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To extend its shelf life, vacuum freeze-dried Toona sinensis (VFDTS) powder was packaged in sealed polyethylene (PE) bags, sealed aluminum foil bags, composite vacuum bags, or aluminum foil vacuum bags. Changes in the sensory quality, moisture content, color, flavor components and total bacterial count of VFDTS powder were monitored during storage under accelerated conditions at (55 ± 1) ℃ and (40 ± 3)% relative humidity (RH). The results showed that VFDTS powder packaged in sealed PE bags exhibited severe moisture absorption and browning with prolonged storage time, and the total bacterial count significantly increased. On the other hand, the moisture content, color and total bacterial count of the powder stored in aluminum foil vacuum bags remained substantially unchanged. A total of 56 volatile compounds were identified in VFDTS powder stored in the different packaging systems, including 13 alcohols, 11 aldehydes, 3 acids, 3 terpenes, 7 ketones, 9 esters, one ether, 2 sulfur compounds, 3 alkanes, and 4 heterocycles. Principal component analysis (PCA) of the volatile compounds showed clear separation among VFDTS powder samples stored in the 4 packaging systems. Based on variable importance in projection (VIP > 1) from orthogonal partial least squares discriminant analysis (OPLS-DA), a total of 14 differential volatile compounds were selected, and their clustering heatmaps could be used to effectively classify the samples in different packaging systems. The samples in aluminum foil vacuum bags had the best quality, followed by sealed aluminum foil bags and composite vacuum bags, while the greatest changes in moisture, color, flavor and microbiological quality were observed in VFDTS powder in sealed PE bags during storage.

Objective To analyze the effects of fat mass and obesity-associated (FTO) gene, IL-6, and HSP-60 gene polymorphism on the incidence rate and prognosis of breast cancer (BCa) for patients with type 2 diabetes mellitus (T2DM). Results A total of 1551 patients with BCa were included in the experimental group and 1605 women of the same age who participated in physical examination were included in the control group. The clinical data of the 3156 participants were collected through the baseline data questionnaire, and the genotypes of FTO, IL-6, and HSP-60 single-nucleotide polymorphism (SNP) were determined through blood sample detection. The predictive value of the three SNPs for the incidence risk of BCa for T2DM patients was evaluated. The OS of 1168 patients with BCa was obtained through follow-up, and the effects of the three SNPs and T2DM on OS of BCa patients were evaluated. Results The three loci were FTO rs3751812, IL-6 rs1800796, and HSP-60 rs2605039. The BCa incidence rate for T2DM women with wild homozygous SNP genotype was significantly higher than that for non-T2DM women (FTO: χ2=3.530, P=0.013; IL-6: χ2=6.288, P=0.029; HSP-60: χ2=4.926, P=0.005). The three wild homozygous genotypes were independent risk factors that influenced the incidence rate of BCa (all P < 0.05). Patients with HSP-60 rs2605039 (GT+TT) genotype had better OS (P=0.031). Conclusion FTO, IL-6, and HSP-60 gene polymorphisms have certain value in BCa prediction for T2DM patients. Patients with BCa and HSP-60 rs2605039 GT+TT genotype have high OS.

[Purpose/Significance] Soil microorganism is the most active component of soil, which is the key factor of a healthy soil ecosystem Therefore, to investigate the hotspots, frontiers and directions of soil microorganism research is helpful for researchers to understand research topics and directions. [Method/Process] Using CiteSpace V software papers of soil microbiology published by key journals from 2010 to 2019 collected by CNKI were used to analyze the distribution characteristics of authors, institutions and journals, and to draw the knowledge graph of keyword co-occurrence, burst terms distribution and the time line map of keyword burst terms. [Results/Conclusions] There are more than 400 articles about soil microorganism area published in China every year, and the research in this field continues to draw attention. Northwest A&F University and University of Chinese Academy of Sciences are the key research institutions, and Shen Qirong, Yang Dianlin and Mao Zhiquan are the core author groups, which have made outstanding contributions to the development of this field. The research hotspots have evolved over time, and can be divided into four modules: First, indirect indicators of soil microorganisms (soil enzyme activity, soil uptake, soil microbial biomass carbon and nitrogen); second, mature development of research methods (Phospholipid-derived Fatty Acids, denaturing gradient Gel electrophoresis, Biolog-ECO technique, and high throughput sequencing); third, the visual representation of soil microorganisms (microbial community structure and functional diversity); Lastly, the impact of factors (soil type and land use, fertilization, crops, nitrogen deposition, and bio-char). The application of molecular biological methods such as high throughput sequencing and system biological methods, the development and utilization of bio-char, the influence of nitrogen deposition and the improvement measures should be paid more attention as the focus of future research.

Objective To determine the effects of maternal and infant separation(MS) on anxiety and depression-like behaviors, Wnt signaling molecules and receptors in the hippocampus of offspring rats. Methods Sixteen Sprague-Dawley (SD) pregnant rats (240~260 g) were randomly and equally divided into 2 groups. The offspring rats from the control group (CON) received no postpartum intervention, while those of the experiment group (MS) were given maternal-infant separation 3 h per day for 14 consecutive days since delivery. Elevated plus maze test (EPMT) and forced swimming test (FST) were conducted to evaluate the anxiety and depression-like behaviors of 1-month-old offspring rats. Real-time quantitative PCR was used to detect the levels of Wnt molecules in the hippocampus of the offspring rats, and Western blotting was adopted to determine the expression of Wnt2 and Wnt3a proteins as well as the Wnt signaling pathway inhibitory molecules DKK1 and sFRP3 in the hippocampus. Results The total number of arm entries (including open-arm and close-arm entries) was not statistically different between the MS and CON groups. However, the MS group had significantly lower percentages of open-arm entries (P < 0.01) and open-arm time (P < 0.05), and longer stationary time (P < 0.05) in the FST than the CON group, indicating obvious anxiety and depression-like behaviors. Moreover, the mRNA levels of Wnt2 and Wnt3a in the hippocampus were notably decreased (P < 0.01), while the protein levels of DKK1 and sFRP3 were increased remarkably (P < 0.01) in the MS group than the CON group. Conclusion Maternal and infant separation leads to depression and anxiety-like behaviors in offspring rats, which may be associated to the disorder of Wnt signaling molecules in the hippocampus.

