OBJECTIVETo observe the change in the amount of sialic acids on hepatocellular carcinoma (HCC) cell membrane.

METHODSSurgical specimens of HCC and liver cirrhosis tissues were obtained from 28 patients to prepare carcinoma cell and hepatocyte suspensions by collagenase digestion. For assay of α2, 3 and α2, 6-sialic acids, the cells were suspended in the staining buffer containing either fluorescein isothiocyanate-Maackia amurensis lectin (FITC-MAL) or fluorescein isothiocyanate-Sambucus nigra bark lectin (FITC-SNA) and incubated for 1 h, respectively. Flow cytometric analysis was carried out to measure the mean fluorescence intensity (MFI) on the cell surface.

RESULTSIn both FITC-MAL- and FITC-SNA-incubated HCC cells, the MFI on the cell surface was greater than that of the hepatocytes.

CONCLUSIONBoth of α2, 3 and α2, 6- sialic acids increases significantly on the hepatocyte membrane after the carcinomatous change.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Membrane ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Sialic Acids ; metabolism

OBJECTIVETo explore practical protocols for cloning bone marrow-derived hepatic stem cells in vitro.

METHODSThe cell fraction rich in CD117(+) cells and CD184(+) cells was separated from fresh bone marrow by density gradient centrifugation and cultured for 0, 7 and 14 days in high-glucose DMEM supplemented with or without 10% autologous serum or in serum-free high-glucose DMEM. All the media were supplemented with different concentrations of hepatocyte growth promoting factors (HGPF), thrombopoietin (TPO) and interleukin-3 (IL-3). The quantitative changes of CD117(+) cells and CD184(+) cells were measured by flow cytometry.

RESULTSThe optimal effect for cell cloning was achieved with high-glucose DMEM with 10% autologous serum group supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3. At day 7 of cell culture in this media, the quantity of CD117(+) cells and CD184(+) cells increased by 6.55 and 6.20 folds, and by 11.62 and 20.57 folds at day 14, respectively.

CONCLUSIONIt is practical for cloning bone marrow-derived hepatic stem cells in high-glucose DMEM with 10% autologous serum supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3.

Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Clone Cells ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; physiology ; Humans ; Liver ; cytology ; Proto-Oncogene Proteins c-kit ; metabolism ; Stem Cells ; cytology ; Thrombopoietin ; pharmacology

OBJECTIVETo establish a mouse model of biliary obstruction.

METHODSSixty-four Balb/c mice were divided into experimental group and control group. Obstructive jaundice was induced in the mice in the experimental group by common bile duct ligation. The level of the common bile duct diameter, WBC, LYM MID, LYM%, MID% and ALT, AST, TBIL, DBIL, IBIL, ALP and CHOL were measured 12 h and 1, 2 ,3, 4, 5, and 7 days after the ligation. The morphological changes in the liver were also observed.

RESULTSThe level of common bile duct diameter, WBC, LYM, MID, LYM%, MID% and ALT, AST, TBIL, DBIL, ALP and CHOL all underwent changes with time following certain patterns.

CONCLUSIONThe jaundice manifestation of this model is similar to that of patients with biliary obstruction, and this model may provide a reliable model for studying the mechanism of obstructive jaundice.

Animals ; Cholestasis, Extrahepatic ; pathology ; Common Bile Duct ; pathology ; surgery ; Disease Models, Animal ; Female ; Ligation ; Liver ; pathology ; Mice ; Mice, Inbred BALB C

OBJECTIVETo determine the optimal cytokine combinations with hepatic growth factor (HGF) that results in the most significant simultaneous in vitro expansion of cc-kit(+)Lin(-) cells derived from the bone marrow.

METHODSC-kit(+)Lin(-) cells were isolated from mouse bone marrow using a high-gradient magnetic cell sorting system (MACS) and expanded in the presence of stem cell factor (SCF), FLt-3 ligand (FL), leukemia inhibitor factor (LIF) thrombopoietin (TPO) and different concentrations of HGF for 7days in a liquid culture system. The total cell number and Annexin-V-positive cell number were counted, and the antigen expressions were studied with fluorescence-activated cell sorting (FACS).

