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 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.

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 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

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

OBJECTIVETo explore the effect of Penqiangyan Granule (PG) on the immunity of patients with chronic pelvic inflammatory disease (CPID) of blood stasis and Shen-deficiency syndrome (BSSDS) type.

METHODSSixty patients were randomly assigned to two groups: the treatment group treated with PG and the control group with Penyanjing Granule, 30 cases in each group. The treatment course was 4 weeks for both groups. The clinical efficacy, plasma levels of CD4 and CD8, and the serum levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-2 (IL-2) were measured before and after treatment.

RESULTSAfter treatment, the total effective rate in the treatment group and the control group was 96.7% and 63.3% respectively with significant difference between groups (P < 0.05); in the treatment group the plasma CD4, CD4/CD8 and serum IL-2 increased obviously, while the plasma CD8 and serum TNF-alpha decreased markedly (P < 0.05), all were significantly different with those in the control group (P < 0.05).

CONCLUSIONPG can improve the immune function and alleviate inflammation in CPID patients of BSSDS type.

Adolescent ; Adult ; CD4-CD8 Ratio ; Chronic Disease ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-2 ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Pelvic Inflammatory Disease ; diagnosis ; drug therapy ; immunology ; Phytotherapy ; Syndrome ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; blood ; Yang Deficiency ; diagnosis ; drug therapy ; immunology ; Young Adult

Asthma is a common disease in children. Adherence to inhaled glucocorticoid therapy directly affects the outcome of asthma. The article mainly reviews the adherence status, assessment method, patient barriers to non-adherence and intervention strategy,so as to provide references for clinical practices and scientific research of nursing staff.

Objective To discuss the treatment mode of mass NBC casualties by field medical unit in NBC environment.Method Field medical unit rehearsed in a simulated NBC environment.Result The procedure for the rehearsal and treatment of field medical unit in NBC environment was determined.The importance of high-tech medical equipment for the obstructed patient flow was made clear.Conclusion High-tech medical equipment should be emphasized on to enhance the treatment capability of field medical unit.