AIM:There is no standard isolation method of mesenchymal stem cells.Percoll density gradient centrifugation is treated as the traditional method among numerous methods but it is so complicated.This study compares whole bone marrow culture to traditional Percoll density gradient centrifugation in order to establish a better method in vitro.METHODS:Experiments were conducted at Central Laboratory of Affiliated Hospital of Qingdao University Medical College from September 2006 to June 2007.Bone marrow was supported by patients who had autologous stem cell transplantation by endocrinology department for diabetes treatment.The experiments had known by the patients and authorized by the hospital ethic committee.The two different methods,whole bone marrow culture and Percoll density gradient centrifugation,were compared in the number of cell clone,cell morphology,cell superficial mark,the differentiation into fat cell.RESULTS:① The number of clone cell by whole bone marrow culture was more than by Percoll density gradient centrifugation(P

OBJECTIVE: Pancreatic stem cell transplantation is a new approach for diabetes treatment. It may overcome the shortage of human donor islets and avoid the long-term applying of immunity depressors. In this article, we review the recent progress of the sources, induced differentiation and the molecular markers of pancreatic stem cells. DATA SOURCES: A computer-based online search of PUBMED was undertaken to identify relative articles published in English from June 1997 to June 2006 with the keywords of "pancreatic, stem cell, differentiate, marker". Meanwhile, we searched Wanfang database for the related Chinese articles published between June 1997 and June 2006 with the same key words in Chinese. STUDY SELECTION: The articles were firstly selected and the quotations cited by every article were looked over. Inclusive criteria: The articles focusing on the source, induced differentiation and molecular markers of pancreatic stem cells. Exclusive criteria: Repeated experiments or Meta analysis articles. DATA EXTRACTION: According to the inclusive criteria, 31 articles were finally selected among the 82 ones collected from databases.Fifty-one old data or duplicated researches were excluded. Among the 31 selected articles, 6 presented the main conceptions, 15 were related to the sources of pancreatic stem cell and its induced differentiation, 9 were focused on the surface molecular markers and 2 discussed the major obstacles at present. DATA SYNTHESIS: Stem cells are pluripotential cells with the ability to self-renew; they are classified into two major classes: embryonic stem cell and adult stem cell. Pancreatic stem cells, belonging to the adult stem cells, could differentiate into tube cells, insulin-secreting cells, exocrine cells and other specific pancreatic tissue cells, which possess the fundamental properties of long-term proliferation and self-renewal. The studies on the sources, induced differentiation and molecular markers of pancreatic stem cells are extremely useful for the treatment of diabetes. Recent studies have shown that besides gene engineering, insulin-secreting cells are mainly generated from the differentiation of embryonic stem cells and adult stem cells such as pancreatic duct epithelial cells, nestin-expressing pancreatic stem/precursor cells, bone marrow mesenchymal stem cells and haematopoietic stem cells. The investigation on the molecular markers of pancreatic stem cell surface is essential to the basic and clinical research. PDX-1, nestin, CK-19, CK-20, Ngn3 and PGP9.5 are the major markers used in the identification, isolation and purification of pancreatic stem cells. CONCLUSION: The investigations on islet cells and pancreatic stem cells transplantations are promoting. Significant advancements in search for the sources and identification of pancreatic stem cells have been achieved. However, many obstacles should be overcome before any clinical breakthroughs can be expected. With the significant improvement in stem cell research and techniques, it is possible to obtain enough pancreatic stem cells from in vitro culture for transplantation.

Objective To assess the effects of tibolone treatment on the hemodynamic changes of the ophthalmic artery(OA) and internal carotid artery(ICA) as sonographic markers of atherosclerosis in menopausal women.Methods A total of 30 menopausal women were treated with 2.5mg of tibolone(Livial) daily as treatment group.At the same time,30 women with natural menopause who were not given any treatment served as control group.Noninvasive measurements of the resistance index(RI) and peak systolic blood flow velocity of ICA and OA were made with ultrasound before the treatment and at 3rd,6th,9th and 12th month of treatment,respectively.Results RI of ICA and OA decreased significantly after tibolone treatment(P

Objective To observe the effects of hormone replacement therapy (HRT) on endothelial function in menopausal women. Methods A total of 30 menopausal women were treated with 2.5 mg of Tibolone (Livial) daily. At the same time, 30 women with natural menopause without any treatment served as the control group. Endothelium-dependent (EDD), endothelium-independent (NID) vasodilatation function, and estradiol (E2) were examined by the non-invasive high-resolution ultrasonography before the treatment and at 12th, 24th, 36th and 48th week of treatment, respectively. Results After hormone treatment, E2 increased significantly and EDD was improved significantly (P<0.05), and E2 was positively related with EDD (r=0.8092, P<0.001). No change of EDD was observed in the control group whereas a significant increase was observed in the treatment group. Conclusion Endothelium-dependent vasodilatation dysfunction is prominent in menopausal women. Tibolone can help improve the condition.