RESULTSIn each group, c-kit(+)Lin(-) cells were expanded effectively and rapidly by 2 to 8 folds. Addition of 10 ng/ml HGF into SCF+FL+LIF+TPO resulted in the most significant expansion of c-kit(+)Lin(-) and total cells by 8.00 and 45.43 folds, respectively, with cell apoptosis rate of 17.42 %. But as the concentration of HGF increased, the c-kit(+)Lin(-) cells and the apoptosis rate decreased.

CONCLUSIONHGF at10 ng/ml shows optimal synergistic effect with SCF, FL, LIF and TPO in expansion of c-kit(+)Lin(-) cells, and excessive HGF may induce cell differentiation.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Count ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Flow Cytometry ; Hepatocyte Growth Factor ; pharmacology ; Mice ; Mice, Inbred BALB C ; Proto-Oncogene Proteins c-kit ; metabolism

Objective To investigate the expression of Th17/Th1cells in peripheral blood and its relationship with rapid virologic response (RVR) to the therapy with pegylated interferon (PEG-IFN)combined with rib(a)ivirin (RBV) in patients with chronic hepatitis C (CHC).Methods Thirty one CHC patients treated with PEG-IFN combined with RBV were enrolled,and the demographic and clinical data including age,gender,body mass index (BMI),results of biochemical parameters in serum and the peripheral blood samples at baseline were collected. Eleven healthy volunteers were enrolled as control group.The RVR in 4th week were observed by prospective experimental research method. The ratio of CD3 +,CD8 -,IL-17 +,Th17 and CD3 +,CD8-,IFN-γ+,Th1in peripheral blood mononuclear cells (PBMCs) were detected at baseline by flow cytometry.Results Compared to the healthy control group,the level of Th1cells was decreased and the level of Th17 cells was increased in patients with CHC.In non-RVR group of CHC,compared to the healthy control group,the level of Th1cells was decreased significantly,and the level of Th17 cells was increased significantly ( P ＜ 0.05 ). Conclusions There was Th1/Th17 subgroups imbalance in no-RVR CHC patients treated with PEG-IFN combined with RBV,the level of Th1cells was reduced,and the level of Th17 cells was increased.The high level of TH17 cells and low level of Th1cells at baseline may not be conducive to the RVR.

OBJECTIVETo ascertain whether mouse c-Kit(+)Lin- bone marrow cells have the potential of hepatic stem cells.

METHODSc-Kit(+)lin- bone marrow cells were isolated and purified by magnetic-activated cell sorting (MACS) from BALB/C male donor mice, and immediately transplanted into age-matched BALB/C syngeneic female mice with 35-Gy total liver irradiation. The recipients were sacrificed 1 month after the transplantation for pathological observation of the liver morphology. The presence of Y-chromosome was examined in the liver cells of the recipient by in situ hybridization (ISH), and alpha-fetoprotein (AFP) and albumin in the cells were detected by immunohistochemistry.

RESULTSThe hepatocytes positive for Sry gene on Y-chromosome were identified 1 month after transplantation, and immunohistochemistry for AFP and albumin confirmed that the donor mice-derived cells were hepatocytes.

CONCLUSIONc-Kit(+)lin- bone marrow cells have the potential of hepatic stem cells, which can reside and differentiate into hepatocytes in the liver after transplantation. c-Kit(+)lin- bone marrow cells can be used as the source cells of cell transplantation for liver disease.

Animals ; Bone Marrow Transplantation ; methods ; Cell Differentiation ; Female ; Hepatocytes ; cytology ; metabolism ; Immunohistochemistry ; Male ; Mice ; Mice, Inbred BALB C ; Multipotent Stem Cells ; metabolism ; transplantation ; Proto-Oncogene Proteins c-kit ; metabolism ; Random Allocation ; Whole-Body Irradiation ; alpha-Fetoproteins ; metabolism

Animals ; Bone Neoplasms ; pathology ; Cell Line, Tumor ; Cell Movement ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Osteosarcoma ; pathology ; secondary

Academies and Institutes ; Disease ; Humans ; Integrative Medicine ; trends ; Research ; trends ; Science ; trends ; Taiwan

OBJECTIVETo evaluate the therapeutic effect of in vitro induced autologous bone marrow-derived liver stem cell transplantation for posthepatitic cirrhosis.

