OBJECTIVE: To establish a HPLC method for the determination of content of entecavir and its related substances. METHODS: The HPLC condition was consisted of a Luna C18 column with a column temperature at 25 ℃, using gradient eluate method, the mobile phase was acetonitrile,with a flow rate of 1.0 mL?min-1 and detection at 254 nm. RESULTS: The calibrated linear curve of entecavir was within 0.033 45～0.167 2 mg?mL-1(r=0.999 98). The average recovery was 100.09%,RSD=0.20%, The content of related substance was 0.54%～1.48%. CONCLUSION: This accurate and reliable HPLC method is applicable for the quality control of entecavir and its related substances.

AIM: To evaluate the protective effect of luteolin on endothelial dysfunction induced by tert-butyl hydro-peroxide (t-BOOH). METHODS: We observed the effect of luteolin on t-BOOH-induced contractions in the aorta rings with or without endot helium, which were incubated with luteolin (10-6 to 10-4 mol/L) for 30 min before determining the concentration-response to t- BOOH. Cultured endothelial cell line (ECV304) was pretreated with different concentrations of luteolin (10-6 to 10-4 mol/L) for 30 min and then exposed to 10 -5 mol/L t-BOOH for 24 hours. Cell morphology was observed , and cell viability was determined by MTT assay. Meanwhile, RT-PCR was used to measure the expression of eNOS and COX-1. RESULTS: Increasing concentrations of t-BOOH produced concentration-dependent cont ractions in aorta rings isolated from rats, luteolin effectively attenuated the contraction in a concentration-dependent manner, and the relaxation response was greater in intact endothelium segments. In MTT and RT-PCR assays, luteolin effectively reduced the cytotoxicity of t-BOOH to endothelium cell and increased the expression of nitric oxide synthase (eNOS) mRNA, which was greatly down-regulated by t-BOOH. CONCLUSION: Luteolin effectively protects the endothelium from the impairment of oxidative stress, and the protection could be related to its negative modulation towards t-BOOH-induced contractions in the aorta.

The number of hypertension patients in China has increased year by year. Traditional Chinese medicine (TCM) has obvious advantages in the treatment of hypertension. Document resources of TCM treatment on hypertension play an important role as references and guidance. Agile construction thinking, architecture based on MVC, and data demand driven will make the building of information service system with efficiency and high quality. The content provided by this information service system will promote the scientific research and clinical practice of hypertension.

Aim To investigate the periphery analge-sic effect of myricetin on a rat model of inflammatory pain and the mechanism. Methods Rat models of in-flammatory pain were induced by complete Freund ’ s adjuvant ( CFA) injection in left hindlimb plantar cen-ter. The thermal withdrawal latency ( TWL) was meas-ured before and after CFA or myricetin treatment. Elec-trophysiological method was used to identify the effect of myricetin on the action potential frequency and the voltage dependent potassium channel currents in small DRG neurons. Results Rats with CFA injected showed thermal hyperalgesia ( P <0. 05 ) and TWL in-creased significantly after myricetin intraperitoneally in-jected ( P <0. 05 ) . Current clamp recording showed the action potential frequency of small DRG neurons in rats was inhibited by myricetin ( P<0. 01 ) and voltage calmap recording showed the inhibitory effect of myr-icetin was enhanced by calcium depended potassium channel currents ( P<0. 05 ) . Conclusion Myricetin exerts periphery analgesic effect by enhancing calcium depended potassium channel currents and inhibiting excitability of small neurons of dorsal root ganglion.

Four small GTPase genes which may be relative to sclerotial development were firstly cloned from medicinal fungus Polyporus umbellatus using rapid amplification of cDNA end PCR (RACE) method. The results showed that full-length cDNA of PuRhoA was 698 bp contained 585 bp ORF, which was predicted to encode a 194 amino acid protein with a molecular weight of 21.75 kD with an isoelectric point (pI) of 6.44; the full length cDNA of PuRhoA2 was 837 bp in length and encoded a 194 amino acid protein with a molecular weight of 21.75 kD and an isoelectric point (pI) of 6.33; the full length cDNA of Puypt1 was 896 bp in length and encoded a 204-aa protein with a molecular weight of 22.556 kD and an isoelectric point (pI) of 5.75; the full length cDNA of PuRas was 803 bp in length and encoded a 212-aa protein with a molecular weight of 23.821 kD and an isoelectric point (pI) of 5.2. There are fani acyl transferase enzyme catalytic site and myrcene-transferase enzyme catalytic site in PuRhoA1 while the PuRhoA2 only possess myrcene-transferase enzyme catalytic site. Puypt1 contains the Rab1-Ypt1 conserved domain of small GTPase family and PuRas contains the fani acyl transferase enzyme catalytic site. According to the phylogenetic analysis all these four small GTPase clustered with basidiomycete group. Quantitative real-time PCR analysis revealed that Puypt1, PuRas and PuRhoA1 transcripts were significantly higher in the beginning of sclerotial formation than that in the mycelia, whereas the transcripts levels of PuRhoA2 gene were particularly lower in sclerotia than that in mycelia, suggesting that these four genes might be involved in P umbellatus selerotial development.

