Ensemble classification for predicting the malignancy level of pulmonary nodules on chest computed tomography images.

Early identification and classification of pulmonary nodules are essential for improving the survival rates of individuals with lung cancer and are considered to be key requirements for computer-assisted diagnosis. To address this topic, the present study proposed a method for predicting the malignant phenotype of pulmonary nodules based on weighted voting rules. This method used the pulmonary nodule regions of interest as the input data and extracted the features of the pulmonary nodules using the Denoising Auto Encoder, ResNet-18. Moreover, the software also modifies texture and shape features to assess the malignant phenotype of the pulmonary nodules. Based on their classification accuracy (Acc), the different classifiers were assigned to different weights. Finally, an integrated classifier was obtained to score the malignant phenotype of the pulmonary nodules. The present study included training and testing experiments conducted by extracting the corresponding lung nodule image data from the Lung Image Database Consortium-Image Database Resource Initiative. The results of the present study indicated a final classification Acc of 93.10±2.4%, demonstrating the feasibility and effectiveness of the proposed method. This method includes the powerful feature extraction ability of deep learning combined with the ability to use traditional features in image representation.

Baicalin suppress growth and virulence-related factors of methicillin-resistant Staphylococcus aureus in vitro and vivo.

A Staphylococcus aureus (S.aureus) was isolated from pigs suffered in pneumonia that can't be cured by antibiotic such as methicillin and vancomycin. It was demonstrated that baicalin, an active natural compound extracted from the traditional Chinese medicinal, possess antimicrobial activity. In the present study, we evaluate it efficacy in vitro and vivo against this isolated methicillin-resistant S.aureus (MRSA). Our findings demonstrated that baicalin can inhibit S. aureus growth in a dose-dependent manner and attenuate the biofilm formation. Scanning electron microscopies showed that cell membrane was damaged and accompany with contents leaks after treated with high concentration of baicalin. In addition, baicalin exerted inhibitory effects on the expression of S.aureus virulence-related factors. Moreover, baicalin treated mice had enhanced survival after a lethal dose of S.aureus infection compared with untreated mice. Simultaneously, the pathological tissue damage and bacterium burden were decrease in baicalin treated mice. These data demonstrated that baicalin displayed a high effectiveness in vitro and vivo against MRSA infection, suggesting that baicalin may potentially be used to treat MRSA infection.

New Lignans from roots of Kadsura coccinea.

Four new dibenzocyclooctadiene lignans, named heilaohusus A-D (1-4), one new arylnaphthalene lignan named heilaohusu E (5), and seven known analogues (6-12) were isolated from the roots of Kadsura coccinea. Their structures and configurations were elucidated by a combination of HR-ESI-MS, 1H NMR, 13C NMR, HSQC, HMBC, NOESY and CD spectra. Among the known compounds, compounds 6 and 8-12 were isolated from this plant for the first time. All of compounds were evaluated for their cytotoxicity activities, compounds 3, 6 and 7 showed weak cytotoxicity against four human cancer cell lines (HepG-2, HCT-116, BGC-823 and Hela) with IC50 values range from 13.04 to 21.93 µM. Compounds 1 and 7 demonstrated potential anti-RA (rheumatoid arthritis) activity against RA-FLS cell line with IC50 values of 14.57 and 11.70 µM, respectively.

Influenza A virus infection induces liver injury in mice.

Respiratory infections such as SARS-CoV in humans are often accompanied by mild and self-limiting hepatitis. As a respiratory disease, influenza A virus (IAV) infection can lead to hepatitis, but the mechanism remains unclear. This study aimed to investigate the occurrence of hepatitis by establishing a model for infected mice for three different subtypes of respiratory IAVs (H1N1, H5N1, and H7N2). Histological analysis was performed, and results showed increase serum aminotransferase (ALT and AST) levels and evident liver injury on days 3 and 7, especially on day 5 post infection. Immunohistochemistry (IHC) results indicated a wide distribution of IAV's positive signals in the liver of infected mice. Real-time PCR results further revealed a similar viral titer to IHC that presented a remarkedly positive correlation with histology injury. All these data showed that the mouse model suitably contributed valuable information about the mechanism underlying the occurrence of hepatitis induced by respiratory influenza virus.

MicroRNA-134 plasma levels before and after treatment with valproic acid for epilepsy patients.

BACKGROUND: Temporal lobe epilepsy is the second most common neurological disorders characterized by recurrent spontaneous seizures. MicroRNAs play a vital role in regulating synaptic plasticity, brain development and post-transcriptional expression of proteins. In both animal models of epilepsy and human patients, miR-134, a brain-specific microRNA has recently been identified as a potential regulator of epileptogenesis. METHODS: microRNA identified as targets for the actions of valproic acid (VPA) are known to have important effects in brain function. In this study, 59 new-onset epilepsy patients and 20 controls matched by sex and age were enrolled. Patients with a score < 3 were allocated into the mild group, 3-5 into the moderate group and >5 into the severe group. The plasma miRNA-134 level was quantitatively measured using real-time PCR. RESULTS: Plasma miRNA-134 level in new-onset epilepsy patients was significantly up-regulated when compared with that in healthy controls, and then considerably down-regulated after oral intake of valproic acid medication. The up-regulated plasma miRNA-134 levels may be directly associated with the pathophysiology and severity of epilepsy. CONCLUSION: Plasma miRNA-134 in epilepsy may be considered as a potential peripheral biomarker that responds to the incidence of epilepsy and associates with use of anti-epilepsy drugs.