To investigate the effect of drying methods on the volatile flavor substances of Toona sinensis, the volatile components of vacuum freeze-dried, heat pump-dried, hot air-dried, and microwave vacuum-dried Toona sinensis samples were analyzed by gas chromatography-ion mobility spectrometry (GC-IMS) and gas chromatography-mass spectrometry (GC-MS) coupled with relative odor activity values, and the differences of volatile components among samples were further explored by combining the stoichiometric principal component analysis and partial least square discriminant analysis. The results showed that 78 peaks were detected and 59 compounds were identified by GC-IMS. 94 kinds of volatile compounds were identified by GC-MS, and the main volatile components were sulfides, olefins and aldehydes. The content of total volatile compounds was significantly reduced after drying. The key aroma compounds in Toona .sinensis samples were identified by ROAV method, such as (E,Z)-di-1-propenyldisulfide, 2-mercapto-3,4-dimethyl-2,3-dihydrothiophene, hexanal, 2-hexenal and eugenol. The PCA and PLS-DA results showed that the samples with different drying methods were not only clearly separated from the raw samples, but also significantly different from each other. The raw samples and vacuum freeze-dried samples were grouped, and the other three dried Toona .sinensis samples were grouped. According to the variable importance in projection score, a total of 10 (VIP>1) labeled volatile compounds were selected. The drying method had significant effect on the flavor characteristics of Toona sinensis, and the vacuum freeze-dried group was the closest to the raw sample, while heat pump-drying was the most suitable dehydration method for Toona sinensis, considering the practical application. This study provides some theoretical references for flavor quality control in the thermal processing process. [ABSTRACT FROM AUTHOR]

The key aroma components in fresh Toona sinensis were analyzed and identified using solid-phase microextraction and solvent assisted flavor evaporation, in combination with odor activity value (OAV) and gas chromatography olfactory mass spectrometry (GC-O-MS). The two extraction methods identified 90 volatile compounds, including 12 sulfur compounds, 11 aldehydes, 34 terpenes, 2 ketones, 4 alcohols, 4 esters, 1 phenol, 1 acid, 15 alkanes, 4 cycloalkanes, and 2 other compounds. Through OAV calculation, nine aroma active components were identified, including (E,E)-, (E,Z)-, and (Z,Z)-bis(1-propenyl) disulfide, trans-2-mercapto-3,4-dimethyl-2,3-dihydrothiophene, methyl-thiirane, hexanal, 3-methyl-butanal, (E)-2-hexanal, and (E)-2-octenal. Seven aroma active compounds were identified using GC-O-MS, including (E,E)-, (E,Z)-, and (Z,Z)-bis(1-propenyl) disulfide, trans-2-mercapto-3,4-dimethyl-2,3-dihydrothiophene, methyl-thiirane, hexanal, and (E)-2-hexanal. The OAV and GC-O-MS analyses confirmed that (E,E)-, (E,Z)-, and (Z,Z)-bis(1-propenyl) disulfide, trans-2-mercapto-3,4-dimethyl-2,3- dihydrothiophene, methyl-thiirane, hexanal, and (E)-2-hexanal were the key aroma components in fresh Toona sinensis. [ABSTRACT FROM AUTHOR]

Chronic exposure to risk factors at the beginning stage of atherosclerosis development can damage endothelium and ultimately initiate endothelial dysfunction. Endothelial dysfunction is one of the major mechanisms in atherosclerosis development. High-density lipoprotein (HDL) is a lipoprotein with complicated function and is composed of a variety of proteins and lipids. Epidemiological studies showed an inverse correlation between HDL and atherosclerosis. The protective role of HDL on endothelium might related to its function in reversing cholesterol transport, reducing peripheral cholesterol accumulation, preventing foam cells formation, dilating vessels, antioxidant and anti-inflammation, all of which antagonize development of atherosclerosis. But HDL obtained from patients with cardiovascular disease which has known dysfunctional HDL may induce endothelial dysfunction. This review addresses the relationships between functional HDL or dysfunctional HDL with the endothelial function. [ABSTRACT FROM AUTHOR]

