BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.

Objective:To explore the effect of high glucose on formation of endogenetic toxins and expression of intercellular cell adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1(VCAM-1) of human brain microvascular endothelial cells.Methods:Cultured HBMEC was used as target cells,anhydrous glucose was added to the cell medium to prepare the injury model of human endothelial cell.Methyl thiazolyl tetrazolium method was used to examine the cell activity in a high glucose environment.Immunocytochemistry method was used to detect the expression of ICAM-1 and VCAM-1 of HBMEC.Results:The expressions of ICAM-1 and VCAM-1of HBMEC were enhanced by high glucose after pretreating HBMEC for 24,48 and 72h(P

Objective:To observe the effect of three-stage sequential therapy on the functions of hypothalamic-pituitary-adrenal(HPA)axis and levels of the glucocorticoid receptor in patients with steroid-dependent asthma.Methods:55 patients with SDA were divided randomly into two groups,29 patients were treated with three-stage sequential recipes plus Pulmicort Turbuhaler(400?g/d)in the treatment group,while the other 26 patients with Pulmicort Turbuhaler(800?g/d)in the control group. The therapeutic courses were both 12-14 weeks.The plasma concentrations of ACTH and COR and the levels of PBMC GCR were determined before and after treatment.Results:The plasma concentrations of ACTH and COR and the levels of PBMC GCR in the two groups were all increased remarkably and there were significant difference between the two groups after treatment(P

ObjectiveTo investigate the laws of distribution and combination of traditional Chinese medicine (TCM) syndromes of patients with diastolic heart failure (DHF); To research the relevance between TCM syndromes and DHF.Methods A four diagnostic information survey was conducted among 124 DHF patients in order to analyze their basic information and information about TCM four diagnostic methods. Statistical analysis method was used to determine the syndrome types of the DHF patients and the features of syndrome combination.ResultsMost DHF patients were in their sixties or seventies, the number of female patients was larger than that of male patients, and their heart function was in II or III grade. Qi deficiency, phlegm turbidity, Qi stagnation, blood stasis, water stagnation, Yang deficiency and Yin deficiency were the common syndromes. Two-card combination and three-card combination were more common, mainly with Qi deficiency, phlegm turbidity or blood stasis as the main patterns of manifestation. Conclusion This research could provide a clinical basis for the establishment of the standard for the TCM syndromes DHF.

BACKGROUND: Chronic congestive heart failure occurs at the late stage of many heart diseases. Cognitive disability exists in chronic congestive heart failure which has been reported in most clinical studies, and measures that can successfully improve heart function can improve cognitive function.OBJECTIVE: To probe into the effect of chronic congestive heart failure after myocardial infarction on spatial learning and memory ability,and to observe the correlation between heart failure and cognitive function.DESIGN: A randomized controlled observation based on the experimental animals.SETTING: Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine), Ministry of Education.MATERIALS: This experiment was performed at the Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine),Ministry of Education. in November 2001. Totally 100 healthy male rats aged 10 to 12 weeks, weighing 200 g to 220 g purchased from Beijing Weitong Lihua Experimental animal Technical Co. Ltd were enrolled and randomly divided into the operation group (n=60) and sham-operation group (n=40).METHODS: Model rats in the operation group were infarcted by ligation of the left anterior descending coronary arteries to produce acute myocardial infarction models. Those in the sham-operation group left a loose tie without ligation. The rats that showed myocardial infarction-like changes through electrocardiogram were taken as the rats for operation (n=38), and those survived rats without abnormal changes and without ligation were taken as the sham-operation group (n=25). The two groups were subdivided into 10-day gro up, 30-day group and 60-day group. Morris water maze test was performed 10, 30 and 60 days after the operation followed by hemodynamic monitoring. Among the 10-day group, 30-day group and 60-day group, rats that did not complete the whole Morris swimming test due to heart failure with poor power and died 2 days after the operation or after hemodynamic monitoring were set as heart failure end period group.Rats in the same group as those died with similar body mass were set as the control group.MAIN OUTCOME MEASURES: ① Escape logarithm latency and the good rate if cognitive ability; ② stroke volume, cardiac output per minute,heart rate and cardiac index of hemodynamics, in which cardiac index was the main index for responding heart functionRESULTS: Totally 33 rats met the criteria in the operation group and 25rats in the sham-operation group. All the animals entered the stage of result analysis. ①Results of hymodynamic index: Stroke volume, cardiac output per minute and cardiac index of the rats in the 10-day group, 30-day group and 60-day group were significantly lower than those of the homochronous sham-operation group (P ＜ 0.01), and those indexes were significantly lower in the heart failure end period group than in the end period control group and each period of the operation (P ＜ 0.01-0.05). As compared with homochronous control group, the cardiac index in 10-day group, 30-day group, 60-day group and end period group was 51.21%,50.58%,55.84% and 33.91% respectively, and all were 60% lower than the normal value. ② Results of Morris water maze: Logarithm latency of 10-day operation group after training of 8 times was obviously longer than that of the 10-day sham-operation group [F(1,28) =5.997,P=0.021]. As for cognitive function, it was obviously worse in the 10-day operation group than that of the 10-day sham-operation group (x2=8.142 ,P ＜ 0.005). There was no significant difference between the 30-day operation group and the 30-day sham-operation group, and there was no significant difference in the logarithm latency as well between the 60-day operation group and the 60-day sham-operation group. As for searching strategies, the good rate of general cognitive ability in the 60-day operation group was worse than that in the 60-day sham-operation group (x2=4.988 ,P ＜ 0.05). Logarithm latency prolonged significantly in the heart failure end period group than in the end period control group [F( 1,6)=19.567 ,P=0.004], and the good rate of general cognitive ability was worse in the heart failure end period than that in the end period control group(x=1 1.82,P ＜ 0.01).CONCLUSION: Spatial learning and memory ability after heart failure in rats with myocardial infarction was injured obviously 10 days after the operation, and it recovered to the normal level 30 days after the operation.Cognitive injury occurred to some extent 60 days after the operation, and the cognitive function was obviously worse at the end period of congestive heart failure.

To explore the effects of Shengmai injection and Xuesaitong injection, compound Chinese herbal medicines for replenishing qi and activating blood, on ventricular fibrillation threshold, heart structure and connexin 43 (Cx43) expression in rats with myocardial infarction (MI).

0.05).Conclusion: Three-stage sequential method could recover the functions of HPA axis of asthmatic rat models with steroid intervention.

To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-kappaB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs.

To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis.

Objective: To observe the expression of cell adhesion molecules ICAM-1 and VCAM-1of cultured rat brain microvascular endothelial cells(MVEC),expecting to explore the mechanisms of new QingKaiLing injection protecting brain from injury of inflammatory cascade in cerebral ischemia diseases.Methods: Rat cerebral MVEC were extracted by separating microvessel sections and collagenase enzymatic digesting,an in vitro ischemia reperfusion model was established(Kreb,95%N2+5%CO2),the protein and mRNA expression of ICAM-1 and VCAM-1 were detected by using immunocytochemical stain and RT-PCR method.Results:The expression of adhesion molecules of model group were significantly higher than those of noral group(P