Object To isolate and identify the chemical constituents of Saussurea japonica (Thunb.) DC. and to obtain some rules of chemical shift effects on hydroxylation and O-glycosylation. Methods The constituents were isolated by silica gel column chromatography and HPLC (RP-18), and their structures were elucidated through spectroscopic analysis (NMR and FABMS). The rules were obtained by comparison with those of known analogous lignans. Results Three lignan glycosides were isolated and identifiedcas (+)-1-hydroxypinoresinol-4″-?-D-glucopyranoside (Ⅵ), (+)-lariciresinol-4-?-D-glucopyranoside (Ⅷ), and (+)-1-lariciresinol-4′-?-D-glucopyranoside (Ⅸ), as well as the rules for effects of the hydroxy group at C-1 (upshift about ??-4.2) and O-glucosylation (downshift about ??+3.0) on 13CNMR chemical shifts of (+)-pinoresinol and (+)-lariciresinol. Conclusion All these compounds were obtained from the plants of Saussurea DC. for the first time and the rules should be as some important evidences to assign the position of hydroxylation and glycosylation.

The cell-surface expression and functional status of the CD95/Fas antigen on primitive hematopoietic progenitors isolated from human cord blood (CB) were studied. The CD34+ cells freshly isolated from CB displayed low CD95 expression. The combinations of cytokines such as SCF + FL could up-regulate the expression of CD95 in vitro culture and tumor necrosis factor-alpha (TNF-alpha) and interon-gamma (IFN-gamma) further increased the CD95 expression induced by positive cytokines. The functional status of CD95-mediated apoptosis were analyzed by incubation of CD34+ CB cells in the presence of anti-CD95 monoclonal antibodies (McAbs). The effects of anti-CD95 McAbs were measured by viable cell counting, flow cytometry, LTIC and CFU-C assays. A decrease of viable cells, CFU-C and LTIC numbers were observed in the presence of anti-CD95 McAbs and TNF-alpha or IFN-gamma. However, growth factor deprivation or the early-acting cytokine such as SCF and FL cross-linking to CD95 caused low apoptosis of CD34+ cells. The correlation of increased intracytoplasmic levels of bcl-2 and the presence of CD95 on fresh CB CD34+ cells suggested that bcl-2 might be involved in protecting against CD95-mediated apoptosis of CB CD34+ cells.
Antigens, CD34 ; Antigens, CD95/*metabolism ; Apoptosis ; Fetal Blood/*cytology ; Hematopoiesis ; Hematopoietic Stem Cells/*metabolism ; Leukocytes, Mononuclear/metabolism ; Proto-Oncogene Proteins c-bcl-2/*metabolism

Objective To investigate the effects of stromal cell-derived factor-1 (SDF-1) on the adherence of mononuclear cells (MNC). Methods Mononuclear cells were isolated from cord blood and the adherent rate was assayed by using MTT with fibronectin coated in different concentration of SDF-1. Then the expression of adherent molecule VLA-4 by FACS as well as the adherent rate when MNC were cultured in vitro. Results The adherent rate of fresh cord blood MNC was (110.2±2.10) %, (128.3±2.09) %, (141.6±2.62) %, (160.8±3.84) %, (162.3±3.60) %, (165.4±2.73) %, (165.1±2.33) %. When the mass concentration of SDF-1 was 50, 100, 150, 200, 250 ng/ml. When MNC were cultured in vitro,the expression of VLA-4 was 26.37te was (160.2±6.38) %, (194.6±6.09) %, (147.5±6.86) %, (144.7±4.96) %, (118.1±3.64) %. The correlation coefficient between the adherent rate and the expression of VLA-4 is 0.826. Conclusion The adherent rate of fresh cord blood MNC increases along with the concentration of SDF-1, however, it tends to be stable when the concentration of SDF-1 reaches 100 ng/ml. When MNC were cultured in vitro with hemopoietic growth factors, the expression of VLA-4 and the adherent rate of MNC increased in the initial stage, however, the expression of VLA-4 and the adherent rate of MNC both decreased gradually along with time extending. There is positive correlation between the adherent rate of MNC and the expression of adherent molecule VLA-4.

