Object To study the chemical constituents of Hibiscus mutabilis L.Methods Isolation and purification were carried out on silica gel, or polyamide column chromatography etc. Constituents were identified and structurally elucidated by physicochemical properties and spectral analysis.Results Ten compounds were obtained, nine of them were determined as tetracosanoic acid (Ⅰ), ?-sitosterol (Ⅱ), daucosterol (Ⅲ), salicylic acid (Ⅳ), emodin (Ⅴ), rutin (Ⅵ), kaempferol-3-O-?-rutinoside (Ⅶ), kaempferol-3-O-?-robinobinoside (Ⅷ) and kaempferol-3-O-?-D-(6-E-p-hydroxycinnamoyl)-glucopyranoside (Ⅸ).Conclusion All compounds are isolated from the plant for the frist time except Ⅱ and Ⅵ.

Objective To construct the uItrasound microbubbIes co-carrying recombinant adenovirus containing stromal cell derived factor 1 (SDF-1α) and bone morphogenetic protein 2 (BMP2),and to study the maximum efficiency of carrying adenovirus and the optimum proportion of double gene combined with ultrasound contrast agents.Methods Microbubbles were combined separately with recombinant adenovirus co-expression of enhanced green fluorescent protein and SDF-1α(pAd-EGFP/SDF-1α) as well as red fluorescence protein and BMP2 (pAd-RFP/BMP2) via biotin-streptavidin method,and the maximum efficiency of carrying DNA in microbubbles was detected.Three microbubbles with binary vectors were prepared by blending the two above-mentioned pAd at different ratio (1 ∶1,1 ∶ 2,2 ∶ 1) into the microbubbles.The microbubbles with binary vectors were evaluated though physiochemical properties,fluorescence microscope and flow cytometry to test the carrying rate of DNA in microbubbles.Results There was no significant difference in PH,average diameter and concentrations between targeted microbubbles and control group (P ＞0.05).The carrying efficiency of DNA increased with virus loads in microbubbles,but lowered if further increasing virus amount after reaching saturation.When the proportion of binary vectors and microbubbles was 1 ∶ 1,its efficiency of carrying SDF-1α gene and BMP-2 was approximately equal,and flow cytometry demonstrated that the positive rate of microbubbles labeled by both fluorescein isothiocyanate(FITC) and rhodamine was (65.6 ± 0.5)％.However,it was (59.0 ± 2.3)％ when their proportion was the 2 ∶ 1,which was significantly lower than those when other two proportions (1 ∶ 1 and 1 ∶ 2).Under the fluorescence microscope,the targeted microbubbles were equally surrounded by bright green or red fluorescence.Conclusions Ultrasound microbubbles of double genes carrying EGFP/SDF-1a and RFP/BMP2 is made successfully via biotin-streptavidin method.The optimal proportion of combining microbubbles with double gene is 1 ∶1,which can reveal the optimum load rate and stable combination.

Objective To explore the relevance of high density lipoprotein cholesterol (HDL-C) level and resistance of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) in advanced non-small cell lung cancer (NSCLC).Methods In a total of 87 NSCLC patients harboring EGFR mutation treated with EGFR-TKI, the correlations of pre-treatment levels of soluble HDL-C between efficacy and resistance of EGFR-TKI were analyzed by Kaplan-Meier method and Cox proportional hazard model.Results Patients with normal HDL-C (≥0.94 mmol/L) had a better disease control rate (DCR) than those with low HDL-C (＜0.94 mmol/L) [94.5 ％ (52/55) vs 65.7 ％ (21/32), P ＜ 0.001].The results of univariate analysis showed that patients with normal HDL-C had longer progress free survival (PFS) (HR =0.27, 95 ％ CI 0.164-0.444, P ＜ 0.001).The results of multivariate Cox proportional hazard model analysis with sex, pathologic histology, smoking history and mutation type showed that only HDL-C group had a significant difference (P ＜ 0.001).The same results were showed when stratified by sex and smoking history (female: P ＜ 0.001, male: P =0.002;Smoker: P ＜ 0.001, non-smoker: P =0.014).Conclusion Normal pre-treatment serum levels of HDL-C are associated with a better outcome and longer progression-free survival in NSCLC patients treated with EGFR-TKI, which may be a predictive marker for a better response.

