OBJECTIVE:To establish a method for the contents determination and cluster analysis of free and combined anthra-quinone of aloe emodin,rhein,emodin,rhubarb phenol and physcion in commercially available Yiqing granule from different man-ufacturers. METHODS:HPLC was performed on the column of Eclipse plus C18 with mobile phase of methanol-0.1%Phosphoricac-id solution (gradient elution) at a flow rate of 0.8 ml/min,the detection wavelength was 254 nm,the column temperature was 25 ℃,and the injection volume was 20 μl. RESULTS:The linear range was 0.001 6-0.128 0 μg for aloe emodin(r=0.999 9), 0.003 4-0.273 6 μg for rhein(r=0.999 9),0.003 7-0.299 2 μg for emodin(r=0.999 9),0.006 7-0.536 0 μg for rhubarb phenol (r=0.999 9)and 0.001 7-0.134 4 μg(r=0.999 9)for physcion;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 97.77%-101.47%(RSD=1.30%,n=6),98.09%-100.26%(RSD=0.76%,n=6),96.42%-100.38%(RSD=1.58%,n=6),96.63%-102.50%(RSD=2.17%,n=6) and 97.10%-103.70%(RSD=2.47%,n=6),respectively;the content range of total anthraquinone was 0.042-0.218 mg/g, 0.029-0.448 mg/g, 0.022-0.167 mg/g, 0.032-0.284 mg/g and 0.006-0.060 mg/g,combined anthraquinone was 0.010-0.111 mg/g,0.013-0.092 mg/g,0.011-0.097 mg/g,0.030-0.246 mg/g and 0.001-0.034 mg/g,and free anthraquinone was 0.022 3-0.143 mg/g,0.015-0.356 mg/g,0.008-0.071 mg/g,0.006-0.075 mg/g and 0.003-0.032 mg/g. Cluster analysis showed there were 4 batches belonged to Category 1,2 batches belonged to Category 2 and 4 batches belonged to Category 3 for the total anthraquinone in 5 components;4 bathes belonged to Category 1,2 batches belonged to Category 2 and 4 batches belonged to Category 3 for the combined anthraquinone;and 6 bathes belonged to Category 1 and 4 batches belonged to Category 2 for the free anthraquinone. CONCLUSIONS:The method is simple and accurate,and can provide reference for improving the quality and diarrhea efficacy control of Yiqing granule;there were great differences in Rheum palma-tum and combined anthraquinone from different manufacturers.

ObjectiveTo investigate the clinical epidemiology of intrahepatic cholelithiasis in Guangxi area. MethodsAn retrospective analysis was made on 8?585 cases of intrahepatic cholelithiasis proved by exploration in a period of 19 years. Data were collected and analyzed by computer PEMS.ResultsThe intrahepatic cholelithiasis accounted for more than one third of all biliary stone disease treated during the same period. The prevalence of intrahepatic cholelithiasis in peasants victim increased from 23.4% out of all gall stones in a period of 1981～1985 to 55.8% between 1991～1999. The constituent ratio of intrahepatic lithiasis in males was nearly the same to that in females. The peak age range of patients with intrahepatic lithiasis was 31～40, and the mortality was the highest among all biliary stone disease. ConclusionsIntrahepatic cholelithiasis is by no means a vanishing disease,especially in rural area.

Objective To explore the effects of surface functionalized multi-walled carbon nanotubes (FMWCNTs) on the cytotoxicity of human peripheral blood mononuclear cell (PBMC).Methods Five different types of MWCNTs (hydroxylated,carboxylated,aminated,nickel-plated and pristine MWCNTs (P-MWCNTs)) with the same diameter and length were evaluated the dispersion and characterizations in physiological salt solution by transmission electron microscopy.PBMC were isolated by density gradient centrifugation from human peripheral blood,and 5 types of MWCNTs were ultrasonically dispersed in serum-containing medium respectively.After incubation with PBMC for 12,24,48 or 72 h,cytotoxicity was detected by CCK-8 kits.Results All the MWCNTs had well dispersion,especially the F-MWCNTs.Cytotoxicity results showed that all types of MWCNTs could induced PBMC death,and presented dose-dependence manner and a certain degree of time-dependence manner.Compared with the P-MWCNTs,F-MWCNTs changed cytotoxicity statistically,with the hydroxylated,carboxylated,aminated MWCNTs weakened,aminated MWCNTs significant (P＜0.05),nevertheless the nickel-plated MWCNTs increased.Compared with the P-MWCNTs (25 μg/ml),cell viability of PBMC after 24 and 48 h incubation with the same dose of nickelplated MWCNTs both decreased,and the differences was statistically significant (P＜0.01,P＜0.05).Conclusions The functional group modification affects not only the MWCNTs dispersion in medium,but also the cytotoxicity of the MWCNTs on PBMC.