ObjectiveTo explore the effects of brucine on multiple myeloma (MM) and to compare the effects between brucine and bontezomib on MM.MethodsMTT method was used to determine the median inhibitory concentratiom (IC50) of brucine and bortezomib on the MM cell line U266.The supernatant of cultured U266 cell line was added to the culture system for inducing the differentiation of osteoblast cell line MC3T3-E1.After aseptic assay,RT-PCR was used to determine the RNA levels of alkaline phosphatase (ALP),osteocalcine (OC),osteoprotegerin (OPG) and recepter activator of NF-κB ligand (RANKL).Results IC50 of bortezomib on U266 cell line for 48 h was 22.4 nmol/L,and that of strychnine was 0.16 nmol/L.The mRNA levels of ALP,OC and OPG in osteoblast co-intervened by brucine combined with the supernatant of MM cells were higher than those in supernatant of U266 cells,while the level of RANKL mRNA was lower (P ＜0.05).The degree of increasing or reducing was greater than the level of control group intervened only by bortezomib (P ＜0.05).ConclusionThe therapeutic effects of brucine on MM might be carried out through the regulation of osteoclast by osteoblast,and the experiment confirmed that the therapeutic effect of brucine on MM was superior to that of bortezomib.

Objective To study the effect of new generation histone deacetylase inhibitor LBH589 single-drug or combined with the mouse serum which contains the radix echinopsis on multiple myeloma (MM) cell line U266 and their mechanism.Methods The acetylation level of α-tublin which was a substrate of HDAC6 was detected by Western blot.The chemical force between Heat shock protein 90 (HSP90) and its client proteins was detected by immune precipitation (IP).Results The different concentrations of LBH589 single drug (0,20,50 nmol/L),and 50 nmol/L combined with the mouse serum which contained the radix echinopsis (1 g/ml) were able to inhibit the proliferation of U266 cell.With the increase of drug concentration and the extension of time,the acetylation levels of α-tublin and HSP90 increased gradually (at 24 or 48 hours) in a dose dependent (P ＜ 0.05).The inhibition of LBH589 combined with the mouse serum was stronger than that of LBH589 single drug (P ＜ 0.05).Conclusion LBH589 could inhibit the growth of MM cells and their cell cycles,and induce the apoptosis of MM cell line U266.

The deficiencies of the 6th International Union Against Cancer (UICC)/American Joint Committee on Cancer (AJCC) staging system of nasopharyngeal carcinoma (NPC) were included as follows: ①There is a lack of hazard discrimination between some T categories; ②The maximum dimension of lymph node was not an independentprognosis in NPC; ③The subsets defi ned by T and N classifi cations that make up a given group typically have different prognosis. The introduction of new types of therapeutic interventions or new technologies may require modifi cations of the classif ication and staging systems. Compared to computerized tomography (CT), magnetic resonance imaging (MRI) is superior to in the detection of early involvement such as paranasopharyngeal space, oropharynx, and retropharyngeal lymphatic metastasis, and demonstrate deep primary tumor infi ltration such as skull base, intracranial erosion more easily, so MRI should be optional method in T stage of NPC. Positron emission tomography/ computerized tomography (PET/CT) is superior to MRI in the detection of cervical lymph node metastasis and distant metastasis. With the development of diagnostic and therapeutic techniques revolutionized in NPC, staging systems should be modified. Research based on the data of 924 NPC patients revealed:①It appeared more prognostic value to modify 4 substages into 3 substages in T-category; ②N-category was more concise by deleting nodal greatest dimension palpation;③ It showed that nodal parameters including level, laterality and extranodal neoplastic spread (ENS) are independent prognostic factors for NPC. N-staging criteria with RTOG guidelines for lymph node levels was set up which adapted to the requirement of conformal radiation therapy. Incorporating ENS and retropharyngeal lymph nodes into the N-staging system for NPC increases the predictive power.

