Quorum sensing systems of pathogens are central regulators for the expression of virulence factors. Increasing evidence implies that targeting the quorum sensing system of many pathogenic bacteria is a promising therapeutic approach to control infections. In this work,we isolated 47 strains of actinomycetes from the mud sample of Jiaozhou Bay. Quorum sensing inhibitory activity was monitored by Chromobacterium violaceum CV026. As a result,the culture broth extract of actinomycetes WA-7 was found to have significant quorum sensing inhibitory activity. This strain was assigned to the genus Streptomyces based on its 16S rDNA sequence. Further investigation revealed that the extract could inhibit the quorum sensing-controlled violacein and proteases production of C. violaceum in a concentration-dependent manner.

Objective To explore the clinical significance of serum ProGRP in diagnosis of SCLC.Methods Serum ProGRP were detected in twenty-three SCLC patients,forty NSCLC patients and forty-three patients with benign pulmonary disease whose were definite diagnosis collected from April 15,2016 to July 19,2016,and a contemporary cohort of 40 healthy controls were while recruited.Results The levels of ProGRP in SCLC group were significantly higher than those in NSCLC group,benign pulmonary disease group and healthy control group(P <0.05).The levels of ProGRP in extensive disease group were significantly higher than those in 1imited disease group(P <0.05).There was no significant difference in the positive rates of ProGRP between the sex group and age group (P > 0.05 ).The boundary value of diagnosis of SCLC through ProGRP identified through ROC curve was 65.66 ng/L.The sensitivity and specificity of the ProGRP for the diagnosis of SCLC were respectively 90.9% and 89.9%.Conclu-sion Serum ProGRP can be use as a sensitive and specific index for diagnosis of SCLC,and the level of ProGRP can also be used for the clinical staging of SCLC.

Animals ; Diet ; Fatty Acids, Monounsaturated ; administration & dosage ; Insulin Resistance ; Physical Conditioning, Animal ; Rats

OBJECTIVETo evaluate the role of serum tumor necrosis factor-alpha (TNF-alpha) and adiponectin on insulin resistance in different fat diet rats.

METHODSThirty weanling female rats were randomly divided into 3 group (n = 10): a low-fat soybean oil (LFS; 22% of total energy fed as fat), high-fat soybean oil (HFS; 40% of total energy fed as fat), or high-fat soybean oil and swimming training at the same time (HFS + T). After fed for 10 weeks, the level of TNF-alpha, adiponectin in serum of rats were observed.

RESULTS(1) The body weight, percentage of body fat of HFS group increased compared with that of LFS group (P < 0.05), however those of HFS + T group were decreased (P < 0.05). (2) The level of serum insulin and ISI in HFS group were increased by LFS group (P < 0.05), in HFS+ T group the levels decreased. (3) And the serum level of TNF-alpha and IL-6 in HFS group were higher than those in LFS group (P < 0.05), the serum levels of adiponectin in HFS group were lower than those in LFS rats, and in HFS+ T group the levels of TNF-alpha and IL-6 were lower than those in HFS group (P < 0.05), the adiponectin level was higher than that in HFS group, and there were no significant difference between LFS group and HFS + T group.

CONCLUSIONExercises training could improve sugar and fat metabolism disorders, which also contributes to improving insulin resistance caused by high-fat diet.

Adiponectin ; blood ; Animals ; Diet, High-Fat ; adverse effects ; Dietary Fats ; administration & dosage ; Female ; Insulin ; blood ; Interleukin-6 ; blood ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Alphavirus ; drug effects ; genetics ; metabolism ; Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; High-Throughput Screening Assays ; methods ; Luciferases ; genetics ; metabolism

BACKGROUND:Hip fracture has become one of the main reasons for morbidity, death and heavy medical costs in the elderly. With aging, the incidence of hip fractures in the elderly is increased day by day. OBJECTIVE:To investigate and analyze the incidence and distribution of hip fractures in middle-aged and old population from Changning District of Shanghai, China. METHODS:Medical records of patients with hip fracture ICD-9 CODE (femoral neck fracture and intertrochanteric fracture) admitted from July 2008 to June 2013 were col ected at the Changning District Public Health Information Center and Shanghai Shen Kang Medical Al iance Center. Population distributions of Changning District in 2010 and 2011 were provided by the Disease Control Center of Changning District. Then, the crude morbidity of hip fracture was calculated in terms of different ages and genders. For patients aged over 50 years, every 5 years served as an age group. RESULTS AND CONCLUSION:The incidence rate of hip fractures in the population aged over 50 years was 197.04/100 000, 88.51/100 000 in male and 304.73/100 000 in female. The annual standardized incidence rate of hip fracture was higher in female than male (P<0.05). The crude incidence rate of hip fracture was increased dramatical y in the population over 70 years, especial y in females. It indicates that the incidence rate of hip fractures is closed related to age, gender and osteoporosis.

