A high performance liquid chromatography-electrospray ionization mass spectrometry- charged aerosol detection ( HPLC-MS-CAD) method was established for the simultaneous quantitative analysis of four Lignans in Magnoliae Flos extract. The components were separated on a YMC-Pack ODS-A column (250 mm× 4. 6 mm, 5 μm) by gradient elution with methanol and water as the mobile phase at aflow rate of 1. 0 mL/min. Then the elution solution was routed into MS equipment at a flow rate of 0. 3 mL/min and CAD detector at a flow rate of 0. 7 mL/min by a split ratio of 3:7 for the further detection. The column temperature was 25 ℃ and the detection wavelength was 278 nm. A method was developed for the quantitative analysis of muti-components by single maker ( QAMS) to determine pinoresinol dimethylether, magnoli, 1irioresinol B dimethylethe and epi-magnoli A . Magnoli was selected as internal standard and the relative correction factors ( RCF) of the four Lignans were calculated. The contents of the four Lignans in Magnoliae Flos extract were determined by both external standard method and QAMS. The QAMS method was evaluated by comparison of its assay result and that of external standard method. Under the selected chromatographic condition, the limits of detection of pinoresinol dimethylether, magnoli, lirioresinol B dimethylethe and epi-magnoli A were 0. 34, 0. 55, 0. 50 and 0. 58 mg/L, respectively, while the linear range were within 6. 8-270 mg/L, 11-546 mg/L, 2. 0-101 mg/L and 2. 3-116 mg/L. The recoveries ( n=9 ) were 98. 2%-99. 5%, and the correlation coefficient were 0 . 9995-0 . 9998 . No significant differences were found between the quantitative results of external standard method and QAMS method. The developed method is accurate, feasible, and can be used for quality evaluation of Magnoliae Flos .

Objective To analyze the chromosome karyotypes,prenatal diagnosis indications and pregnancy outcomes of high-risk pregnant women in Guangzhou.Methods 2 475 cases pregnant women with screening high risk were operated amniocen-tesis or cordocentesis from January 2010 to September 2012,then amniotic fluids and cord bloods were cultured and the cell were collected for chromosome preparation,G banding,karyotype analysis.We completed follow-up works lastly.Results 38 cases were detected chromosomal abnormality(including 12 cases Down′s syndrome,9 cases sex chromosome abnormality,7 cases transloca-tion,5 cases Edwards′syndrome,2 cases inversion,2 cases deletion,1 cases triploid),the abnormal rate was 1.54%.132 cases were detected chromosomal polymorphism(60 cases 1,9,16qh+ ,30 cases inv(9),25 cases D/Gs+ ,17 cases Y polymorphism).Research on prenatal diagnosis indications,there were 449 cases advanced age,668 cases Down′s screening with high risk,158 cases with ab-normal B ultrasound screening,38 cases with adverse pregnancy history.Conclusion The highest percentage abnormal karyotype is Down′s syndrome.Down′s screening high risk is the main reason for prenatal diagnosis.It is very important to do prenatal diagnos-tic and system B ultrasound for the high-risk pregnant women.

Objective To analyze the plasty of the tip framework and the nasal tip in insufficient-lining cases with auricular and septal cartilage.Methods The tip framework was formed by using septal extension graft (SEG) and columellar strut (CS) to construct the main framework of nasal tip,and then according to the size of the graft the next processes were chosen for supporting structure.When the graft size met the tip position,the caudal margin of the framework could be beyond the maximum stretch position of the lower lateral cartilage and reached or was close to the expected tip position.When the graft size was insufficient,auricular cartilage was used to hold the caudal margin of the framework to extend or to heighten the framework further.Refined plasty of nasal tip was as follows:when the end of framework was higher than or equal to the dome above 3 mm,auricular cartilage strip was used to reconstruct the dome;when less than 3 mm,multilayer onlay grafts were used to heighten the dome.Results From May 2013 to May 2015,42 cases were followed up from 6 months to 18 months.18 cases belonged to the type 1,including 13 cases using ear cartilage to reconstruct dome,5 cases using onlay graft;24 cases belonged to the type 2,including 14 cases using onlay graft and 10 cases using ear cartilage to reconstruct the dome.They all got good results.Conclusions To the insufficient lining cases,the appropriate method can be chosen to form stable tip framework according to the preoperative design and the graft size.Then onlay grafts or dome reconstruction technique is used to form the tip.Most of the insufficient-lining cases can get satisfactory results.

Objective:To analyse and prepare bispecific antibody(BsAb, anti human bladder cancer/anti VEGF) which could enhance the effectiveness of tumor cell killing activity.Methods:Monoclonal antibodies against human bladder cancer and VEGF were used to prepare bispecific antibody.Results:IC50 of BsAb was 10 -9.5 ,while those of antibodies against human bladder cancer and VEGF were 10 -8.9 and 10 -8.3 .Conclusion:The effectiveness of BsAb was significantly higher than that of monoclonal antibodies. [

Objective To investigate the immune localization of bispecific gene antibody (anti human bladder carcinoma/anti VEGF),and to determine whether bispecific gene antibody can inhibit growth of human experimental bladder carcinoma. Methods Immunohistochemistry and immune electron microscope were used to study the immune localization in 60 samples of bladder cancer (experiment group),and 20 samples from BPH cases served as control group.Bispecific antibody was injected into the sites in nude mice,which was adjacent to the xenograft tumor of human bladder transitional cell carcinoma.The tumor size was measured at different times.The microvascular density(MVD) and apoptotic index(AI) in the tumors were examined. Results Of the 60 bladder cancer samples 53 (88.3%) were positive for antigen,while only 1 (5.0%) of the 20 in control group was positive.Two weeks after treatment,the tumor size of experiment group was ( 21.47 ?6.57)mm 2,while that of control group was (59.85?15.43)mm 2.MVD of experiment group was 2148?109,while that of control group was 4056?367.AI of experiment group was 17.26,while that of control group was 7.09.The differences between the 2 groups were significant (each P

