Objective To screen the gene expression profiles of human bladder cancer cell line EJ,in which RNA interference was used to downregulate the EZH2 expression and to investigate the EZH2 molecular mechanism in cancer.Methods The vector expressing EZH2 shRNA was constructed and transfected into EJ cell line with Lipofectamine 2000.We used the gene chip to screen differential expression genes in EJ cells with transfection with EZH2 shRNA compared with untransfected EJ cells.The data were up-loaded to Passway miner serve((http://www.biorag.org/passway.htm))Results We found 436 differentiation genes,in which 257 genes upregulated and 179 genes downregulated.Bioinformation analysis showed that 115 genes locate in the known biological ways,including EGF,P53,proliferation,cell cycle,Notch and Wnt passway.EZH2 target genes such as WNT1,MYT1,CNR1,WT1 were upregulated,some of which were linked to tumor stem cell and cell differentiation.Conclusion The linkage between EZH2 expression and cell proliferation,which is induced by genes encoding cell differentiation.The research focusing on EZH2 regulating genes will explore its molecular mechanism in oncog-enesis.

Objectives To study the anatomic characteristic of medial patellar retinaculum and its function in patellar stability. Methods Anatomic dissection of the medial patellar retinaculum was performed on 26 knees of 13 embalmed cadaveric specimens,and its composition and conducted functional studies of the several parts were observed respectively. Results The medial patellar retinaculum comprises patellofemoral ligament, patellotibial ligament and patellomeniscal ligament. The medial patellofemoral ligament which is the thickest part in the medial patellar retinaculum contributes to the major restraints to the lateral patellar displacement.

Objective To construct a eukaryotic expression vector of huma n insuli n-like growth factor-1 (hIGF-1) gene,and to investigate the transfection and ex pression of pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid in myoblast cells. Methods pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid was co nstructed. Myobl ast C2C12 cells were transiently transfected with lipofectamine. RT-PCR a nd immunohistochemical methods were used to detect the expression of hIGF-1 gene . MTT test was used to detect the biological activity of the conditioned medium after the transfection. Results There were significant increas es of hIGF-1 mRN A and protein in the myoblast cells transfected with pcDNA3.1(-)/hIGF-1. The con ditioned medium after the transfection showed the biological activity which coul d stimula te the proliferation of fibroblast cells. Conclusion The pcDNA 3.1(-)/hIGF-1,a eukaryotic expression plasmid for hIGF-1 gene is constructed. High levels of hI GF-1 mRNA and protein expression can be obtained in the myoblast cells transfect ed with pcDNA3.1(-)/hIGF-1 eukaryotic expression plasmid. The expressed protein has the biological activity.

Objective To explore the effects of specific T-cell immunity against prostate cancer(PC) cells induced by dendritic cells(DCs) derived from fetal organs.Methods Mononuclear cells(MNCs) were obtained from the fetal bone marrow and liver.Then MNCs were cultured in medium with induction of rhGM-CSF,rhIL-4 and rhTNF-?to get DCs.Lysates of DU145 containing HSP-peptide complex were prepared by 50%-70%(NH4)2SO4 saturation.T lymphocytes from fetal spleen were co-cultured with DC loading DU145 antigen for 72 h,whereby CTL was obtained.The cytotoxicity of CTL against DU145,PC3 and EJ was detected by MTT assay.Results Mature DCs were induced from fetal organs,which expressed CD1a,CD_(86),HLA-DR and CD_(83) at high levels.DC stimulated with tumor lysates transformed T cells to specific CD~+_8 CTL.Phenotype of CD~+_8 cell was(14.09?(2.46))% before transformation,and(62.76?2.64)% after transformation,respectively(P

Objective To study the biomarker panel of superficial bladder transitional cell carcinoma(SBTCC)and analyze the biological pathway in tumorigenesis by Shotgun proteomics strategy.Methods Normal urothelium cells and cancer cells were harvested by laser capture microdissection from clinical specimen and the proteomic expression profile was identified by two-dimensional liquid chromatography tandem mass spectrometry.The isoelectric point,molecular weight,grand average of hydropathicity,transmembrane helices were analyzed by using proteomics tools.Gene ontology was used to comment the identified proteins.The pathway analysis was performed by ArrayTrack software,and visualized by GenMAPP.Results There were 440 and 218 proteins expressed in cancer cells and normal cells respectively,among them 388 proteins were differerntially expressed.All the database about identified proteins was deposited in an accessible form to researchers at http://www.Proteome-SBTCC.org.cn and http://www.Proteome-NHTE.org.cn.There were 267(68.8%)differentially expressed proteins which had GO biological process comments.The biological pathwavs of these proteins included MAPK signaling pathway,focal adhesion,oxidative phosphorylation,ECMreceptor interaction,etc.Conclusion Shotgun strategy proteomies database of normal transitional epithelium and SBTCC is successfully constructed.And the basis for the understanding of cell biology and discovery of biomarker panel for SBTCC iS provided.

