With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.
Glycoproteins ; chemistry ; Glycosylation ; Polysaccharides ; chemistry ; Ultrafiltration ; instrumentation

With development of bio-technique, more and more proteins were applied as clinical approaches. However, the protein homogeneity, especially the N-glycosylation limited the further research and application of these protein drugs. The analysis method for N-glycans is believed to be critical in protein drugs development. To enhance the N-glycans isolation efficiency and accelerate the pretreatment, a new strategy was built on ultrafiltration-devices. New methods increased the isolation efficiency of N-glycans containing N-acetylglucosa mine with 10%-20%. The degrading of N-glycans containing sialic acids was also minimized with this method. 20%-100% more N-glycans with sialic acids were isolated. The pretreatment was finished within 30 min. Coupled with HPLC-HRMS, an effective and reliable strategy designed for protein drugs N-glycans analysis were developed.

Objective To investigate the association between the expression of heparanase(Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues.Methods(1)Matrigcl invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells.(2)Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines(JEG-3 cells and JAR cells)were detected by immunocytochemistry and western blot.Results(1)The invasive cell number was significantly larger in JEG-3 cells than in JAR cells(191?17 vs 106?13,P

Objective To analyze postoperative effect of a new rectopexy technique with Douglas pouch elevation for complete rectal prolapse (CRP) in adults.Methods From January 2010 to May 2011, 52 CRP cases were treated by rectopexy with Douglas pouch elevation.In terms of different surgical techniques, patients were divided into two groups : Group A (n =28) received laparoscopic rectopexy with Douglas pouch elevation;and Group B (n =24) received laparoscopic rectopexy with Douglas pouch elevation combined with procedure for prolapse and hemorrhoids (PPH).Rectal prolapse, constipation, and fecal incontinence in the two groups were assessed respectively before surgery, and on the 6th, 12th, and 24th month after surgery, postoperative complications were evaluated with severe grading of surgical complications.Results Symptoms of rectal prolapse disappeared at half a year after surgery in all patients, and relapsed to different extent afterwards.However, two years after operation, the status of rectal prolapse in Group B tended to be stable.The trend of constipation after surgery was consistent with that of rectal prolapse.In addition, fecal continence improved gradually on half a year after surgery in the two groups and recovered to the optimal status in the first postoperative year.However, in the second year, the results of fecal incontinence reduced slightly in Groups A, and Group B became stable.Two years after surgery, four cases in Groups A relapsed while there was no recurrence in Group B.Difference of Grade I to Grade Ⅲ complications among the two groups was statistically insignificant (x2 =0.05, P ＞ 0.05).Conclusions The clinical effect of laparoscopic rectopexy with Douglas pouch elevation associated with the procedure for prolapse and hemorrhoids (PPH) is better than that without PPH for female and male CRP patients with severe symptoms.

OBJECTIVE To analyze the relationship of Acinetobacter baumannii in Ningbo No.2 Hospital by pulsed field gel electrophoresis(PFGE) and amplified fragment length polymorphism(AFLP).METHODS The sensitivity of 13 kinds antibacterials were analyzed according to American National Committee for Clinical Labotatory 2004′s Standard.The genotypes were analyzed by PFGE and AFLP.The relationship was analyzed with cluster analysis methods.RESULTS Twenty seven strains of A.baumannii were divided into two groups by AFLP method or into four groups by PFGE method.Cluster analysis showed the concordance.CONCLUSIONS Amplified fragment length polymorphism has more clinical practice.

