Objective To discuss the possibility of differentiation of mesenchymal stem cells(MSCs)from umbilical cord blood(UCB)into hepatocyte-like cells in the in vitro culture.Methods MSCs were isolated from UCB,cultured and passaged.The surface markers were examined by flow cytometry.When cells were cultured to the third passage,they were inoculated into a 6-well plate.A two-stage induction method was used:MSCs in the first phase were cultured in DMEM/F12 medium supplemented with dexamethasone(final concentration of 0.5 μmol/L,the same below),hepatocyte growth factor(HGF,10 ng/ml),epidermal growth factor(10 ng/m1)and 1×insulin-transferrin-Se(ITS)for two weeks,then in DMEM/F12 supplemented with 0.5 μmol/L dexamethasone,10 ng/ml HGF,1×ITS,10 ng/ml Oncostatin M for another two weeks.Morphological changes were observed under a microscope.The gene expression correlated with hepatocytes was detected by using RT-PCR.Immunofluorescence staining was used to identify the expression of specific protein related to hepatocytes(AFP,Albumin,CK-18).Ultrastructure was detected under an electron microscope.Results In the cultured MSCs from UCB,CD34/CD45/CD14,CD54,CD49f and HLA-DR were not detected,there was low expression of CD106 and strong expression of CD29,CD44 and CD13.The gene expression of AFP,albumin,CK-18 and TAT was discovered and three kinds of protein AFP,albumin and CK-18 were positively showed in cytoplasm after 4 weeks'induction.The hepatin granules and fatty drops in cytoplasm of cells induced for 4 weeks were found under an electron microscope.Conclusion The MSCs fromUCB can differentiate into hepatocyte-like ceils in the in vitro culture under some conditions.

BACKGROUND: Reproductive activity of hepatocytes is limited. There are numerous studies concerning stem cells differentiation into hepatocytes, including embryonic stem cells, bone marrow cells, pancreas stem cells, neural stem cells, various sources of mesenchymal stem cells.OBJECTIVE: To explore the possibility of induction of mesenchymal stem cells (MSCs) from umbilical cord blood (UCB) into hepatocyte-like cells in vitro culture.DESIGN, TIME AND SETTING: A cytological in vitro study was performed at the Institute of Hematology, Medical College of Jinan University from October 2005 to April 2006.MATERIALS: Fetus cord blood was obtained from spontaneous delivery and caesarean delivered healthy pregnant women at the Department of Obstetrics and Gynecology, First Affiliated Hospital of Jinan University. The parturients signed the informed consents.METHODS: UCB-MSCs were incubated in vitro, and digested in trypsin-EDTA. The third passage of cells at 5 × 10~4 cells/cm~2 wereinoculated. Original medium was removed 48 hours later. Cells were washed in phosphate-buffered saline. In the first phase, cells were incubated in F12 medium supplemented with dexamethasone, hepatocyte growth factor, epidermal growth factor and 1 ×its for 2 weeks. In the second phase, cells were incubated in F12 medium containing dexamethasone, hepatocyte growth factor,oncostatin M and 1 × ITS for two weeks.MAIN OUTCOME MEASURES: The following parameters were measured: expression of surface marker of UCB-MSCs using flow cytometry, expression of related gene and protein of hepatocytes following induction respectively using RT-PCR and immunofluorescence staining.RESULTS: No CD34/CD45/CD14 of hematopoietic markers were detected, either no the CD54, CD49f, HLA-DR were found. The low expression of CD106 and high expression of CD29, CD44, CD13 were found. The gene expression of a-fetoprotein, albumin,CK-18 and TAT were discovered, and three kind of protein a-fetoprotein, albumin, CK-18 were positively observed in cytoplasm after 4 weeks of induction using immunofluorescence staining.CONCLUSION: UCB-MSCs are able to differentiate into hepatocyte-like cells in vitro culture following combination with many growth differentiation factors, such as dexamethasone, hepatocyte growth factor, epidermal growth factor, tumorigenesis M and ITS.

