Experimental membranous glomerulonephritis(MGN) in Japanese white rabbits was induced by administration of C-BSA according to Border's method. The results showed that the lipid peroxides(LPO) content in plasma and renal cortex were significantly increased compared with that in the controls. On the other hand, the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) in whole blood and renal cortex were significantly reduced compared with that in the controls. The results suggested that experimental MGN existed a mechanism of cellular injury manifested as lipid peroxidation in biomembrane, and such injury might be related to toxic effects of reactive oxygen on biomembrane. Tetramethylpyrazine was used to treat experimental MON. The results showed that the drug might have reduced the amount of proteinuria and the extent of lipid peroxidative injury. At the same time, the drug might haod raised the activities of SOD and GSH-px, and improved the lesion ot some extent. It is suggested that tetramethylpyrazine had the effect of anti-lipid peroxidative injury.

Objective:To establish the reference values on blood lymphocyte immunophenotype by means of standardized quality control and proper statistical analysis.Methods:The whole bloods of 93 healthy Chinese adults were stained by fluorescent conjugated monoclonal antibodies of Simultest TM IMK lymphocyte Kit and then were analysed by FACS Calibur flow cytometer.The data were acquired and were analysed by SimulSET.Statistical analysis would be conducted after conforming the date of standardized quality control.Results:CD + 3,CD + 3CD + 4,CD + 3CD + 8 cells showed normal distribution but CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 showed skewness distribution .the 95% reference of confidence interval of CD + 3?CD + 3CD + 4?CD + 3CD + 8 cells were established by the method of normal distribution and they were respectively 49 28%～80 52%?25 29%～48 11%?11 93～44 77%;the 95% reference of confidence interval of CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 were established by the method of percentiles and they were respectively 4 74%～19 84%?10 63%～42 86%?0 65～3 05.Conclusion:To establish the reference values,it is considered that the methods of normal distribution and percentiles should be use comprehensively.It is also suggested that the reference values should be confirmed by means of standardized quality control and proper statistical analysis,so the reference values obtained from various laboratories of the same race but in different regions with the same reagents and instruments performed compatibility.

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 investigate the differentially expressed genes of gastric mucosa of precancerous lesion of chronic atrophic gastritis (CAG) in rats intervented by promoting blood flow therapy of TCM,and to analyze its mechanism at genetic level. Methods:The model of precancerous lesion of CAG rats was induced by spring insertion and intragastric administration of NaCl solution and hot paste. To detect the differentially expressed genes of precancerous lesion of CAG rats by gene chip technology,and the main differentially expressed genes were indentified by real-time PCR. Results:Compared with blank group,HSP70 and ptk2 significantly reduced in natural recovery group,the HSP70 expression increased signifi cantly after being treated with supplementing qi for promoting blood flow,and ptk2 expression increased obviously after being treated with regulating qi and promoting blood flow. The results of real-time PCR in expression level of HSP70mRNA and ptk2mRNA were the same as that of the gene chip technology. Conclusion:Supplementing qi for promoting blood flow can upregulate HSP70,thus inhibit precancerous lesion of CAG in rat gastric mucosal cell apoptosis and promote mucosal repair; Regulating qi and promoting blood flow can up-regulate ptk2,sequentially regular remodeling of actin cytoskeleton and promote repairing and healing of injury gastric mucosal. These may be one of mechanism that supplementing qi for promoting blood flow and regulating qi and promoting blood flow treat precancerous lesion of CAG.

