OBJECTIVETo establish a new extrahepatic growing hepatocellular carcinoma cell line.

METHODSA specimen from extrahepatic growing hepatocellular carcinoma was cultured in vitro. Cancer cells were studied morphologically and subjected to karyotype analysis, DNA analysis, and tumor formation evaluation.

RESULTSMorphological observation and functional analysis showed that their features were similar to those of HCC. Chromosomes with a variation of 76 approximately 104 were able to secret AFP in vitro and to form bile canaliculi with microvilli.

CONCLUSIONEGHC-9901 cell line has characteristics of the extrahepatic growing hepatocellular carcinoma.

Adult ; Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Chromosome Aberrations ; Humans ; Liver Neoplasms ; genetics ; pathology ; Male ; Mice ; Mice, Nude ; Tumor Cells, Cultured ; alpha-Fetoproteins ; analysis

Objective The aim of the study was to observe the survival and differentiation of embryonic neuroepithelial cells in lateral ventricle of rats after transplantation. Methods Embryonic rats(E12d) were obtained from the pregnant Sprague\|Dawley rats;neuroepithelial cells were dissociated from embryonic neural tube and treated with 0\^25% trypsin,then transplanted into the lateral ventricle of father rat.The hosts were anesthetized and transcardially perfused with PBS followed by 4% paraformaldehyde fixative by 10?d,14?d,21?d and 28?d transplantation respectively.Coronal sections of the brain were cut with in cryostat microtome.Using the techniques of histology staining,Nissl staining and immunocytochemistry, survival of neuroepithelial cells transplant and neuro\|specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression of some stem cells were examined. Results Some grafts were found survival which attached to the ventricle by day 10.Some grafts could migrate into the third ventricle and grew into a mass of cells.Some stem cells were able to differentiate into pyramidal cells.By immunocytochemistry,a mass of NSE\|positive cells and GFAP\|positive cells could be detected in the core of graft region.Around intracerebral grafts,some astrocytes showed immunoreactive for glial fibrillary acidic protein.Conclusion\ These findings demonstrate that embryonic stem cells dissociated from neural tube can survive and differentiate into neurons and astrocytes.

AIM: To compare the protective effects of tongxinluo, a Chinese medicine, and carvedilol and valsartan on myocardium microvascular endothelial function and integrity after late reperfusion of acute myocardial infarction (AMI) in rabbits. METHODS: Forty-eight rabbits were randomly assigned to the following groups: (1) sham operated rabbits; (2) ischemia-reperfusion (I-R) controls; (3) tongxinluo (1.0 g?kg~ -1?d~ -1); (4) carvedilol (5 mg?kg~ -1?d~ -1); (5) valsartan (10 mg?kg~ -1?d~ -1) and (6) ticlopidine + aspirine (30 and 20 mg?kg~ -1?d~ -1, respectively) groups. After 3 d of drug treatment, the left coronary artery in the rabbit was ligated for 2 h and loosed subsequently for another 2 h. The serum levels of nitric oxide (NO_2~-/NO_3~-) and endothelin (ET) at baseline before AMI, 2 h after both AMI and reperfusion were examined. Also, the number of circulating endothelial cells (CEC), MI size and percentage myocardium focal bleeding incidence were determined 2 h after reperfusion. RESULTS: (1) The baseline level of NO_2~-/NO_3~- was significantly higher in tongxinluo group than that in other groups (all P

The animal experiment studies have demonstrated that hyperthermia is a strong teratogen to many kinds of animal with high incidence of neural tube defects(NTD).Epidermiological investigation showed that hyperthermia was closely related to the acencephaly and excenphaly in human being,but little was known about the distinct mechanism of NTD induced by hyperthermia.In order to study the developmental mechanism of NTD induced by hyperthermia,we observed the effects of hyperthermia on the neuroepithelial cells in primary culture.The neural tubes of the hamster embryos on 10 d after fertilization were obtained.the neuroepithelia were dissociated and then seeded at the density of 1×106 cells per well.The cells were divided into experimental and control groups randomly.The experimental groups were exposed to 42 C for 20 min,whereas groups exposed to 37 Cserved as control.After treatment of hyperthermia,the cells were incubated continuously at 37 C in a 95%air/5%CO2 humidified incubator,the culture was terminated after different intervals.Phase-contrast microscopy.scanning electron microscopy.transmission electron microscopy,MTT assay,agarose gel electrophoresis analysis,TUNEL detection,immunocytochemistryand image analysis were made.The results of the experimental groups indicated that the floating cells and apoptotic cells increased in number,the neurites of the cells were shortened even disappeared,the survival cells decreased in number,the ultrastructure of the cells demonstrated distinct abnormal changes,the function of mitochondria was impaired,the expressions of both bcl-2 and bax were abnormal.The above results suggest that hyperthermia may induce apoptosis of neuroepithelial cells,duringwhich bcl-2 and bax genes may play important regulatory roles.