Objective:To explore the effect of health education under KABP model on negative emotions and pregnancy outcome in patients with high-risk pregnancy.Methods:Choosing 180 patients with high-risk pregnancy as the research objectsfrom June 2014 to May 2016 in our hospital,we divided the patients into two groups using the random number table method,each for 90 cases.The control group was given comprehensive nursing;the observation group was given health education under KABP model on the basis of control group.We compared the differences of negative emotions and pregnancy outcome between the two groups.Results:Before nursing,there was no significant difference of the Self-Rating Anxiety Scale (SAS) score and Self-Rating Depression Scale (SDS) score between the two groups (P ＞ 0.05).After intervention,the fetal distress,neonatal asphyxia and postpartum hemorrhage rate of observation group were significantly lower than control group,while the rate of natural delivery of observation group was significantly higher than control group (P ＜ 0.05).Nursing satisfaction of observation group was significant higher than control group (P ＜ 0.05).Conclusion:The health education under KABL model helps to significantly improve negative emotions and pregnancy outcome in patients with high-risk pregnancy,thus significantly improve nursing quality.It is worthy of clinical popularization and application.

BACKGROUND: There are no studies on differentiation of bone marrow mesenchymal stem cells (BMSCs) into neurons directly induced by neural cells. OBJECTIVE: To establish the co-culture system between BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron in the co-culture system. DESIGN, TIME AND SETTING: The in vitro cytology control experiment was performed at the Central Laboratory of Hospital Affiliated to Medical College of Qingdao University from December 2006 to December 2007. MATERIALS: BMSCS were harvested from bone marrow of patients with diabetes meliuts, who underwent autologous stem cell transplantation at the Hospital Affiliated to Medical College of Qingdao University. Neural cells were collected from brain tissues of infants die of asphyxia during delivery. The third passage of neural cells was used in this study. Transwell double-deck culture dish was purchased from Corning Costar, with a pore diameter of less than 3.0 ?m. Cells could not traverse, but the medium could traverse. METHODS: Neural cells were incubated in Transwell double-deck culture dish at a density of 1?106 in the co-culture group. BMSCs were incubated in the upper layer in LG-DMEM medium for 4-5 days. BMSCs were incubated in both layers in the control group. MAIN OUTCOME MEASURES: The morphological changes of BMSCs were observed and the special markers of neurons cells in BMSCs were examined by immunofluorescence. RESULTS: BMSCs in the co-culture group grew slowly, showing radial processes and connected each other. Positive rate of neuron specific enolase was (32.7?11.5)%. BMSCs in the control group were flat and wide, and negative for neuron specific enolase. CONCLUSION: Microenvironment provided by neural cells promotes the differentiation of BMSCs into neurons.

BACKGROUND: There is no ideal method about adult bone marrow mesenchymal stem cells (BMSCs) differentiate into islet-secreting cells and appreciation in vitro at present. Transgene requests strict conditions and complex program. The induction using chemicals is a present-used method. OBJECTIVE: To investigate the culture conditions for inducing adult BMSCs into islet-like cells in vitro. DESIGN, TIME AND SETTING: The cytology in vitro experiment was performed at the Central Laboratory of Affiliated Hospital of Medical College of Qingdao University from January to October 2007. MATERIALS: Bone marrow was collected from diabetic patients undergoing autologous stem cell transplantation at Department of Endocrinology, Affiliated Hospital of Medical College of Qingdao University after signing the informed consent. Epidermal growth factor and basic fibroblast growth factor were obtained from Peprotech Asia. B27 adjunct was purchased from Gibco. Nicotinamide, 2-mercaptoethanol, glutamine and dithizone were bought from Sigma, USA. METHODS: The adult BMSCs were isolated by Percoll from adult bone marrow aspirates and cultured in LG-DMEM, were suspended by Trypsin and passaged for subsequent passages. At the third to fifth passages, BMSCs were incubated at a density of 1?108 L-1 and were induced differentiation into islet-like cells through three developmental stages. In the first stage, BMSCs were incubated in HG-DMEM supplemented with 2-mercaptoethanol, glutamine for 2 days. In the second stage, BMSCs were incubated in HG-DMEM supplemented with epidermal growth factor, basic fibroblast growth factor, B27 and glutamine for 6 days. In the third stage, BMSCs were incubated in HG-DMEM supplemented with nicotinamide and 2-mercaptoethanol for 6 days. BMSCS in the control group were only incubated in the HG-DMEM. MAIN OUTCOME MEASURES: Morphological changes in BMSCs were analyzed under a phase contrast microscopy. Duodenal Homeobox 1 (PDX-1) expression was detected during the second and differentiation stage by immunofluorescence assay. Islet ?-like cell clusters from the 3rd stage were identified by positive dithizone staining. The insulin secretions under the stimulation of high or low glucose were detected by electrochemiluminescence immunoassay. RESULTS: The undifferentiated BMSCs exhibited adherent long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells expressed the PDX-1 gene at 8 days and formed islet-like cell clusters that exhibited positive dithizone staining at 14 days. After high and low glucose treatment, no insulin was detected in the control group; insulin content significantly increased at 14 days (t=3.638-9.387, P

