Liver tissue engineering is one of the important subjects in tissue engineering field.The major goal of the research is to treat end-stage liver failure and liver based inborn errors of metabolism.Two strategies are usually employed:seed cell transplantation for liver tissue repair and regeneration of "liver tissue" consisted of an extraeorporeal bioreactor loaded with cenular component.The seed cells is one of the key ingredient of liver tissue engineering,which include adult hepatocytes,various kinds of liver stem cells,and immortalized cell fines,etc.However,to date,there is no optimal cell resource for application.Many problems should be solved before these cells can be widely used in clinic.This review focuses on the research regarding the seed cells and their potential for the application in liver tissue engineering.

As a safe and effective treatment method of fulminic hepatic failure and end stage liver disease, hepatocyte transplantation has been widely verified in many animal experiment and clinical research. It has many advantages. For instances, It is safe and easy to perform. It causes little trauma and is convenient for retransplantation. The hepatocytes are less immunogenetic and the cells from one donor liver can be used for several patients, and so on. The cell source, route of transplantation, theoretical basis and clinical application of the method are reviewed in this paper.

According to traditional theory tumor growth arised from the proliferation of all tumor cells, therefore management mainly aimed at the majority of the cells within cancer tissue. As a result, relapse and metastasis developed often, which caused treatment failure. It was stem cell that makes it possible to treat cancer radically and prevent it from relapsing and metastasizing by specifically killing targeted cancer stem cells and even some old concepts such as tumorigenesis mechanism, cell signaling pathways etc may need to be reevaluated, that raised great challenge to traditional oncotherapy. In this text the origin and existing evidence of cancer stem cell, it's relationship to common stem cell, clinical significance and prospect were reviewed.

This study was aimed to investigate effect on the expression of cell inflammatory factor IL-1α, IL-1β and IL-6 of modified San-Jia-San (SJS) decoction on cultured hippocampal neurons injury induced by β-amyloid 25-35 protein (Aβ25-35). Primary cultured hippocampal neurons were divided into the normal group, model group, huperzine A group, low-dose SJS group, middle-dose SJS group, and high-dose SJS group. After selective culture for 7~10 days and the absorption of culture fluid, the blank culture fluid, normal cerebrospinal fluid (normal saline group), huperzine A cerebrospinal fluid, and low-, middle- and high-dose SJS cerebrospinal fluid were added, respectively. The culture fluid was added up to an equivalent medium. The incubation was 2 h under the temperature of 37℃. Ex-cept the normal group, Aβ25-35 dealt with aging (the final concentration was 5 μmol/L) was added to other groups to establish AD cell model. An equivalent amount of culture fluid was added to the normal group. After 24 h of incuba-tion, the cell culture supernatant was collected. Enzyme-linked immunosorbent assay (ELISA) was used in the content detection of IL-1α, IL-1β and IL-6. The results showed that modified SJS decoction can decrease the content of IL-1α, IL-1β and IL-6 in the cell culture supernatant. It was concluded that the modified SJS decoction can effectively inhibit the expression of cell inflammatory factor IL-1α, IL-1β and IL-6. It had certain anti-inflammatory effect to protect hippocampal neurons.

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.

Objective To investigate the effect of Bunao capsule on learning,memory and antioxidative abilities of rats with Alzhheimer’s disease(AD)induced by D-galactose combined with amyloid β-protein(Aβ25-35 ),and provide experimental basis for the prevention and treatemtn of AD. Methods A total of 90 SD male rats were randomly divided into model control group,piracetam group,sham operated group,Bunao capsule(0.79,1.58,3.15 g·kg-1 )groups(n= 15 each).The rat models were established by intraperitoneal injection of D-galactose and injection of Aβ25-35 into the bilateral lateral cerebral ventricle.Then rats were given corresponding drugs by gavage in different groups for 8 weeks.The learning and memory abilities were meseured by Morris water maze test.The morphology of brain cells was observed by HE staining.The activities of glutathione peroxidase(GSH-Px)and superoxide dismutase( SOD),and the malondialdehyde( MDA)contents in the brain tissues were measured by spectrophotometry. Results The target quadrant residence time was(20.39±7.75)s and(20.82±5.09)s in Bunao capsule (1.58,3.15 g·kg-1 )groups,which were significantly increased as compared with that in model control group[(12. 35 ± 6.95)s](P<0.01).Brain nerve cell morphology in Bunao capsule(1.58,3.15 g·kg-1 )groups was obviously improved as compared with that in model control group,and was close to that in sham operated group.The activities of GSH-Px and SOD were significantly increased,and MDA contents decreased in Bunao capsule groups as compared with those in model control group (P<0.01). Conclusion Bunao capsule can dose-dependently improve the learning,memory and antioxidative abilities of AD rats.The mechanism may involve upregulation of antioxidative enzyme activities and removal of oxidative products.

AIM:To investigate the changes of histone modifications during the activation of primarily cultured rat hepatic stellate cells ( HSCs) and the relationship between histone modification patterns andα-smooth muscle actin (α-SMA) expression, and to explore the roles of histone modifications in the activation of HSCs.METHODS:The rat HSCs were isolated by in situ perfusion of collagenase combined with density gradient centrifugation, cultured in vitro and identi-fied by immunofluorescence staining.The morphological features of the cells were observed under inverted microscope.The changes of desmin and α-SMA during the activation of HSCs were detected by immunofluorescence staining and Western blotting.The levels of histone 3 lysine 4 dimethylation (H3K4me2), histone 3 lysine 9 dimethylation (H3K9me2), his-tone 3 lysine 9 acetylation (acH3K9) and histone 4 lysine 12 acetylation (acH4K12) in quiescent HSCs and activated HSCs were determined by Western blotting.RESULTS: The morphology of HSCs shifted from a quiescent phenotype to highly activated myofibroblast during the culture.Immunofluorescence staining and Western blotting showed that the expres-sion levels of α-SMA and desmin were increased over time and reached maximum at 15 d.According to the results of cell morphology and immunofluorescence staining, the cells cultured for 24 h and 15 d were quiescent and activated HSCs, re-spectively.Compared with quiescent HSCs, there were higher H3K4me2 and lower H3K9me2, acH3K9 and acH4K12 modification levels in activated HSCs ( P<0.01 ) .CONCLUSION: Histone modifications show anomalous expression during the activation of primarily cultured rat HSCs.Histone modifications may contribute to the transdifferentiation of HSCs and the development of hepatic fibrosis.