Objective To observe the effect of the injection of myoblast carrying human insulin-like growth factor-1(hIGF-1) on the expression of endogenous IGF-1 mRNA and IGF-1 level in mice skeletal muscle following injury.Methods Seventy two male C3H mice(20~30g,7~11w)were randomly divided into three groups(24 mice in each group) with four mice normal controls.Applied a falling hit from certain height at the medial calf of right lower limbs in three groups,the injured skeletal muscle model was successfully simulated.Three days following injury,the mice in group A and B were injected with 1?106 myoblasts either carried with or without hIGF-1 gene respectively and the mice in group C were injected with 100?l saline at the injured muscle.Three mice in each group were sacrificed randomly at day 2,5,10,15,20,30 after contusion.The expression level of mIGF-1 was assessed by immunohistochemical staining and real time PCR.Results mIGF-1 mRNA expression and mIGF-1 factor secretion were observed in all three groups;the amount of mIGF-1 mRNA expression and mIGF-1 secretion in group A were significant higher than that in group B and C.Conclusion Myoblast carrying hIGF-1 transplantation could promote endogenous IGF-1 secretion in injured skeletal muscle.

Objective To observe the survival of myoblasts carrying hIGF-1 gene transplanted into the mice, and the expression of hIGF-1 in the transplanted mice. Methods Eighty four male C3H mice(20～30g,7～11w)were divided into four groups: group A, B, C and D (20 mice each group), and the remaining four mice were used as normal control. At the middle of the right gastrocnemius muscle, the mice in group A and B were injected with 1?10~6 myoblasts either carried with or without hIGF-1 gene. Muscle contusion at the middle of the right gastrocnemius muscle of the mice in group C and D was produced. At day 3 following injury, they were injected with 1?10~6 myoblasts either carried with hIGF-1 gene (group C) or without hIGF-1 gene (group D). At the day 2, 5, 10, 20, 30 after injection, four mice of each group were sacrificed randomly. BrdU staining in all mice were performed to evaluate cells surviving, and the expression level of hIGF-1 in group A and C was assessed by immunohistochemical staining and real-time-PCR. Results The BrdU staining in both normal and injured mice transplanted with myoblast carried with or without hIGF-1 were positive. hIGF-1 was expressed and secreted in both group A and C. Conclusion The myoblast carrying hIGF-1 gene transplanted into normal or injured mice can survive for a certain period of time, and can secrete hIGF-1.

Neurons descending from the griseum centrale mesencephali, nucleus Darkschewitsch and nucleus interstitialis of Cajal to the nucleus raphe magnus and adjacent reticular formation (nucleus reticularis gigantocellularis and nucleus reticularis pontis caudalis) were identified in 9 adult cats with the retrograde HRP method. In the griseum centrale mesencephali, the labeled neurons were found bilaterally but slightly more ipsilaterally. In the nucleus Darkschewitscb and nucleus interstitialis of Cajal, the labeled neurons were consistently found in its rostral part ipsilateral to the injected side at the level of the posterior commissure. In addition, in 5 of the 9 cases, a few labeled neurons were observed in the nucleus raphe dorsalis.

The projections of diencephalon onto the rostral part of the mesencephalic periaqueductal gray matter (PAG) were investigated by using the method of retrograde transport of HRP in the rat. The results have led to following conclusions:1. The diencephalic afferents to the rostral part of PAG originate mainly from medial preoptic area, anterior hypothalamic area, dorsomedial and ventromedial hypothalamic nuclei, dorsal premamillary nucleus, lateral hypothalamic region, Forel's field and zona incerta ipsilaterally, and a few of them originate contralaterally.2. The anterior hypothalamic area, ventromedial hypothalamic nucleus, dorsal premamillary nucleus and zona incerta project more to the ipsilateral nucleus: lateralis of PAG than to the nucleus medialis and contralateral nucleus lateralis of PAG.3. In the anterior hypothalamic area, a large number of neurons projecting to PAG was located in the lateral anterior nucleus.4. In the lateral hypothalamic region, the lateral preoptic area and anterior division project to both nuclei lateralis and medialis, while its tuberal and mamillary divisions probably project only to nucleus lateralis.

Objective To observe the effect of different interventions on muscle healing following contusion by electromyographical signal, and functionally to study the significance of electromyographical technique in evaluating the healing quality of skeletal muscle after injuries. Methods A standard contusion model was developed at the right gastrocnemius by specific device in 108 male rats. The animals were divided into three groups:group IGF-1,local injection by exogenous IGF-1 at injured site; group Chinese medicine treated with the injection of Huangqidanshen, and group natural healing for control. EMG signals of gastrocnemius, such as fibrillation potentials, positive sharp waves as well as the latency and wave-amplitude of compound muscle action potentials (CMAPs) were probed, recorded and compared at day 2、5、7、10、14、21、28、35 and 56 after injury respectively. Results (1) Fibrillation potentials and positive sharp waves were discovered at day 2 to 5 after contusion, and reached its peak by day 7 to 10, and then normalized gradually around the 35th day with muscle healing in group natural healing. Above abnormal signals disappeared much more early in group IGF-1, then that in group Chinese medicine. (2) The latency and wave-amplitude of CMAP shown a gradual recovery process with muscle healing. The recovery tendency was most rapid in group IGF-1, slower in group Chinese medicine, and latest in group natural healing. (3) The wave-amplitude of CMAP approached to normal in group IGF-1 by 8 weeks following contusion, a little bit later in group Chinese medicine groups, and lastly in group natural healing. Conclusions(1) Electromyographical study verified that IGF-1 and Huangqidanshen could improve healing and functional recovery following muscle injury. IGF-1 was a much more strong intervention for healing promotion. (2) EMG is an useful tool in evaluating the healing process and healing quality of skeletal muscle.