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Metabolomics on Pudilan Xiaoyan Oral Liquid in treatment of Influenza A/H1N1-induced pneumonia based on GC-MS

QIAN Wen-juan;YANG Rui;XIE Tong;YAO Wei-feng;KANG An;DI Liu-qing;SHAN Jin-jun

Chinese Traditional and Herbal Drugs,Vol 49,No. 10

【Abstract】 Objective To screen the pneumonia-related abnormal metabolites in lung tissue of mice infected by Influenza A/H1N1, and to monitor the regulation effect of Pudilan Xiaoyan Oral Liquid (Pudilan) and to explore potential anti-pneumonia mechanism. Methods ICR mice were randomly divided into four groups with ten mice in each group: normal group, model group, Pudilan group, and Ribavirin group. The mice were infected with H1N1 virus intranasally and by gavage once every day for six consecutive days. 2 h after the last dose, the mice were sacrificed and lungs were collected. Metabolomics based on GC-MS was applied to analyze the changes of metabolites in the lung tissue of each group. The potential biomarkers of H1N1-induced pneumonia were screened out under three conditions of P < 0.05, VIP > 1.0, and Fold Change > 1.5. Metabolic pathways related to the treatment mechanism of Pudilan were analyzed. Results The infection of H1N1 virus led to infiltration of inflammatory cells in the lungs of mice and various degrees of pneumonitis and metabolic disorders. Pudilan and ribavirin both could ameliorate the symptoms of pneumonia and restore various metabolites back to the normal levels. Conclusion The treatment effect of Pudilan on H1N1-induced pneumonia is related to its regulatory effects on 14 potential biomarkers and 12 associated metabolic pathways.

Non-targeted metabolomics in septic mice infected with Klebsiella pneumoniae

ZHANG Jia-xuan;SUN Lang;PANG Jing;HU Xin-xin;NIE Tong-ying;LU Xi;WANG Xiu-kun;YANG Xin-yi;YOU Xue-fu;LI Cong-ran

Acta Pharmaceutica Sinica,Vol 53,No. 07

【Abstract】 UHPLC-QTOF-MS was applied to non-targeted metabolomics study of mice infected with K.pneumoniae ATCC ® BAA 2146 to discover potential biomarkers and metabolic pathways that are associated with sepsis. Fifty-eight metabolites were identified by principal components analysis (PCA) and partial least squares discriminant analysis (OPLS-DA), which was combined with variable projection importance (VIP) and nonparametric test. Eighteen of the 58 metabolites were further found to be involved in 8 metabolic pathways, including nicotinate and nicotinamide metabolism, pyrimidine metabolism, vitamin B6 metabolism, taurine and hypotaurine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism and glycerophospholipid metabolism.

Development of an ELISA for identification of immunodominant protein antigens of Mycoplasma hyopneumoniae

Yaoqin Zhou;Feng You;Jie Zhong;Haoju Wang;Honglei Ding

Chinese Journal of Biotechnology,Vol 34,No. 01

【Abstract】 We developed a method to identify serological humoral immunodominant proteinic antigen of Mycoplasma hyopneumoniae (Mhp). After constructing the recombinant plasmid pGEX-6P-1-mhp366 and transforming it into Escherichia coli BL21 (DE3), the recombinant GST-Mhp366 protein was expressed successfully. The lysates of the recombinant GST-Mhp366 and genetic engineering GST were added into glutathione coated plates and reacted with 17 positive sera or 13 negative sera. Meanwhile, the optimization of experimental conditions, including coated antigen, blocking buffer, dilutions of sera and second antibody were determined. The optimal concentration of the coated antigen was the original bacteria lysates without dilution, and the optimal blocking buffer contained 10% FBS and 2.5% skim milk in PBS. Besides, the working concentration of serum samples and the HRP-tagged rabbit anti-pig IgG secondary antibody were 1:500 and 1:40 000, respectively. Thus, an indirect ELISA was established for identification of immunodominant protein antigens of Mhp. Meanwhile, this method was confirmed by the identified serological humoral immunodominant proteinic antigen Mhp156 and Mhp364. This method can be used for identification of the candidate vaccine antigens on a genome-wide scale. Furthermore, it can lay the foundation for identifying the candidate vaccine antigens through colostra and the nasal mucosal secretions.

Research Progress on Mechanism of Traditional Chinese Medicine in Treating Mycoplasma Pneumonia in Children

TAN Dan;JIANG Zhiyan

Chinese Archives of Traditional Chinese Medicine,Vol 36,No. 06

【Abstract】 Mycoplasma pneumoniae pneumonia is a common respiratory disease in children, and macrolide drugs are the prior choice for clinical treatment. Macrolide drugs have been widely used for children’s respiratory diseases, leading to the increased resistance rate and gastrointestinal adverse reactions. Hence, more and more doctors begin to use traditional Chinese medicine (TCM) based on syndrome differentiation due to its obvious advantages. Studies have revealed that the efficacy of TCM in anti-MP is mainly related to the direct inhibition of MP, immune regulation, protection and repair of epithelial cells, microcirculation improvement, and reduction of adverse effects induced by western medicine.

Sequencing and comparative genomics analysis of Klebsiella pneumoniae plasmid p1512-KPC

JIANG Zhaofang;ZHAO Yachao;LI Manli;ZHOU Dongsheng;ZHU Tianchuan;TAN Xiaoyan;TONG Yigang;LONG Jun

Acta Microbiologica Sinica,Vol 59,No. 02

【Abstract】 [Objective] Comparative genomics analysis of the fully sequenced plasmid p1512-KPC from the multi-drug resistant clinical Klebsiella pneumoniae 1512 isolate was performed. [Methods] We used 16S rRNA gene sequencing to identify the bacterium. Seven housekeeping genes ( gapA, infB, mdh, pgi, phoE, rpoB, and tonB) were used for the multilocus sequence typing (MLST) scheme. Resistance genes were screened by PCR amplification. The plasmid was transferred into recipient Escherichia coli EC600 using conjugal transfers and electroporation experiments. The activity and type of carbapenemase were detected using Carba NP, and the minimum inhibitory concentration (MIC) was determined using VITEK 2 compact system. Sequencing and bioinformatics analysis were used to characterize the plasmid p1512-KPC. [Results] The 1512 isolate was an ST11 multi-drug resistant K. pneumoniae strain showing Ambler class A carbapenemase. PCR demonstrated strain 1512 harbored blaKPC-2, dfrA1, and sul1 genes. The blaKPC-2 gene was located in non-self-transmissible plasmid p1512-KPC. Sequencing and bioinformatics analysis revealed that p1512-KPC, 117.69 kb in length, harbored IncFII replicon and replicon repB belonging to Rep_3 family unknown incompatibility groups. In addition, p1512-KPC contained blaKPC-2, blaCTX-M-65, blaTEM-1, and rmtB genes, among which blaKPC-2, blaCTX-M-65, and blaTEM-1 were carried by ΔTn 6296, Tn6367, and ΔTn 2, respectively. [Conclusion] The resistance of K. pneumoniae 1512 to penicillin, cephalosporin, carbapenem, monobactam, and aminoglycoside was mediated by non-self-transmissible plasmid p1512-KPC. Moreover, the comparative genomics analysis provided a deeper insight into the diversification and evolution of those plasmids that harbored IncFII replicon and replicon repB belonging to Rep_3 family unknown incompatibility groups.

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