AIM: To explore the possibility of differentiating functional insulin-producing cells from human BM-derived stem cells. METHODS: Mesenchymal stem cells were isolated from human bone marrow. Then these cells were induced with epidermal growth factor, β-mercaptoethanol and high concentration of glucose. The gene expression related to islet β cells was detected by RT-PCR. Insulin in the treated cells was examined by immunocytochemistry. In addition, the levels of insulin secretion and glucose-stimulated insulin release were examined by microparticle enzyme immunoassay. Finally, the induced cells were implanted into the right renal subcapsular space of diabetic mice. Blood glucose levels were monitored 16 d after implantation. The right kidneys of the treated mice were harvested for immunohistochemistry. RESULTS: The key genes related to pancreatic β cells had been confirmed to express by PCR and insulin was detected by immunocytochemistry in differentiated human BM-derived stem cells induced by high glucose, which responded to glucose challenge. Furthermore, implantation of the cells in renal subcapsular space was able to lower the glucose levels in hyperglycemic mice. After 16 days, the implanted cells were determined still to be insulin positive cells by immunohistochemistry. CONCLUSION: These results indicate human BM-derived stem cells are capable of differentiating into functional insulin-producing cells and may represent a pool of cells for the treatment of diabetes.

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.

?AlM:To investigate the sensitive parameters of the anterior chamber changes with Pentacam anterior segment analysis system before and after laser peripheral iridectomy (LPl) in primary angle-closure suspetive (PACS).
? METHODS: Sixty eyes of 33 PACS patients were enrolled in this study. Pentacam examination was performed before and 1d after LPl to measure the central anterior chamber depth ( CACD ) , the peripheral anterior chamber depth ( PACD ) , the anterior chamber volume ( ACV) and the peripheral anterior chamber angle ( ACA) . Statistical analysis used paired t test.
?RESULTS: There was no statistical significance on the changes of ACD. PACD and ACV increased significantly between before and 1d after LPl. ACA was widened from (22. 26o±5. 18o) to (26. 42o±5. 20o), which were increased significantly between before and 1d after LPl.
?CONCLUSlON: LPl can deepen the PACD and increase the ACV in PACS. PACD and ACV are the sensitive parameters of the anterior chamber changes with Pentacam anterior segment analysis system.

Objects To observe whether mesenchymal stem cells exist in umbilical cord blood and if they can differentiate into osteoblasts and adipocytes.Methods Nucleated cells were separated with Hespan,Mesenchymal-like stem cells were identified by surface marker with flow cytometry.Osteoblast was identified by von Kossa staining and alkaline phosphatase staining,adipocyte was identified by Oil Red O staining.Results From the Nucleated fraction of UCB,we demonstrated the presence of a subset of cells that express adhesion molecules CD13+,CD29+,and CD44+,but not antigens of hematopoietic CD34,CD45,CD14 and neither antigens of endothelia CD106.Exposure of these cells to osteogenic inductive agents resulted in an increase in the expression of alkaline phosphatase and the appearance of hydroxyapatite nodules by Von Kossa staining.Incubation with adipogenic inductive agents resulted in morphological change and positive staining with oil Red O.Conclusions Mesenchymal-like stem cells exist in cord blood and have a lower precursor frequency.The cells can differentiate into osteoblast and adipocyte.Cord blood may function as a new source of cells for cellular therapeutics.