Objective To establish a zebrafish IGFBP-2 gene knock-down model by morphilino modified antisense oligonucleotide injection, so as to investigate the abnormal phenotypes of heart and vessels in early stage of zebrafish development and the expression of zebrafish cardiogenesis related genes. Methods The spatiotemporal expression of IGFBP-2 gene in early stage of zebrafish development was testified by whole mount in situ hybridization with antisense RNA IGFBP-2 probe. The IGFBP-2 morpholino (IGFBP-2 MO) that especially inhibited the gene promoter and standard control morpholino (Con-MO) were designed and synthesized by Gene-tools Corporation. Four different concentration gradients (0.05, 0.10, 0.25 and 1.0 mmol/L) were set as IGFBP-MO injection groups with 0.25 mmol/L Con-MO injection group and wild type group as controls. Contribution to the incidence of heart abnormal phenotypes and mortality rate induced by 4 different IGFBP-2 concentrations injection group was recorded and compared with 2 control groups. Heart abnormal phenotypes at different developmental stages in 0.25 mmol/L IGFBP-2 injection group were observed in detail. To validate the effectiveness of IGFBP-2 MO, the expression of enhanced green fluorescence presented by wild type zebrafish embryos at 12hpf which received single injection of IGFBP-2 EGFP recombinant plasmid and those co-injected with Con-MO or IGFBP-2 MO were detected. To investigate the regulation relationship between IGFBP-2 gene and other cardiogenesis related genes, expression of atrium specific marker gene Amhc was detected in IGFBP-2 MO and wild type group by in situ hybridization. Ventricle specific green fluorescence of Vmhc-EGFP transgenic zebrafish embryos whose IGFBP-2 gene was knocked-down were compared with those untreated. Zebrafish peripheral vascular development in the IGFBP-2 MO group was also checked out by micro-angiography. Results Whole mount in situ hybridization revealed that IGFBP-2 gene expressed in turn at eyes, midbrain and then focused on liver in early stage of zebrafish development. The micro-injection of 0.25 mmol/L IGFBP-2 MO resulted in heart malformation in nearly 60% of all injected zebrafish embryos. Heart malformation phenotypes included slow heart beat, pericardial edema, weak ventricle systole contraction and heart tube looping disorder. Some of them represented atria dilation, blood regurgitation and ciculation obstruction. Wild type zebrafish embryos that received single injection of IGFBP-2 EGFP plasmid DNA or co-injected with Con-MO presented strong enhanced green fluorescence at 12hpf, meanwhile, the fluorescence was barely seen in the embryos co-injected with IGFBP-2 MO. This strongly validated the gene specific knock-down effect of IGFBP-2 MO. Amhc was down-regulated at 48hpf in IGFBP-2 MO group. Vmhc-EGFP transgenic zebrafish down-regulated by IGFBP-2 gene also resulted in attenuated expression of ventriclar-specific green fluorescence protein at 48hpf. Intersegmental blood vessels of IGFBP-2 MO group by micro-angiography at 60hpf demonstrated an sparsate and chaos image, which suggested that IGFBP-2 gene expression was involved in the regulation of normal vascular development. Conclusions Micro-injection of IGFBP-2 MO is an efficient way to knock-down IGFBP-2 gene in zebrafish embryos. IGFBP-2 gene expression down-regulation leads to heart and vessels maldevelopment and have an impact on the expression of cardiogenesis related genes of zebrafish embryos as well. In short, IGFBP-2 plays a critical role in the normal cardiovascular development of zebrafish embryos.

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.

Objective To investigate the anti-apoptotic effect of gene A20 in treatment of trau-matic brain injury (TBI). Methods Thirty-five Sprague-Dawley rats were made severe TBI models and assigned randomly to experimental group and control group (35 rats in each group). After severe TBI, the rats in experimental group were injected with liposome-pcDNA3.1-A20 and those in control group injected with liposome pcDNA3.1-A20 at 30 minutes after severe TBI. The animals in both groups were sacrificed to remove the brain of five rats from each group at 12, 24, 48, 72 and 168 hours for sec-tioning. The expression of A20 and neurocyte apoptosis were defined by immunohistological method and TUNEL accordingly. The other ten rats were testified for neurological function at 1,2, 3 and 4 weeks af-ter TBI. Results The expression of A20 in experimental group was higher than that in control group, with statistical differences (P < 0. 01). The peak neurocyte apoptosis was found at 72 hours after TBI. The number of apoptosis cells in experimental group was lower than that in control group at 12, 24, 48 and 72 hours afte TBI (P < 0.01 or 0.05). At the 4th week after TBI, the neurological function in exper-imental group was better than that in control group (P < 0.05). Conclusion Gene therapy with A20 may have anti-apoptosis effect and exert neuroprotective effect on severe TBI.