Objective To observe the association between adiponectin gene polymorphism, serum adiponectin levels with the incidence of coronary artery disease (CAD) and the severity of coronary artery stenosis in patients with essential hypertension (EH). Methods A total of 414 patients with EH (234 cases with CAD and 180 cases without CAD) and 185 control subjects were recruited in this study. Serum adiponectin levels were measured by enzyme-linked immunosorbent assay (ELISA). Adiponectin single-nucleotide polymorphisms rs266729, rs7649121, rs1501299 and rs3774262 were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results Serum adiponectin levels were significantly lower in EH with CAD group than those in control group and EH without CAD group. Adiponectin SNP rs7649121 AT genotype reduced the risk of CAD compared with AA genotype (adjusted OR=0.566, 95% CI 0.346-0.925, P=0.023). Logistic regression analysis showed that age and LDL-C were risk factors of CAD, and adiponectin was the protective factor for CAD in EH patients. The severity of coronary artery stenosis was negatively related to the level of adiponectin. Adiponectin levels were not affected by the adiponectin gene polymorphism. Conclusion The decreased serum adiponectin level was the independent risk factor for CAD in EH patients, which was negatively related to the severity of coronary artery stenosis. Adiponectin SNP rs7649121 may contribute to the risk factors of CAD in EH patients. [ABSTRACT FROM AUTHOR]

Yeast surface display (YSD) is a technology that fuses the exogenous target protein gene sequence with a specific vector gene sequence, followed by introduction into yeast cells. Subsequently, the target protein is expressed and localized on the yeast cell surface by using the intracellular protein transport mechanism of yeast cells, whereas the most widely used YSD system is the α-agglutinin expression system. Yeast cells possess the eukaryotic post-translational modification mechanism, which helps the target protein fold correctly. This mechanism could be used to display various eukaryotic proteins, including antibodies, receptors, enzymes, and antigenic peptides. YSD has become a powerful protein engineering tool in biotechnology and biomedicine, and has been used to improve a broad range of protein properties including affinity, specificity, enzymatic function, and stability. This review summarized recent advances in the application of YSD technology from the aspects of library construction and screening, antibody engineering, protein engineering, enzyme engineering and vaccine development.

Protein complexes are involved in a variety of biological activities. Accurate and comprehensive characterization of the structures and interactions of protein complexes is crucial in determining their biological functions. Chemical cross-linking coupled with mass spectrometry (CXMS) is an emerging investigative technique for protein complexes. CXMS enables the sensitive high-throughput analysis of protein complexes without the requirements of molecular weight and purification. These attributes have spurred the increased use of CXMS for the structure and interaction characterization of purified protein complexes and complicated cell lysate samples. CXMS utilizes chemical cross-linking reagents to covalently connect two reactive amino acids in or between proteins that are spatially close to each other. Subsequently, the cross-linked proteins are digested into cross-linked peptides, followed by LC-MS/MS analysis, as well as database searching to provide cross-linking information for the composition, interaction, and structural site distance restrictions of protein complex identification. Therefore, identification of cross-linked sites has a decisive influence on the characterization of protein complexes. This identification is limited by the unsatisfactory quality of the cross-linked peptide spectrum. Insufficient b/y fragment ions and poor continuity of amino acid sequence matching lead to low coverage and accuracy of cross-linked site identification. Based on the complementary feature of mirror-cutting digestion, an orthogonal digestion strategy based on LysargiNase combined with trypsin was developed in this study. Trypsin is the most commonly utilized digestion enzyme in proteomics, with extremely high enzyme activity and specificity. Trypsin generates C-terminally charged peptides after lysine (K) and arginine (R). LysargiNase is a mirror protease complementary to trypsin that cleaves before the K and R residues. This generates peptides with an N-terminal positively-charged residue. Owing to the different physical and chemical micro-environments of the cross-linked peptides digested by LysargiNase and trypsin, the behavior of their detection ability in MS analysis is diverse. Using the orthogonal digestion strategy, both simple and complicated cross-linked samples were analyzed in this study. For the analysis of bovine serum albumin (BSA), 291 pairs of non-redundant cross-linked sites were obtained, of which 216 pairs of cross-linked sites were provided by trypsin digestion, whereas 75 pairs of cross-linked sites were exclusively supplied by LysargiNase digestion. Except for the 35% increase in the number of identified cross-linked sites, 32% of the spectra of the commonly identified cross-linked peptides have better quality with more b-type fragment ions and consecutive sequence matching. Furthermore, for the Escherichia coli sample, 726 pairs of cross-linked sites were obtained in total, among which, 624 and 274 pairs were identified from trypsin and LysargiNase digestion, respectively. LysargiNase digestion yielded 120 individual cross-linked sites, which resulted in a 16% increase in single trypsin digestion. Consistent with the BSA sample, the quality was improved in 35% of the spectra of commonly identified cross-linked peptides. Corresponding to the identified cross-linked peptides, 242 structural constraints with 607 pairs of intra-cross-linked sites and 29 sets of protein-protein interactions with 119 pairs of inter-cross-linked sites were obtained. The collective results demonstrated that, mirror cutting-assisted orthogonal digestion strategy could significantly increase the number of identified fragment ions and amino acid sequences matching the continuity of the spectra by contributing b-and y-type ions, respectively. This improved the accuracy and coverage of cross-linked peptide identification. The findings additionally demonstrate the superiority of our method in the accurate identification of the cross-linked peptide spectra and the increased number of identified cross-linked sites. In a word, this method is expected to provide new insights for the large-scale and highly accurate characterization of protein complexes.

Coronary artery disease (CAD) is a chronic disease but causes the highest mortality and morbidity among the cardiovascular diseases worldwide. Correlations between CAD and gut microbiota have been observed. This suggests that the gut microbiota could become a vital diagnostic marker of CAD, and restoring the gut habitat may become a promising strategy for CAD therapy. The elevated level of trimethylamine-N-oxide (TMAO), a gut microbiota-derived metabolite, was found to be associated with the increased risk of cardiovascular disease and the all-cause mortality. Preclinical studies have shown that it has pro-arteriosclerosis properties. It is likely that regulating the production of TMAO by gut microbiota may become a promising strategy for anti-atherosclerosis therapy. This review summarizes the clinical and preclinical researches on the intervention of CAD by regulating the gut microbiota and the microbiota-derived metabolite TMAO, with the aim to provide new target for the therapy of CAD.