OBJECTIVE:To study the effects of Tanshinone capsules on glucose and lipid metabolism and serum sex hormone level in rats with polycystic ovary syndrome (PCOS). METHODS:90 female SD rats were randomly divided into normal group (distilled water),model group (distilled water),positive control group (metformin 200 mg/kg) and Tanshinone capsules low-dose,medium-dose and high-dose groups(30,60,90 mg/kg),with 15 rats in each group. Those groups were induced PCOS mod-el except for normal group;after modeling,those groups were given relevant medicine intragastrically for consecutive 28 d,once a day. The ovulation rate was compared before intervention and 28 d after intervention. Body weight,fat wet weight,fasting insulin (FINS),fasting blood-glucose (FPG),AUC of glucose,HOMA-IR,ISI,serum levels of FSH,LH and T were detected. RE-SULTS:Compared with normal group,anovulation,body weight,fat wet weight,FINS,FPG,AUC of glucose,HOMA-IR, FSH,LH and T were all increased significantly in model group,while ISI decreased significantly(P<0.05). Compared with mod-el group,ovulation rate and ISI of positive control group and Tanshinone capsules groups were increased significantly after interven-tion, while body weight (except for Tanshinone capsules low-dose group), fat wet weight (except for Tanshinone capsules low-dose group),FINS,FPG,AUC of glucose,HOMA-IR,FSH,LH and T were all decreased significantly(P<0.05),especial-ly in Tanshinone capsules high-dose group and positive control group. CONCLUSIONS:Tanshinone capsules can regulate glucose and lipid metabolism and serum sex hormone secretion disorder in PCOS rats.

OBJECTIVE:To investigate the preventive and therapeutic effect of Shuganning injection on alcoholic liver fibrosis (ALF) in model rats,and provide experimental basis for its clinical application for alcoholic liver disease. METHODS:50 rats were enrolled and intraperitoneally given mixed liquid of 60% alcohol-corn oil-pyrazole to reduce ALF model. Another 10 rats were enrolled and intraperitoneally given normal saline,as normal control group. After 16 weeks,survived model rats(n=40)were ran-domly divided into model group,positive control group(Anluo huaxian pill 0.75 g/kg,ig),Shuganning injection high-dose,medi-um-dose,low-dose groups(4.8,2.4,1.2 mL/kg,ip),8 in each group. Normal control group and model group were intraperitone-ally injected equal volume of normal saline (5 mL/kg),administration groups were given relevant medicines,once a day,for 8 weeks;and modeling was contiuously conducted at the same time. After administration,body mass of rats was weighed,and the levels of liver function indexes [aspartate aminotransferase(AST),alanine aminotransferase(ALT)] and liver fibrosis indexes [hyal-uronic acid(HA),laminin(LN),type Ⅲ procollagen(PⅢNP),type Ⅳ collagen(Ⅳ-C)] in serum of rats were detected. Liver index of rats was determined and pathological changes of liver tissue were observed. RESULTS:Compared with normal control group,body mass of rats in model group was significantly decreased(P<0.05);liver index,and liver function index,liver fibro-sis index levels in serum were significantly increased (P<0.05 or P<0.01). Liver tissue showed steatosis,hepatocyte vacuoliza-tion,a large number of fibrous tissue deposition around portal areas and other pathological changes. Compared with model group,above-mentioned changes were improved significantly in administration groups (P<0.05 or P<0.01). CONCLUSIONS:Shugan-ning injection can obviously improve liver tissue damage of model rats with ALF,showing certain preventive and therapeutic effect on alcoholic liver disease.

OBJECTIVETo compare the acute toxicity and content of daphnoretin among extracts of unprocessed indian string-bush root and its two different processed products, and to provide a basis for discussion of the mechanism of two processed methods.

METHODExtracts of unprocessed indian stringbush root and processed indian stringbush root with "sweat" and "artificial sweat" were prepared. The mice were intragastrically administrated once with these three extracts, the mortalities of mice were observed, and the median lethal dose (LD50) of different extracts were calculated with Bliss method. The determination of daphnoretin in these three samples was performed by high performance liquid chromatography.

RESULTThe LD50 of indian stringbush root extracts, indian stringbush root processed with "sweat" and with "artificial sweat" were 46.678, 72.190, 67.953 g x kg(-1), respectively. The contents of daphnoretin in unprocessed indian stringbush root, indian stringbush root processed with "sweat" and with "artificial sweat" were 0.189%, 0.407% and 0.345%, respectively.

CONCLUSIONThe toxicity of indian stringbush root processed with both "sweat" and "artificial sweat" is lower than that of the original rude drug. But the decreasion of toxicity of processed products is not by the reduced daphoretin content.

Animals ; Chromatography, High Pressure Liquid ; Coumarins ; chemistry ; toxicity ; Female ; Lethal Dose 50 ; Male ; Mice ; Plant Extracts ; chemistry ; toxicity ; Plant Roots ; chemistry ; Toxicity Tests ; Wikstroemia ; chemistry