Objective To investigate the anti-HBV constituents from Alternanthera philoxeroides.Methods The constituents were isolated with silica gel and gel permeation chromatography,and purified by HPLC.Their structures were elucidated by spectroscopy.The antivirus effects of the isolated compounds were tested by ELISA method in vitro.Results Ten compounds were isolated and elucidated as followings:oleanolic acid(Ⅰ),oleanolic acid 3-O-?-D-glucuronopyranoside(Ⅱ),oleanolic acid 28-O-?-D-glucopyranoside(Ⅲ),chikusetsusaponin Ⅳ a methyl ester(Ⅳ),4,5-dihydroblumenol(Ⅴ),N-trans-feruloyl 3-methyldopamine(Ⅵ),N-trans-feruloyl tyramine(Ⅶ),3?-hydroxystigmast-5-en-7-one(Ⅷ),24-methylenecycloartanol(Ⅸ),and cycloeucalenol(Ⅹ).The values of inhibition percent of compounds Ⅰ-Ⅲ,Ⅴ-Ⅶ revealed a significant distinction compared to the control group.Compounds Ⅱ and Ⅴ showed significant inhibition against HepG2 cells transected with cloned hepatitis B virus DNA,their inhibitive ratios were 85.38% and 87.37% at 50 ?g/mL,respectively.Conclusion Compounds Ⅳ-Ⅶ are isolated from this plant for the first time and phenolic amides have been determined as the new structure type from the plants of Alternanthera Forsk.Compounds Ⅱ and Ⅴ from A.philoxeroides show the more significant anti-HBV activities.

Objective To explore the first-aid treatments and nursing measures of children with Adams-Stokes syndrome.Methods Totally 6 data of patients with Adams-Stokes syndrome who was successfully rescued were analyzed and discussed.Results The main cause of Adams-Stokes syndrome were Severe ventricular tachycardia.The third degree atrioventricular block,Severe myocarditis,Dilated cardiomyopathy,the common inducing factors of Adams-Stokes syndrome were Hypokalemia,psychological factors,overeating,constipation.Conclusion The premise of the successful rescue on Adams-Stokes syndrome children are regulated observation、knowing the patient' s condition well,the medicine and goods for first-aid treatment all ready,and timely finding,correct estimate,giving emergency treatments timely,appropriate nursing are the keys of its successful rescue.

Objective To establish the quality standard of Zhimiling Suppository (Cortex Phellodendri, Radix Sophorae Flavescentis, Catechu, Borneolum Syntheticum, Alumen). Methods Catechu, Borneolum Syntheticum, Alumen in the preparation were identified by physics and chemstry method. Berberine Hydrochloride was determined by HPLC. Results The standard curve for Berberine Hydrochloride was linear in the range of 0.080 2～0.802 ?g, the average recovery was 100.4%, RSD was 0.85% (n=6). Conclusion The methods are simple, accurate and reproducible.

Objective To compare the field-matching techniques and dosimetric characteristics in the target area of testicular seminoma including the abdominal and pelvic cavities between direct aperture optimization intensity modulated radiation therapy (DAO-IMRT) and 3D conformal radiation therapy (3D-CRT),and to analyse the advantages and disadvantages of DAO-IMRT.Methods DAO-IMRT and 3D-CRT plans were designed with Pinnacle treatment planning system for 7 testicular seminoma patients,and the characteristics of both the irradiation methods were analyzed and evaluated by means of the parameters like dose homogeneity indexes,dose volume histograms and etc.Results DAO-IMRT had the hot and cold volumes in the target areas both significantly smaller than those by 3D-CRT,while the conformity index and homogeneity index superior to those by 3D-CRT.The mean doses (Dmean) by DAO-IMRT of the organs at risk (OAR) except the left femur head were all lower than those by 3D-CRT,aud the V15 values of liver,double kidney,small intestine and rectum by DAO-IMRT were statistically lower than those by 3D-CRT (P＜0.01).3D-CRT had the V5 values of OAR all lower than those by DAO-IMRT,in which the differences of double kidney,small intestine,bladder,left femur head and normal tissues were significant statistically (P＜0.05).The monitor units by DAO-IMRT roughly doubled when compared with those by 3D-CRT,and the difference was also significant (P＜0.01).Conclusion DAO-IMRT with easy operation and high reliability can avoid the connection of fields in the target area of testicular seminoma,and is superior to 3D-CRT in dosimetry.