Objective To study the changes in the central nervous system in adult mice after various degrees of acute heat stress. Methods Adult mice placed in insulated cages were exposed to 24℃, 34℃, 37℃, 38.5℃, 40℃ or 42℃ for 60min respectively with a constant humidity of 60%. The behavior response was carefully monitored. Rectal temperature was measured before the exposure and after the exposure. Electroencephagrams were taken. Then the hippocampal neurons of these animals were examined with transmission electron microscope. Results Heat stress at 34℃ for 60 min only raised the rectal temperature, and heat stress above 37℃ for 60 min not only raised rectal temperature, but also induced water loss and irritability and attempt to escape. Abnormal EEG with increased amplitude and retarded rhythm could be observed. However, when mice were exposed to 24℃ and 34℃ such behavior did not appear. After exposure to 42℃, EEG frequency increased and pathological changes in hippocampus neurons were found. The main ultrastructural changes included degeneration of hippocampal neurons, expansion of space around capillary, decrease in number of synaptic vesicle, and decomposition of synapse. Conclusion Mice were very sensitive to heat stress. An increase in core temperature could induce behavioral thermoregulation. EEG and electron microscopic study revealed changes in the central nervous system after heat stress. Following exposure to high environmental temperature under 40℃, acute dysfunction of brain was reversible. At 42℃, damage to the brain tissue occurred, and most mice died of heat stroke.

4-Chloromercuribenzenesulfonate ; pharmacology ; Adult ; Case-Control Studies ; Female ; Graves Disease ; metabolism ; physiopathology ; Humans ; Hydrogen Peroxide ; metabolism ; Iodine ; metabolism ; Male ; NADPH-Ferrihemoprotein Reductase ; metabolism ; Thyroid Function Tests

Objective To study the joint effect of small dosages of Aspirin and compound Danshen dropping pills on type 2 diabetes(T2DM) patients'and impaired glucose tolerance(IGT) patients' hypercoagulation blood.Methods The study followed diagnostic standards for T2DM by World Health Organization (WHO) in 1999,and randomly selected two groups of clinical patients:one had 63 patients with IGT and the other contained 68 patients with T2DM.Both groups took compound Danshen dropping pills with the total dosage of 10 pills each time,3 times per day,for continuum of 3 months.After that both groups stopped medication for 7 days,then began taking Aspirin after meal,with the dosage of 100 mg per day,for a continuum of 3 month.After that both groups stopped the medication for 7 days,then both group continued to take same dosage of ASA with addition of compound Danshen dropping pills of total amount of 10 pills per each time,3 times per day,for acontinuum of 3 months.Each medication period (ev ery 3 months) was monitored for platelet count (PET),prothrombin time (PT),kaolin partial thromboplastin time (APPT),The effective rate of anticoagulation for both groups were then calculated.Results The difference of PT、APTT between before and after taking single dosage of ASA for patients with impaired glucose tolerance (IGT) was statistically significant (P＜0.05),while patients with T2DM had no significant difference before and after taking same single dosage of ASA.Similarly, both groups with single dosage of compound Danshen dropping pills had no significant difference between before and after taking the medication.However,the difference of PT、APTT between before and after taking the combination of Aspirin and Danshen dropping pills of both groups were statistically significant (P＜0.05).IGT group with single dosage of ASA had an effective rate of 71.4％ for anti coagulation,while T2DM group with the same dosage reached 23.8％.The IGT group with combinatory dosage of ASA and Danshen dropping pills reached an effective rate of 100％ for anticoagulation,while T2DM group with the same dosage was 86.7％.The difference between those effective rates of anticoagulation was statistically significant in different medicine takeways of two groups (P＜0.05).Conclusion The combinatory use of small dosage of Aspirin with compound Danshen dropping pills had evident effect to improve IGT and hypercoagulation blood of T2DM patients who had good control of their blood glucose.