Objective To investigate the effect of necrostatin-1 on locomotor recovery after spinal cord injury (SCI) in mice, and to explore the role of apoptosis and M1 type-microglia/macrophage-mediated pro-inflammation in the protective effect. Methods Male C57BL/6 mice were randomly divided into four groups: control group, necrostatin-1 group, SCI model group, necrostatin-1-treated group after SCI, with 20 mice in each. For SCI model group, mice were anesthetized with 10 g/L pentobarbital sodium with a dose of 8 mL/kg. After skin disinfection, T8 laminectomy was performed under operating microscope, and the T8 spinal cord was clearly revealed. The injury model was established with a device designed by our own with the parameter at 0.2 mm-width for 20 seconds. Manual urination was performed once a day. For necrostatin-1-treated group after SCI, 7.8 mg/kg of necrostatin-1 was intravenously administrated at the 1, 2, and 3 days after SCI. For necrostatin-1 group, necrostatin-1 was intravenously injected for three days. Basso Mouse Scale(BMS) score and standardized rump-height index were used to evaluate locomotor function at 1-, 3-, 5-, 7-, 10- and 14-day after injury. To observe cell apoptosis in injured cord, TUNEL staining was performed at 1-, 3-, 7-, and 14-day after injury. Western blot and immunohistochemical staining were performed to detect the expression of inducible nitric oxide synthase (iNOS), a classical marker of M1 type microglia/macrophage. Real time quantitative PCR was used to detect mRNA levels of TNF-α, interleukin-1β (IL-1β), IL-18, IL-4, IL-5, and IL-10. Results Necrostatin-1 significantly promoted the locomotor recovery in mice after SCI, reduced cell apoptosis around the SCI area; decreased the protein expression of M1 type microglia/macrophage marker iNOS and the number of iNOS-positive microglia/macrophage, and down-regulated the transcription levels of pro-inflammatory cytokines TNF-α, IL-18, and IL-1β, while promoting the transcription of anti-inflammatory cytokines IL-4, IL-5, and IL-10. Conclusion Necrostatin-1 significantly promotes locomotor function recovery after SCI in mice by reducing the number of apoptotic cells and inhibiting M1 microglia/macrophages-mediated pro-inflammatory factors.

Objective: To assess the association of 5A/6A polymorphism in the promoter region of MMP3 gene with the stability of extracellular matrix of atherosclerotic plaque., Methods: Clinical data of 776 consecutive patients undergoing percutaneous coronary intervention (PCI) was reviewed. MMP3 gene polymorphisms and serum level of MMP3 for the second admission were collected. The target gene fragment containing MMP3 promoter region was transfected into HepG2 vector cells. The influence of the polymorphism on the expression of the MMP3 gene was determined in vitro., Results: Compared with the first admission data, the proportion of mutant MMP3 genotypes (5A/5A+5A/6A) was significantly higher in patients with acute myocardial infarction (AMI) compared with the control group (37.6% vs. 24.9%, P<0.01). 64.1% of the patients carrying the 5A allele had AMI, whereas only 50.11% of those carrying the 6A allele had AMI (P<0.01). The proportion of wild-type MMP3 genotype (6A/6A) was significantly higher in the stenotic group compared with the non-restenosis group (79.5% vs. 66.5%, P<0.01). Restenosis has occurred in 9.5% of patients harboring the 5A allele compared with 16.2% in those carrying the 6A allele (P<0.01). In addition, serum level of MMP3 in the restenosis group was significantly lower than that of the non-restenosis group (P<0.01). In vitro studies confirmed that the expression of pGL2-Basic/6A was significantly lower than that of pGL2-Basic/5A., Conclusion: The 5A/6A polymorphism in the promoter region of the MMP3 gene may influence its transcriptional activity and impact on the degradation or push-up of extracellular matrix, resulting in a difference in the stability of atherosclerosis plaques, which in turn may induce different pathological processes in AMI or restenosis after stenting.

Numerical simulation is a powerful technology for photoacoustic imaging (PAI) in both theory studies and practical applications. In this paper, a simulation platform for PAI was designed and implemented based on Matlab. The simulation platform utilized finite element method (FEM) and k -space pseudospectral method to calculate the forward and inverse problem of PAI. And a graphical user interface (GUI) was realized. Structural design, work process and other operating details of the platform was also provided. By compared with theoretical temporal waveform of photoacoustic signal and reconstruction results of COMSOL, the validity and reliability was verified. And a reliable simulation tool was proposed for PAI.

Objective To establish a method for primary culture of adult dermal fibroblasts. Methods Fresh skin tissues were digested with dispase II to separate the epidermis from dermis. Then fibroblasts were isolated from the dermis by tissue-adhesive and type I collagenase digestion method respectively. The growth of cells was observed under an inverted phase contrast microscope. the expression of vimentin in cultured cells was detected by immunofluorescence cytochemical staining to identify cell type and assess the purity of fibroblasts. The proliferation activity of fibroblasts was analyzed by MTT assay and drew growth curve. Cell cycle distribution was detected by flow cytometry. Results The fibroblasts were cultured for 48 hours, and more than 95% adherent cells were obtained by collagen protease separation. At the same time, the cells began to grow from the adherent wall at the edge of the tissue block. A large number of fibroblasts adherently grew on day 5. The cells grew rapidly, and on day 7, the layers of proliferating cells were covered in petri dishes. A small number of epithelial cells were mixed in fibroblasts obtained from the tissue-adhesive method, while the vimentin positive rate of cells isolated by collagenase digestion method was nearly 100%, almost all of which were fibroblasts. Conclusion Both collagenase digestion and tissue-adhesive method are rapid, economical and effective methods for obtaining dermal fibroblasts. Collagenase digestion method can prevent epithelial cell contamination more effectively, and the isolated dermal fibroblasts have good proliferative capacity.