Aim To develop a sensitive,rapid and ac-curate LC-MS /MS method for the simultaneous deter-mination of cytochrome P450 probe substrates,inclu-ding caffeine and its metabolite paraxanthine for CYP1 A2,omeprazole and its metabolite 5-hydroxyome-prazole for CYP2C1 9,dextromethorphan and its metab-olite dextrorphan for CYP2D6,midazolam and its me-tabolite 1 ′-hydroxymidazolam for CYP3A4.Methods Probe drugs with the IS diazepam were extracted using ethyl acetate.Gradient elution was performed on an Agilent Eclipse Plus-C1 8 column (50 mm ×2.1 mm, 3.5 μm).The mobile phase consisted of 0.01 % for-mic acid(1 mmol·L -1 ammonium formate)and aceto-nitrile.The flow rate was 0.3 mL·min -1 ,and the in-jection volume was 1 0 μL.The analyte was detected u-sing electrospray ionization(ESI)in positive multiple reaction monitoring(MRM+)mode.The reaction se-lected ions were 1 95.0 /1 38.1 m /z for caffeine, 1 81 .1 /1 24.1 m /z for paraxanthine,346.1 /1 98.1 m /z for omeprazole,362.1 /21 4.1 m /z for 5-hydroxyome-prazole, 272.2 /1 47.1 m /z for dextromethorphan, 258.1 /1 57.1 m /z for dextrorphan,326.1 /291 .1 m /z for midazolam,342.1 /324.1 m /z for 1 ′-hydroxymid-azolam and 285.1 /1 54.0 m /z for diazepam as internal standard.Results The linear ranges of caffeine,pa-raxanthine,omeprazole,5-hydroxyomeprazole,dextro-methorphan, dextrophan, midazolam and 1 ′-hydroxymidazolam were 1 .95 ~2 000,0.98 ~250, 0.48 ~2 000,0.98 ~250,0.98 ~2 000,0.48 ~1 25,1 .95 ~2 000 and 1 .95 ~250 μg·L -1 respec-tively.The RSD of all probe drugs was less than 1 5%and matrix effects in plasma on the ionization of probe drugs were negligible.Conclusion This sensitive and rapid LC-MS /MS method is suitable for determination of the drug/metabolite concentrations in plasma,so as to study the metabolism of CYP1 A2, CYP2C1 9, CYP2D6 and CYP3A4 in depth.

BACKGROUND:Lots of in vitro experiments have explored the toxicity of upconversion nanoparticles, but its toxicity in vivo is little reported. OBJECTIVE:To investigate the toxicity of upconversion nanoparticles in mouse organs. METHODS:After tracheotomy, 36 Balb/c mice were randomly divided into three groups, fol owed by instil ed with 28 mg/kg upconversion nanoparticle (experimental group), the same volume of normal saline (control group), and nothing (sham operation group), respectively. The functional changes of the lung, liver and kidney were detected at 1 day, 1 and 2 weeks postoperatively, and meanwhile, the morphological changes of the lung, liver, kidney, and heart were observed. RESULTS AND CONCLUSION:At 1 day postoperatively, the pH values in the experimental group were lower than those in the control and sham operation groups (P<0.05), while the glutamic-pyruvic transaminase level was higher than that in the control and sham operation groups (P<0.05). The oxygen partial pressure in the sham operation group was higher than that in the other two groups at 1 day postoperatively (P<0.05). The oxygen partial pressure and glutamic-pyruvic transaminase level did not significantly differ among groups at 1 and 2 weeks postoperatively. The carbon dioxide differential pressure and kidney function showed no significant differences among groups at different time points after surgery. At postoperative 1 day, in the experimental group, hyperplasia and inflammation were most obvious, distorted alveolar cavity and congestion of blood vessels were visible. In the control group, obvious hyperplasia and inflammation were found, the alveolar cavity was crimped and the gap between alveoli was broadened. The sham operation group had normal alveoli with no inflammations. Lung lesions in the experimental and control groups became mild with time at postoperative 1 and 2 weeks. One day postoperatively, hepatocyte swel ing and vacuolar degeneration were severer in the experimental group. Moderate hepatocyte swel ing and vacuolar degeneration occurred in the control group. The sham operation group showed mild hepatocyte swel ing and vacuolar degeneration. The morphology of the liver in each group returned to normal at 2 weeks postoperatively. Fortunately, the heart and kidney structure showed no overt changes in each group. These findings suggest that upconversion nanoparticles cause transient damage to the mouse lung and liver.

