Objective: To study the optimum extraction conditions of flavonoids from the leaves of Hibiscus mutabilis L. Methods: The orthogonal design L 9(3 4) was used. The 70% alcohol refluent method was compared with the decoction or 75% alcohol warm infusion process. Results: The 70% alcohol reflux was the best method.Conclusion: A 1B 3C 3D 3 is the best extraction condition of flavonoids from the leaves of Hibiscus mutabilis L..

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 establish the synthetic method of the substrate for angiotensin-converting enzyme(ACE),and investigate the assay method of sernm ACE. Methods The substrate was prepared by the use of N-hydroxysuccinimide ester in peptide synthesis,and serum ACE was determined by spectrophotometric assay using suhstrate. Results The substrate of furanacryloyl-phenylalanyl-glycyl-glycine with purity of 99.5% was obtained.The K_m value for the substrate was 0.25 mmol/L,the normal reference interval for serum ACE activity was 46.7 U/L to 113.3 U/L,and the within-run and between-run coefficient of variation(CVs)ranged from 3.9 to 4.9%.The serum ACE activity in patients with squamous carcinoma and adenocarcinoma of lung were significantly decreased(P<0.05),while that in patients with lung small cell carcinoma was not significantly decreased(P>0.05). Conclusion The synthesized substrate could be reliably used as a marker for diagnosis of different lung cancers in clinics.

In this study, calcium phosphate cements (CPC) were prepared by mixing cement powders of tetracalcium phosphate (TTCP) with a cement liquid of phosphate acid saline solution. Tetracycline (TTC)-CPC, chitosan-CPC and chitosan-TTC-CPC were investigated with different premixed schedule. It was demonstrate that both TTC and chitosan worked on the phase transition and crystal characteristics. TTCP mixed with phosphate acid saline solution had similar features of Fourier transform-infrared spectrometry (FT-IR) no matter it was mixed with chitosan or TTC or both. TTC premixed with cement liquid or powder had significant different features of FT-IR and 876 cm(-1) seemed to be a special peak for TTC when TTC was premixed with cement liquid. This was also supported by XRD analysis, which showed that TTC premixed with cement liquid improved phase transition of TTCP to OCP. Chitosan, as organic additive, regulates the regular crystal formation and inhibits the phase transition of TTCP to OCP, except when it is mingled with cement liquid premixed with TTC in field scanning electron microscope. It was concluded that the premixed schedule influences the crystal formation and phase transition, which may be associated with its biocompatibility and bioactivities in vivo.

OBJECTIVE:To establish a method for the determination of related substances in anastrozole. METHODS:HPLC was performed on the column of Welch Materials XB C18 with mobile phase of acetonitrile-water(35:65,V/V)at a flow rate of 1.0 mL/min,detection wavelength was 210 nm,column temperature of 25 ℃,and the injection volume was 20 μL. RESULTS:Only one impurity(impurity A)was detected in anastrozole,the linear range was 0.1-1.6 μg/mL(r=0.9996);limit of quantitation was 0.1 μg/mL,limit of detection was 0.02 μg/mL;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 98.95%-105.29%(RSD%=1.78%,n=9). CONCLUSIONS:The method is simple,accurate,and can be used for the determination of related substances in anastrozole.

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.

Objective To evaluate the methods and the efficacy of interventional therapy for huge aneurysm.Methods Seven patients with huge aneurysm including 2 with pulmonary aneurysm, 2 with renal aneurysm, 1 with humeral artery aneurysm, 1 with right common iliac artery aneurysm, 1 with right internal iliac artery aneurysm. Among these, 5 were true aneurysm, and 2 were pseudoaneurysms caused by congenital, trauma, arteriosclerosis. Three patients were treated with endovascular covered stent graft and 2 patients with embolization containing metallic coils. Two patients were treated with partial aneurysm and feeding artery trunk embolization with metallic coils. Results All 7 patients were successful carried out the interventional therapy with successful rate of 100%. Six aneurysms were completely obstructed with disappearance of symptoms and signs. One died of aneurysm rupture. No other complication occurred.Conclusion Interventional therapy for huge aneurysm is an effective method.

