Objective: To establish a rapid method for detecting acetylbritannilactone (ABL) by online sweeping-micellar electrokinetic chromatography (MEKC) and to elevate the sensitivity of the detection. Methods: The combination of online sweeping technique with MEKC was used to determine the content of ABL in the extract of Inula britannica in plasma of rats. Results: ABL was completely separated within 15 min in running buffer and sample buffer. The optimal conditions were as follows: on uncoated fused quartz silica capillary, with separation voltage of 23 kV, capillary temperature of 25 °C, and detection wavelength of 195 nm. The regression equations revealed good linear relationships between the peak area and concentration of ABL (r = 0.998), with the detection limits of 0.005-0.15 mg/mL. The relative standard deviations of migration time and peak areas for intra- and inter-batch were < 2.45% and < 2.26%, respectively. The recovery rate of this method was 96.3%-97.2%. Conclusion: This method provides some advantages in separation speed, testing sensitivity, and operating convenience, with low sample and reagent consumption. The online sweeping-MEKC is an effective method for pharmacokinetic study and analysis on tracing biological samples. © 2014 Tianjin Press of Chinese Herbal Medicines.

Objective:To study the effect of comfort care to alleviate postoperative pain of tibial eminence fracture by fixation. Methods:55 patients with tibial eminence fracture between Apr 2013 and Mar 2015 in our hospital were selected and divided into observation group with 26 patients and control group with 29 cases according to the nursing methods. The control group was given the orthopaedic routine nursing method, and the observation group was given the comfortable nursing method to compare two groups of patients with postoperative pain and psychological status before and after the intervention (HAMA anxiety score and HAMD depression scale). Results:There were significant differences between two groups of patients with pain score after the intervention (t=9.545, P<0.05), and HAMA and HAMD scores decline. HAMA and HAMD scores in the observation group were significantly lower than that in the control group (t=11.593, t=10.628;P<0.05). Conclusion:Comfort care can significantly relieve postoperative pain of the tibial condyle spine fractures and improve postoperative anxiety and depression. It has good clinical application value.

Objective To value the effect and safety of butyl phthalide sequential treatment in cerebral infarction of branch sclerosis of arterial congee appearance. Methods Sixty patients with acute cerebral infarction were randomly divided into treated group( n = 31)and control group( n = 29). According to the condition of illness,all patients were given aspirin,atorvastatin calcium,and the injection of ozagrel sodium intravenous;controlled the blood pressure,blood sugar,blood lipid,and treated complications posstively;take the early rehabilitation of nerve treatment afte the illness was in stable condition. Butyl phthalide was used in the patients of treated group(100 ml,twice per day,intravenous drip,during 14 days period therapy,and then 0. 2 g oral,third per day),besides the routine therapy. The degree of neural function defect score( NIHSS)and activities of daily living score(BI)between two groups were observed before and after treatment. Corresponding adverse consequences were recorded. Results Compared with pretreatment,the NIHSS of postreatment at the 14th day in treat and control groups were decreased(treated group:(4. 36 ± 3. 11)vs.(11. 42 ± 3. 20);control group:(6. 12 ± 2. 67)vs.(11. 64 ± 3. 43),P < 0. 05,and the treated group was significantly lower than control group(F inner groups = 2. 125,P < 0. 01;F between groups = 18. 63,P < 0. 01;F cross groups = 25. 34,P< 0. 01;P < 0. 05). The BI of postreatment in two group were increased(treated group:(86. 72 ± 8. 44)vs. (26. 54 ± 13. 36);control group:(75. 96 ± 9. 86)vs.(26. 38 ± 13. 02)),and the treated group was significantly lower than control group(F inner groups = 29. 27,P < 0. 01;F between groups = 32. 48,P < 0. 01;F cross groups= 42. 41,P < 0. 01;P < 0. 05). There was no the adverse reactions. Conclusion Butyl phthalide sequential treatment can improve the NIHSS and BI of cerebral infarction of branch sclerosis of arterial congee appearance and have a better therapy effect.

Objective To investigate the inhibitory effects of periplocin from cortex periplocae (CPP) on human lung cancer cell line QG56 and to discuss its mechanism. Methods QG56 cells were cultured in vitro. The final concentrations of CPP in control group were 1.25, 2.50, 5.00, 10.00 and 20.00μg/L. QG56 cells were treated with ascending concentration of CPP for 24 h, 48 h and 72 h. The cell proliferation was measured using MTT method. The morphological changes of QG56 cells were observed under inverted microscope. Flow cytometry (FCM) was used to detect the effects of CPP on cell cycle and cell apoptosis. The expression of apoptosis associated gene bax mRNA in QG56 cells was detected by RT-PCR. The expres-sion of bax protein before and after treatment of CPP was examined by SP immunocytochemistry. Results The inhibitory ef-fect of CPP on the proliferation of QG56 cells was increased with the increasing concentrations of CPP and the prolonged du-ration of treatment. The morphological changes were displayed in QG56 exposed to CPP. The results of FCM showed that CPP caused cell cycle arrest at G0/G1 phase. The apoptotic rate of QG56 cells was significantly increased after CPP treatment for 48 h (P<0.05). The expression of bax mRNA was increased in QG56 exposed to CPP. The result of immunocytochemis-try indicated that CPP up-regulated the expression of bax protein. Conclusion CPP showed significant inhibitory effect on human lung cancer cell lines QG56 through inducing cell cycle arrest and apoptosis.