Objective To study the relationship of expression of metastasis suppressor gene nin23-H1 protein in gastric carcinoma tissue and tumor infiltration and metastasis.Methods The expression of metastasis suppressor gene nm23-H1 protein in 50 gastric carcinoma tissues was detected by SP immunohistocbemistry tech-nology.Results Correlations were presented among expression of nm23-H1 protein, infiltrate grade of gastric carcinoma, lymph node metastasis and TNM stage.Conclusions Expression of nm23-H1 may suppress infiltra-tion and metastasis of gastric carcinoma, Low expression of nm23-H1 protein may be of great reference value in judgement of tumor progression, metastasis and reeidivation.

Objective To Study the expression and clinical meaning of nm23-H1,c-Met and survivin protein in gastric carcinoma tissue.Methods Expression of nm23-H1,c-Met and survivin protein in 50 paraffin block samples of gastric carcinoma were detected by S-P immunohistochemistry technology.The relationship of expression to tumor differentiation degree,infiltration depth,lymph node metastasis,TNM stage,tumor metastasis and reeidivation were analyzed.Results The positive expression rates of nm23-H1,c-Met and survivin protein were 46%,72% and 80%,low nm23-H1 protein expression and high c-Met protein expression correlated tightly with TNM stage and lymph node metastasis of gastric carcinoma,high survivin protein expression correlated tightly with differentiation degree,TNM stage and lymph node metastasis of gastric carcinoma,there was a inverse correlation between nm23-H1 protein expression and survivin,but a direct correlation between c-Met and survivin.Conclusions Low nm23-H1 protein expression and high c-Met and survivin expression has important guidance significance of making a early diagnosis and judging prognosis.Selective gene therapy guided by these may be helpful.

Objective To investigate the biological effects of bcl-2 gene on neurons. Methods The recombinant expression plasmid pc DNA3-bcl-2 was constructed from pSFFV-bcl-2,then it was introduced into PC12 cell line by liposome method.Western blotting and immunohistochemistry in situ were applied to exam the exogenous gene expression.The two groups of cells(Group A,PC12 transfected by pcDNA3-bcl-2 and Group B,PC12 transfected by pcDNA3) were exposed to cisplatin with the concentration of 10*!?mol/L,50*!?mol/L, and 100*!?mol/L 72 hours later,the survival cells were estimated.Cell cycle indexes between these two groups were also studied by FCM. Results The recombinant expression plasmid pcDNA 3-bcl-2 was constructed successfully and PC12 cell line transfected by the plasmid could express Bcl-2 protein effectively.Compared with the control(25 79%),there was a significant decrease of cells from the S phase in PC12 with bcl-2 gene(8.81%).After exposed with 10*!?mol/L,50*!?mol/L,and 100*!?mol/L cisplatin,the surviving cells in group A were 276?13,185?11 and 108?10 respectively,which increased much more than in group B while they were 100?9,12?3 and 2?2 accordingly.Conclusions bcl-2 can protect PC12 cells against cytotoxic insults of cisplatin,and it suggested that it might act via cell cycle controlling.