DNAzymes Dz13 target the c-jun possess antiviral activity against influenza A viruses.

The emergence of anti-influenza A virus drugs resistant strain highlights the need for more effective therapy. Our earlier study demonstrated that c-jun, a downstream molecule of JNK, might be important in viral infections and inflammatory responses. In the present study, we explored the function of DNAzymes Dz13 that target c-jun in influenza A virus infected mice. Dz13 displayed non-toxic side effects on A549 cells and BALB/c mice. Moreover, Dz13-treated mice had enhanced survival after influenza compared with untreated mice. Simultaneously, the pulmonary inflammatory responses and viral burden were decreased in Dz13 treated mice. Furthermore, proliferation levels of infection-induced CD4+ and CD8+ T cells were impaired. These data demonstrated that Dz13 could reduce viral replication and inflammatory response in vivo, suggesting that Dz13 may potentially be used to treat influenza A viral infection.

First sporadic case of pathogenic Escherichia coli infection in Black swan in China.

A strain of bacteria was isolated from the diseased black swan (Cygnus atratus) died from enteritis diarrhea, and designated tentatively as B-1 strain. Morphological and biochemical tests, as well as phylogenetic analysis derived from 16S rRNA and fimC gene sequencing both strongly indicated that B-1 strain is identical to Escherichia coli. Furthermore, the polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) profile of the isolate was different from that of two reference strains. Antibiotic sensitivity testing of B-1 strain was carried out by the standard Kirby-Bauer disc diffusion method. Animal experiments demonstrated that B-1 strain is pathogenic to mice and chickens. This is first sporadic case of pathogenic E. coli infection in Black swan in China.

Development of an indirect competitive ELISA for simultaneous detection of enrofloxacin and ciprofloxacin.

Modified 1-ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method was employed to synthesize the artificial antigen of enrofloxacin (ENR), and New Zealand rabbits were used to produce anti-ENR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (ELISA) standard curve was established. This assay was sensitive and had a linear range from 0.6 to 148.0 µg/kg (R(2) = 0.9567), with the half maximal inhibitory concentration (IC(50)) and limit of detection (LOD) values of 9.4 µg/kg and 0.2 µg/kg, respectively. Of all the competitive analogues, the produced pAb exhibited a high cross-reactivity to ciprofloxacin (CIP) (87%), the main metabolite of ENR in tissues. After optimization, the matrix effects can be ignored using a 10-fold dilution in beef and 20-fold dilution in pork. The overall recoveries and coefficients of variation (CVs) were in the ranges of 86%-109% and 6.8%-13.1%, respectively. It can be concluded that the established ELISA method is suitable for simultaneous detection of ENR and CIP in animal tissues.

Monoclonal antibody-based ELISA and colloidal gold immunoassay for detecting 19-nortestosterone residue in animal tissues.

This article presents the generation of monoclonal antibodies (mAbs) with high specificity against 19-nortestosterone (NT) through cell fusion techniques and the development of a mAb-based indirect competitive ELISA (icELISA) method and colloidal gold-based immuno-chromatographic assay to detect NT residues in beef and pork samples. A modified carbodiimide method was employed to synthesize the artificial antigen, and BALB/c mice were used to produce anti-NT mAbs. On the basis of the checkerboard titration, an indirect competitive ELISA standard curve was established. This assay was sensitive and had a linear range from 0.03 to 38 ng/mL in phosphate buffered saline (PBS), with IC(50) and LOD values of 0.52 ng/mL and 0.01 ng/mL, respectively. Of all the competitive analogues, the produced mAb exhibited a high cross-reactivity to 17α-nortestosterone (83.6%), the main metabolite of NT in animal tissues. Except for moderate cross-reactivities with trenbolone (22.6%) and ß-boldenone (13.8%), the other interference to the assay was negligible (<0.05%). In contrast, the strip test had a visual detection limit of 1 ng/mL in PBS, 2 µg/kg in beef, and 2 µg/kg in pork, respectively, and the results can be judged within 10 min. The ELISA and GC-MS results showed close correlation in beef (R2=0.9945) and in pork (R2=0.9977). Therefore, the combination of two immunoassays provides a useful screening method for quantitative or qualitative detection of NT residues in animal-origin products.

Embryonic and developmental toxicity of the ionic liquid 1-methyl-3-octylimidazolium bromide on goldfish.

