Objective To explore the effective method for in vitro culture of the dendritic cells(DCs) and the specific anti-leukemic cell effect mediated by dendritic cells pulsed with acute myelogenous leukemia antigen. Methods Bone marrow mononucleas cells (BMMNCs) isolated from AML patients were induced to undergo differentiation with 500 ng/ml A23187 and pulsed with AML antigen. After 96 h, DCs and T cells were co-cultured for 5 to 7 days. The cytotoxic activity of cytotoxic T lymphocyte(CTL) to AML were detected with MTT colorimetry. Results BMMNCs isolated from AML patients treated with 100 ng/ml rhGM-CSF in combination with 500 ng/ml A23187 for 96 h exhibited typical morphology of DCs with rapidly increased expression of CD1a and CD83 (P<0.01). Dendritic cells pulsed with acid eluted tumor antigen peptides group displayed the strongest cytotoxic activity of CTL to AML. There was a significant difference between two groups(P<0.01). Conclusion BMMNCs isolated from AML patients can be successfully induced to DCs with rhGM-CSF and A23187 and dendritic cells pulsed with acid eluted tumor antigen peptides group had the strongest cytotoxic activity of CTL to AML.

Objective To explore the protective effect of selenium, an antioxidant, on fluoride-induced renal injury in rats and find out the optimal level of selenium against fluoride toxicity and its valid molecular target.Methods All 80 male weanling SD rats were randomly divided into 8 groups by body weight as follows: normal control group(drinking tap water), fluoride exposed group (drinking water containing 50 mg/L of NaF), low, middle,high selenium exposed groups(drinking water containing 0.375, 0.750, 1.500 mg/L of Na2SeO3) and low, middle,high Se-fluoride groups (drinking water containing both 50 mg/L NaF and three doses of Na2SeO3 as abovementioned, respectively). After 6 months, the rats were killed then the oxidation level and nuclear factor κB(NF-κB)expression level in kidney were measured. Results The weight of the fluoride exposed group[(695.95 ± 55.89 )g]was significantly deceased than the controls[(782.69 ± 56.12)g, P ＜ 0.01]. Glutathione peroxidase(GSH-Px)activity of fluoride exposed group[(55.86 ± 5.09)U/mgprot] was not significantly different but decreased. Tatal antioxidant capacity (T-AOC) activity in fluoride exposed group [(7.54 ± 1.35)U/mgprot] significantly decreased than the controls[(9.03 ± 0.37 )U/mgprot, P ＜ 0.05]. In addition, a significant increase of malondialdehyde ( MDA )in fluoride exposed group[(3.86 ± 0.31 )mnol/mgprot, P ＜ 0.05] was observed than the controls[(3.14 ± 0.32)nmol/mgprot, P ＜ 0.05]. GSH-Px activity of high Se-fluoride group[(74.99 ± 8.41 )U/mgprot] was significantly higher than the fluoride exposed group[(55.86 ± 5.09)U/mgprot, P ＜ 0.05] and its MDA level[(3.17 ± 0.20)nmol/mgprot] was lower than the fluoride exposed group[(3.86 ± 0.31 ) nmol/mgprot, P ＜ 0.05]. NF-κB expression levels of fluoride group, high selenium group and low Se-fluoride group(0.360 ± 0.015,0.367 ± 0.007,0.376 ± 0.006,respecyively) were obviously increased compared with the controls(0.312 ± 0.022, P ＜ 0.05); it was significantly lower in high Se-fluoride group(0.312 ± 0.005) than in fluoride exposed group(0.360 ± 0.015, P ＜ 0.05). Conclusions Na2SeO3 of 1.5 mg/L is the optimal dose against chronic fluorosis on kidney injury under this experimental condition.NF-κB is likely to be a target molecule of the selenium as an antagonist on fluorosis.

Aim To observe the effect of hemodialysis on pharmacokinetics of sparfloxacin in thepatients contracting chronic renal failure. Methods Sparfloxacin concentrations inserum and urine of hemodialysis and non-dialysis patients were measured with a highperformance liquid chromatography method after administration a single oral dose of 200mg sparfloxacin. The pharmacokinetic parameters were computed with the programPKBP-N1.Results The main pharmacokinetic parameters in hemodialysis group wereT1/2(ka) - (1. 25 ?0. 57) h, T1/2(?) = (11. 88?4. 13) h, Tpeak = (4. 18 ? 0. 78) h,Cmax = (0.80 ? 0. 17) mg? L-1 and AUC0-= (6. 90 ? 3. 25) mg?h?L-1, while innon-dialysis group were T1/ 2(ka) = (1. 12 ? 0. 42) h, T1/ 2(?) = (15. 93 ? 5. 20) h, Tpeak =(3. 88 ? 0. 75) h, Cmax = (0. 69 ? 0. 37) mg?L -1, AUC0-= (10.05 ? 4. 13) mg?h?L-l. The original sparfloxacin discharge rats in urine within 24 h were (8. 98 ? 3. 92) % and(10. 58 ? 5. 64) % separately. T1/2(?) and AUC in hemodialysis group were markedly lowerthan in non-dialysis group (P

OBJECTIVEAn accurate and reliable analytical method for-simultaneous determination of six active components (scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C) in plants of Erycibe was developed.

METHODScopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C in the samples were well separated in analytical HPLC by gradual elution with methanol-0.1% formic acid solution. The chromatographic condictions: Agilent Poroshell 120 EC-C18 column, flowing rate being 1 mL x min(-1), detecting wavelength at 345 nm.

