Aim To develop a convenient and effective method to isolate and culture primary rabbit aortic vascular smooth muscle cells(VSMCs).Methods The thoracic aortas were removed by dissection under sterile conditions.Aortic smooth muscle cells were excised and cleaned of fat and connective tissue,and the isolated vessel media was cut into 1 mm3 pieces.The explants were digested with different concentrations of collagenase typeⅠ,and incubated at 37℃ for different time,then undispersed explants were placed onto a sterile 100-mm plastic tissue culture dish with growth medium.Results VSMCs could emigrate from the explants digested 6 h by collagenase typeⅠ(1.5 g?L-1)for 24 hours,the cells would passage for another 4~5 days.Confluency could be reached within 3~4 days after subculturing.VSMCs were identified by immunoreactivity with ?-actin and by the smooth muscle cell-specific,hills and valley-like morphology.Conclusion It was an effective method to culture primary VSMCs from the explants digested for 6h by collagenase type Ⅰ(1.5 g?L-1),which could shorten primary culture time.

Aim To develop a convenient and effective method to isolate and culture primary rat aortic endothelial cells (RAECs). Methods The thoracic aortas were removed by dissection under sterile conditions. Aortas were turned over to expose the luminal surface, and the surfaces were digested with different concentrations of collagenase typeⅠ, incubated at 37℃ for different times, then, cut into pieces and placed luminal side down onto collagen-coated flask with growth medium. Results RAECs could emigrate from explants digested 1h by collagenase typeⅠ(2.0 g?L-1) for 24 h and cells would passage for another 4~5 days. Reached confluency within 3~4 d after subculturing. RAECs were identified by immunoreactivity with Factor-Ⅷ and by the endothelial cell-specific, cobblestone-like morphology. Conclusion It is an effective method to culture primary RAECs from explants digested for 1h by collagenase type Ⅰ(2.0 g?L-1),that can shorten primary culture time.

Objective To analyze the characteristics of toxin, the PCR-ribotyping(RT) and the multilocus sequence typing(MLST) of Clostridium difficile strains isolated from China-Japan Friendship Hospital in order to provide a basis for monitoring the outbreak of nosocomial Clostridium difficile infection.Methods A total of 321 samples were collected from the patients with suspected Clostridium difficile infection(CDI) in China-Japan Friendship Hospital(CJFH) during 2012 to 2013.All Clostridium difficile strains were isolated and identified by the standard phenotypic culture method.Cytotoxicity test was performed to detect toxin B.Toxin genes (tcdA and tcdB) and binary toxin genes (cdtA and cdtB) harbored by those strains were analyzed.RT and MLST were used for homologous analysis.Clinical data of the patients were collected to analyze the isolation rate of Clostridium difficile in different populations.Results Forty-eight strains of Clostridium difficile were isolated from 46 patients with diarrhea and three of them were isolated from the same patient.The incidence of CDI among all patients, outpatients and inpatients were 14.3%(46/321), 12.8%(5/39) and 14.5%(41/282), respectively.Toxin B was detected in all of the strains as indicated by the cytotoxicity test.Strains of sequence type 1(ST1) showed the strongest cytotoxicity of all the isolated Clostridium difficile strains.Ten out of the 48 strains (20.8%) were tcdA(-)/tcdB(+) strains, which belonged to either ST37 or ST81.The results of RT and MLST were consistent in assigning the strains into nine types, in which the predominant type was ST1/RT027 accounting for 27.1% (13/48).All of the ST1/RT027 strains presented a toxin gene profile of tcdA(+)/tcdB(+) and cdtA(+)/cdtB(+).Most of the ST1/RT027 strains were isolated from the Traditional Chinese Medicine Department of Respiratory, where smallnosocomial outbreaks of ST1/RT027 strain infection might happen.Conclusion CDI diagnosed in CJFH mainly belongs to nosocomial infection.Most of the isolated strains harbor tcdA(+)/tcdB(+) genes.Surveillance for the outbreaks of CDI caused by ST1/RT027 strains over producing toxins A and B should be strengthened in hospitals.

Objective:To investigate the clinical value of monitoring serum complement C1q/tumor necrosis factors-associated protein 3 (CTRP3) and lipoprotein-associated phospholipase A2(Lp-PLA2) levels in patients with coronary heart disease, especially patients with acute myocardial infarction (AMI).Methods:This case-control study included 99 patients with angina pectoris aged (60.4±10.4) years, 105 patients with AMI aged (61.7±14.3) years, and 60 healthy individuals aged (43.6±9.5) years. Serum CTRP3 was detected by ELISA, and Lp-PLA2 was detected by automatic biochemical analyzer. Logistic regression analysis was performed to determine the correlation between CTRP3, Lp-PLA2 in angina pectoris and AMI patients. The diagnostic efficiency of each index was analyzed by receiver operating characteristic (ROC) curve.Results:Serum Lp-PLA2 was significantly higher in AMI group than in angina pectoris group ([313.1±68.1] U/L vs [205.8±71.4] U/L, P<0.001), while CTRP3 was significantly lower in AMI group than in angina pectoris group ([64.2±18.5] μg/L vs [84.8±25.0] μg/L, P<0.001). Logistic regression showed that serum CTRP3 was negatively correlated with AMI ( OR=0.964, 95% CI 0.935-0.993, P=0.019), and Lp-PLA2 was positively correlated with AMI ( OR=1.020, 95% CI 1.008-1.032, P=0.001). ROC analysis showed that the AUC (95% CI) of AMI diagnosed by CTRP3 was 0.753 (0.685-0.821), P<0.001; the AUC (95% CI) of AMI diagnosed by Lp-PLA2 was 0.884 (0.833-0.935), P<0.001; the AUC (95% CI) of diagnosis efficacy by combined indices was 0.910 (0.870-0.950), P<0.001. Conclusions:Lower serum CTRP3 and higher serum Lp-PLA2 levels are associated with increased risk for AMI. Combined detection of both indices can improve the diagnostic efficacy of AMI.

