Therapeutic Effect of Huangqi Chifengtang on Cerebral Infarction Rat Model and Its Effect on Expression of ZO-1， Claudin-5， P-gp and MRP1 Protein in Brain Tissue / 中国实验方剂学杂志

ObjectiveTo explore the therapeutic effect and mechanism of Huangqi Chifengtang on middle cerebral artery embolism（MCAO） rat model. MethodThe 90 SPF male rats were randomly divided into sham operation group， model group， high， medium and low dose groups of Huangqi Chifengtang （8.10，4.05，2.025 g·kg-1） and positive drug group （Naoxintong 0.32 g·kg-1）. MCAO rat model was established and intervened with Huangqi Chifengtang， and the neurological scores of each group were scored. The area of cerebral infarction was calculated by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Serum interleukin-6 （IL-6） and interleukin-1β（IL-1β） were detected by enzyme-linked immunosorbent assay （ELISA），The contents of matrix metalloproteinase-9（MMP-9）， vascular endothelial growth factor（VEGF） and vascular endothelial growth factor receptor 2 （VEGFR2）， the pathological changes of brain tissue in each group were observed by hematoxylin eosin （HE） staining. Western blot was used to detect zonule atresia protein-1（ZO-1）， tight junction protein-5（Claudin-5） and P-glycoprotein （P-gp） and multidrug resistance protein 1（MRP1）. ResultCompared with the sham operation group， the neurological function score and cerebral infarction rate of the model group were significantly increased（P<0.01）， and the levels of IL-6， IL-1β and MMP-9 in serum were significantly increased（P<0.01）， the levels of ZO-1 and Claudin-5 protein expression decreased significantly（P<0.01）， and the levels of P-gp and MRP1 protein expression increased significantly（P<0.01）. Compared with the model group， the neurological function score of rats in each administration group decreased significantly at 14 days （P<0.05，P<0.01）， the pathological changes of brain tissue effectively improved， the rate of cerebral infarction significantly reduced （P<0.01）， and the expression level of IL-6， IL-1β and MMP-9 in serum decreased significantly （P<0.05，P<0.01）， the content of VEGFR2 increased significantly （P<0.01）， and the content of VEGF increased significantly in high， medium dose and positive drug groups （P<0.05，P<0.01）. Although it decreased in low dose group， there was no significant difference. The levels of ZO-1 and Claudin-5 protein expression in brain tissue of high dose group and positive drug group increased significantly （P<0.05，P<0.01）， the level of MRP1 and P-gp protein expression decreased significantly （P<0.05）. ConclusionHuangqi Chifengtang can play a therapeutic role in rats with cerebral infarction by improving the pathological changes of brain tissue， reducing inflammatory reaction， promote angiogenesis and regulating the function of blood-brain barrier（BBB）.

Construction of SNP-STR Multiplex Amplification System with Genetic Markers and Its Forensic Application / 法医学杂志

Objective To select and develop a SNP-STR multiplex amplification system with genetic markers compatible with current STR databases. To understand its genetic polymorphisms in Sichuan Han population and its application value in DNA mixture analysis. Methods Based on the STR genetic markers in commercial kits, SNPs adjacent to these STR markers were selected to be SNP-STR genetic markers. A SNP-STR multiplex amplification system with genetic markers based on allele-specific amplification was constructed using allele-specific amplification primers. The genetic polymorphism of the system in the Sichuan Han population was investigated and the efficiency of systems with different numbers of loci to detect the two individual DNA mixture samples was evaluated. Results An allele-specific multiplex amplification system constituted of 13 SNP-STR genetic markers was selected and constructed. In Sichuan Han population, the heterozygosity of each locus ranged from 0.76 to 0.88, and the combined discrimination power reached 0.999 999 999 999 999 968. In the analysis of the two individual DNA mixture samples： for single-locus amplification, the genotype of the minor components can still be detected when the mixture ratio reaches 1 000∶1; for multiple loci multiplex amplification, the maximum mixture ratio can reach 500∶1. As the number of loci in the system increased, the detection efficiency of the minor components in the DNA mixture decreased. Conclusion SNP-STR genetic markers have a higher polymorphism than STR. The multiplex amplification system made of SNP-STR genetic markers has a better analysis efficiency for mixed samples than traditional STR multiplex amplification system.

