This paper first briefly introduces the basic information of aminoglycosides for pharmaceutical use such as origin, classification, chemical structures, diversity of composition and the research originators, etc. Then it reviews the development process of the technologies for the analysis of components of aminoglycosides, discusses the advantages and disadvantages of current methodologies and explains the advantage of liquid chromatography coupled with pulsed amperometric detection method for analysis of the components of aminoglycosides. At last, it introduces some new analysis technologies and ideas and forecasts the direction of future development.

OBJECTIVE: To establish an LC/MSn method for identification of the related substances in etimicn sulfate detected under the chromatographic condition described in Chinese Pharmacopoeia 2010. METHODS: The HPLC separation was carried out on a Welch Ultimate LP-C18 column (4.6 mm × 300 mm, 5 μm) with mobile phase consisting of 0.2 mol · L-1 trifluoroacetic acid (containing 0.1% propionic acid ) -methanol (84: 16) at a flow rate of 1.0 mL · min-1. Thirty percent of the eluent was detected by ion trap mass spectrometry, and the parent ions and the corresponding product spectra of all the related substances in etimicin sulfate were determined and elucidated. RESULTS: Addition of 0.1% propionic acid into the mobile phase significantly enhanced the sensitivity of MS detector without altering the chromatographic behavior such as retention time and elution order of the related substances. Twenty-eight related substances were separated and detected by the LC/MSn method in a typical sample. Nine of them were identified with the help of corresponding impurity reference substances and 14 of them were elucidated by MS fragment information, while the other five were not identified due to limitated information. CONCLUSION: The established method can be applied to the identification of the related substances in etimicn sulfate detected under the chromatographic condition described in Chinese Pharmacopoeia 2010, which is helpful to the quality improvement and process optimization of etmicin sulfate.

AIMTo investigate whether tazobctam has crystallization water.

METHODSIn order to establish a method for water content determination in tazobctam, many methods including Karl Fischer titration, weight loss on drying, TGA, DSC and X-ray diffraction experiment of an orthorhombic form were studied.

RESULTSThe water content of the same batch tazobactam determined by different mthods was different.

CONCLUSIONThe results showed that one mole tazobatam contains half mole water of crystallization and the strength of attraction between tazobatam molecule and water molecule is rather strong.

Crystallization ; Enzyme Inhibitors ; chemistry ; Molecular Conformation ; Molecular Structure ; Penicillanic Acid ; analogs & derivatives ; chemistry ; Water ; X-Ray Diffraction ; beta-Lactamase Inhibitors ; beta-Lactamases ; chemistry

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.
Animals ; Bombyx ; virology ; Gene Deletion ; Gene Expression Regulation, Viral ; Nucleopolyhedrovirus ; genetics ; physiology ; Transcription, Genetic ; Viral Proteins ; genetics ; metabolism ; Virus Replication

To study the related substances in purified alliin, HPLC-MS/MS method carried out on a Phenomenex NH2 column was used for screen and identification of the related substances with full scan MS spectra determination of their [M + H] + ions and then the analyses of the retention time, product MS spectra and/or chemical reference standards. The full scan HPLC-MS chromatogram showed that there were seven major related substances in the purified alliin and their m/z of the [M + H] + ions with increasing retention were 116, 133, 147, 152, 175, 178 and 178, separately. And they were identified as proline, asparagine, glutamine, methiin, arginine, isoalliin and cycloalliin (both were isomers of alliin), respectively. The major related substances in purified alliin are amino acids, homologen and the isomers of alliin.
Chromatography, High Pressure Liquid ; methods ; Cysteine ; analogs & derivatives ; analysis ; Tandem Mass Spectrometry ; methods

