Guocai WU
Chinese Journal of Tissue Engineering Research 2010;14(49):9303-9306
BACKGROUND: The connection between Natural killer(NK)-cells and allogeneic bone marrow transplantation(allo-BMT)has aroused increasing attention.OBJECTIVE: To explore the effect of NK cells on graft rejection,hematopoietic and immune reconstitution in mouse undergoing allo-BMT.METHODS: Lethally and nonlethally irradiated BALB/c(H-2d)mice were transplanted with C57BL/6(H-2b)bone marrow plus donor peripheral T cells and/or NK cells.RESULTS AND CONCLUSION: Compared with lethally irradiated and allo-BMT group without infusion of NK cells,the survival rate in lethally irradiated and allo-BMT group with infusion of NK cells significantly enhanced; leukocytes count,expression level of CD19+and CD34+cell count recovered rapidly; expression level of H-2b*cell obviously increased.Expression level of CD34"cell in the group with infusion of NK cells was obviously lower than that of the group without infusion of NK cells at 28 days after transplantation,but there was no significant difference between the 2 groups at 60 days(P > 0.05).In nonlethally irradiated and allo-BMT group without NK cell infusion,expression level of H-2b*cell significantly decreased at 30 days after transplantation,and reduced to before transplantation level at 60 days; while expression of H-2b+cell yet could be detected with more than 80% at 60 days after transplantation in group infused with high and low concentration of NK cells.In alIo-BMT mice,alloreactive NK cell inhibits graft rejection,enhances engraftment,promotes the reconstitution of hematopoiesis and immunity,and increases survival rates.
8.Central arterial augmentation index in Uygur health adults from Hetian,Xinjiang Detection by Sphygmocor pulse wave analysis system
Xiaohui LIANG ; Xinjuan XU ; Yanping MA ; Lihua SUN ; Yitong MA ; Ding HUANG ; Yining YANG
Chinese Journal of Tissue Engineering Research 2010;14(52):9877-9880
BACKGROUND:Arterial stiffness is an independent risk factor for cardiovascular disease.Augmentation index(Alx)is an important marker which reflects the systemic arterial stiffness.OBJECTIVE:To investigate Alx and the affecting factors in Xinjiang Uygur health adults.METHODS:A total of 559 healthy adults(251 males and 308 females)with a mean age of(44.95±11.87)years were selected from Hetian,Xinjiang Uygur Autonomous Region.The following indexes of all subjects were measured,including questionnaire,physical examination,blood biochemistry and C-reactive protein(CRP).Central systolic blood pressure(cSBP)and central Alx were measured by Sphygmocor.Pearson correlation and partial correlation analysis were used for two-group analysis of correlation,and multiple regression analysis was used to analyze influential factors for Alx.RESULTS AND CONCULUSION:Alx was greater in female than in male and was positively related to age in both sexes(male:r=0.548,female:r =0.405,P < 0.05),as well as adjusted other influential factors(male:r=0.182,female:r =0.234,P < 0.05).Multiple regression analysis showed that cSBP,systolic blood pressure(SBP),sex,age and CRP were main factors which affected Alx in Uygur healthy adults,cSBP,SBP,age,body mass and CRP were main factors which affected Alx in the male,while cSBP,SBP,age,height and CRP were main factors which affected Alx in the female.Results revealed that Alx was greater in female than in male which was positively related to age in both sexes.Age,cSBP,SBP and CRP were main factors that affected Alx in Xinjiang Uygur healthy adults.
9.Distribution features of stress-strain,principle stress vectors and displacements of normal femur during gait
Xueling BAI ; Peng SHANG ; Chengtao WANG
Chinese Journal of Tissue Engineering Research 2010;14(52):9873-9876
BACKGROUND:Studies demonstrated that,the probability of middle and distal end of femur bone fatigue or fracture reaches maximum when the moving loads increase or under an impact load.OBJECTIVE:To analyze the stress-strain distribution,principle stress vectors and displacements of femur during gait.METHODS:The contour curves of femur and muscles attached to femur were extracted slice by slice based on the serial CT data from a healthy male volunteer,which were used to build musculoskeletal system models.And the straight-line muscle models of the lower extremities were constructed to evaluate muscles forces and joints forces by using a multi-objective optimization method based on gait analysis.The finite element models of femur during gait were built by taking muscle forces and joint forces as boundary conditions in ANSYS 10.0 to obtain the stress-strain distribution and displacements.RESULTS AND CONCLUSION:The maximal displacement occurred on the femur head,and the maximal stress occurred in the middle and distal end of femur during the whole gait.There exists bigger tensile stress in the location of the femur neck and middle and distal end of corpus ossis femoris.The result validates the fact that femur fracture often happens at the middle and below of femur stem from the viewpoint of biomechanics.
10.Sequential bilateral lobar transplantation A case report
Zhenrong ZHANG ; Deruo LIU ; Yongqing GUO ; Bin SHI ; Zhiyi SONG ; Yanchu TIAN ; Yanning SHOU ; Chaoyang LIANG ; Haitao ZHANG ; Jingyu CHEN ; Zaiyong WANG ; Tong BAO ; Qianli MA ; Guangliang QIANG ; Hongxiang FENG
Chinese Journal of Tissue Engineering Research 2010;14(53):10067-10070
BACKGROUND: Lung transplantation can improve quality of life of patients who get terminal pulmonary disease and also it can help to get better survival.Now it has become one of the best therapeutic methods for terminal pulmonary disease.However,limited donors leave the development of lung transplantation in dilemma.The emergence of living lobar transplantation and cadeveric lobar transplantation let this procedure much easier.OBJECTIVE: To evaluate the clinical probability of bilateral lobar transplantation.METHODS: Sequential bilateral lobar transplantation was performed for one 26 years old cystic fibrosis female.Cardiac pulmonary bypass was used during operation.Anti-rejection(Tacrolimus,mycophenolate,etc)and anti-infection was used postoperatively.RESULTS AND CONCLUSION: The recovery course postoperatively was smooth,and the recipient got out of hospital 7weeks later.Bilateral lobar transplantation could offer satisfied short-term pulmonary function.The long term results should be further evaluated.