The embryonic developmental toxicity of the ionic liquid (IL) 1-methyl-3-octylimidazolium bromide ([C(8)mim]Br) on the goldfish Carassius auratus was evaluated in this study. First, the 72 h 50% lethal concentrations (72 h-LC(50)) for [C(8)mim]Br in goldfish embryos at the stages of cleavage, early gastrula, closure of blastopore, and heart beating were determined by preliminary acute toxicity tests. After that, fish embryos in different developmental stages (cleavage, early gastrula, closure of blastopore, and heart beating) were exposed to 10.4, 20.8, 41.6, and 104 mg/L of [C(8)mim]Br until their hatching stage. The results of the acute toxicity tests showed that 72 h-LC(50) values at the early cleavage, early gastrula, closure of blastopore, and heart beating stages of development were 208.96, 187.1, 245.03, and 298.33 mg/L, respectively. In the subchronic tests, [C(8)mim]Br exposure prolonged the duration of embryo dechorionation and decreased the hatching rates of the treated embryos compared to control embryos. In addition, [C(8)mim]Br treatment also caused remarkable increases of embryonic malformation and mortality ratio in most treatment groups. Finally, we also found that the embryonic and developmental toxicity of [C(8)mim]Br on fish embryos was dose-response and developmental stage-specific. These results indicate that [C(8)mim]Br has toxic effects on the early embryonic development of goldfish, and the risk to aquatic ecosystem by ILs leaking into the water body must be evaluated in the future.

The toxic effects of ionic liquids on the activities of acetylcholinesterase and cellulase in earthworms.

The earthworm Eisenia foetida was exposed to different concentrations of imidazolium ionic liquids with varying chain lengths according to the method of OECD [OECD, 1984. (The Current Organization of Economic and Cooperative Development Acute Earthworm Toxicity Test) Guidelines for the Testing of Chemicals, No. 207. Earthworm Acute Toxicity Tests]. The acute and subchronic toxic effects of [C(8)mim]Br on the activities of acetylcholinesterase (AChE) and cellulase in earthworms were determined under an artificial soil condition. Using filter paper contact tests, the 48 h-LC(50) values of [C(4)mim]Br, [C(6)mim]Br, [C(8)mim]Br, [C(10)mim]Br and [C(12)mim]Br on the earthworm were 73.33, 28.25, 2.69, 0.37 and 0.02 microg cm(-2), respectively. The 7 d-LC(50) of [C(8)mim]Br was 206.8 mg kg(-1) artificial soil (dry weight) and the 14 d-LC(50) was 159.4 mg kg(-1) artificial soil (dry weight), under the condition of artificial soil. After 1 d and 3 d of acute exposure, the activity of AChE was markedly inhibited when compared to the control, while it was increased at 7d. The cellulase activity was elevated significantly in the treatment groups of 20-160 mg kg(-1) after 3 and 7d of acute exposure. The activity of cellulase was also promoted under the subchronic exposure condition in the 10 and 20 mg kg(-1) groups. The experimental results suggest that [C(8)mim]Br may interfere with the nervous function of the earthworms and increase their cellulase activity. These results indicate that [C(8)mim]Br-exposure can affect the metabolized enzyme activity of earthworms at low concentrations and can even cause worm death at high doses, both of which have potential impacts on the soil environment.

Effects of the 1-alkyl-3-methylimidazolium bromide ionic liquids on the antioxidant defense system of Daphnia magna.

This study examined the antioxidant responses of Daphnia magna following exposure to different concentrations of the ionic liquid (IL) 1-octyl-3-methylimidazolium bromide and the 50% LC(50) concentrations of methylimidazolium bromide ILs with different alkyl-chain lengths. Activities of antioxidant defense enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase) and levels of the antioxidant glutathione and the lipid peroxidation by-product malondialdehyde were measured using traditional methods or commercial kits. The concentration and the alkyl-chain length of ILs were found to strongly influence the antioxidant system of D. magna following IL exposure, and exposure to higher IL concentrations and to ILs with longer alkyl chains generally increased the enzyme activities and biomarker levels examined. Therefore, the present study suggests that oxidative stress is involved in the mechanism of IL-induced toxicity in D. magna.

Identification of neutralizing epitopes on the VP2 protein of infectious bursal disease virus by phage-displayed heptapeptide library screening and synthetic peptide mapping.

Infectious bursal disease virus (IBDV) is the causative agent of infectious bursal disease, which is one of the most important and widespread infectious diseases in commercial chickens. Conformational epitopes have been reported in the highly variable region of the VP2 protein of IBDV. In the present study, a random heptapeptide library was screened by using monoclonal antibodies (mAbs), YNW17 and YNW29, directed to the VP2 of IBDV and two peptide motifs, D-X-P-R and A-R-G, were identified. The motifs are present on the N and C terminal sequences of the highly variable region of VP2. Synthetic overlapping peptides covering the motifs on VP2 were analyzed by Dot- ELISA with the mAbs and two epitopes 197CDSSDRPRVYTIT209 and 329ARGSLAVTI337 identified. The above epitopes were also recognized by chicken anti-IBDV sera and shown to inhibit the binding of their mAbs to recombinant VP2. Both mAbs and sera from mice immunized with the conjugated epitope-peptides were able to neutralize serotype I IBDV. These results indicated that the epitopes are two neutralizing linear B-cell epitopes and would be useful for the development of peptide-based IBD vaccines.