The diagnostic technology of acute stroke by microwave imaging has the advantages of being non-ionizing, fast, small, and low-cost. Therefore, this technology is expected to become an auxiliary or alternative means to CT and MRI technology. As the signal transmitting and receiving device of the microwave imaging system, the antenna has an important influence on the performance of the imaging system. At present, there are many antennas with different performances used in imaging systems, but there is a lack of clear evaluation criteria for them. In this paper, several typical antennas were introduced, their advantages and disadvantages from the perspective of bandwidth and near-field were analyzed, and the common requirements of imaging systems for antennas and the performance indicators of various types of imaging systems were summarized. Moreover, the development trend of antenna technology for microwave imaging was pointed out to provide a reference for the study of stroke microwave imaging technology.

Escherichia coli was metabolically engineered to produce poly(glycolate-co-lactate-co-3-hydroxybutyrate) using glucose and xylose as carbon sources. The combinatorial biosynthetic route was constructed by the overexpression of a series of enzymes including D-tagatose 3-epimerase, L-fuculokinase, L-fuculose-phosphate aldolase, aldehyde dehydrogenase, propionyl-CoA transferase, β-ketothiolase, acetoacetyl-CoA reductase, and polyhydroxyalkanoate synthase. Overexpression of polyhydroxyalkanoate granule associated protein significantly improved biopolymer synthesis, and the recombinant strain reached 3.73 g/L cell dry weight with 38.72% (W/W) biopolymer content. A co-culture engineering strategy was developed to produce biopolymer from a mixture of glucose and xylose, achieving 4.01 g/L cell dry weight containing 21.54% (W/W) biopolymer. The results of this work offer an approach for simultaneously utilizing glucose and xylose and indicate the potential for future biopolymer production from lignocellulosic biomass.
3-Hydroxybutyric Acid ; Escherichia coli ; Glucose ; Glycolates ; Lactates ; Metabolic Engineering ; Polyesters ; Xylose

The purpose of this study is to provide a culture for mouse bone marrow-derived macrophages (BMDM) and peritoneal macrophages (PM) and to characterize their molecular and cellular biology. The cell number and purity from the primary culture were assessed by cell counter and flow cytometry, respectively. Morphological features were evaluated by inverted microscope. Phagocytosis by macrophages was detected by the neutral red dye uptake assay. Phenotypic markers were analyzed by real-time fluorescent quantitative PCR. Our results show that the cell number was much higher from culture of BMDM than PM, while there was no significant difference regarding the percentage of F4/80+CD11b+ cells (98.30%±0.53% vs. 94.83%±1.42%; P>0.05). The proliferation rate of BMDM was significantly higher than PM in the presence of L929 cell conditioned medium, by using CCK-8 assay. However, PM appeared to adhere to the flask wall and extend earlier than BMDM. The phagocytosis capability of un-stimulated BMDM was significantly higher than PM, as well as lipopolysaccharide (LPS)-stimulated BMDM, except the BMDM stimulated by low dose LPS (0.1 μg/mL). Furthermore, Tnfα expression was significantly higher in un-stimulated BMDM than PM, while Arg1 and Ym1 mRNA expression were significantly lower than PM. The expression difference was persistent if stimulated by LPS+IFN-γ or IL-4. Our data indicate that bone marrow can get larger amounts of macrophages than peritoneal cavity. However, it should be aware that the molecular and cellular characteristics were different between these two culture systems.
Animals ; Bone Marrow Cells ; physiology ; Cells, Cultured ; Culture Media, Conditioned ; Lipopolysaccharides ; metabolism ; Macrophages ; classification ; physiology ; Mice ; Phagocytosis

Objective To track and evaluate the implementation of the Radiation Shielding Requirements in Room of Radiotherapy Installations—Part 1: General Principle (GBZ/T 201.1–2007) among relevant personnel in medical radiation institutions, and to provide a scientific basis for revising the standard. Methods According to the Guidelines for Health Standards Tracking Evaluation (WS/T 536–2017) and the implementation protocol of standard evaluation, an online survey was conducted among 212 relevant workers from 146 medical radiation institutions across 18 provinces in China. The data were aggregated and analyzed with the use of Microsoft Excel 2010. Results A total of 215 questionnaires were returned, of which 212 were valid. Among the valid respondents, 77.8% believe that this standard is universally applied; 96.2% believe that this standard can meet work needs; 63.7% have participated in relevant training on this standard; 74.1% use this standard once or more per year; and 10.8% believe that this standard needs to be revised. Conclusion Medial radiation workers have a high rate of awareness of the basic information and content of the standard, but the understanding and application of the standard content need to be improved. We recommend that relevant departments further strengthen the promotion of and training on the standard, revise some content based on actual situation, and improve workers’ ability to use the standard.