Objective: To investigate the predictive value of quick sequential organ failure assessment (qSOFA) score on the prognosis of adult patients with infection in intensive care unit (ICU)., Methods: A retrospective analysis was conducted on the clinical data of the infected patients in the ICU of the 401st Hospital of the People's Liberation Army from August 1st, 2000 to December 31st, 2017. The clinical data included patients' gender, age, basic diseases, etc.; the worst values of vital signs and laboratory test results within 24 hours of admission were recorded, the scores of the qSOFA, sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation II (APACHE II) were calculated separately; the outcome of ICU was recorded. The predictive values of three scoring systems were evaluated by receiver operating characteristic curve (ROC)., Results: Excluding patients with incomplete clinical data, cancer and immunosuppressive patients, a total number of 1 059 patients were enrolled in this study, with 679 males and 380 females, the average age was 72.57±16.06, the ICU mortality was 35.32% (374/1 059). The ROC curve analysis showed that the areas under ROC curve (AUC) of APACHE II, SOFA, qSOFA scores to predict the prognosis of infected patients were 0.713, 0.744 and 0.662, respectively. Although the AUC of qSOFA in predicting prognosis was significantly lower than that of other two scoring systems (both P < 0.05), but it still had some predictive ability. According to the Youden index, the best cut-off point for qSOFA was 2 to evaluate the prognosis of the infection, and the sensitivity was 71.65%, the specificity was 53.87%, the positive likelihood ratio was 1.55, the negative likelihood ratio was 0.53, the positive predictive value was 0.426, the negative predictive value was 0.799, and the accuracy was 59.62%. The mortality of the infected patients was increased with qSOFA score, and the mortality difference among patients with different qSOFA scores was statistically significant (χ 2 = 84.605, P = 0.000). The patients were divided into two groups according to the cut-off value of qSOFA, and the mortality in qSOFA score ≥2 group was higher than that in qSOFA score < 2 group [odds ratio (OR) = 2.767, 95% confidence interval (95%CI) = 2.116-3.617, P = 0.000]., Conclusions: qSOFA, SOFA and APACHE II scores have the capability of predicting the outcome for the infected patients. qSOFA score is expected to be a quick and simple tool to judge the prognosis of ICU infection patients because of its advantages of quick acquisition.

Objective: To observe the impact of improving the compliance of ventilator bundle on morbidity of ventilator-associated pneumonia (VAP) in intensive care unit (ICU) patients undergoing mechanical ventilation (MV) guided by context of Joint Commission International (JCI) settings, and to study the oral care efficacy of suction tube sponge brush., Methods: A prospective study was conducted. The patients who needed MV admitted to Department of Critical Care Medicine of the First Affiliated Hospital of Xiamen University from January 2013 to December 2016 were enrolled. In the context of JCI settings, necessary measurements were taken to enhance the compliance of ventilator bundle each year. In 2013, the preventive measures were set up and the education was strengthened. In 2014, the compliance of hand hygiene and bedside elevation was strengthened. In 2015, a control study was conducted to evaluate the effect between the traditional cotton dipped in chlorhexidine and the suction tube sponge brush rinsed with chlorhexidine on oral health impact parameters. The suction tube sponge brush rinsed with chlorhexidine oral care was introduced to improve compliance. In 2016, electronic bundle checklist for daily self-audits was conducted. The annually morbidity of VAP through the software of hospital and ICU was collected and calculated. The annual incidence of VAP was indicated by the VAP cases per 1 000 MV days. Based on the VAP incidence rate in 2013 as 1, the VAP incidence-rate ratio (IRR) of each year was calculated., Results: During the study period, a total of 2 733 patients admitted to the ICU, including 1 403 patients undergoing MV. Ninety-four of the 1 403 patients with community-acquired pneumonia (CAP), aspiration pneumonia, back elevation ban, incomplete information, and withdrew from the study were excluded. 1 399 patients undergoing MV were enrolled in the final analysis, with total MV days of 11 012 days, and 94 patients occurred VAP. The annual incidence of VAP was progressively declined from 2013 to 2016, and the VAP cases per 1 000 MV days were 17.0, 10.0, 5.9, 3.5 cases, respectively. Based on the VAP incidence rate in 2013, the IRR of VAP from 2014 to 2016 was also progressively declined, which was 0.59 [95% confidence interval (95%CI) = 0.35-0.98], 0.35 (95%CI = 0.18-0.64), and 0.21 (95%CI = 0.09-0.41), with statistical significance (all P < 0.05). In 2013, ICU patients had the lowest rates of bedside elevation and hand hygiene compliance, which were 28.57% and 54.29%, respectively. Compared with 2013, by the implementation of two quality control circle (QCC) projects for bedside elevation and hand hygiene, the rates of bedside elevation and hand hygiene compliance were improved significantly in 2014, which were 82.35%, 91.18%, respectively (both P < 0.05). In 2015, the compliance of chlorhexidine oral care which was the worst performed in 2014 had been improved by the method of QCC, and the rate of the compliance was significantly higher than that in 2013 (87.10% vs. 62.86%, P < 0.05). Compared with 2013, bundle compliance was significantly increased in 2016, except for the sterile operation of the suction tube [daily wake and weaning: 95.00% vs. 71.43%, bedside elevation for over 30degree angle: 92.50% vs. 28.57%, hand hygiene: 97.50% vs. 54.29%, chlorhexidine mouth care once per 6-8 hours: 95.00% vs. 62.86%, turned back and posture drainage: 97.50% vs. 80.00%], the differences were statistically significant (all P < 0.05). The incidences of bad breath, dirt residue and plaque were significantly lower in the group of oral care by using suction tube sponge brush with chlorhexidine (30 cases) compared with the group of traditional cotton pad with chlorhexidine (30 cases; bad breath: 10.0% vs. 40.0% %, dirt residue: 16.7% vs. 70.0%, plaque: 3.3% vs. 30.0%, all P < 0.05). There was no significant difference in the incidence of oral ulcers between the oral brush group and the traditional group (10.0% vs. 30.0%, P > 0.05)., Conclusions: Ventilator bundle can effectively reduce the morbidity of VAP in the context of JCI settings, and the oral care by using suction tube sponge brush and chlorhexidine can effectively improve oral hygiene.