OBJECTIVE:To prepare Baicalin monolithic osmotic pump tablets and investigate its in vitro drug release behavior. METHODS:Using accumulative release rate as evaluation index,baicalin solid dispersion was prepared to improve solubility,sin-gle factor test and orthogonal test were used to optimize preparation technology(the amount of penetrating agent and pore-forming agent,weight gaining of coating film) of monolithic osmotic pump tablets using baicalin solid dispersion as intermediate. Release rate and mechanism of samples prepared by optimized technology were investigated in 3 kinds of release medium (water,0.1 mol/L HCl,simulated gastric fluid). RESULTS:The optimal preparation technology was that penetrating agent sodium chloride was 30 mg;pore-forming agent polyethylene glycol 400 was 20% amount of excipient cellulose acetate;weight gaining of coating film was 2%. RSD of 12 h accumulative release rate was 1.06%(n=3)for 3 batches of Baicalin monolithic osmotic pump tablets pre-pared by optimized technology. 12 h accumulative release rate of them in 3 kinds of medium were similar to each other,being all more than 80%. Release equation was in line with zero-order drug release model (r=0.9985). CONCLUSIONS:Prepared Ba-icalin monolithic osmotic pump tablets after optimization can release drug at controlled rate.

Objective To explore the methods of therapy in lung cancer patients with bone metastases, and evaluate the effects and side effects of Strontium-89 palliative therapy in lung cancer patients with bone metastases. Methods About 56 cases of bronchiogenic cancer patients with bone metastases who did not receive any radiotherapy, according to 1.48×10~7 Bq/person/time, using standard intravenous injection ~(89)Sr as treatment. Follow-up 6 months, assess according to the following parameters: pain and frequency of pain were given quantized value and got pain score, using T test for comparing the pain score. According to before and after treatment bone imaging showed the size of focus and change of the number, upgrade focus therapy effect. Examine (CEA) and (NSE), using T test for changes before and after treatment. Using T test for changes of LEU and platelet after treatment. Results After treatment for 6 months, for 77 % patients are alleviating pain (43/56), the pain went off of 13 patents, account for 23 percent of the total. The pain score from 7.3±3.6 before treatment decrease to 5.3±3.4 after treatment, dropping obviously. After treatment, the focus regressed in 14 cases, decreased in 5 cases, total efficiency is 34 %. Before and after treatment, CEA from (33.64±18.15)μg/L obviously decreased to (t=4.26, P<0.01) to (21.36±11.65) μg/L, NSE from (27.16±10.12) μg/L obviously decreased to (t=4.26, P<0.05) to (12.56±4.23) μg/L. After treatment, LEU and platelet decreased to the lowest, LEU decreased about 27.9 %, platelet decreased about 19.7 %, after 3 months,normal rate of blood picture is 75 %(42/56). Conclusion The method of strontium-89 palliative therapy in lung cancer patients with bone metastases is good, safe and has little side effects, it can improve the quality of patients life.

ObjectiveTo explore the endocytic pathway of TAT-LHRH modified chitosan/DNA nanoparticle (TLCDN) that exhibits high transfection efficiency and targeting to HepG2.MethodsPlasmid DNA was labeled with fluorescein,and the resulting fluorescent DNA was complexed with chitosan or TAT-LHRH modified chitosan to form chitosan/DNA nanoparticle (CDN) and TLCDN by the complex coacervation method.Internalization of TLCDN or CDN by HepG2 cells were measured in the presence of three kinds of inhibitors of endocytic pathway,Chlorpromazine,Filipin or Dynasore,using High-Content Analyzer to collect and analyze the data.ResultsChlorpromazine led to more decreased uptake of CDN than that of TLCDN,although not statistically significant.Filipin demonstrated significant inhibitory effect on the uptake of TLCDN while promoted the uptake of CDN.Dynasore resulted in a similar decrease in the uptake of both nanoprticles.ConclusionIt was demonstrated that CDN was taken up by HepG2 cells mainly through the clathrin-dependent endocytic pathway and TLCDN was more likely to be internalized by HepG2 cells through the caveolin-mediated endocytic pathway although the clathrin-dependent endocytic pathway was also involved.

Objective To prepare stable aqueous dispersions of chitosan/multi-walled carbon nanotubes (CS/MWCNTs) composites,and observe the effects of CS/MWCNTs on the growth of human umbilical vein endothelial cells (HUVEC).Methods CS/MWCNTs composites were prepared by electrostatic interactions between negatively charged MWCNTs and positively charged low-molecular-weight CS.The prepared CS/MWCNTs were characterized by transmission electron microscopy and Zetasizer nano-analyser.The cellular uptake of the fluorescently labeled CS/MWCNTs was observed by laser confocal microscopy after incubating with HUVEC for 24 h at different concentrations.In vitro cytotoxicity and cellular reactive oxygen were also detected.Results When the mass ratio of low-molecular-weight CS to MWCNTs was equal or greater than 10∶1,the CS/MWCNTs can be stabilized in solution.Cellular uptake experiments showed that the CS/MWCNTs could enter into the cells and locate mainly in the cytoplasm.Cytotoxicity study showed that the CS/MWCNTs composites was less toxic than MWCNTs alone at high concentration (10 and 20 μg/ml).However,there was no significant differencein the level of cellular reactive oxygen between the two groups (P＜0.05).Conclusions CS/MWCNTs composites showed low cytotoxicity and high stability,which would be a promising carrier for drug delivery.

