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.

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 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.

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 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.

OBJECTIVE: To evaluate the therapeutic effect of transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine for the treatment of limb ischemia. METHODS: Twenty-three patients with limb ischemia were treated. G-CSF was used to stimulate the bone marrow. The mononuclear cells were separated from the aspirated bone marrow fluid in the stem cell studio. The cell amount was above 1x10(9). The transplantation was performed by the way of intra-muscular multi-injection. Traditional Chinese medicine for replenishing qi to activate blood was prescribed from the first day after operation. The pain, poikilothermia, ulcer or necrosis and ankle/brachial index (ABI) of the ischemic limb were evaluated before and after the treatment. RESULTS: The pain score and poikilothermia score decreased one month after the transplantation, with distinct differences as compared with the scores before the treatment (P<0.05). The ABI increased gradually after the treatment, and one month after the treatment, it was 0.15 higher than that before the treatment. CONCLUSION: Transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine can decrease the symptoms and signs of severe lower limb ischemia effectively, and improve the circulation of the ischemic area.