Objective To investigate the effect of the asthmatic mice's contents of nuclear factorerythroid-2-related factor 2,Nrf2 and HO-1 reated with curcumin in lung tissue.Methods All 45 mice were divided into three teams randomly,the normal mice,the asthmatic mice and the curcumin mice group.We tested the expression of Nrf2 and HO-1 of three mice teams in lung by immunolfluorescence technique.We tested the protein contents of Nrf2 and HO-1 of the three teams in the lung tissue by Western blot.We tested the binding activity of Nrff2 and ARE of the three teams in lung by EMSA.Results The expression of Nrf2 and HO-1 in curcumin group was significantly higher than that in asthma group and normal control group by immunolfluorescence technique.There was no sigrnificant difference in content of cytoplasm Nrf2 protein in lung tissue between three groups by western blot(P ＞ 0.05).The content of nuclear Nrf2 and HO-1 protein in the lung tissue of the curcumin group was significantly higher than that in the other two groups,the difference is statistically significant (Nrf2,P ＜0.05;HO-1,P ＜0.01).The binding activity of Nrf2-ARE in lung tissue of curcumin group was significantly higher than that of asthma group and control group,with statistical significance(P ＜ 0.05).Conclusion Curcumin can increase the protein contents of the Nrf2 and HO-1 of mice and enhance the expression of them.

Objective To investigate the effect of the asthmatic mice's contents of NF-κB、IκB、p-IκB treated with curcumin.Methods Thirty Balb/c mice were divided into three groups randomly,the normal mice,the asthmatic mice and the curcumin mice.The bronchial hyperresponsiveness of the three groups were exanined by lung function.The NF-κB 、IκB 、p-IκB content of three teams were tested.Results The content of cytoplasm NF-κB in asthmatic mice was lower than the control (P ＜ 0.01).However,the content of cytoplasm NF-κB in curcumin team was more than the asthmatic mice (P ＜ 0.05).In the other hand,The content of nuclear NF-κB in asthmatic mice was more than the control(P ＜0.01).However,the content of nuclear NF-κB in curcumin mice was lower than the asthmatic mice (P ＜ 0.05).The content of cytoplasm p-hκB in asthmatic mice was more than the control and the content of IκB was lower than the control(P ＜ 0.01).Howerer,the content of IκB in curcumin team have much more than the asthmatic mice and the content of p-IκB have much lower than the control(P ＜ 0.01).Condusion Curcumin alleviates the airway hyperresponsiveness of the asthmatic mice through the suppression of NF-κB transcribe to the nuclear via alleviating the phosphorylation of I-κB.

Objective To understand the correlation of gene detection of Mycoplasma pneumoniae and clinical refractory Mycoplasma pneumoniae pneumonia.Methods (1) For children with Mycoplasma pneumoniae pneumonia in our hospital with serum Mycoplasma pneumoniae antibody positive,we collected the pharyngeal swab specimens over the same period,applied nested PCR to amplify 23SrRNA gene and undergoing electrophoresis and find out 97 cases of both positive,conducted DNA sequencing analysis of macrolide resistant gene to isolate the mutants,compared clinical manifestations of drug-resistance gene group with no drug-resistance gene mutation group.(2) Ninety-seven cases of mycoplasma pneumonia (MPP) patients were devided into the general MPP group (68 cases) and refractory MPP group (29 cases),retrospectively analyzed clinical manifestation,laboratory examination and differences of imaging performance.Multivariate logistic regression analysis for the performance of refractory Mycoplasma pneumoniae pneumonia was carried out to examine whether there is relevance between the mutant of drug-resisting gene and refractory Mycoplasma pneumonia.Results (1) Seventeen of 97 cases (17.5％) were found out without mutations,the other 80 cases (82.5％) exist drug-resistance gene mutations.(2) Mutation of drug-resistance gene group showed high CRP values,heating time,hospitalization time,macrolide drug application time,application of macrolides fever time and longer cough,by statistical analysis with statistical significance,higher incidence of lobar pneumonia.(3) Compared to general MPP group,refractory MPP group showed high peripheral blood neutrophil percentage percentage,CRP,calcitonin) and lactate dehydrogenase (LDH) values,heating time,hospitalization time,macrolide drug application time,application of macrolides fever time and longer cough.There was significant difference (P ＜ 0.05);macrocyclic lactones drug application time and resistance gene mutation and refractory Mycoplasma pneumonia were correlated.Conclusion MPP drug-resistant genes are widespread.Drug resistance gene mutations group shows long clinical symptoms duration,slow recovery rate,higher CRP value,higher rates of lobar pneumonia.Compared with ordinary MPP group,there are higher drug resistance mutation rate,inflammatory indexes and lactate dehydrogenase value,large ring lactone class drugs after a longer time of cough and fever in RMPP group.Drug application time and resistant gene mutations are associated with RMPP.