OBJECTIVE:To establish a RP-HPLC method for the determination of nitrochlodipine in rabbit plasma.METH-ODS:Chromatographic conditions and extractive conditions were optimized with orthogonal design,internal standard:felodipine,solid phase extract column:C18,column size:NOVA-PackC18(4.6mm?250mm),detection wavelength:237nm.RESULTS: The optimal chromatographic conditions were:mobile phase acetonitrile-water(55∶45),flow rate 1.0ml/min,column temperature 45℃.The extraction conditions were A1 B1 C3.The calibration curve was linear within the range of 6.25～400?g/L(r=0.9 996) and the minimum limit of detection was 2.5?g/L.The average RSD of within-day ranged from 3.81% to 5.58% and that of day-to-day from 4.60% to 8.76%.The average recovery ranged from 100.7% to 101.8%.CONCLUS-ION:The method is simple,rapid,highly accurate and precise.It can be used for the quantitative determination of nitrochlodipine in plasma.

Objective: To study the expression of L selectin and its possible role in pulmonary injury in rat models undergoing cardiopulmonary bypass (CPB) and to investigate the protecive effect of fucoidin during the procedure. Methods: Models of CPB and pulmonary perfusion in rats were used to investigate the expression of L selectin pre and post operation. Its expression in antagonist group was also studied. Other indexes, including the concentration of SOD, MDA and MPO in rat lung tissues as well as PaO 2/FiO 2 and histological changes, were studied to illustrate the possible mechanism of L selectin in lung reperfusion injury. Results: The expression of L selectin increased after creating CPB but decreased after pulmonary infusion with fucoidin during the procedure, and lung injury relieved as well. Conclusion: L selectin might lead to lung injury during CPB, blocking its expression may relieve lung injury and enhance the recovery of lung function.

Purpose:To study the activity of toremifene and its synergistic effect with anti-tumor drugs on human lung adenocarcinoma cell line A549,which might be expected to provide a new mode of therapy for the clinical management of lung cancer.Methods:The cytotoxic effects of these agents on human lung cancer cell line A549 were measured by a tet- razolium-based volorimetric assay (MTT assay).Results:Toremifene can inhibit the growth of A549 cell directly.The A value of the low dose toremifene combined with anti-tumor drugs were lower than that of anti-tumor drugs alone.Toremifene combined with VCR,ADM,DDP and VP-16 showed better effects.Conclusions:Toremifene combined with chemotherapy drugs shows significant synergistic anti-tumor effect on A549 cell.This might provide experimental evidence for lung cancer therapy.

Objective To estimate the biological effects of leptin on human HaCaT keratinocytes and explore their molecular mechanisms.Methods Cell counting kit-8 (CCK-8) was used to evaluate the proliferation of cultured HaCaT cells treated with different concentrations of leptin for 24 and 48 hours.Some HaCaT cells were classified into four groups to remain untreated,be treated with leptin (100 μg/L) and piceatannol (a specific inhibitor of STAT3 phosphorylation) alone or in combination for 24 hours,respectively,followed by the evaluation of cell proliferation using CCK-8 kit.Flow cytometry was performed to assess cell cycle of HaCaT cells treated with leptin of 100 μg/L,Western blot to determine the phosphorylation level of Erk1/2 and STAT3 in HaCaT cells treated with leptin of 100 μg/L for different durations.Statistical analysis was done by Student's t-test for unpaired data using GraphPad Prism 5 software.Results The proliferation of HaCaT cells was accelerated to different degrees after treatment with leptin of 50 and 100 μg/L for 24 and 48 hours,and the accelerating effect was in a dose-dependent manner within 24 hours (r =0.9989,P ＜ 0.05).Piceatannol apparently inhibited the promotive effect of leptin on the proliferation of HaCaT cells.There was an obvious elevation in the percentage of cells at S phase ((57.70 ± 5.88)％ vs.(42.50 ± 7.55)％,P ＞ 0.05),but a significant decrease in that at G0/G1 phase ((39.70 ± 1.57)％ vs.(45.20 ± 1.44)％,P ＜ 0.05),with a significant increase in proliferation index (0.603 ±0.0157 vs.0.564 ± 0.0144,P ＜ 0.05) in HaCaT cells treated with leptin of 100 μg/L for 24 hours compared with the untreated controls.Western blot showed that leptin of 100 μg/L markedly enhanced the phosphorylation level of STAT3 in HaCaT cells.Conclusion Leptin may upregulate the proliferation of HaCaT cells through activation of STAT3 pathway.

OBJECTIVE:To detect the contents of chlorogenic acid and rutoside in Saussurea involucrate,and to optimize the decoction and extraction technology of S. involucrate from different producing areas. METHODS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate from different producing areas were determined by HPLC. L9(34)orthogonal experiment was used to optimize the water amount,decoction times and decoction time using comprehensive score of extraction transport rate of chlorogenic acid and rutoside as index. The verification test was also conducted. RESULTS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate were 0.380%-0.546% and 0.334%-0.617%;the optimal decoction technology was as follows as the amout of crude material of S. involucrate 100 g,soaking for 20 min,decocting for 3 times,12,10 and 10 fold of water,decocting 45,30 and 30 min,respectively. The extraction transfer rates of chlorogenic acid and rutoside were 96.2%(RSD=2.66%,n=3)and 89.3%(RSD=3.31%,n=3)in verification test. CONCLUSIONS:For S. involucrate from different producing areas,the contents of effective components are different;optimized decoction and extraction technology is stable and feasible.

