The influences of varied concentrations of AG on the 8H-TdR(3H-Thymidine ) incorporation inhibition rate and the release of PGI2 and TXA2 were studied in cultured human and rat lymphocytes . The results indicated that the AG increased the 3H-TdR incorporation inhibition rate on the concentrations of 1.5? 10-7mol/L and 3?10-7mol/L. Using the RIA method, authors find that AG on the concentrations of 4 ? 10-7mol/L, 4 ? 10-3mol/L and 4 ? 10-11mo/L can significantly decrease the cultured lymphocytes TXA2 release while has no clear influences on PGI2 release

This paper reported the estimation of free calcium concentration in rabbit resting lymphocytes, and the effect of (-) Daurisoline (8701) on the cyto-plasmic free calcium concentration by using fluorescent indicator Fura -2/Am estimating technique. Results were compared with the effect of the Ca2-ionophore A23187.The results indicated that the free calcium concentration of resting lymphocytes was 109. 6nmol ? L-1, after 10-5 mol? L-1,10-4.5 mol ? L-1,and 10-4 mol ? L-1-)Daurisoline administration, the cyto-plasmic free calcium concentration increased to 178. 8%, 204. 4%and 817. 5% respectively, while same concentration A23187 only led the increase of 164. 6% , 198.4% and 327% respectively.Results suggested that ( - )Daurisoline may be a Ca2- agonist and may induce the cytotoxicity effect on lymphocytes at high concentr ation.

Objective To investigate the role of osteopontin (OPN) in cholesterol gallstone formation.MethodsGallbladder bile was obtained from patients with cholelithiasis (n=36,the experimental group) and from donors of liver transplantation (n=19,the control group).OPN,calcium ion and lipid were analysed quantitively.The nucleating role of OPN in bile was evaluated using nucleating time (NT) approach.ResultsOPN inhibited cholesterol nucleation in a dose dependent manner.OPN (50 μg/ml and 100 μg/ml) prolonged NT by 48.90％ (91.51％) and 17.07％ (32.93％) in lithogenic and control bile,respectively.OPN (100 μg/ml) also inhibited the nucleating effect induced by calcium ion.Furthermore,a combination of OPN (50 μg/ml) and calcium prolonged NT by 75.78％ and 33.96％ in lithogenic and control bile,respectively.A combination of OPN (100 μg/ml) and calcium prolonged NT by 125.9％ and 62.26％ in the 2 groups.The contents of osteopontin and calcium were significantly lower in lithogenic bile than control bile (P＜0.05).On the other hand,the cholesterol saturation index and the contents of cholesterol,phospholipid and bile acid were significantly higher (P＜0.05).ConclusionsOPN inhibited cholesterol gallstone formation.It may be involved in the pathogenesis of cholelithiasis.

Objective To investigate the dynamic changes of osteopontin (OPN) in the guinea pig model of cholesterol gallstone disease.Methods Forty-four guinea pigs were randomly assigned to cholesterol gallstone group and control group.The animals were sequentially killed on Day 7,14,28,42,56 and 70 after operation.The expressions of OPN mRNA in gallbladder and liver tissues were detected by real-time PCR.The changes of OPN,mucin,α1-acid glycoprotein (AAP) and apolipoprotein A1 (APOA1) in bile and blood plasma were detected by ELISA kits.Results The expression of OPN mRNA in gallbladder and liver tissues increased gradually and reached the peak in the 6th week,and then decreased.The increased expression of OPN in bile in the gallstone group started from the 1st week and reached the peak in the 6th week (P ＜ 0.05),which gradually decreased to the baseline in the 10th week (P ＞ 0.05).The expressions of OPN in bile and blood demonstrated similar trends,while the peak time in blood samples was much earlier (4th week).The changes of APOA1 in bile and blood were similar to OPN,although there was no advanced peak value in blood.The levels of mucin and AAP in bile and blood increased after operation,and were kept at high level throughout the study.Conclusions OPN is involved in the whole process of cholesterol gallstone formation,which may be associated with other nucleation-active factors.

Objective To investigate the role of osteopontin (OPN) in the pathogenesis ot cholesterol gallstone formation in bile.Methods The nucleation time of OPN in model bile and human gallbladder bile was studied by the nucleation time assay,the effect of OPN on cholesterol crystal growth in model bile was examined by the cholesterol crystal growth assay.The effect of OPN on vesicle was detected by the transmission electron microscopy in model bile and gallbladder bile; then the content of OPN and calcium were detected via the commercial kits in human bile.Results Osteopontin prolonged nucleation time in a dose dependent manner in model bile and human bile,and this effect was correlated with calcium.Compared with control group,the nucleation times were prolonged by 1.50and 1.93 times in lithogenic bile at the concentration of osteopontin 50 μg/ml and 100 μg/ml (P＜0.01),respectively. Nucleation time were prolonged by 1.17 and 1.33 times in normal bile (P＜0.01) and by 1.29 and 1.48 times in model bile (P＜0.01),respectively.The rate of cholesterol crystals growth was not influenced by calcium ions,but inhibited by osteopontin in a dose dependent manner in the model bile.Furthermore,the formation,aggregation and fusion of vesicles were delayed by osteopontin in bile samples as indicated by the transmission electron microscopy.The concentration of osteopontin [(0.53± 0.08) mg/ml vs. (0.65 ± 0.14) mg/ml,P＜0.05] and the calcium ions [ (0.71 ± 0.17) mmol/L vs. ( 0.84 ± 0.08 ) mmol/L,P ＜ 0.05 ] were lower in lithogenic bile than in control.Conclusions Osteopontin can inhibit the cholesterol gallstone formation in model and human gallbladder bile as the anti nucleating factor.