Objective: To explore the effect of VNTR-ZNF and -14C/T variants of the promoter region of the ABCA1 gene on the transcription activity of genes in vitro., Methods: The recombinants were constructed by ligating DNA fragment containing VNTR-ZNF ACCCC inserted/deleted allele with or without -14C/T substitution fragments with a PGL2-basic vector containing luciferase reporter gene. The recombinants were then transfected into HepG2 cells using the cationic lipid method. After 48 h, transfected cells were collected and used to detect the luciferase activity., Results: Luciferase activity of PGL2-ZNF-ACCCCDel was greater than that of PGL2-ZNF-ACCCCIns. Luciferase activity of PGL2-ZNFDel-14C was greater than that of PGL2-ZNFDel-14T, PGL2-ZNFIns-14C, PGL2-ZNFIns-14T., Conclusion: Compared with the insertion type, the ACCCC-deleted type of VNTR-ZNF can significantly enhance the transcription activity of ABCA1. And co-transfection of -14 C allele can further enhance this activity.

Objective: The performances of Lactobacillus acidophilus NCFM, Lactobacillus plantarum121 and Lactobacillus pentosus ML32 in removing benzo(a)pyrene [B(a)P] in simulated starch conditions were studied. To provide a novel way to reduce the potential risk of B(a)P to human, several factors which affect the binding of the 3 Lactobacillus strains to the chemical toxin were investigated in starch system., Methods: The percentage of B(a)P bound by Lactobacillus strains was determined with HPLC after incubation at 37℃ for 4 h. The B(a)P-removing capabilities of Lactobacillus strains in various simulated starch systems were evaluated, including the concentrations, types and gelatinization of starch, incubation time, pH and starch-hydrolyzed products., Results: The B(a)P-binding percentages of the Lactobacillus strains increased as starch concentrations were elevated from 2% to 10%. Starch types, either from corn, potato or sweet potato, had little effects on the ability of these bacterial cells to bind B(a)P. The gelatinization of starch favored Lactobacillus strains to bind more B(a)P. The percentage of these bacterial cells to bind B(a)P grew fast at 37℃ for the first 4 h-incubation time, and then slowly increased. The three Lactobacillus strains bound more B(a)P when starches were acidified to pH of 3 to 4 or alkalified to pH of 8 to 9. The viable cells of three Lactobacillus strains removed more B(a)P via their binding than their dead cells did. Moreover, both glucose and maltose, the end products after starch is hydrolyzed, improved significantly the performance of the 3 bacterial strains to remove B(a)P., Conclusion: All of the three Lactobacillus strains perform good ability to bind B(a)P in the presences of starch. Their B(a)P-removing ability would be improved with an increased starch concentration, and the gelatinization of starch as well as the supplements of glucose and maltose. Thus, the selected Lactobacillus strains in our current work should be promising as a biological agent to reduce the occurrence of B(a)P in starch-based food products.

Objective: To investigate the single nucleotide polymorphism (SNP) of toxin-antitoxin-chaperone (TAC) system of Mycobacterium (M.) tuberculosis with different genotypes and its biological significance., Methods: A total of 183 clinical M. tuberculosis isolates were collected for spoligotyping. The sequences of higA,higB and Rv1957 were obtained by using PCR and DNA sequencing. The sequences were compared for possible mutations. Functional consequences of nonsynonymous SNPs were predicted by using I-Mutant 2.0 servers., Results: Among the 183 M. tuberculosis isolates, 138(75.41%) belonged to the Beijing family, while 45(24.59%) belonged to the non-Beijing family. A total of 149(81.42%) isolates showed polymorphisms in the TAC system. We discovered 6 nonsynonymous SNPs and 2 synonymous SNPs. All the synonymous mutations occurred in higA gene, while nonsynonymous SNPs were found in the higA,higB and Rv1957 genes either. All the synonymous mutations and 4 nonsynonymous SNPs were restricted to the Beijing family strains and only 2 nonsynonymous SNPs were observed in the non-Beijing family strains. Of the 6 nonsynonymous SNPs studied, 4 were predicted to have ability to affect the stability of respective protein., Conclusion: The SNPs in the coding sequences of TAC system in clinical isolates can be relatively high and the Beijing family strains are with higher polymorphism, which might benefit to adapt to different host environment.