Objective: To investigate the activity of coagulation and fibrinolysis systems in bronchoalveolar lavage fluid (BALF) in lung fibrosis in rats, and to study their role in diagnosis of lung fibrosis. Methods: Thirty-six Sprague-Dawley rats were randomly divided into 2 groups (n=18 in each group ). Lung fibrosis and control models were made by tracheal instillation bleomycinA_5 (BLMA_5) (5 mg/kg) and saline. On days 7, 14 and 28, the recalcification time points of normal pooled plasma for studying procoagulation activity (PCA), the levels of von Willebrand Factor (vWF), the activities of Antithrombin-Ⅲ(AT-Ⅲ) , plasminogen activity inhibitor-1 (PAI-1) and urokanise-type plasminogen activator(uPA) in BALF were measured respectively. Results: (1) In BLM group,the recalcification time points in BALF were (56?10), (78?3) and (172?11) seconds respectively, they were (190?10), (186?8) and (184?6) seconds respectively in control group. The difference was significant(P

Objective To observe the effect of ganglioside GM3 on the proliferation and cell cycle of multiple myeloma (MM) cell line U266. Methods The experimental groups were treated with 20, 40, 80, 160μmol/L GM3, while the control group was not given GM3. The growth inhibition effect of GM3 on U266 cells after 48 hours were measured by using methyl thiazolyl tetrazolium (MTT) method. The cell cycle was tested by flow cytometry with PI labeling. Results The results indicated that the GM3 displayed anti-proliferative effects on U266 cells in a dose-dependent manner. The cell inhibition rates were (13±4) %, (26± 4) %, (47 ±6) %, (55 ±10) % in different dose of GM3 (20, 40, 80, 160 μmol/L), respectively. There was a difference between the experimental group and the control group (F= 93.063, P< 0.05). At the same time, the cell cycle analysis results showed that the U266 cell ratio in S phase was increased from (22.6 ±3.7) % to (71.5±3.8) %(P<0.01) and in G2/M phase was decreased from (42.6±2.5) %to (0.8±0.6) %(P<0.05) while in G0/G1 phase was not significantly changed. Conclusion GM3 can inhibit the proliferation of MM cell line U266 by blocking them in the S phase.

Objective:To investigate the effect of small interfering RNA (siRNA) silencing nicotinamide phosphoribosyltransferase (NAMPT) gene expression on proliferation and apoptosis of human multiple myeloma U266 cells and its mechanism.Methods:In vitro, NAMPT gene-specific siRNA was synthesized to transfect U266 cells. The experiment was divided into si-NAMPT group (transfected siRNA-NAMPT U266 cells) and si-NC group (transfected negative control siRNA U266 cells). The proliferation of U266 cells after transfection was detected by the methyl thiazolyl tetrazolium (MTT) method, and the apoptosis was detected by flow cytometry. Western blot was used to detect the expression levels of NAMPT, protein kinase B (AKT), phosphorylation-AKT (p-AKT), glycogen synthase kinase-3β (GSK-3β), phosphorylation-GSK-3β (p-GSK-3β), and β-catenin proteins in each group after transfection.Results:Compared with the si-NC group, the proliferation inhibition of U266 cells (absorbance value at 570 nm) increased after transfection for 48 h and 72 h in the si-NAMPT group (48 h: 0.78±0.06 vs. 1.62±0.11; 72 h: 1.23±0.14 vs. 2.37±0.18), and the differences were statistically significant ( t = 3.54, P = 0.034; t = 4.72, P < 0.01). The early apoptotic rate of cells in the si-NAMPT group increased compared with si-NC group [(53.42±0.25)% vs. (25.98±3.18)%], and the difference was statistically significant ( t = 4.41, P < 0.01). Compared with the si-NC group, the levels of p-AKT, p-GSK-3β, NAMPT and β-catenin proteins were significantly reduced in the si-NAMPT group (all P < 0.05). Conclusion:Silence of NAMPT gene can significantly inhibit U266 cell proliferation and induce cell apoptosis, and it may play a role by inhibiting the AKT-GSK-3β-β-catenin signaling pathway.