Objective Analysis of myocardial microvascular perfusion in patients with chronic total coronary occlusion (CTO) who underwent a coronary artery bypass graft (CABG) use real-time myocardial contrast echocardiography (RTMCE),to provide an effective method of detecting viable myocardium and a reference for the choice of CABG indications.Methods Twenty-seven patients with CTO underwent RTMCE 1 week before CABG,they underwent follow-up echocardiography and coronary artery 256-slice multislice computed tomography aagiography 1 year after CABG.Myocardial viability was defined as a postoperative ultrasound wall motion significantly improved ≥ 1 point.Semi-quantitative analysis of contrast images,myocardial viability was defined as myocardial perfusion score ≤ 2 points.Viable myocardium by quantitative assessment of myocardial blood flow (MBF) was determined by analyses of receiver-operating characteristic (ROC) curves.Results Patients with LVEF increased significantly after CABG (P ＜ 0.01),Of 259 segments with wall motion abnormality,149 (58％) showed wall motion significantly improved ≥ 1 point after CABG,considered viable myocardium,110 (42％) were not observed in wall motion improved,considered to be non-viable.The viable myocardial segments were significantly greater than non-viable myocardial segments in A,β,A × β value (P ＜ 0.01).Compared with the semi-quantitative analysis,quantitative analysis of MBF increased the sensitivity and accuracy of RTMCE for detecting viable myocardium (P ＜ 0.05).Conclusion RTMCE could accurately assess myocardial viability and provide a strong reference for clinical decision making and judging prognosis.

OBJECTIVE:To develop a method for simultaneous determination of geniposide,baicalin,aloe-emodin,rhein, emodin,chrysophanol and physcion in Zhizi jinhua dispersible tablets. METHODS:HPLC method was adopted. The determination was performed on Dimonsil C18 column with mobile phase consisted of methanol-0.05%phosphoric acid(gradient elution)at the flow rate of 0.8 mL/min. The detection wavelength was set at 254 nm,and the column temperature was 25℃. The sample size was 20 μL. RESULTS:The linear ranges of geniposide,baicalin,aloe-emodin,rhein,emodin,chrysophanol and physcion were 0.0323-0.323 μg (r=0.9998),0.1374-1.374μg(r=0.9999),0.00372-0.0372μg(r=0.9997),0.0069-0.069μg(r=0.9995),0.00332-0.0332μg (r=0.9997),0.00864-0.0864 μg(r=0.9997) and 0.00122-0.0122 μg(r=0.9995),respectively. The limits of quantitation were 0.0321,0.1374,0.00372,0.0067,0.00330,0.00864,0.00122 μg,respectively. The limits of detection were 0.0095, 0.0041,0.0012,0.0020,0.0010,0.0026,0.0003 μg,respectively. RSDs of precision,stability and reproducibility tests were all lower than 3%. The average recoveries were 96.54%-99.52%(RSD=1.17%,n=6),97.23%-101.23%(RSD=1.36%,n=6), 97.22%-101.25%(RSD=1.83%,n=6),97.32%-100.23%(RSD=1.09%,n=6),97.99%-102.71%(RSD=1.73%,n=6), 96.99%-100.23%(RSD=1.21%,n=6),96.99%-103.01%(RSD=2.31%,n=6),respectively. CONCLUSIONS:The methods is simple and reproducible. It can be used for the content determination of 7 components in Zhizi jinhua dispersible tablets.

Objective To detect the expression of Mcl-1 gene in gastric cancer cell lines SGC7901 and MGC-803, and to screen the most effective Mcl-1-targeted siRNA sequence. Methods Mcl-1 expression was evaluated in human gastric cancer cell lines SGC7901 and MGC-803 by RT-PCR. Four segments of siRNAs (siRNA1, siRNA2, siRNA3 and siRNA4) targeting Mcl-1 mRNA and a no-sense control segment were designed by bioinformatic technology . Mcl-1 specific siRNAs were transfected transiently into SGC7901 and MGC-803 cells by using lipofectamine 2000 . After transfected 24 , 48 and 72 h , quantitative real-time PCR was applied to detect the mRNA expression of Mcl-1 and western-blot analysis was applied to detect the protein expression. Results Mcl-1 gene was expressed in both SGC7901 and MGC-803 cells. Overall, siRNA1 exhibited the best inhibitory effect after being transfected for 48h. The inhibition rate of mRNA level in SGC7901 group and MGC-803 group was 73.8%and 67.6%, and the inhibition rate of protein level in SGC7901 group and MGC-803 group was 79.3%and 96.1%. Conclusion Mcl-1 specific siRNA sequences were successfully designed, and siRNA1 was selected as the most effective sequence, which can simultanandeously inhibit the expression of Mcl-1 in GC7901 and MGC-803 cells.