Objective To observe the effect of combined treatment with rhTRAIL(recombinant human TNF-related apoptosis inducing ligand) and selective Cox-2 inhibitor Celecoxib on gallbladder carcinoma in vitro and to explore the possible mechanism of the effect. Methods Western blot analysis was used to detect the expression of c-FLIP and death receptors after treatment by Celecoxib. Apoptosis of gallbladder cell line SGC-996 after the combined treatment with Celecoxib and rhTRAIL was detected in three ways: (1) phase microscopy of the cells, (2) detection of effector caspase-3 and caspase-7 activity, and (3) determination of the proportion of apoptotic cells labeled by Annexin V-PI flow cytometric analysis using CELLQUEST software. Results Celecoxib down-regulated the expression of c-FLIPs and up-regulated the expression of DR5 in a dose- and time-dependent mode on cell line SGC-996. Apoptotic levels in the combined treatment group in cell line SGC-996 were significantly higher than those in the single drug treatment group and control group. Conclusion Celecoxib markedly sensitized rhTRAIL-induced apoptosis through the down-regulation of c-FLIPs and up-regulation of DR5 in gallbladder carcinoma cell line SGC-996.

Lymphokine-osteoclast activating factor(OAF) is a potent mediator of bone resorption. PGs which mediate bone resorption are necessary for OAF. It has been reported that mononuclear is an important factor in bone resorption; and circulating monocytes may be osteoclast precursors,but it is not clear at present. We used CT and PTH which are potent physiological calcium-regulator hormones to stimulated mono-nuclear to study it's role for bone resorption. Mononuclear was isolated from normal human peripheral blood by Ficoll-Hypaque. After cultured in C02 incubator at 37℃ for 24h. 0.05g/L PTH and 100U/L CT were added and incubated in the same condation. After 2.5min, 5min, 10min, and 15min cAMP was measured by RIA. In the sam way PGE was determined. Our experiment exhibited that circulating mononuclear responds to CT and PTH with an increase in cAMP firstly, and enhances PGE production by CT and PTH stimulated mononuclear secondly. The results suggest that mononuclear participated in the regulation of bone resorption by PGE, OAF and other factor.

In order to obtain some evidences of the role of parathyroid hormone (PTH) in regulating calcium metabolism, and explore the mechanisms by which PTH regulates calcium in the heart, 45Ca was used to study the influence of bovine PTH1-34 fragment (bPTH1-34) on calcium transsacolemma. The results indicate that bPTH1-34(l?I0-7mol ? L-1) increases beating rate of heart cells under normal state and it markedly increases the rapid (5 min) and the slow (120 min) phases of 45Ca influx in heartcells (under normal, KC1 10-5 mol?L-1 high K+depolarised, NA 10-7mol?L-1receptors activated states), but it could also reduce 45Ca efflux from the cultured heart cells. It suggests that the mechanisms caused by bPTH1-34 which enhances the cytosolic calcium concentration and myocardial beating rate may be related with cAMP level elevated in heart cells.

Objective To develop an albumin aptamer that may potentially serve as a selective ligand for albumin removal from experimental samples.Methods A single-stranded 59nt DNA library that contains 21 random oligo nucleotides was synthesized in vitro.An albumin aptamer A6 was developed by SELEX technique using bovine serum albumin (BSA) as target.The enrichment of aptamer and evaluation of its binding properties were monitored by flow cytometry.The secondary structure of A6 was predicted by MFord software.Results The aptamer A6 strongly bound to BSA with a Kd of 77.4 nmol/L,and had minimal cross reactivity with control proteins including ovalbu min,IgG,and trypsin.Conclusions Aptamer A6 may be a potential tool in albumin removal.

Objective To explore whether quercetin-loaded PEG-PE micelles(M-Q) can synergize the growth-in-hibitory activity of adriamycin prepared ( ADR) by reversing the drug resistance of MCF-7 ADRr breast cancer cells in vitro.Methods M-Q was prepared by adding saline to lipid film containing quercetin and PEG-PE.The size of M-Q was characterized by dynamic light scattering ( DLS) .The inhibition of MCF-7 ADRr cells was evaluated by MTS assay after incubation with M-Q and ADR.Results The incorporation efficiency of quercetin by the micelles was above 74%.The average size of M-Q was 11.11 nm.Compared with the quercetin dissolved in ethanol , M-Q more effectively reversed the drug resistance of MCF-7 ADRr cells in vitro.Conclusions PEG-PE micelles may potentially deliver quercetin to cancer cells for reversal of drug resistance .