Objective To monitor the co-infection status of Borrelia burgdorferi sensu lato (R.b.s.1) and spotted fever group Rickettsia (SFGR) in tourist areas of Heilongjiang province.Methods Polymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B.b.s.1 and ompA of SFGR in ticks,dynamically collected from tourist areas of Heilongjiang province in 2010.Amplification products from positive ticks were sequenced,and phylogenetic analysis was conducted by Mega 5.0 software package.Results 849 ticks were collected from two tourist points,with the dominant ticks in Tiger Mountain and Jingpo Lake were Ixodes persulcatus and Haemaphysalis concinna.Regarding the Ixodes persulcatus from Tiger Mountain,the infection rates of B.b.s.1 and SFGR were 26.15％ and 10.05％.The infection rate of SFGR was 13.33％ in Haemaphysalis concinna and the B.b.s.1 was tndiscovered in the same ticks from Jingpo Lake.However,the co-infection could only be detected in Ixodes persulcatus of both tourist areas.Surveillance data showed that the major ticks were more likely to be appeared in July at Tiger Mountain and in June at Jingpo Lake.Data from the sequence analysis on B.b.s.1 showed that the B.b.s.1 in tourist areas could be classified into three different genotypes,other than B.garinii and B.afzelii.We first detected B.valaisiana-like group genotype in northeast of China.Results from the sequence analysis of SFGR positive products showed that the two DNA sequences of newly detected agents were completely the same as Rickettsia sp.HL-93 which was detected in Hulin and Rickettsia sp.H820 found in northeast,China.Conclusion The co-infection of B.b.s.1 and SFGR was detected in ticks from the tourist areas of Heilongjiang province,and data from the sequencing of specific fragment showed that various kinds of genotypes existed in this area.However; the rates of co-infectionitis-different according to environment,time and population that contributed to the kinds of and the index of ticks existed in the surveys points,also the infection rate of the ticks was studied.

OBJECTIVETo observe the characteristics and difference of gene expression in the pituitary adenomas and para-tumor normal pituitary tissues.

METHODSUsing serial analysis of gene expression (SAGE), two SAGE libraries were generated. Forty clones from each SAGE library were sequenced, and the results were analyzed by SAGE2000 software and compared with the SAGE map at NCBI.

RESULTSA total of 655 gene tags, representing 43 genes, were extracted from the 40 sequence files of the para-tumor normal pituitary tissues and 737 gene tags, representing 53 genes, were extracted from the 40 sequence files of the pituitary adenomas. Of these tags, 13 were not reported before. The genes related to pituitary hormone secretion and energy metabolism were highly expressed in the two kinds of tissues. Some growth factors and cytokines were also expressed, including those involved in the immunological system. But there were also much difference of gene expression in the two tissues. Thirty-one and five tags were only detected in para-tumor normal pituitary tissues and pituitary adenomas, respectively.

CONCLUSIONSGenes involved in hormones secretion and energy metabolism were highly expressed in the pituitary adenomas and para-tumor normal pituitary tissues. Many growth factors and cytokines were also expressed in pituitary. There was also much difference of gene expression in the two kinds of tissues. SAGE can be used not only in understanding the quantity information of gene expression, but also in finding new genes.

Adenoma ; genetics ; metabolism ; Base Sequence ; Cloning, Molecular ; Expressed Sequence Tags ; Gene Expression ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Neoplastic ; Gene Library ; Humans ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Pituitary Gland ; metabolism ; Pituitary Neoplasms ; genetics ; metabolism

OBJECTIVETo search for the disease-associated haplotype in the PRKCZ gene, a susceptibility gene for type 2 diabetes in Han population of North China, by case-control study and linkage disequilibrium (LD) analysis using single nucleotide polymorphisms (SNPs).

METHODSSNPs located in the PRKCZ gene were chosen from public SNP domain by bioinformatic methods and single base extension (SBE) method was used to genotype the loci in 173 sporadic type 2 diabetes patients and 152 normal individuals to perform case-control study and LD analysis. Haplotype block were constructed in these populations.

RESULTSSeveral SNPs in the PRKCZ gene were found to be associated with the disease. The SNPs formed different haplotype block pattern in case and control groups. The frequencies of the haplotypes formed by 5 SNPs were statistically different between the two groups.

