OBJECTIVE To investigate Mycoplasma pneumoniae load index(MPLI)usefulness in the diagnosis of Mycoplasma pneumoniae(MP) infection in children.METHODS 110 throat swabs were collected with paired sera from the department of pediatry in our hospital from March to July 2008.The former fluorescence quantitative(FQ)-PCR assay was used to analyze throat swabs,microparticle agglutination assay(MAG) and MP-IgM assay for the sera.MP-IgM positive once or twice was used as a gold standard for MP infection,on which sensitivity(SEN),specificity(SPE),positive predictive value(+PV),negative predictive value(-PV) and ROC were calculated.RESULTS According to the gold standard,the values of SEN,SPE,+PV,-PV were 73.1-100.0%for a single MP-IgM,26.9-94.0% for a single MAG,46.2-94.0% for double MAG,26.9-95.2% for FQ-PCR alone and 26.9-98.8% for MPLI,respectively.CONCLUSIONS MAG shows limit in its clinical usage for the detection of MP antibodies.MPLI can diagnose acute MP infection earlier than MP-IgM.

Objective To investigate three-dimensional localization for sentinel lymph node (SLN) of breast cancer,and by which we can remove the SLNs directly.Methods The ipsilateral axillary lymph nodes of 40 patients were inspected by B-ultrasound and axillary artery and subscapular artery bifurcation point and its trend with Doppler B-ultrasound in the preoperation,then located them in the surface.We found SLNs using methylene blue as the mapping agent with endoscope during the operation,determined which lymphatic group the sentinel lymph node belonged and the spatial location and the surface projection according to the anatomical location.Results We found the three-dimensional location of SLNs in the group of 39 in 40 patients with endoscope,of which 34 cases located in central group,accounting for 87.18％ ;while 4 cases located in the subscapular group,accounting for 10.26％,and their spatial location was as follows:set the root of subscapular artery in this location as a starting point,the subscapular artery as a diameter,and made a diameter of 5cm circle to the bottom,then let the latissimus dorsi as the end,and made a quasi-cylinder through the circle to the axillary central.The height of the quasi-cylinder got up to the surface of the intercostal brachial nerve.Then set the nerve as the diameter of circle of quasi-cylinder,and the centre of circle was crosspoint of subscapnlar artery's surface projection with intercostal brachial nerve.The height of quasi-cylinder varies with somatotypes of the patients,its height was less than or equal to 5cm.What's more,the fiften enlarged lymph nodes located by B-ultrasound in the preoperation were all in the quasi-cylinder,and they were SLNs.Conclusion SLN lies in quasi-cylinder consisting of spatial location of subscapular group and central group lymph nodes.If the enlarged lymph nodes found by B-ultrasound are in above mentioned quasi-cylinder,they can be considered as the SLNs.Make a 5cm-incision parallelling the intercostal brachial nerve and intersecting the surface projection of subscapular artery in the surface of quasi-cylinder,then dissect toward the origin of the subscapular artery,you can find SLNs.

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

To evaluate the feasibility of delivering NO through N_2O path in anesthesia machine,NO at 800 ppm was delivered through nitrous oxide path in anesthesia machine SIEMENS Ventilator 710 or NARKOMED 2B,to ventilate a test lung. The minute volume of ventilation (MV) was set from 2 to 6L/min, and NO concentration from 10 to 80 ppm. NO and NO_2 concentrations were measured by hemiluminescence and electro-chemical fuel cell technique. SIEMENS Ventilator 710:Non-rebreathing model,its inspiratory limb was prolonged to 7 meters,the NO level was adjusted at 80ppm before passing through soda lime, NO and NO_2 samples were taken before absorber,after absorber, 1, 2,4 and 6 meter site in inspiratory limb. NARKOMED 2B:Oxygen flows were at the rates epual to the MV mentioned above, NO concentration was at 10,20,40,60 and 80 ppm,the inspiratory gas mixture was sampled immediately passing through absorber, NO_2 levels were less than 5.0ppm in all conditions listed above. With SIEMENS Ventilator 710, the highest level of NO_2 was 3.45 ppm before absorber and 2.43 ppm after absorber,with NARKOMED 2B,that was 3.6 ppm. As MV increased,NO_2 level before absorber decreased (P

0.05).Tn-C、CD44v6 were negative expression in group C.Conclusion Tn-C、CD44v6 protein are useful in early diagnosis of follicular thyroid carcinoma.