METHODSBetween Jun 2008 and Mar 2009, 12 patients with posthepatitic cirrhosis and portal hypertensive underwent azygousportal disconnection and splenectomy in our department. The patients were then divided into two groups to receive autologous bone marrow-deprived liver stem cell infusion via the hepatic artery after in vitro induction for 7 days (n=6) or saline (n=6). The therapeutic effects of the operations on the liver functions and liver fibrosis index were evaluated.

RESULTSAll the patients recovered uneventfully and no side effect of the operation was found. After the operation, the patients receiving bone marrow-deprived liver stem cell infusion showed better hepatic function improvement than those receiving saline infusion (P<0.05).

CONCLUSIONTransplantation of in vitro induced autologous bone marrow-derived liver stem cell via the hepatic artery is safe and effective for treatment of posthepatitic cirrhosis.

Adult ; Bone Marrow Cells ; cytology ; Female ; Hepatitis, Viral, Human ; complications ; Humans ; Liver Cirrhosis ; etiology ; therapy ; virology ; Male ; Middle Aged ; Stem Cell Transplantation ; Transplantation, Autologous

Objective To investigate the effects and cost of four formulations of niclosamide ethanolamine salt in Oncomela-nia hupensis snail control in the field in marshland and lake regions,so as to provide the evidence for drawing up the plan of mol-luscicide using in schistosomiasis endemic areas. Methods One drainage channel and one channel without water in the same area with snails in Jiangling County,Jingzhou City were selected as the research fields. The drainage channel was divided into 9 sections,except one section as a blank control group where the natural death rate of snails was observed only,and the remaining 8 sections were taken as the observation groups,where different dosages of 4％niclosamide ethanolamine salt powder,5％ni-closamide ethanolamine salt granules,25％ niclosamide ethanolamine salt suspending agent,26％ metaldehyde and ni-closamide ethanolamine salt suspending agent,and 50％niclosamide ethanolamine salt wettable powder were used re-spectively. The channel without water were divided into 4 sections,except one section as a blank control group,the oth-er 3 segments were taken as the observation groups,where 4％niclosamide ethanolamine salt powder,5％niclosamide ethanolamine salt granules,and 50％ niclosamide ethanolamine salt wettable powder were used respectively. Before and after spraying molluscicide for 7 days and 15 days,the system sampling method was used to observe the effects of snail control. Meanwhile,the unit cost method was used to calculate the costs of the different mulluscicide formulations abovementioned in unit area(1 m2). Results In the field at the drainage channel,the snail mortality rates of the groups spraying 4％niclosamide ethanolamine salt powder(50 g/m2),5％niclosamide ethanolamine salt granules(40 g/m2),25％ niclosamide ethanolamine suspending agent,26％ metaldehyde and niclosamide ethanolamine salt sus-pending agent,and 50％niclosamide ethanolamine salt wettable powder(2 g/m2 and 4 g/m2)for 7 days were 79.52％-97.87％,while the rates after spraying for 15 days were 71.00％-96.30％,and compared with those before spraying, the differences were statistically significant(all P<0.01). For the groups spraying with 2 g/m2 or 4 g/m2 suspending agent as well as wettable powder for 7 days,the snail mortality rates were significantly different(both P<0.05). In the field at the channel without water,the snail mortality rates of the 3 observation groups after spraying molluscicide for 7 days were 97.14％-100％,while for 15 days were 94.32％-100％,and compared with the rates before spraying,all the differences were statistically significant(all P<0.01). The unit costs per 1 m2 of the molluscicide abovementioned were ranged from 0.280 Yuan to 0.416 Yuan. Conclusions In marshland area inside embankment,the molluscicide formulations of the powder and granule are suitable for the environments without water or with instability water level , while the molluscicide formulations of the suspended agents and wettable powder are suitable for the water environment. Though the unit cost of powder is the lowest,the molluscicide in this formulation flies away seriously.