Objective To explore the mucosal healing quality of hydrotalcite combined with esomeprazole in gastric ulcer.Methods Forty-two patients visiting from June 2014 to December 2015 and diagnosed with gastric ulcer were selected and divided into combination therapy group and single therapy group with 21 patients in each group.The patients of combination therapy group received esomeprazole combined with hydrotalcite,and the patients of single therapy group received esomeprazole alone.The total therapeutic course was eight weeks.At the same period,21 health check-up participants were enrolled as normal control group.The healing of gastric ulcer was observed under white light endoscopy.The morphological changes of gastric pits and microvessel of mucosal at peripheral mucosa around ulcer and normal gastric mucosal were observed under narrow band imaging magnifying endoscopy.The gastric mucosa tissues of the two groups before and after treatment,and normal gastric mucosa of healthy control group were taken.The amount of deposition and composition of collagen fibers,the expression level of factor Ⅷ,the level of transforming growth factor (TGF)-beta1 and the content of hydroxyproline were analyzed by Masson,immunofluorescent and immunohistochemistry staining as well as enzyme linked immunosorbent assay.Chi square test,one-way analysis of variance (ANOVA),least significant difference (LSD) method,Dunnett's T3 and Kruskal-Wallis test and other method were used for comparison.Results After treatment,18 patients of combination therapy group (21 patients) had regular microvessel nets (85.7％),which was significantly more than those of single therapy group (12 cases,57.1％) and healthy control group (10 cases,47.6 ％),and the differences were statistically significant (x2 =4.200,P=0.040).In the comparison of maturity of regenerative mucosa between combination therapy group and single therapy group after treatment,the ration between collagen type Ⅰ and Ⅱ,deposition of collagen fibers,the number of factor Ⅷ positive cells,the level of TGF-beta1 and the content of hydroxyproline were 36.05 and 23.14;269 375.63.± 171 608.63 and 137 693.14±98 330.93;34.91±8.40 and 28.24±6.93;104 498.71±40 487.96 and 70 757.11±19 323.95;(1 897.80±879.35) and (1 230.57±536.05) μg/L,respectively;while in healthy control group,the above parameters were 36.81,245 696.90 ± 224 687.00,23.10 ± 8.40,94 048.04 ±41 306.55 and 1 681.20 ± 423.61 μg/L,and the differences were statitically significant among these three groups (H=7.375,F=3.465,11.680,5.190,5.160;all P＜0.05).Those parameters of combination therapy group were significantly higher than those of single therapy group (H=2.416,LSD method;all P＜0.05).Conclusion Hydrotalcite combined with esomeprazole in the treatment of gastric ulcer could significantly improve microvessel morphology,maturity degree of regenerative mucosal structure and function,and the mucosal healing quality was also superior to single esomeprazole group.

Objective To investigate the relationship between the content of stromal cell-derived factor-1 (SDF-1) in myocardium in different period of myocardial infarction and left ventricular function.Methods Twenty three Chinese miniature pigs were randomly divided into the experimental group and the control group.The swines in experimental group were prepared as acute myocardial infarction model by ligating anterior descending coronary artery and were randomly divided into 6 subgroups according to the different time points after infarction.The left ventricular end-diastolic diameter (LVDd),left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were measured respectively.Global circumferential strain (GCS) and global radial strain (GRS) of left ventricle were measured.The content of SDF-1 were also measured by real-time quantitative PCR.Results Compared with the control group,SDF-1 levels were significantly elevated,and LVEF,LVFS,GCS and GRS were reduced.However,LVDd were significantly increased.The content of SDF-1 and GCS has a negative correlation (r =-0.580,P =0.000).Conclusions The content of SDF-1 in myocardial tissue have a certain relationship with GCS of left ventricular myocardium.

Objective:To assess the effectiveness and safety of radiotherapy combined with surgery for gastric cancer compared with single surgery. Methods:We retrieved relevant randomized controlled trails in Cochrane Library (Issue 2, 2009), PubMed (Jan 1966-Jun 2009), EMBASE (Jan 1974-Jun 2009), Chinese Biomedical Literature Database (Jan 1978-Jun 2009), Chinese Science and Technology Periodicals Database (Jan 1989-Jun 2009) ,China National Knowledge Infrastructure (Jan 1994 to Jun 2009) and Wanfang database (Jan 1997-Jun 2009). Two researchers assessed the quality of included randomized controlled trials (RCT) and extracted data independently. The RevMan 5.0 software was used for Meta-analysis. Results:Nine RCTs including 1 548 patients were selected for Meta analysis. Five RCTs were related with comparison of preoperative radiotherapy plus surgery with single surgery. Two RCTs were about the comparison of surgery plus perioperative radiotherapy with single surgery. Two RCTs were the comparative studies between surgery plus postoperative radiotherapy and single surgery. The Meta analysis results showed that: (1)compared with surgery alone, preoperative radiotherapy combined with surgery can increase 3 years [OR = 1.78, 95% confidence interal (CI):1.14-2.78], 5 years (OR = 1.67, 95%CI :1.22-2.29),and 10 years (OR = 1.64, 95%CI 1.03-2.60) survival rate,and reduce the tumor recurrent rate(OR = 0.59, 95%CI :0.37-0.92) and metastasis rate (OR= 0.44, 95%CI :0.27-0.73); (2) The tumor recurrent rates (OR = 0.19,95%CI :0.03-1.14 )and tumor metastasis rates (OR = 0.09, 95%CI :0.00-1.77)had no difference between single surgery group and peri-operative radiotherapy plus surgery group;(3) Postoperative radiotherapy compared with surgery had no significant effects on 1 year (OR = 0.83, 95%CI :0.60-1.15) and 3 years (OR = 0.75, 95%CI :0.51-1.11) survival rate compared with single surgery, but the 5 years (OR = 0.57,95%CI :0.34-0.95) survival rate of the patients who received surgery alone was higher than those who received combined therapy. No difference of the tumor recurrence rate(OR=0.59,95%CI :0.33-1.05) and tumor metastasis rate (OR=0.90,95%CI:0.51-1.59) were observed between the two groups. Conclusion:Preoperative radiotherapy combined with surgery is more rational and effective than surgery alone in the treatment of gastric cancer. However in terms of the clinical effects of perioperative or postoperative radiotherapy combined with surgery, more RCTs with larger samples and higher quality need to be carried out to make further verification.