BACKGROUND: It is of great significance to understand the effects of different components of blood pressure on the occurrence of cerebrovascular diseases and administer proper decompression treatments in various situations.OBJECTIVE: To study the relationship of increased pulse pressure with the occurrence and severity degree of cerebral infarction by analyzing the data of pulse pressure recorded in hospitalized patients with acute cerebral infarction.DESIGN: Case-control analysis.SETTING: First Affiliated Hospital of Harbin Medical University.PARTICIPANTS: A total of 300 patients hospitalized at the First Affiliated Hospital of Harbin Medical University between June 2002 and January 2003 for ischemic cerebral infarction were enrolled, including 196males and 104 females aged (57.9±11.9) years. Another 199 persons who came to the same hospital for physical examination at the same period were set as normal group, including 110 males and 89 females aged (55.9±12.4)years.METHODS: ① Measurement of blood pressure: Systolic pressure and diastolic pressure of each person were recorded with mercury sphygmomanometer at the site of brachial artery of the right upper extremity and pulse pressure was calculated. ② Evaluation of neurological impairment:Upon hospital admission, each patient with ischemic cerebral infarction was evaluated with the stroke scale made by the National Institute of Health (NIHSS), in which a higher score means more severe infarction.MAIN OUTCOME MEASURES: ① Comparison of blood pressure parameters of subjects in the two groups. ② Distribution of different pulse pressure levels in the two groups. ③ Comparison of pulse pressure of subjects of different age in the two groups. ④ Results of non-parameters tests of neural function loss in cerebral infarction patients with different pulse pressure levels.RESULTS: ① Mean systolic pressure: It was significantly higher in the case group than in control group [(152±22), (133±19) mm Hg, t' =10.494,P ＜ 0.01]. Mean diastolic pressure: It was higher in the case group than in control group [(93±14), (81±11) mm Hg, t' = 10.129, P ＜ 0.01]. Meanpulse pressure: It was higher in the case group than in control group [(59.61±11.86), (51.93±14.10) mm Hg, t' =5.612, P ＜ 0.05]. Pairwise corre lation analysis showed that Pearson correlation coefficient between pulse pressure and systolic pressure was 0.789 (P ＜ 0.01); Pearson correlation.coefficient between pulse pressure and diastolic pressure was 0.169 (P ＜ 0.01). Therefore, there was a close correlation between pulse pressure and systolic pressure. ② Pulse pressure was distributed between 60 mm Hg and 69 mm Hg in most subjects in the case group, which accounted for 27.7%.It was 40-49 mm Hg in control group, which accounted for 35.7%. It indicated that the incidence rate was high when the pulse pressure was over 60 mm Hg in cerebral infarction. ③ Pulse pressure increased with age.The level of pulse pressure in 40-69 years case group was higher than that in 40-69 control group [(54±16), (45±9)mm Hg, t=4.86, P=0.000]. ④ Patients with cerebral infarction of different levels of pulse pressure were given non-parameter tests of neurological impairment evaluation. The resuits of Kruskal-Wallis test were χ2=4.779, P=0.572 ＞ 0.05; results of median test were as x2=8.365, P=0.213 ＞ 0.05. The results of the two non-parameter tests suggested that there was no significant differences in hospitalization evaluation, that is, although the pulse pressure increased obviously in cerebral infarction, the degree of increase had no correlation with the severity of neurological impairment.CONCLUSION: The increase of pulse pressure is related to the occurrence of cerebral infarction and is also an important factor for evaluating cerebral infarction. However, pulse pressure change is not related to the severity of cerebral infarction.