An offline silver-impregnated silica gel solid-phase extraction (Ag-SPE) approach combined with a programmed temperature vaporization-gas chromatography-flame ionization detector (PTV-GC-FID) was proposed for routine analysis of mineral oil saturated hydrocarbons (MOSH) in chocolates. The MOSH in chocolates were extracted by n-hexane and 1 mL of extract was purified by offline Ag-SPE column. The SPE columns packed with 0.3％ Ag-activated silica gel were used to separate MOSH from triglycerides and olefins in chocolates. The eluent of MOSH fraction was only 5 mL and then concentrated to 0.2 mL through nitrogen blowing with little evaporation loss. The PTV parameters were as follows: the initial temperature was set at 45℃ and held for 1 min (split ratio was 200∶1), then warmed up to 360℃ at linear gradient of 250℃ minSymbolm_1 and held for 27 min (split valve was closed for 2 min followed by split ratio of 100∶1). The GC injection volume was 40 μL. The GC column was heated from 35℃ (3 min) to 350℃ at 25℃/min, and then raised to 370℃ (10 min) at 5℃/min. The flow rate of the carrier gas was 1.3 mL/min (and pressure was 60 kPa), FID temperature was set at 380℃. The limit of quantification (LOQ) and the recoveries of the method were 0.5 mg/kg and 84.9％-108.6％, respectively, with the relative standard deviations (RSD) of 0.2％-1.5％. Twenty-five commercial chocolate samples were analyzed with the proposed method, and it was found that the MOSH in three samples were not detected, and the concentrations of MOSH in other 22 samples were 1.09-8.15 mg/kg (the concentrations of MOSH with C16-C35 component were 0.56-4.43 mg/kg). The results suggested that it was necessary to routinely detect mineral oil contamination in chocolates for food safety.

Aim To study the change of mutant pre- vention concentration (MPC) in carbapenem-resistant Acinetobacter baumannii (CRAB) treated with moxi- floxacin ( MFX ) and/ or cefoperazone/ sulbactam (CFS) in vitro, and provide a theoretical support for preventing the bacterial resistance. Methods To cal- culate the fractional inhibitory concentration (FIC) in- dex, the minimum inhibitory concentration (MIC) of 20 clinical isolates of CRAB treated with MFX and/ or CFS was determined by checkerboard microdilution as- say. In addition, to calculate the selection index (SI), the MPC of 20 clinical isolates of CRAB treated with MFX and/ or CFS was determined by agar plate di- lution assay. Results Our study showed that there was synergistic/ addictive action, rather than antago- nism action against clinical isolates of CRAB when treated with MFX + CFS. The SI of the 20 isolates trea- ted with MFX or CFS alone was 4 ~128 and 8 ~64 re- spectively, but reduced to 1 ~8 and 4 ~16 when trea- ted with MFX + CFS, which decreased by 2 ~16 and 2 ~4 times respectively compared with the single treat- ment. Conclusion These results suggest that the combination treatment of MFX + CFS against clinical isolates of CRAB might lower the MPC of the isolates treated with MFX/ CFS alone, narrow the mutant selec- tion window, and prevent the generation of drug - re- sistant mutants.

Objective To construct pcDNA 3.1(+)/HPV 18 E 6 fusion gene and a single-codon mutation pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R fusion gene in eukaryotic expression vector and study the effects on proliferation and apoptosis of cervical carcinoma cell line Hela. Method HPV 18 E6 gene sequence and the single-point mutation HPV 18 E6 F49R or HPV 18 E6 F127R were amplified from total RNA of Hela cell line by reverse transcription- polymerase chain reaction ( RT- PCR), then the gene sequences were respectively inserted into pcDNA 3.1(+) vector to reconstruct recombinant plasmids which were transfected transiently into Hela cells. MTT and RT-PCR were used to test the expression levels of HPV 18 E6 and the growth of HeLa cells after transfected about 48 h. The proliferation and apoptosis of Hela cells were detected respectively by cell counting and AO/EB fluorescent vital staining. Results The pcDNA 3.1(+)/HPV 18 E6, pcDNA 3.1(+)/E6 F49R and pcDNA 3.1(+)/E6 F127R eukaryotic expression vectors were successfully constructed. The gene of HPV 18 E6 was discriminably detected in the HeLa cells which were transfected with the recombinant plasmids. After several days, the proliferation of Hela cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R plasmid were obviously inhibited and the apoptotic rates were significantly increased, then the proliferation of cells transfected with pcDNA(+)/HPV 18 E6 was rather increased slightly, and we could observe the phenomena of early apoptosis and the formation of thekaryopyknosis by fluorescent microscope in the cells transfected with pcDNA 3.1(+)/E6 F49R or pcDNA 3.1(+)/E6 F127R. Conclusion The eukaryotic expressing vectors encoding HPV 18 E6 F49R and HPV 18 E6 F127R provide fundamental basis for the further study on HPV 18 E6 mechanism as well as prevention and treatment of uterine cancer.