Objective To evaluate the diagnosis and surgical treatment of adult congenital choledochal cyst(ACCC). Methods Clinical data of 345 ACCC cases from 1974 to 2007 were analyzed retrospectively. Results There were only 110 patients (31.9%) with typical triad (abdominal pain, palpable mass and jaundice). B-ultrasound, CT, ERCP, PTC and MRCP were methods for diagnosis. Cancer arising from choledochal cysts was found in 19 cases who had undergone internal drainage and its incidence of cholangiocarcinoma is much higher than in the general population. All cases underwent surgical treatment, in which total excision of choledochal cyst and Roux-en-Y hepaticojejunostomy was main operation, 337 cases recovered well and 8 (2. 3%) died. C..ondusions Accurate diagnosis of ACCC was difficult depending on clinical symptoms only. B-ultrasound was the choice of atraumatic diagnosis. All patients with an established diagnosis of ACCC should undergo exploratory laparotomy. Total excision of eholedochal cyst and Roux-en-Y hepaticojejunostomy is most effective treatment for ACCC.

Objective To establish the reference intervals of total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C)and low density lipoprotein (LDL-C)in the healthy population in Hengshui region.Methods 16 403 healthy individuals were collected from the various counties and cities in this region and detected the above 4 blood lipid indexes (TC,TG,HDL-C, LDL-C)by adopting the automatic biochemical analyzer.The subjects were grouped according to different genders and ages and the detection results were performed the statistical analysis.Results TC 3.17-7.20 mmol/L;TG 0.08-3.16 mmol/L;HDL-C 0.94-2.30 mmol/L;LDL-C 1 .14 -4.59 mmol/L.Conclusion The differences of blood lipid detection results exist in males and fe-males in Hengshui region,the results are different among different age groups,it is suggested that different genders and different age groups should establish their own normal reference intervals in order to provide the diagnostic basis for clinic.

BACKGROUND:Arsenic trioxide is considered to inhibit the proliferation of vascular smooth muscle cel s and promote cel apoptosis. Therefore, we wondered whether the arsenic can inhibit the hyperplasia of vascular smooth muscle cel s, an arsenic-coated stent can be compatible with the vascular tissue, and a better vascular intimal coverage as early as possible can reduce intimal hyperplasia.
OBJECTIVE:To observe the vascular histocompatibility of the arsenic-coated stent.
METHODS:Fourteen white rabbits were randomized into two groups and respectively subject to the implantation of arsenic-coated 316 L stainless steel stents and bare 316 L stainless steel stents into the abdominal aorta. After 28 days, the distal and proximal parts of the vessel at the implantation site were ligated and the ligated vessel was taken for hematoxylin-eosin staining and light microscope observation.
RESULTS AND CONCLUSION:(1) Gross observation:the vessel at the stent site was a little larger than the adjacent vessels in the outer diameter, which was expanded but had no visible thrombus. After cutting the stent, the neointima formed smoothly on the stent surface. (2) Light microscope observation:the stent was located in the middle of the vessel, the medial smooth muscle was pressed, and vascular intimal smooth muscle hyperplasia was found around the stent, thereby thickening the vascular intima. The vascular neointima formed and covered the stent, and there was a thin black layer between the stent and the vascular tissue, which consisted of arsenic and its compounds. These findings suggest that the arsenic-coated stents can be covered with vascular tissues, possessing good vascular histocompatibility.

Objective To establish the reference interval of serum urea nitrogen (BUN) ,creatinine(Cr) ,uric acid(UA) for the healthy adults of Hengshui City .Methods 16 403 cases of healthy adults of different areas in the city were enrolled in the study .By using automatic biochemical analyzer ,3 kidney function indicators(BUN ,Cr ,UA) were determined .According to different sex and age groups ,statistical analysis was performed .Results The reference intervals of 3 kidney function indicators for healthy people in Hengshui City were as follows ,BUN :2 .75-7 .05 mmol/L for male and 2 .24-6 .58 mmol/L for female;Cr:49 .69-87 .96 μmol/L for male and 35 .82-72 .22 μmol/L for female;UA :209 .51 -500 .06 μmol/L for male and 146 .25 -389 .69 μmol/L for female . Conclusion The reference intervals established could basically reflect the concentrations of 3 kidney function indicators for the healthy adults in Hengshui City ,which could provide a reference .