Iodoacetamide-induced aquaporin 1 expression in fibroblasts is energy-dependent / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the impact of energy metabolism at the cellular level on the expression of the water channel protein aquaporin 1 (AQP1).</p><p><b>METHODS</b>Balb/c mouse fibroblasts were incubated with iodoacetamide (IA) in vitro, and the changes in AQP1 expression were detected by immunoblotting and immunohistochemistry at 0, 4, and 6 h.</p><p><b>RESULTS</b>IA induced the expression of AQP1 at 4 and 6 h accompanied with cell death. Reverse transcription PCR showed an increased expression of AQP1 mRNA in the cells. AQP1 expression was also upregulated by the inhibitor of microtubule and cytochrome C oxidase.</p><p><b>CONCLUSION</b>A pretranslational regulation occurs in IA-induced AQP1 expression in mouse fibroblasts, and the up-regulated AQP1 accumulation is characterized by mitochondria-related energy dependence.</p>

Preliminary molecular epidemiology of the Staphylococcus aureus in lower respiratory tract infections: a multicenter study in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Staphylococcus aureus (S. aureus) remains as an important microbial pathogen resulting in community and nosocomial acquired infections with significant morbidity and mortality. Few reports for S. aureus in lower respiratory tract infections (LRTIs) have been documented. The aim of this study was to explore the molecular epidemiology of S. aureus in LRTIs in China.</p><p><b>METHODS</b>A multicenter study of the molecular epidemiology of S. aureus in LRTIs was conducted in 21 hospitals in Beijing, Shanghai and twelve other provinces from November 2007 to February 2009. All the collected S. aureus strains were classified as minimum inhibitory concentration (MIC), mecA gene, virulence genes Panton-Valentine Leukocidin (PVL) and γ-hemolysin (hlg), staphylococcal cassette chromosome mec (SCCmec) type, agr type, and Multilocus Sequence Typing (MLST).</p><p><b>RESULTS</b>Totally, nine methicillin-sensitive S. aureus (MSSA) and 29 methicillin-resistant S. aureus (MRSA) strains were isolated after culture from a total of 2829 sputums or bronchoalveolar lavages. The majority of MRSA strains (22/29) had a MIC value of ≥ 512 µg/ml for cefoxitin. The mecA gene acting as the conservative gene was carried by all MRSA strains. PVL genes were detected in only one S. aureus strain (2.63%, 1/38). The hlg gene was detected in almost the all S. aureus (100% in MSSA and 96.56% in MRSA strains). About 75.86% of MRSA strains carried SCCmec III. Agr type 1 was predominant (78.95%) among the identified three agr types (agr types 1, 2, and 3). Totally, ten sequence type (ST) of S. aureus strains were detected. A new sequence type (ST1445) was found besides confirming ST239 as the major sequence type (60.53%). A dendrogram generated from our own MLST database showed all the bootstrap values ≤ 50%.</p><p><b>CONCLUSION</b>Our preliminary epidemiology data show SCCmec III, ST239 and agr type 1 of S. aureus as the predominant strains in LRTIs in Mainland of China.</p>

Drug-resistant genes carried by Acinetobacter baumanii isolated from patients with lower respiratory tract infection / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Acinetobacter baumanii (A. baumanii ) remains an important microbial pathogen resulting in nosocomial acquired infections with significant morbidity and mortality. The mechanism by which nosocomial bacteria, like A. baumanii, attain multidrug resistance to antibiotics is of considerable interest. The aim in this study was to investigate the spread status of antibiotic resistance genes, such as multiple β-lactamase genes and aminoglycoside-modifying enzyme genes, from A. baumanii strains isolated from patients with lower respiratory tract infections (LRTIs).</p><p><b>METHODS</b>Two thousand six hundred and ninety-eight sputum or the bronchoalveolar lavage samples from inpatients with LRTIs were collected in 21 hospitals in the mainland of China from November 2007 to February 2009. All samples were routinely inoculated. The isolated bacterial strains and their susceptibility were analyzed via VITEK-2 expert system. Several kinds of antibiotic resistant genes were further differentiated via polymerase chain reaction and sequencing methods.</p><p><b>RESULTS</b>Totally, 39 A. baumanii strains were isolated from 2698 sputum or bronchoalveolar lavage samples. There was not only a high resistant rate of the isolated A. baumanii strains to ampicillin and first- and second-generation cephalosporins (94.87%, 100% and 97.44%, respectively), but also to the third-generation cephalosporins (ceftriaxone at 92.31%, ceftazidine at 51.28%) and imipenem (43.59%) as well. The lowest antibiotic resistance rate of 20.51% was found to amikacin. The OXA-23 gene was identified in 17 strains of A. baumanii, and the AmpC gene in 23 strains. The TEM-1 gene was carried in 15 strains. PER-1 and SHV-2 genes were detected in two different strains. Aminoglycoside-modifying enzyme gene aac-3-Ia was found in 23 strains, and the aac-6'-Ib gene in 19 strains. aac-3-Ia and aac-6'-Ib genes hibernated in three A. baumanii strains that showed no drug-resistant phenotype.</p><p><b>CONCLUSIONS</b>A. baumanii can carry multiple drug-resistant genes at the same time and result in multi-drug resistance. Aminoglycoside-modifying enzyme genes could be hibernating in aminoglycoside sensitive strains without expressing their phenotype.</p>