CONCLUSIONThe haplotype formed by 5 SNPs in the PRKCZ gene may be associated with type 2 diabetes in Han population of China, which is confirmed from statistics to be a susceptibility gene for the disease.

Alleles ; Asian Continental Ancestry Group ; Case-Control Studies ; Diabetes Mellitus, Type 2 ; genetics ; Ethnic Groups ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Linkage Disequilibrium ; Polymorphism, Single Nucleotide ; Protein Kinase C ; genetics ; Protein Kinase C-delta

OBJECTIVETo study the function of 5 single nucleotide polymorphisms (SNPs) of the PRKCZ gene, a susceptibility gene for type 2 diabetes in Han population of North China, in the pathogenesis of the disease.

METHODSBioinformatic methods and reporter gene activity determination were used to analyze the function of the 5 SNPs.

RESULTSThe reporter gene activities of different alleles of 2 SNPs, rs427811 and rs809912, were obviously different, which implies that these 2 SNPs might be susceptibility loci of the disease.

CONCLUSIONThe PRKCZ gene is further confirmed to be a susceptibility gene for type 2 diabetes in Han population of North China. Two SNPs in the gene play a role in the pathogenesis of the disease by affecting the expression level of PRKCZ gene.

Alleles ; Asian Continental Ancestry Group ; Diabetes Mellitus, Type 2 ; genetics ; Ethnic Groups ; Genetic Predisposition to Disease ; Humans ; Polymorphism, Single Nucleotide ; Protein Kinase C ; genetics ; Protein Kinase C-delta

OBJECTIVE: To analyze the differentially phosphorylated proteins in DENV-2-infected human umbilical venous endothelial cells (HUVECs) and explore the possible pathogenic mechanism of DENV-2 infection. METHODS: The total proteins were extracted from DENV-2-infected HUVECs and blank control HUVEC using SDT lysis method. The phosphorylated proteins were qualitatively and quantitatively analyzed using tandem mass spectrometry (TMT). The identified differentially phosphorylated proteins were analyzed by bioinformatics analyses such as subcellular localization analysis, GO enrichment analysis, KEGG pathway analysis and protein-protein interaction (PPI) analysis. Western blotting was used to detect the expressions of phosphorylated Jun, map2k2 and AKT1 proteins in DENV-2-infected HUVECs. RESULTS: A total of 2918 modified peptides on 1385 different proteins were detected, and among them 1346 were significantly upregulated (FC > 1.2, P < 0.05) and 1572 were significantly downregulated (FC < 0.83, P < 0.05). A total of 49 phosphorylated conserved motifs were obtained by amino acid conservative motif analysis. The most abundant differentially phosphorylated peptides in protein domain analysis included RNA recognition motif, protein kinase domain and PH domain. Subcellular localization analysis showed that the differentially modified peptides were mainly localized in the nucleus and cytoplasm. GO enrichment and KEGG pathway analysis showed that the differential peptides were mainly enriched in the regulation of stimulation response, biosynthesis of small molecules containing nuclear bases, and migration of phagosomes and leukocytes across the endothelium. PPI and KEGG joint analysis showed that the up-regulated and down-regulated differentially phosphorylated proteins were enriched in 15 pathways. In DENV-2-infected HUVECs, Western blotting detected differential expressions of phosphorylated proteins related with the autophagy pathway, namely JUN, MAP2K2 and AKT1, and among them p-JUN was significantly down-regulated and p-AKT1 and p-MAP2K2 were significantly upregulated (P < 0.01). CONCLUSION DENV-2 infected HUVECs show numerous differentially expressed proteins. The downregulation of p-JUN and upregulation of p-MAP2K2 and p-AKT1 suggest their potential roles in regulating autophagy, which is probably involved in the mechanism of DENV-2 infection.
Humans ; Autophagy ; Cell Death ; Cell Nucleus ; Human Umbilical Vein Endothelial Cells/virology* ; Dengue ; Proteome