AIM: To investigate the differences in the distribution of SRY-related HMG box 5 (SOX5) gene single nucleotide polymorphisms (SNPs) among stable chronic obstructive pulmonary disease (COPD) patients, COPD with pulmonary hypertension (PH) patients and healthy controls, and to explore the association of the SOX5 SNPs in COPD-related PH.METHODS: From April 2013 to April 2015, 250 patients with stable COPD were enrolled continuous-ly in Ningxia People’s Hospital according to COPD treatment guidelines (2013 edition).All the patients received echocar-diography, and were divided into COPD with PH group [pulmonary artery systolic pressure (PASP)≥50 mmHg, n =103] and COPD without PH group (PASP <50 mmHg, n =147).The healthy persons (matched for age, sex, race and smoking index, n =127) were selected as control group at the same period.Genotyping of SOX5 gene rs10842262 and rs11046966
loci was performed using MassARRAY genotyping system ( Sequenom).Genotype frequencies were calculated.RE-SULTS: Age, sex and smoking index showed no significantly difference between control group and COPD group, neither between COPD with PH group and COPD without PH group.Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci between control group and COPD group was of significant difference (P<0.05).Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci showed no significant difference between COPD with PH group and COPD without PH group.CONCLUSION: SOX5 gene rs10842262 and rs11046966 loci may play an important role in COPD, but not in COPD-related PH.

Spiramycin (SP) belongs to the 16-member macrolide antibiotics. It contains three components,namely SP I, SP II and SP III, which differ structurally in the acylation moieties on the C3 of the lactone. The SP I component contains a hydroxyl group at C3. SP II, and SP III are formed by further acetylation or propionylation of the C3 of SP I, by the same 3-O-acyltransferase (3-O-AT) . The study focused on simplifying spiramycin components. Theoretically, disruption/deletion of the 3-O-AT gene will reduce/stop the acylation of SP I to SP II and SP III. In this study, degenerated primers were designed according to the conserved regions of 3-O-acyltransferase, MdmB and AcyA in the medicamycin and carbomycin producers of S. mycarofaciens and S. thermotolerans, respectively, and an 878bp DNA fragment was amplified from the spiramycin-producer of S. spiramyceticus F21. Blast analysis of the 878bp DNA fragment suggested that it encoded the 3-O-acyltransferase (3-0-AT, sspA) gene for spiramycin biosynthesis. The flanking regions of this 878bp DNA fragment were then amplified by single-oligonucleotide-nested PCR, and a total of 4.3 kb DNA was obtained (3457nt among the 4.3kb fragment was sequenced, and deposited in GenBank DQ642742),covering the whole putative 3-O-acyltransferase gene, sspA. The sspA was then deleted from the S. spiramyceticus F21 genome by double cross-over homologous recombination, mediated by temperature-sensitive plasmid pKC1139. A comparison was done of the components of spiramycins produced by the sspA-deleted mutant strain with that of the parent strain by HPLC analysis, which showed that sspA-deleted mutant produced SP I (72%), SP II (18%), and SP III (9.6%), whereas parent strain produced SP I (7.8%), SP II (67%), and SP III (25%), respectively, demonstrating the role of ssp A in the acylation of SP I into SP II and SP III. The ssp A-deleted mutant strain obtained in this study may be used for the production of SP I, or may serve as a good starter for the construction of spiramycin derivatives.
Acyltransferases ; genetics ; Aminoglycosides ; biosynthesis ; Gene Deletion ; Genes, Bacterial ; genetics ; Genetic Engineering ; methods ; Streptomyces ; enzymology ; genetics

Objective: To realize the classification and identification of Cynomorii Herba from different producing areas based on fourier transform infrared spectroscopy (FTIR) and chemometrics. Method: FTIR spectrum data of 106 batches of Cynomorii Herba from 12 cities in 5 provinces were collected by transmission method and preprocessed. The FTIR fingerprints of Cynomorii Herba were established, and spectrum analysis was performed. The FTIR similarities of Cynomorii Herba from different producing areas were calculated by correlation coefficient method. The first derivative (1D) spectrum of average FTIR of Cynomorii Herba from different producing areas were obtained. The soft independent modeling of class analog (SIMCA) model based on principal component analysis (PCA) was established by the preprocessed 1D spectrum data. The orthogonal partial least squares (OPLS) model was established by top 6 principal components. Result: The FTIR fingerprint trend and main absorption peaks of Cynomorii Herba from different producing areas were basically the same,and 16 common characteristic absorption peaks were recognized. Similarity and 1D spectrum of FTIR fingerprint of Cynomorii Herba from different producing areas showed significant and unique characteristics. The established SIMCA model can realize the classification and identification of Cynomorii Herba from different provinces,while OPLS model can realize accurate classification and identification of Cynomorii Herba in different cities. The classification and identification of Cynomorii Herba from 12 city producing areas showed obvious geographical clustering characteristics. Conclusion: The established method based on FTIR and chemometrics can realize the classification and identification of Cynomorii Herba from 12 cities.

Objective::To establish a pre-column derivatization reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of 17 amino acids in Cynomorii Herba from different producing areas and conduct a multivariate statistical analysis. Method::RP-HPLC with pre-column derivatization was employed, with phenyl isothiocyanate (PITC) as derivatization reagent. Separation was performed on a WondaSil C₁₈-WR column (4.6 mm×150 mm, 5 μm), with 0.05 mol·L^-1 sodium acetate solution (pH 6.5) as mobile phase A, and acetonitrile-methanol-water (3∶1∶1) as mobile phase B for gradient elution at a flow rate of 0.8 mL·min^-1. The detective wave length was set at 254 nm, and the column temperature was maintained at 35 ℃. Principal component analysis (PCA) and systematic cluster analysis (HCA) models were established for multivariate statistical analysis and quality evaluation. Result::17 Kinds of amino acid were detected in Cynomorii Herba, 7 of which were essential amino acids. The 17 amino acids showed good linearity in respective concentration range, r = 0.999 0-0.999 9.The average recoveries were between 98.03%-103.89%with RSD<3.5%. The results of PCA and HCA were basically the same, and both methods can be used to clearly distinguish Cynomorii Herba from 12 municipal producing areas into 6 regions. PCA can be used to classify Cynomorii Herba according to different municipal or provincial production areas, and HCA can be used to classify it according to provincial production areas. It showed that the amino acid contents in Cynomorii Herba from different municipal and provincial producing areas had differences, and the content distribution showed obvious geographical clustering characteristics. PCA showed that Cynomorii Herba from Gansu province and Inner Mongolia had higher amino acid contents and better quality as compared with other producing areas. Conclusion::The established method can be used for content determination of 17 amino acids in Cynomorii Herba from different producing areas, and provide a reference for its comprehensive quality evaluation.