OBJECTIVE To analyze the existence state of drug-resistant gene,integron and Tn21/Tn501 transposon and the relationship of Acinetobacter baumannii in Ningbo No.2 Hospital by genetic mark.METHODS Twenty seven strains of A.baumannii were clinically isolated.Drug-resistant gene,integron,and Tn21/Tn501 transposon were analyzed using polymerase chain reaction(PCR).The relationship was analyzed with cluster analysis methods.RESULTS The positive rates of blaTEM,blaOXA-23 cluster,blaADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sulⅠ,and intⅠ1 were 81.5%,44.4%,85.2%,85.2%,66.7%,81.5%,85.2%,and 85.2%,respectively.The others were negative.The result of multi-gene cluster analysis showed that the clone transmission existed.CONCLUSIONS The clone transmission of A.baumannii exists in Ningbo No.2 Hospital.The outbreak of A.baumannii may be possible because there are more than 3 clone transmission strains.

Objective:To improve the quality standard for Kangshiming mixtures. Methods:The identification of Angelicae Sinen-sis Radix, Astragali Radix and Phellodendri Chinensis Cortex was carried out by TLC. The contents of puerarin and paeoniflorin in the preparations were determined by HPLC. Results:The spots displayed in TLC were clear without interference from the negative control. The linear range for puerarin was 1. 64-49. 20μg·ml-1(r=0. 999 9) and 4. 22-63. 30μg·ml-1(r=0. 999 6) for paeoniflorin. The average recovery was 99. 63%(RSD=2. 14%,n=9) and 99. 05%(RSD=2. 70%,n=9) for puerarin and paeoniflorin, respectively. Conclusion:The method is accurate, reliable and specific, and can be used in the quality control of Kangshiming mixtures.

Objective To assess the application of automated four-dimensional (4D) volume color Doppler ultrasonography in normal fetal heart screening. Methods Four-dimensional volume color Doppler was acquired at the level of four chamber view level in 120 fetuses from 18 to 23 weeks of gestation. Three diagnostic planes including color Doppler information were retrieved by automated multiplanar imaging from the 4D volumes. The left ventricular outflow view (cardiac plane 1), the right ventricular outflow view (cardiac plane 2) and ductal arch view (cardiac plane 3) were acquired. Tomographic ultrasound imaging (TUI) was added to display all the three diagnostic planes. The data were then analyzed to determine whether cardiac planes 1-3 were displayed correctly in each volume. Results Automated 4D volume color Doppler ultrasonography displaying rate of cardiac plane 1, cardiac plane 2, cardiac plane 3 were 100%, 96.67% and 91.67%, respectively. Cardiac plane 1 was displayed at least three TUI plane, cardiac plane 2 was displayed at least two TUI plane, and cardiac plane 3 was displayed at least two TUI plane. Conclusion It is possible to acquire important planes of the fetal heart with automated 4D volume color Doppler ultrasonography and the visualization rate of the three diagnostic planes is fine.

Objective The study analyse the multi-drug resistance and epidemiological investigation in Acinetobacter baumannii.Methodes Using,multi-drug resistance gene test,three-dimension test,membranes active efflux test studied multi-drug resistance and epidemiological investigation.The genetype were analyzed by Amplified Fragment Length Polymorphism(AFLP).The strains' homology were researched by multi-gene cluster analysis methods.Results Cluster analysis show that the result of drugsensitive test,multi-drug resistance gene test and AFLP are accordant.The study find at least 3 clones by multi-gene cluster analysis methods.AFLP show that all of the multi-drug-resistance of A.baumannii evolutes from a clone.Conclusions The study combined with clinical practice find that ICU in Ningbo No.2 Hospital now is the core of the arises,evolution,transmission of A.baumannii.Strengthening management of ICU is very important to pretect the prevalence of Acinetobacter baumannii in nosocomial infection.

As a treasure resource of novel drug lead compounds, how to rapidly and high-efficiently screen and isolate active components from natural products is critical. Thanks to its high resolution, high automation and flexible integration, online two-dimensional liquid chromatography has great potential for screening active ingredients from complex matrices by integrating a highly specific bio-recognition process into a two-dimensional liquid chromatography system before, on or after the column separation. This review comprehensively summarized recent developments, applications and shortcomings of online two-dimensional liquid chromatography for natural product screening from different integration modes, including pre-column, on-column and post-column screening methods.