Background Acute bacterial conjunctivitis is a common ocular infectious disease.It can be cured by topical administration of antibiotics,but these antibiotic eye drops often was used several times per day at present.Azithromycin is thought to offer less using times and shorten the duration of treatment.Objective The purpose of this clinical trail was to evaluate the clinical efficacy and safety of topical administration of 1％azithromycin eye drops for the treatment of acute bacterial conjunctivitis.Methods This study was approved by Ethic Committee of Beijing Tongren Hospital and followed the Declaration of Helsinki.Written informed consent was obtained prior to entering into this trial.A randomized,double-blind and placebo-controlled study was designed.One hundred and eighty patients with acute bacterial conjunctivitis were enrolled in Affiliated Hospital of Qingdao Medical College and Qingdao Municipal Hospital from may,2011 to September,2011.Azithromycin eye drops at the concentration of 1％ was topically administered on 89 eyes of 89 patients twice per day initial and once per day later for 9 days in the trial group,and placebo eye drops was used on 91 eyes of 91 patients in the same way in the control group.The inflammation response of all eyes was examined and scored under the slit lamp microscope,and germiculture was carried out.All adverse responses were assessed during the follow-up.Clinical efficacy was evaluated with the clinical cure rate as the primary indicator and bacterial clearance rate as the secondary indicator.The safety of drug was determined,including the ocular irritation sign,lens transparency and intraocular pressure.The differences of the examining results mentioned-above were compared with Chi Square test.Results One hundred and eighty patients completed the clinical trials according to the program without lost and exclusion cases.The clinical cure rate in 89 subjects in the trial group and 91 control individuals was 76.40％ (68/89) and 43.96％ (40/91),respectively,with significant difference between them (x2=19.73,P＜0.01).The bacterial eradication rate was 85.71％ (24/28) in the trial group and 60.53％ (23/38) in the control group,showing a significant difference (x2 =4.99,P＜0.05).Both 1％ azithromycin eye drops and placebo were well-tolerated with a low incidence of adverse events.Conclusions One percent azithromycin eye drops is effective and safe for the treatment of acute bacterial conjunctivitis with less adverse reaction.

Objective To discuss the feasibility on building lewis lung carcinoma mouse models through different methods and improve the methods. Methods The method of culture LLC cells in vitro, trypsin digestion method, Ⅳ collagenase method and homogenate method were compared to make the different dose of cell suspension injected into C57BL/6 mice. The feasibility of the improved method was determined through observing the cell count, the tumor formation ratio, the tumor formation time, tumor volume, weight and life habit. Results The method of culture LLC cells in vitro could get needed cells and its tumor formation ratio was 100 %. Trypsin digestion method and homogenate method could get less cells and its tumor formation ratio was about 80 %～90 % and 60 %～75 %. Whereas 1V collagenase method could get most cell count and its tumor formation ratio was 100 %. Conclusion IV collagenase method is a preferred method which is simple,high efficiency and make a strong base on the cancer experimental study.

Objective To explore the relationship between Arg913Gln(G→A) polymorphism of solute carrier family 12 member 3 (SLC12A3) gene and diabetic nephropathy (DN) in type 2 diabetes mellitus (T2DM) in Han population of Shanghai. Methods Two hundred and fifty-eight Han ethnic people in Shanghai with T2DM (T2DM group) were divided into non-DN group (DN0 group, n=95) and DN group (n=163) according to 24 h urine albumin excretion rate (AER), and those in DN group were subdivided into microalbuminuria group (DN1 group, n=95) and macroalbuminuria group (DN2 group, n=68). Besides, 82 people with normal results of oral glucose tolerance test (OGTT), without diabetes mellitus and nephropathy were served as controls. PCR-sequencing was used to detect the genotypes of Arg913Gln polymorphism of SLC12A3 gene. Genotypic and allelic frequencies and clinical characteristics were compared among groups. Results Three genotypes (GG, GA and AA) were detected. The frequencies of GA+AA genotype and A allele in T2DM group were higher than those in control group, while there was no significant difference between groups (P>0.05). There was no significant difference in genotypic or allelic frequencies among subgroups of T2DM group (P>0.05). The level of triglyeeride (TG), AER, level of fasting insulin (FINS) and HOMA-IR in patients with GA+AA genotype were significantly higher than those in patients with GG genotype in T2DM group (P<0.05). Conclusion Arg913Gln(G→A) polymorphism of SLC12A3 gene is not significantly associated with T2DM and DN in Han population of Shanghai. The AER of people with GA+AA genotype is significantly higher than that with GG genotype. Arg913Gln (G→A) polymorphism of SLC12A3 gene may predict the risk of increase of albuminuria in patients with T2DM in Han population of Shanghai.