RESULTGood linearities of scopolin, chlorogenic acid, scopoletin, isochlorogenic acid A, isochlorogenic acid B and isochlorogenic acid C were in the range of 0.026 8-2.68, 0.027 0-2.70, 0.008 1-0.81, 0.018 8-1.88, 0.017 6-1.76, 0.019 6-1.96 μg, respectively (r > 0.999 6). The average recoveries of the six components were 98.1%, 98.7%, 100.8%, 100.4%, 99.7%, 101.1%; the relative standard deviations were 2.67%, 2.86%, 2.62%, 1.98%, 2.76%, 2.19%.

CONCLUSIONThe method is simple, feasible and reproducible and can be used for the quality control of plants of Erycibe.

China ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Convolvulaceae ; chemistry ; Coumarins ; analysis ; Drugs, Chinese Herbal ; analysis ; Glucosides ; analysis ; Scopoletin ; analysis

0.05).Conclusion There are no significant effects on bone metabolism and growth of children with small dose of IGs per day for a longer time.

Objective To evaluate the clinical significance of sinus heart rate turbulence(HRT)and the heart rate variability(HRV)in the patients with essential hypertension(EH).Methods HRV and HRT examination were carried out in sixty patients with EH.A subgroup of patients receive metoprolol+nifedipine(n=26)or nife- dipine alone(n=34)were investigated seperately to evaluate the effect of ?-blocker on the HRV and HRT.Fifty healthy persons were served as control.In the HRT determination,turbulence onset(TO)was defined as sinus heart rate acceleration,after ventrieular mature beat while turbulence slope(TS)as sinus heart rate deceleration slope after ventricular premsture beat.Normal value of TO was2.5 ms/RR period. SDNN,RMSSD,LF/HF in HRV were analysed from 24 hours ambulatory electrocardiography(before and after 1 month medication).Results ① In hypertensive group,52 patients showed positive TO(86.6%),48 patients pos- itive TS(80.0%)and 46 patients TO+TS(76.7%),compared with hypertension group only 2(4%)positive TO and 3(6%)positive TS in the healthy control.Forty-two cases SDNN(70.0%),41 cases RMSSD(68.3%)and 38 cases LF/HF(63.3 %)were positive in hypertensive group,while only 1(2 %)SDNN,1(2 %)RMSSD and 3(6 %) LF/HF in control. ② Metoprolol didn't change the positive percentage of parameters in HRT but parameters showing Heart rate variablity in HRV was decreased significantly found between TO,TS and TO+TS.Conclusion HRT and HRV is two indices for determination the dysfunction of autonomic nervous system in hypertension.?-re ceptor blocker inhibit sympathic activity,miligate the decreasing of HRV,but the bipbase accelerative and decelera- tive phenomena of HRT didn't change,HRT seems to be a more sensitive index for monitoring of autonomic sysfunc tion in hypertension.

Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.

(3S)-Linalool synthase (LIS) is a key enzyme involved in the monoterpene biosynthetic pathway. Based on our previous transcriptome study, the expression level of LIS gene was exceedingly related to glycyrrhizic acid (GA) biosynthesis. Therefore, we used hairy root culturing to further investigate the effect of LIS on the GA biosynthesis. A LIS gene (GenBank accession number: MZ169552) was cloned from Glycyrrhiza uralensis. The plant binary overexpression vector pCA-LIS was constructed by gene fusion. G. uralensis hairy roots overexpressing LIS were induced by the Agrobacterium rhizogenes ATCC15834. The expression levels of LIS were analyzed by real-time quantitative PCR (RT-qPCR) and the contents of GA in hairy root lines were determined by UPLC. It was found that in the hairy root lines overexpressing LIS, the expression levels of LIS were significantly higher than that in the wild type, while the contents of GA were remarkably lower than those in the wild type and negative control. These findings indicate that the expression level of LIS is negatively correlated with the accumulation of GA. In this study, LIS was cloned from G. uralensis for the first time and the negative regulatory effect of LIS on GA biosynthesis was confirmed by reverse genetics. This work provides support for further improvement of the molecular regulatory network of GA biosynthesis in G. uralensis.

OBJECTIVETo evaluate the efficacy and progression-free survival of erlotinib after progression of disease to gefitinib in patients with advanced pulmonary adenocarcinoma who previously obtained a disease control with gefitinib.

METHODIn this retrospective study, 12 patients with advanced or metastatic pulmonary adenocarcinoma,who were previously obtained a partial response or a stable disease with gefitinib,were treated with erlotinib after gefitinib failure. Erlotinib efficiency, progression-free survival and overall survival were analyzed.

RESULTSNice (75%)achieved stable disease and three (25%) achieved progression disease with erlotinib treatment after gefitinib failure. No complete response or partial response was observed. The disease control rate was 75%. The median progression-free survival and overall survival of erlotinib were 180 days and 831 days.

CONCLUSIONErlotinib seems to be an optional treatment after gefitinib failure for advanced pulmonary adenocarcinoma patients,who previously responded to gefitinib.

Adenocarcinoma ; drug therapy ; Aged ; Aged, 80 and over ; Antineoplastic Agents ; therapeutic use ; Drug Tolerance ; Erlotinib Hydrochloride ; Feasibility Studies ; Female ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Quinazolines ; therapeutic use ; Retrospective Studies ; Treatment Outcome

Animals ; Blood Pressure ; Lymphatic Vessels ; physiopathology ; Male ; Norepinephrine ; metabolism ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; metabolism ; physiopathology