Objective: To investigate the therapeutic effect of Ad-hVEGF165 on the endothelial cells dysfunction induced by homocysteine (Hcy) and related molecular mechanisms., Methods: Human umbilical vein endothelial cells CRL-1730 were treated with Hcy at different concentrations (0, 0.05, 1.00 mmol/L) for 24 h. The same concentration of Hcy, Ad-Track and Ad-hVEGF165 were added to the cells in the following groups: blank group, Hcy0.05 group, Hcy1.00 group, Ad-Track group, Hcy0.05+Ad-Track group, Hcy1.00+Ad-Track group, Ad-hVEGF165 group, Hcy0.05+Ad-hVEGF165 group, Hcy1.00+ Ad-hVEGF165 group for 48 h. The mRNA and protein expressions of eNOS and DDAH2 were detected by real-time PCR and Western blot. The correlations of mRNA and protein expressions between endothelial nitric oxide synthase (eNOS) and dimethylarginine dimthylaminohydrolase (DDAH)2 were evaluated by Pearson correlation analysis., Results: Compared with blank group and Ad-hVEGF165 group, the mRNA and protein expressions of eNOS were decreased in Hcy0.05 group and Hcy0.05+Ad-hVEGF165 group (both P < 0.05), and the mRNA and protein expressions of DDAH2 in cells treated with 0.05 mmol/L and 1.00 mmol/L Hcy were reduced as well (all P < 0.05). DDAH2 mRNA and protein expression are increased (all P < 0.05) in Ad-hVEGF165 group compared with the blank group and Ad-Track, Hcy0.05 + Ad-hVEGF165 and Hcy0.05 group compared with Hcy0.05+Ad-Track group, Hcy1.00+Ad-hVEGF165 and Hcy1.00 group compared with Hcy1.00+Ad-Track group. The mRNA and protein expressions of eNOS and DDAH2 were uncorrelated under the effect of Hcy (r = 0.057 and 0.449, both P > 0.05) and VEGF (r = 0.284 and 0.432, both P > 0.05)., Conclusion: Recombinant adenovirus Ad-hVEGF165 could reverse Hcy-induced endothelial cells dysfunction via upregulating the expressions of eNOS and DDAH2.

Objective: To investigate the in vitro effects of immune inhibitor tacrolimus on platelet function., Methods: Fresh venous blood was collected from healthy volunteers at ages of 18-25 years old, who are not taking antiplatelet drugs within two weeks. The platelets were isolated from the blood and incubated with different concentrations of tacrolimus (0.06, 0.6, 6, 60, 120, 240 μmol/L) at 37 °C for 2 hours, and then the changes of mitochondrial membrane potential and P-selection of platelets were detected by flow cytometry, the expression of apoptosis related protein by Western Blot, and the change of the platelet aggregation function by platelet aggregation analyzer., Results: Tacrolimus at concentration of 0.06 μmol/L could promote collagen induced platelet aggregation, inhibit thrombin induced platelet aggregation, have no effect on ristocetin and vWF induced platelet aggregation function. Tacrolimus at concentration of 120 μmol/L and 240 μmol/L could reduce the platelet mitochondrial membrane potential and induce the expression of apoptosis protein caspase-3., Conclusion: In vitro experimental results showed that high concentration of tacrolimus could lead to platelet apoptosis. But the current therapeutic dose of tacrolimus at 0.06 μmol/L (which is equivalent to 50 ng/ml blood concentration) could have different effects on platelet aggregation function according to different stimulating agents.

Objective: To study the effect of reactive oxygen species (ROS) on the regulation of platelet apoptosis., Methods: Washed healthy human platelets were pre-incubated with N-caetyl-Lcysteine (NAC), and then stimulated with dibucaine or thrombin. The production of ROS and depolarization of mitochondrial membrane potential (∆ ψm) were detected by flow cytometry. The activation of caspase-3 and expression of Bcl-xL were analyzed by Western blot., Results: (1)The average ROS fluorescence value of NAC+dibucaine group was lower than that of dibucaine group(0.66 ± 0.11 vs 1.06 ± 0.08, P<0.01), while that of NAC+thrombin group was also lower than that of thrombin group(0.45 ± 0.05 vs 0.71 ± 0.11, P=0.001). (2)The percentage of platelets with normal ∆ψm in NAC+Dibucaine group was higher than that of dibucaine group[(86.30 ± 9.37)% vs (13.52 ± 3.01)%, P=0.000], while that of NAC+thrombin group was also higher than that of thrombin group[(93.00 ± 3.03)% vs (76.58 ± 5.28)%, P=0.000]. (3)Fragmentation generated by caspase-3 activation in dibucaine group was much more than that in DMSO control group, while the fragmentation in NAC+dibucaine group was significantly decreased. (4)The expression of anti-apoptosis protein Bcl-xL of NAC+dibucaine group was significantly higher than that of the dibucaine group, while that of NAC+thrombin group was also higher than that of thrombin group., Conclusion: Through the regulation of ROS, NAC could inhibit the platelet apoptosis induced by dibucaine or thrombin.