Objective To develop a sensitive,specific, simple and rapid quantitative real-time PCR (Q-PCR) assay for detection of Staphylococcus aureus with SmartCycler.Methods According to the nuc gene sequences specific to S.aureus, a pair of primers and one TaqMan probe were designed. An internal amplification control (IAC) which is a chimeric double-stranded DNA constructed from a fragment of the Listeria monocytogenes hly gene flanked by the nuc-specific target sequences was added to the reaction system. This IAC was detected using a second TaqMan probe labeled with a different fluorophore. The performance of the nuc-IAC Q-PCR was evaluated using artificially contaminated drinking water and commercial UTH whole milk samples spiked with ATCC 6538.Results The nuc-IAC assay could be used reliably for detection with a sensitivity of 5 copies of linear plasmid DNA per reaction, 10 fg of genomic DNA in 62.5% of the reactions or 50 cfu/ml S.aureus cells with 50% probability. The quantification was linear (r~2≥0.998) over a 6-log dynamic range, with a PCR efficiency over 0.967. The 5×10~2 CFU per 25 ml mimic sample of drinking water or milk could be detected by this assay consistently and quantifiably.Conclusion The nuc-IAC Q-PCR assay for S.aureus is developed. It could not only be applied for the quantitative detection of S.aureus, but also prevent the false negatives and underestimations of contamination loads due to PCR failure.

Aim To investigate estradiol inhibition of neointimal proliferation after rat carotid artery balloon injury. Methods Eight to ten-week-old SD rats (male,n=21,female,n=21) were divided into intact control(n=7),gonadectomy control(n=7) and estradiol (n=7, gonadectomy)groups in each sex. Left carotid artery was not injured with 2.0 F PTCA balloon until estradiol was injected for three days. Rats were killed 2 wk after injury. Neointimal areas and media area, ratios of intimal areas/media areas were measured with computer. Results Male neointimal areas and ratios of intimal areas /media areas in estradiol group were less than those in intact control group significantly(all P0.05). Conclusions Estrdiol inhibits neointimal proliferation after the gonadectomy in rats undergoing carotid artery balloon injury.

AIM: To investigate the immune mechanisms for Periplocin from Cortex Periplocae (CPP) in tumor-bearing mice. METHODS: H_(22) tumor-bearing model BALB/c mice were applied to evaluated in vivo immunoregulatory effect of CPP. The influence of different dose CPP (0.25, 0.50 and 1.00 mg/kg) on immune organs in tumorbearing mice were observed. T cell subsets of mice spleen were detected by flow cytometry. MTT assay was used to determine the influence of CPP on lymphocyte proliferation of mice spleen stimulated by ConA. The levels of TNF-α, IL-2 and IL-12 in serum from mice were detected by means of ELISA. RESULTS: Thymus index and spleen index of H_(22) tumor-bearing model control mice became less than that of normal mice (P < 0.05). Compared to both model and normal control groups, thymus index and spleen index of H_(22) tumor-bearing mice treated with CPP increased obviously (P < 0.05). CPP had no influence on the number of CD8~+ T cells, but up-regulated markedly the number of CD3~+, CD4~+ T cells and the ratio of CD4~+/CD8~+ in tumorbearing mice. In CPP-treated mice, the percentage of CD3~+, CD4~+ T cells were not different from normal mice (P<0.05), the ratio of CD4~+/CD8~+ was higher than that of normal mice (P < 0.05). CPP enhanced obviously lymphocyte proliferation of mice spleen induced by ConA, the SI scores were even higher than that of nornal mice. The levels of TNF-α, IL-2 and IL-12 in serum from CPP-treated mice, increased significantly compared to model control group (P<0.05) in a dose-dependent manner, were similar to or higher than that of normal mice. CONCLUSION: CPP protected immune organs of tumor-bearing mice, increased obviously the percentage of CD4~+ and CD4~+/ CD8~+ among the T cell line, and enhanced lymphocyte proliferation of mice spleen significantly, stimulated the production of TNF-α, IL-2 and IL-12. The results suggested that CPP possessed potent immunoregulatory effect.