Corresponding author: SONG Guang yao Human umbilical venous endothelial cells (HUVEC) were incubated with free fatty acids (FFAs) of various degrees of saturation. C18:1 and C18:2 inhibited the synthesis of nitric oxide (NO) and endothelin (ET) in HUVEC, and ET synthesis was inhibited much more than NO synthesis, and C18:0 increased ET synthesis in HUVEC, suggesting that the effects of FFAs on HUVEC were related to their degree of saturation.

AIM: To preliminarily study the molecular mechanism for the cardiac injury in rat by adriamycin and the mechanism for the acute repair in the body. METHODS: The male Sprague Dawley rats were randomly divided into four groups (n=10 in each): The first group was kept without treament and served as the control ;the second, the third and the fourth received ADR in different doses (10, 20, 40 mg?kg -1 , respectively) by injection of adriamycin. The content of malondialdehyde (MDA)in the serum was estimated with thiobarbituric acid. Cu Zn SOD was measured by its reaction with xanthine oxidase. GSH was measured by its reaction with 5, 5 nitrobenzoic acid. Using semi quantitative reverse transcription polymerase chain reaction (RT PCR),we analyzed the expression of the associated gene. RESULTS: MDA contents in the medium and high ADR dose groups were higher than that in the control group (P

Objective To evaluate the effect of evidence-based nursing in preventing and controlling the pulmonary infection of patients with stroke.Methods The randomized controlled trials (RCT) on the evidence-based nursing in preventing and controlling the pulmonary infection of patients with stroke were collected using the domestic and foreign databases,which were analyzed by RevMan 5.2 software.Results Nine RCTs were included in the study.The results of Meta-analysis revealed that the effect of the amount of OR values and 95％ confidence interval of the incidence and the cure rate of pulmonary infection was 0.42 (0.33～0.54),14.10 (4.57-43.50).Conclusions The evidence-based nursing can reduce the incidence of the pulmonary infection and improve the cure rate.

Objective To study the risk factors and to provide preventive measures of pulmonary infection for patients with severe brain injury.Methods A total of 320 patients with severe brain injury were analyzed with hospital-based case-control study method.A total of 101 patients complicated with lung infection were set as the experimental group.Another 101 patients with no pulmonary infection according to the proportion of 1:1 choice over the same period were set as the control group.The related factors were analyzed with the univariate analysis and the multiple factors Logistic regression analysis.Results Univariate analysis showed that the occurrence of pulmonary infection of patients with severe brain injury was closely related to the old age,low GCS score,long time stay in the bed,long time hormone application,long time antibiotic application,the combined underlying disease,use of mechanic ventilation,long-term smoking history and indwelling gastric tube.Multivariable Logistic regression analysis showed that the application of mechanic ventilator,the combined underlying disease and long time stay in bed were the independent risk factors for pulmonary infection of patients with severe brain injury,but GCS score was a protective factor.Conclusions The occurrence of lung infection were related to the ventilator application,the combined underlying disease,long time stay in bed and low GCS score.Considering these risk factors,the medical staffs should take preventive measures early to control lung infection.

OBJECTIVE:To establish an RP-HPLC method for content determination of s-omeprazole sodium and its related substances.METHODS:The separation of s-omeprazole sodium and the related substances was carried out on a Phenomenex Luna C18 column,the mobile phase consisted of methanol-0.033 mol?L-1 ammonium dihydrogen phosphate-triethylamine (58∶41.8∶0.2,adjusted to pH 7.0 by phosphate acid).The detection wavelength was 302 nm,the flow rate was 1.0 mL?min-1,the column temperature was 25 ℃,and the sample size was 20 ?L.RESULTS:The linear range of omeprazole sodium was 10～500 mg?L-1 (r=0.999 7).The average recovery rate was 100.27% (RSD=0.74%).The average content of the related substances in samples was 0.42%.CONCLUSION:This method is simple,accurate,specific and applicable for content determination of s-omeprazole sodium and its related substances.