AIM: To investigate the effects of salidroside on intracellular free calcium concentration (([Ca~(2+)]i)), apoptosis, mitochondrial membrane potential (MMP) and activity during injury induced by hypoxia/hypoglycemia in cultured SH-SY5Y cells. METHODS: Mitochondrial activity was measured by methylthiazolyl tetrazolium test. MMP, [Ca~(2+)]i and apoptosis were measured by flow cytometry. RESULTS: SH-SY5Y cells were cultured in a hypoxia/hypoglycemia condition for 2, 4, 6 and 12 h, [Ca~(2+)]i and apoptosis rate significantly increased compared with control group (P

OBJECTIVE: To investigate the effects of Radix notoginseng extracts drug-containing serum on the expressions of apoptosis-regulating proteins including Bax, bcl-2 and p21WAF1 in precancerous gastric cells. METHODS: The N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) transformed eternalized human gastric mucosa epithelium GES-1 cell line (MC cell) was used in vitro as a model of gastric precancerous lesion. The medicated canine serum was prepared by feeding to the adult Beagle dog with Radix notoginseng extracts and obtaining the serum after 2-hour medication. MC cells were cultured with medicated canine serum (medicated serum group) or non-medicated canine serum (normal control group) for 72 hours. Expressions of Bax, bcl-2 and p21WAF1 proteins were detected by immunocytochemical assay and the average optical density of the cells was determined by an image analysis system. RESULTS: Compared with those of the normal control group, Bax and p21WAF1 expressions in medicated serum group were significantly enhanced (P<0.01), while the expression of bcl-2 was significantly reduced (P<0.01). CONCLUSION: Radix notoginseng extracts may inhibit the proliferation and promote the apoptosis of precancerous gastric cells through altering expressions of the bcl-2, Bax and p21WAF1 genes.

OBJECTIVE: To study the apoptosis-promoting effect of the serum from Panax notoginseng extracts-fed dog on precancerous gastric cells by means of flow cytometry. METHODS: In the experiment, we adopted eternalized human gastric mucosa epithelium GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) (MC cells) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at two different points of time (2 and 6 hours) after feeding the dog with Panax notoginseng extracts for experiment. The MC cells were cultured in mediums with different concentrations of the medicated serum at 2- or 6-hour point of time for 72 hours. By means of flow cytometry, we examined the apoptosis-promoting effects of the serums on the MC cells. RESULTS: The medicated serums at these 2 points of time had significant effects in promoting MC cell apoptosis. The proportions of apoptotic cells in culture mediums with medicated serums had a significant increase as compared with those in culture mediums with non-medicated serums (serum obtained before administration of extracts to the dog) under the same conditions (P<0.05). The number of MC cells in G(0)/G(1)phase was decreased (P<0.05) and that in G(2)/M phase increased (P<0.05), while no consistent changes were observed in S phase. CONCLUSION: The medicated serums obtained at the two different points of time have significant apoptosis-promoting effects on MC cells. They decrease the number of MC cells in G(0)/G(1) phase and increase the number of MC cells in G(2)/M phase. This is probably responsible for the effects of Panax notoginseng extracts in inhibiting the proliferation of MC cells and promoting its apoptosis.

OBJECTIVE: Through cell cultivation, we studied the inhibiting effects of the serum of the dog fed with Panax notoginseng extracts on precancerous gastric cells, trying to find the best time points or periods when the extracts' function was the strongest after administration of the extracts to the dog. METHODS: The experiments adopted eternalized human gastric mucosa epithelium GES-1 cells and MC cells gained from GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at different points of time after feeding the dog with Panax notoginseng extracts for experiment. By means of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we examined the inhibiting effects of the serum after culturing the GES-1 and MC cells for 72 hours with different concentration (8%, 4%, 2%) of medicated serum obtained from the dog at different points of time, so as to find that, at which points of time the medicated serum obtained, it would be the most effective. RESULTS: The results showed that the GES-1 and MC cells inhibition rates of medicated serum from the points of 2-hour and 6-hour were the highest, and the culture medium containing 8% of medicated serum from these two points had prominent inhibiting effects on both kinds of cells. The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 2-hour was 70.8% (P<0.01) and that of the MC cells was 45.3% (P<0.01). The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 6-hour was 88.5%(P<0.01) and that of the MC cells was 42.4% (P<0.01). CONCLUSION: The points of time with the strongest inhibiting effects are 2 hours and 6 hours after being fed with Panax notoginseng extracts. At these two points, the serum is most effective in inhibiting the proliferation of GES-1 and MC cells.

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.