There are plenty of haematopoietic stem cells in umbilical cord blood, which are regarded as important resources for transplantation therapy. There are some arguments on whether or not mesenchymal stem cells(MSCs) exist in umbilical cord blood. Some researchers have such opinions that there exist a number of MSCs in umbilical cord blood as similar with one from bone marrow in many aspects. Others believe that the content of MSCs in umbilical cord blood is too low to expand in vitro. We summarized the data from last five years researches. On umbilical cord blood MSCs during last to help further research and application of this resource of MSCs.

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: 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.

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.

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 investigate the synaptopathy of hidden hearing loss mice,and to observe the synapses of the cochlear inner hair cell after temporary threshold shift of noise exposure.Methods Mice were divided into normal control group and experiment group,the latter was exposed under noise of 98 dB SPL for 2 h to establish the model of temporary threshold shift.Mice cochleae of the two groups were dissected and prepared with whole mount and immunostaining.Cellular morphology was observed under confocal laser scanning microscope.Cochlear lengths were measured through cochlear frequency map to localize hair cells in different frequency regions.Then,3-D morphometry of synapses was constructed by Amira software to observe pre-synaptic ribbons,post-synaptic receptors and its pathological changes.Results In control group,each cochlear nerve fiber contacted a single inner hair cell by a single synapse,each inner hair cell had 5-30 synapses contacting cochlear nerve fibers.The larger ribbons patched smaller receptors located in the modiolar side,and the smaller ribbons patched larger receptors located in the pillar side.While in experiment group,noise overexposures caused moderate or completely reversible thresholds shift,i,e.,distortion product otoacoustic emission (DPOAE) and auditory brainstem response (ABR) thresholds increased 30-40 dB.Although returned to normal after 2 weeks,ABR wave Ⅰ amplitudes recovered to only 46.1 ％ of pre-exposure amplitudes.There was 41.3％ synapses loss of inner hair cell,but there was no loss of inner hair cells and spiral ganglion neurons.Conclusions Threshold test is not sensitive to degeneration and loss of synapse in mice inner hair cells,while super threshold test is sensitive to it.

Objective:To develop a qualitative analysis model for the fast identification of Dens Draconis and its adulterants by NIR correlation coefficient method. Methods:On the basis of the traditional morphological identification, the spectra were collected u-sing the fiber accessory of a near-infrared spectroradiometer. The reference spectra were set up using the NIR spectra of certified Dens Draconis. The characteristic spectral section was chosen and the appropriate threshold was set to establish a qualitative analysis model for the rapid identification of Dens Draconis and its adulterant. Results:The spectral section of 5 000-4 200 cm-1 was selected as the characteristic spectral section, the correlation coefficient of Dens Draconis and its adulterant was calculated in training set samples, and 92. 67% was used as the threshold. Totally 10 batches of validation set samples were validated the qualitative analysis model, and the prediction accuracy was 90%. Conclusion:The method has good prediction ability, and can be used in the rapid identification of Dens Draconis and its adulterant.

Objective To observe the characteristics of cognitive impairment in patients with primary hypothyroidism.Methods A total of 90 primary hypothyroidism patients untreated with thyroid hormone were selected.All the 90 patients were divided into subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group,and 30 patients in each group.The other 30 healthy volunteers were selected as controls.Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used for the testing their orientation,immediate memory,attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power.One-way analysis of variance was performed to determine significant differences among four groups.Results The MMSE scores in subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group((27.53 ± 2.16),(26.90±1.88) and (24.80 ± 2.10) respectively) were lower than those of the control (28.23 ± 1.33).The MoCA scores of the above hypothyroidism groups ((23.57 ± 3.33),(2 1.60 ± 2.81) and (20.53 ± 3.03) respectively) were also lower than that of the control (26.63 ± 2.31) (P ＜ 0.05).Except for orientation and immediate memory,statistical significances of the other cognitive function were existed between hypothyroidism groups and the healthy controls (P＜ 0.05).With the increase in severity of hypothyroidism,the abnormality of attention,calculation,linguistic competence,visual space and executive ability,naming ability and abstracting power were appearing gradually in hypothyroidism groups (P ＜ 0.05),and the scores were low(P＜ 0.05).Conclusion Defects of attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power are existed in primary hypothyroidism patients.

AimThe aim of this study was to establish a rodent model with similar characters of human metabolic syndrome(MS).Methods Three species mice and Wistar rats were fed with high energy chows(HEC)for 6 to 23 weeks.Animals were weighted every week.Fasting blood glucose(FBG)together with total cholesterol(TC)and low density lipoprotein-cholesterol(LDL-C)were investigated by oxidase test every two week.And fasting blood insulin(FINS)was determined by radioimmunoassay.Homeostasis model assessment of insulin resistance(HOMA-IR)was calculated as FBG×FINS/22.5.At the end of the experiment,oral glucose tolerance test(OGTT)was performed.Then animals were decapitated,and coel-fat and orchio-fat were collected and weighted to calculate the visceral fat coefficient(VFC).Results FBG,serum TC and LDL-C significantly increased(P<0.01)after 6 weeks feed of HEC in KM mice.The mice also formed abdominal obesity and insulin resistant together with impairment of glucose tolerance(P<0.05 or P<0.01).Though similar to the KM mice,C57BL/6 and BALB/c mice couldn't form abdominal obesity while the latter had increased body weight(P<0.05 or P<0.01).Wistar rats formed hyperlipidemia from 1 to 10 week and hyperglycemia from 10 to 23 week together with insulin resistance and impaired glucose tolerance(P<0.05 or P<0.01).Conclusion KM mice feed with HEC for 6 weeks could successfully establish metabolic syndrome mice model which might be suitable for drug-screening,the major characters includes the formation of abdominal obesity(increase of VFC),the increase of serum TC,LDL-C,FBG and HOMA-IR,and the decrease of OGTT.