Objective To prepare a redox-responsive doxorubicin-loaded nanoparticle,and to study its in vitro realease behavior and targeting effect on heptoma cells.Methods Cystamine was grafted on the side chains of hyaluronic acid with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide catalyst,and then β-cyclodextrin (β-CD) was conjugated on the amine groups of the cystamine by Schiff's base reaction to prepare β-CD modified hyaluronic acid (HACD).The HACD/DOX nanoparticles were prepared by encapsulating DOX into HACD using dialysis method.The drug loading,encapsulation efficiency,particle size and distribution,zeta potential and other physical and chemical properties,as well as in vitro drug release behavior of the HACD/DOX nanoparticles were characterized.The cytotoxicity of HACD/DOX nanoparticles to HepG2 cells was studied by cell counting kit-8 (CCK-8) method.The targeting effect of HACD/DOX nanoparticles on HepG2 cells was studied using flow cytometry and confocal laser scanning microscopy (CLSM).Results HACD were successfully synthesized,which could carry DOX to form uniform homogeneous nanoparticles.The drug loading of DOX in the nanoparticles was (16.1±0.2)％ and the encapsulation efficiency was (64.2±0.9)％.The transmission electron microscope images indicated that the shape of the HACD/DOX nanoparticles was homogeneous sphere.The results of granularity analysis showed that the average size of the HACD/DOX nanoparticles was (203.1 ±2.5) nm with a narrow size distribution (PDI =0.202).The zeta potential of the HACD/DOX nanoparticles was (-29.1±0.8) mV.The in vitro release behavior of the nanoparticles exhibited obvious redox-sensitivity.The results of in vitro cytotoxicity showed that the blank carrier material HACD had no obvious toxicity to hepatoma cells,and the HACD/DOX nanoparticles could effectively kill hepatoma cells with the 0.38 μg/ml half maximal inhibitory concentration (IC50) value at 48 h.Flow cytometry and CLSM results demonstrated that the HACD/DOX nanoparticles could target hepatoma cells through the mediating effect of hyaluronic acid.Conclusions The prepared HACD/DOX nanoparticles have suitable particle size,high drug loading and encapsulation efficiency,and can release DOX under the stimulation of reducing agent.These nanoparticles have obvious targeting effect on hepatoma cells,which is expected to be applied as the drug delivery system of hepatocellular carcinoma (HCC) therapy.

Objective To study the clinical application value of humidified high flow nasal cannula(HHFNC) on bronchitis in children.Methods Total 85 cases of bronchitis that needed oxygen therapy in our department from Oct 2015 to Feb 2016 were randomly divided into three groups,HHFNC group,NCPAP group and nasal cannula oxygen group(control group).According to the blood gas results,FiO2 was adjusted to maintain PaO2 in 50~70mmHg(1mmHg=0.133kPa),TcSO2 90% to 95%.The main symptoms and signs(wheezing,shortness of breath,three depression sign,wheezing rale) disappeared time,length of oxygen inhalation and stay,the change of PaO2,PaCO2,respiratory rate were compared among the three groups.Results Compared with control group,the clinical symptoms and signs disappeared time and length of oxygen inhalation and stay were significantly shorter in HHFNC group(P<0.05).The recovery of blood gas and respiratory rate in HHFNC group were better than those in control group(P<0.05).No statistically significant differences existed between HHFNC group and NCPAP group(P>0.05).Conclusion HHFNC can significantly improve the clinical symptoms,signs and blood gas results in children with bronchitis,reduce the length of oxygen inhalation and stay.HHFNC is an effective and well-tolerated treatment for bronchitis in children.