The effects of AM fungi on the growth, nutrient uptake and microelement of Scutellaria baicalensis under different applied phosphorous levels in the pot culture were studied. The results showed that applied phosphorous levels had remarkable effect on AM fungal inoculation. Inoculation effect of AM fungi was the best under 0.12-0.18 g x kg(-1) soil. At those levels, the effect of AM fungi on the growth of S. baicalensis was the best, but either lower or higher phosphorous level showed suppressed infecting of AM fungi to S. baicalensis. After inoculation, the contents of biomass, total P, baicalin, total N of root were increased as well as the microelements of K, Ca, Mg, Fe, Zn, but the contents of total N of shoot, N/P were decreased as well as the microelements of Mn, Cu. In conclusion, the growth of host plant and using rate of phosphorus fertilizer can be promoted by inoculation of AM fungi.

Model-based system analysis is an important tool for evaluating the potential and impacts of biofuels, and for drafting biofuels technology roadmaps and targets. The broad reach of the biofuels supply chain requires that biofuels system analyses span a range of disciplines, including agriculture/forestry, energy, economics, and the environment. Here we reviewed various models developed for or applied to modeling biofuels, and presented a critical analysis of Agriculture/Forestry System Models, Energy System Models, Integrated Assessment Models, Micro-level Cost, Energy and Emission Calculation Models, and Specific Macro-level Biofuel Models. We focused on the models' strengths, weaknesses, and applicability, facilitating the selection of a suitable type of model for specific issues. Such an analysis was a prerequisite for future biofuels system modeling, and represented a valuable resource for researchers and policy makers.

Estrogen-related receptor a (ERRalpha) is a key regulator for energy metabolism and adipogenesis. However, its role in lipolysis is unknown. To study the function of ERRalpha in lipolysis, primary cultured differentiated porcine adipocytes were treated by a specific inverse agonist XCT790 or infected with adenoviral vector expressed ERRalpha for 48 h, in the absence and/or presence of specific protein kinase A (PKA) inhibitor or extracellular signal-related kinase (ERK) inhibitor. Then, we measured the triglyceride (TG) content and the glycerol release into the culture media to analysis the effect of ERRalpha on lipolysis; Further, we analyzed the expression of PPARgamma, perilipin A, p-perilipin A, HSL and ATGL with Western blotting. Here, we found that ERRalpha significantly increased adipocytes differentiation, TG accumulation and glycerol release. Separately or simultaneously block the PKA and ERK pathway do not significantly altered the effect of ERRalpha on glycerol release. ERRalpha significantly up-regulated the proteins expression of PPARgamma, perilipin A, HSL and ATGL, while the p-perilipin A protein level was not significantly changed. These findings imply that ERRalpha could increase lipolysis via up-regulating HSL and ATGL, thereby to supply more FFA as substrate for a larger turnover of cellular triglyceride pool during adipocytes differentiation.

To produce many epitopes of peptide mut, different tandem repeats containing HER-2 mimetic peptide mut were constructed and expressed. The oligonucleotide coding mut sequence was inserted into pET28a(+) to construct the prokaryotic recombinant plasmid pET28a-mutl. The mut coding sequence was repeatedly inserted into the vector in tandem to obtain a series of plasmids pET28a-mut2, pET28a-mut4, pET28a-mut8 and pET28a-mut16. The plasmids were transformed into E. coli. BL21 and induced by IPTG. The recombinant proteins were expressed in E. coli. The binding analysis verified that the interaction between the tandem peptides mut and Herceptin was increased. In conclusion, the tandem peptides consisted of many antigen epitopes, and laid the foundation for the further study of HER-2 peptide vaccine for biotherapy of cancer with HER-2 overexpression.

Hsc70-SNCG fusion protein cDNA fragment containing signal peptide sequence of Igkappa, MMP9, and P37 was inserted into the vector pVAX1 to construct recombinant plasmid pVAX-Igkappa-Hsc70-SNCG, pVAX-MMP9-Hsc70-SNCG, and pVAX-P37-Hsc70-SNCG. Three eukaryotic vectors were constructed and verified by restriction enzyme digestion and sequencing. After transfection with recombination plasmids in QM-7 cells, the transient expression and secretion of three fusion proteins were detected by ELISA and Western Blot. The results suggested that Hsc70-SNCG carrying three different signal peptides could be expressed and secreted by transfected cells, and three signal peptides effectively directed secretion of fusion protein by QM-7 cells. BALB/c mice were immunized by three plasmids using gene gun system. The serum levels of anti-SNCG antibodies in mice were measured by ELISA. The results showed that three secreted plasmids could stimulate humoral immune responses to SNCG in mice, which depended on the secreted expression levels induced by signal peptides.

Lactoferrin in milk is a multifunctional protein. In addition, lactoferrin has antiviral, antifungal and antiparasitic activity. In this study, the N-terminus from porcine lactoferrin (PLF-N) was designed to express the antimicrobial action of recombinant porcine lactoferrin. We cloned a 1077 bp fragment of the PLF gene from mammary gland tissue of the lactating sow at the third day. Comparing nucleotide sequence with four strains of PLF gene published on GenBank, the homology was more than 99%. With the reference template of the cloned fragment of PLF-N and optimizing codon bias, we synthesized the gene of N-terminus encoding porcine lactoferrin (PLF-NS). The high expression gene of PLF-NS was cloned into the fusion expression vector pET30b and expressed in E. coli BL21 (DE3). After induced with Isopropyl beta-D-1-Thiogalactopyranoside (IPTG), the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. The protein had a molecular weight of 42 kDa and accounted for 32% of the total cellular protein. After purification and renaturation, the purity of the expressed protein was 98%. The expressed PLF-NS protein showed obviously antibacterial activity. This method provides an excellent way for high expression of antimicrobial proteins when optimizing codon bias.