Based on reservoir condition and fluid characteristics, the oil-degrading bacterial strain NX-2 was screened from Ma-2 fault block of Huabei oil field. Bacterial metabolism and the capability of improving oil property were evaluated on oxygen-deficient condition. At 87℃ which reservoir temperature is, artificial homogeneous core displacement experiment indicated the enhanced oil recovery of microbe was 7.1% higher than that of waterflooding. In experiment on individual well Ma-410, additional oil production of 669 tons was gained, and decreased water production of this trial well reached more than 3000 tons. These results demonstrated NX-2 could adapt to stratum conditions, enhance oil recovery and improve oil property as well.

Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.

Objective To explore the effect of iron chelators on labile iron pool and expression of apoptosis associated genes in cells of K562, an erythroleukemia cell line.Methods K562 cells were incubated at 37 ℃ in RPMI 1640 containing 10% heat-inactived fetal bovine serum in an saturated humidity and 5% CO_2 incubator. K562 cells were incubated with different concentrations of desferro-(xamine(DFO)). The study groups were divided as following: DFO group, iron+DFO group and the control group. Following indices were detected which included apoptosis by flow cytometry (FCM) assay, expression of Rb, c-myc, bax mRNA by RT-PCR. The intracellular LIP was measured with a fluorimetric assay using the metalsensitive probe calcein-AM.Results 1. The viability of K562 cells incubated with different concentrations of DFO was lower than that of control group at 12 h,24 h and 48 h (P

Objective To investigate the effect of As2 O3 in combination with cinobufacini on proliferation and apoptosis of the K562 cells and provide theoretical basis for clinical application.Methods Cell proliferation was assayed by analyzing the growth and viability of the cells.Apoptosis was assayed by performing cell morphology,Annexin-V/PI staining,DNA-PI staining,and DNA gel electrophoresis.Results After exposure to As2O3 and cinobufacini,the growth of K562 cells was inhibited and the viability of K562 cells was decreased. After treated with 1.0μmol/L As2O3,0.125μg/ml cinobufacini,0.25μg/ml cinobufacini,1.0μmol/L As2O3 + 0.125 μg/ml cinobufacini,1.0μmol/L As2O3 + 0.25μg/ml cinobufacini for 24 and 48 hours,the proliferation inhibition rate were(24 ± 1.3)%,(21 ± 1.5)%,(38 ± 3.1)%,(57 ±2.7)%,(66 ±3.3)% and(49 ±2.9)%,(48 ±2.7)%,(61 ±2.1)%,(77 ±3.8)%,(82 ±4.2)%,the apoptosis rate of K562 cells were(4.8 ± 0.5)%,(5.6 ± 0.7)%,(9.8 ± 0.6)%,(11.9 ± 1.2)%,(15.2±1.5)% and(11.0 ±0.9)%,(12.9 ±1.1)%,(18.4 ±1.5)%,(21.0 ±2.0)%,(28.0 ±1.9)%.The percentage of apoptotic cells was a time- and dose-dependent manner.Typical DNA ladder was shown by DNA gel electrophoresis.Conclusions As2O3 combined with cinobufacini inhibited the proliferation of K562 cells and induced apoptosis of the K562 cells,the combination of the two drugs had better effect.

Objective To evaluate the diagnosis and management of ectopic pancreas.Methods The clinical data of 62 cases of ectopic pancreas,which were diagnosed by endoscopic uhrasonography (EUS)or pathologic findings between July 2006 and December 2007 were retrospectively analyzed.The cases were divided into 4 groups according to different ways of management.Group A included 37 patients,who were diagnosed as having ectopic pancreas(＜19mm)by EUS only and were followed up via phone call every 3 months.Eight patients in group B were diagnosed by EUS as having submucosal lesions suspected as ectopic pancreas,and underwent operation because of large size or difficulty in diagnosis.Eight patients in group C received operation for other diseases and the eetopic pancreases were found in operation.Group D included 9 patients who underwent surgery due to malignant tumors.ResultsEctopic pancreas were most commonly found as a single lesion in gastric antrum(35/62,56.5%)with mean size of 9.2±5.4 mm.All patients in group A were asymptomatic,of which 10 received followed-up endoscopy and no changes in size of the lesion were found.All patients in group B,C and D were diagnosed as ectopic pancreas pathologically.Conclusion Ectopic pancrea is relatively common and asymptomatic,only part of them could be diagnosed clinically.Carcinoma arising from the ectopic pancreas is rare and lesion of small size diagnosed by EUS could be followed up endoseopieally.