0 05) Left ventricular (LV) end diastolic pressure (LVEDP), volume (LVV), weight (LVW) and septal thickness (STh) were all higher and left ventricular pressure maximal rate of rise and fall (?d p /d t ) were lower (all P

To establish a new extrahepatic growing hepatocellular carclnoma cell line. Methods: A speciment that was pathologically identified extrahepatic growing hepatocellular carcinoma were cultured in vitro. The morphology,karyotype analysis, DNA analysis, the tumor formation of heterotransplantation were observed. Results: Morphological observation and functional analysis showed that it had the comnnon features of HOC.It was found to be able to secret AFP in vitro. Conclusion: EGHC-9901 has characteristic of the extrahepatic growing hepatocellular carcinoma cell line.

Objective To explore the optimal condition of direct differentiation into dopaminergic neurons of embryonic stem cells in serum-free and feeder layer cell free medium. Methods We used the method of phase induction to culture embryonic stem cells. At first, embryonic stem cells were cultured in the serum-free medium with bFGF and LIF so as to realize the direct differentiation from embryonic stem cells to neural precursors. Differentiated cells were determined by nestin immunocytochemical staining. On this basis, we transferred embryonic stem cells to the B27 serum-free medium with IL-1 so as to realize the direct differentiation from neural precursors to dopaminergic neurons. Differentiated cells were determined by TH immunocytochemical staining. Results Approximately 85 percent of cell masses were nestin immuno-positive. The differentiation ratio of dopaminergic neurons was 13%, which increased significantly in comparison with natural differentiation ratio of dopaminergic neurons.Conclusion Without serum and feeder layer cell, we can induce embryonic stem cells to differentiate into dopaminergic neurons effectively by adding different growth factors at different phases, which makes the inductive processes more easily.

Objective To carry out construction of chimeras from embryonic stem cells of outbred KM mice. Methods We isolated embryonic stem cells from inner cell mass of KM mice blastocysts. Then we transferred embryonic stem cells into blastocoele of C57BL/6J inbred mice by microinjection in order to construct chimeric mice. Results The cells isolated from inner cell mass have typical characteristics,i e positive alkaline phophatase staining,normal karyotype,forming embryoid body. Now,we have constructed one chimeric mice successfully. Conclusion Embryonic stem cells isolated from inner cell mass can be used for the chimeras production successfully,which forms the substantial base of transgenic animal model by the way of using embryonic stem cells.

Objective To explore the inductive effect of striatal tissue on mouse embryonic stem cells and further analyse the cell source and inductive pattern of this inductive effect. Methods We employed striatal extracts、conditioned medium of striatal astrocytes and conditioned medium of striatal neurons to induce embryonic stem cell to differentiate directly. The differentiated cells were evaluated by morphological observation and TH immunocytochemical staining method. We also performed a quantitative analysis of the results. Results Striatal extracts and conditioned medium of striatal astrocytes had obvious inductive effect on embryonic stem cell.Percentages of three groups were 15%,15.2% and 3% respectively. Conclusions The astrocytes in striatum might have an inductive effect on dopaminergic neuronal differentiation of embryonic stem cells.The determination of inductive factor will be our next research aim.;

Objective To explore the survival,migration and differentiation of embryonic stem cells after transplantation into striatum and provide advantageous data for feasibility and safety of therapeutic transplantation. Methods We transplanted embryonic stem cells(undifferentiated ESCs and ESCs that had already developed into the stage of neural progenitor cells respectively) into striatum of the rat.Then differentiated cells were determined by morphological observation,Nissl's staining,TH and BrdU immunocytochemistry. Results We found that after transplanted 4 weeks,partially differentiated ESCs could survive and migrate into the surrounding host tissue.Some of them differentiated into TH-positive cells which had Nissl's bodies in cytoplasm.Whereas undifferentiated ESCs couldn't differentiate into TH-positive cells effectively and have the tendency to form tumor.Conclusion When conducting transplantation experiments of ESCs,it's better for ESCs to be induced into the stage of neural progenitor cells first and then transplanted.;