Objective Developing an internal quality control substance of Ureaplasma urealyticum(UU)‐DNA for real‐time PCR to establish an internal quality control system and preliminary evaluation its clinical value .Methods Internal quality control sub‐stance was prepared by mixing samples which Ct value were 24-25(positive sample) and 32 -33(weak positive sample) ,respec‐tively .At the same time ,selecting samples that test results were negative as negative control .The target value ,standard deviation (s) and coefficient of variation(CV) of internal quality control substance were defined by“instant method”for the first 20 runs and Levey‐Jennings quality control(QC) chart after the first 20 runs .Using the“Westdard” multi‐rule quality control methods to ana‐lyze the detection results .Exporting OPSPecs chart by quality control rules in Unity Real Time (URT ) system and setting up new quality control rules according with OPSPecs chart .Results 131 times of the detection of quality control substance were performed totally .The first 20 runs were defined by“instant method”and later 111 runs were defined by Levey‐Jennings QC chart ,the results were stable of quality control substance and reasonable quality control rules .Conclusion Preparing of internal quality control sub‐stance of UU‐DNA used in real‐time PCR might be easy and stable .So ,the internal quality control substance of UU‐DNA could be worthy for practical application in this PCR laboratory .Design internal quality control rules based OPSPecs chart in molecular de‐tection is very simple and practical .

Objective To investigate the influence of bloody cervical cell samples on human papillomavirus(HPV) genotyping detection using flowcytometry fluorescence hybridization kit.Methods According to the concentration of hemoglobin(Hb),cervical cell samples were divided into five groups,including group A with Hb of 0 g/L(10 cases),group B with 0 g/L100 g/L.In each group,50% samples were with initial positive results of HPV genotyping detection.Every sample was detected using sample volume of 50,100,200,500 and 1 000 μL different volumes for testing and analysis results.Results For all samples with initial negative results,five samples with initial positive results in group A and five samples with initial positive results,the detected results of different sample volume were without differences.In the 10 samples,with initial positive results of group C,D and E,there were four,four and nine samples respectively,were with false negative detected results of different sample volumes.Conclusion Bloody cervical cell samples could cause false negative results of HPV genotyping detection using flowcytometry fluorescence hybridization kit.It might be necessary to set optimal sample volume according to the concentration of Hb for ensuring the accuracy of genotyping detection.

Objective To study and observe the change of nerve function and erythrocyte immune indexes of patients with severe craniocerebral injury during the perioperative period.Methods Forty patients with severe craniocerebral injury treated with surgery in our hospital from July 2013 to August 2015 were selected as the observation group and 40 healthy persons with the same ages in the same period were selected as the control group.The nerve function related indexes and erythrocyte immune indexes of observation group before and at first,third,fifth,seventh,tenth and fourteenth day after the surgery and those of control group were compared.Results The serum nerve function related indexes of observation group before and at first,third,fifth,seventh,tenth and fourteenth day after the surgery were higher than those of control group.The erythrocyte immune indexes of observation group before and at first,third,fifth,seventh and tenth day after the surgery were worse than those of control group.The erythrocyte immune indexes at fifth and seventh day after the surgery were worse than those before and at first,third,third,tenth and fourteenth day after the surgery.The differences of comparison indexes were all significant (P＜0.05).Conclusion The change of nerve function and erythrocyte immune indexes of patients with severe craniocerebral injury during the perioperative period are obvious.Those indexes should be paid with enough monitoring and intervention.

Objective To ivestigate the clinical feature and therapeutic ways of the gastroduodenal ulcer perforation(GDUP) in aged patients.Methods The clinical data of 58 GDUP patients with age more than 60 gears treated in recently tweenty yesas in our hospital were analysed retrospectively. Results Among the 58 patients,56 patients underwent operation,46cured and 12 died.Of the 12 patients,dead of cardiorespiratory function failture in 5 ; septic shock in 4 ;and renal failturl in 3 .Conclusions Aged patients with GDUP would be operated as early as possible.Repair of the performed ulcer plus modified vagotomy is a better choice.

Objective To investigate the effect of ethanol on radiosensitivity of human breast cancer MCF-7 cells.Methods Human breast cancer MCF-7 cells were divided into four groups including control group,ethanol treatment group,X-ray exposed group,and ethanol combined with X-ray group.Clonogenic assay was used to determine cell survival.Flow cytometry was employed to analyze cell cycle progression.Annexin V-FITC kit was used to determine cell apoptosis induction.Results Ethanol(50 and 100 mmol/L,50 h)had no influence on MCF-7 cell growth( t =0.82,1.15,P ＞0.05 ).The radiosensitivity of MCF-7 cells was reduced when the cells were pretreated with 50 mmol/L ethanol (t =4.15,P ＜0.05)and 100 mmol/L ethanol ( t =10.28,P ＜ 0.05 ) for 2 h. Compared with irradiation with X-ray alone,ethanol treatment decreased G2/M phase arrest(t =7.18,P ＜0.05) and sub-G1population(an indicator of apoptosis induction) ( t =5.39,P ＜ 0.05).A decrease of advanced and early apoptosis in the cells pretreated with ethanol was also confirmed by Annexin V-FITC apoptosis assay( t =4.86,7.59,P ＜ 0.05 ).Conclusions Ethanol causes radioresistance in human breast cancer MCF-7 cells,where the decreases of radiation-induced G2/M phase arrest and apoptosis may be involved.