Objective To develop a new Multiplex Allele-Specific polymerase chain reaction(MAS-PCR) assay to detect the main point mutations in the katG and rpoB genes, which has been reported to account for the majority of clinical Mycobaterium tuberculosis resistant to isoniazid and rifampin. Methods Based on the sequences of katG and rpoB genes, specific primers were designed to carry out the MAS-PCR to detect the most common mutations in codon315 of katG and codons 531,526,516 of rpoB gene. Results The purified DNA preparation of 96 clinical Mycobacterium tuberculosis strains were used to optimize PCR. No mutation was detected in 19 isoniazid-sensitive strains. The 315Ser point mutation was detected in 79.2%(61/77)of isoniazid-resistant strains, the type of mutation includes the most common S315T and the less common S315N, which could’t be detected by PCR-RFLP(polymerase chain reaction-restriction fragment length polymorphism). However, S315G couldn’t be detected by MAS-PCR and that will make a false negative. The mutations in codons 531,526,516 were detected by the MAS-PCR. Compared with the results of direct sequencing of rpoB gene, no mutation was detected in sensitive strains. For rifampin-resistant strains, the total sensitivity was 81.5%(66/81). Conclusions MAS-PCR is a new molecular method with high sensitivity and specificity, which can be used to detect the point mutation in katG and rpoB gene rapidly and economically. It can be used in clinical laboratories to detect drug-resistant tuberculosis strains. Simultaneous detection for katG and rpoB gene mutations in one MAS-PCR system will help to improve the efficiency of this method.

0.05). Conclusions In human liver microsome system in vitro,CYP1A2,2B6 and CYP2A6 contribute to the metabolism of 629.It is very important for bioreduction drugs design and development,and provide the basic experimental and theoretical profiles for extensive application in clinic.

Objective To investigate the spherical diopter and astigmatism change of humans at sitting and supine position.De- sign Prospective case series.Participants 96 eyes of 52 patients (spherical diopter from-2.50 D to-10.00 D,astigmatism diopter from -0.75 D to-4.50 D) were selected.Methods The subjects were examined with NIKON portable retinomax at sitting and supine posi- tion,respectively.Main Outcome Measures The spherical diopter,cylinder diopter and axis change were analyzed statistically.Re- sults Spherical diopter at supine position (-5.31?3.43 D) was a little higher than that at sitting position (-5.27?3.24 D) statistically(P= 0.25),and cylinder diopter at sitting position (-2.27?1.24 D) and at supine position (-2.35?1.19 D) was no statistically difference (P= 0.20).The axis of astigmatism changed from-16?to +18?.Axis change was within 2?in 52.1% eyes,6?-10?in 5.2%,over 10?in 3.1%. The change of axis rotation tended to counter-clockwise in the right eye and clockwise in the left eye.Conclusions Eye rotation at sit- ting and supine position may cause the astigmatism axis change.It may be one of the main factors affecting the results of LASIK.

To understand molecular modules related to polyunsaturated fatty acids (PUFA) synthesis and eventually produce PUFA at high efficiency, we developed a protein complex analysis technology in Synechocystis sp. PCC 6803, and applied it to identify possible partner proteins interacting with the key enzymes that catalyze PUFA biosynthesis. We first constructed a recombinant expression of protein of slr1609 encoding the fatty acid activation enzyme, by fusing 3xFLAG tag with the target protein. Then we verified its expression by Western blotting targeting 3xFLAG tag. To maximize purification of Slr1609 protein complex, we optimized the protein expression conditions of Slr1609 in Synechocystis in a 5 L fermenter by monitoring its gene expression using RT-qPCR. The purification of the Slr1609 protein complexes was demonstrated by a Native-PAGE analysis. Finally, LC-MS/MS proteomic analysis allowed identification of the possible partner proteins interacting with Slr1609.
Bacterial Proteins ; chemistry ; Chromatography, Liquid ; Fatty Acid Synthases ; chemistry ; Fatty Acids, Unsaturated ; biosynthesis ; Proteome ; chemistry ; Proteomics ; Synechocystis ; enzymology ; Tandem Mass Spectrometry

To study on the phylogenetic characterization of the VP1 genes of coxsackievirus A16 (CVA16) causing hand-food-mouth disease (HFMD) isolated from Anhui province in 2014. A total of 413 throat swab specimens from HFMD patients were collected during January to November, 2014 for the isolation and identification of enteroviruses using real-time RT-PCR assays. The VP1 regions of CVA16 isolates were amplified using RT-PCR and sequenced. And the phylogenetic tree was constructed among the VP1 regions of those isolates, the different genotypes and sub-genotypes of CVA16 strains. A total of 97 enteroviruses were isolated from 413 samples, the positive rate was 23.49% (97/413), including seventeen CVA16, seventy six HEV71 and four other enteroviruses. The results of the phylogenetic tree showed that 17.CVA16 strains isolated from Anhui in 2014 clustered within B1b evolution branch of B1 genotype. The nucleotide and amino acid sequence identities were 95.30%-100% and 98.70%-100% among the isolates, respectively, but within B1b branch of 17 strains formed several small transmission chains. The nucleotide acid of 17 CVA16 isolates in Anhui province were closed to the strains isolated from Yunnan, Hunan, Guangdong, Tibet and Jiangsu, especially from Hunan in 2013 and from Shenzhen of Guangdong in 2014, the identity were 96.40%-99.70%. The CVA16 strains isolated from Anhui in 2014 were all belong to genetic subtype B1b of B1 genotype was dominant, and among those isolates, several small virus transmission chains had formed with co-circulating and evolution.
China ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; Enterovirus Infections ; virology ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Viral Proteins ; genetics ; metabolism