First observation of the mating, egg-laying and hatching behaviour of a captive female Kong skate, Okamejei kenojei (Müller & Henle, 1841).

This study provides a novel record of the reproductive behaviour of the Kong skate (Okamejei kenojei) in captivity. These skates were found to mate and deposit eggs at a temperature of 16.5 ± 0.5°C. The results showed that 76.13% of the eggs possessed one yolk, 0.77% of the eggs possessed two yolks and 23.11% of the eggs had no yolk (N = 1043). The deposition of non-yolk and double-yolk eggs was random. A total of 100 eggs were collected. After nearly 92 ± 5 days of incubation, 28 eggs failed to hatch, and 72 skates were successfully hatched with a female-to-male ratio of 1:1 (P > 0.05). The results enrich our knowledge of the reproduction in cartilaginous fishes and can inform management and conservation strategies for this species.

Significant genes in response to low temperature in Fenneropenaeus chinensis screened through multiple transcriptome group comparisons.

Fenneropenaeus chinensis is a migratory marine species with a suitable growth at 18-30°C. To prolong breeding season and reduce mortality in winter, breeding new shrimp varieties with cold tolerance is essential. Genes upregulated and highly expressed at low temperature are reasonable candidate genetic markers for the breeding of cold tolerant strain variants. This study screened genes with these features by comparing multiple low-vs. normal-temperature transcriptome groups. The results showed that nine genes were upregulated and highly expressed at low temperature in more than seven of the nine comparison groups. Six of them were identified as genes encoding transcription factor ATF2, RNA recognition motif domain-containing protein, cytochrome b5-like protein, troponin C, tubulin alpha-1, and 18S/5.8S/28S rRNA, respectively. Cold-inducible upregulations of ATF2, cytochrome b5, and rRNAs were novel findings in this study. The other three novel genes were predicted to encode a membrane-bound extracellular protein and two lncRNAs. Four of the screened genes were verified by real time RT-PCR, and their expression levels were consistent with the sequencing results, demonstrating the accuracy of the transcriptome sequencing data. Function analysis showed that ATF2 might be the master transcription factor regulating the expressions of proteins involved in cellular responses to cold. The other genes played a role in events such as enhancing translation, increasing energy, inhibiting apoptosis, and preserving cell integrity. The expression features of these nine genes suggested that they were of great significance to the cold tolerance of shrimp.

Identification and Characterization of Non-Coding RNAs in Thymoma.

BACKGROUND Thymoma is the most common tumor of the anterior mediastinum, and can be caused by infrequent malignancies arising from the epithelial cells of the thymus. Unfortunately, blood-based diagnostic markers are not currently available. High-throughput sequencing technologies, such as RNA-seq with next-generation sequencing, have facilitated the detection and characterization of both coding and non-coding RNAs (ncRNAs), which play significant roles in genomic regulation, transcriptional and post-transcriptional regulation, and imprinting and epigenetic modification. The knowledge about fusion genes and ncRNAs in thymomas is scarce. MATERIAL AND METHODS For this study, we gathered large-scale RNA-seq data belonging to samples from 25 thymomas and 25 healthy thymus specimens and analyzed them to identify fusion genes, lncRNAs, and miRNAs. RESULTS We found 21 fusion genes, including KMT2A-MAML2, HADHB-REEP1, COQ3-CGA, MCM4-SNTB1, and IFT140-ACTN4, as the most frequent and significant in thymomas. We also detected 65 differentially-expressed lncRNAs in thymomas, including AFAP1-AS1, LINC00324, ADAMTS9-AS1, VLDLR-AS1, LINC00968, and NEAT1, that have been validated with the TCGA database. Moreover, we identified 1695 miRNAs from small RNA-seq data that were overexpressed in thymomas. Our network analysis of the lncRNA-mRNA-miRNA regulation axes identified a cluster of miRNAs upregulated in thymomas, that can trigger the expression of target protein-coding genes, and lead to the disruption of several biological pathways, including the PI3K-Akt signaling pathway, FoxO signaling pathway, and HIF-1 signaling pathway. CONCLUSIONS Our results show that overexpression of this miRNA cluster activates PI3K-Akt, FoxO, HIF-1, and Rap-1 signaling pathways, suggesting pathway inhibitors may be therapeutic candidates against thymoma.

Combined use of circulating tumor cells and salivary mRNA to detect non-small-cell lung cancer.

Liquid biopsy is an emerging technique for noninvasive detection of various cancers. Majority of liquid biopsy tests still, however, use solitary type of biomarkers with unsatisfactory sensitivity and specificity. To this end, a combined approach of circulating tumor cells (CTCs) and salivary mRNA biomarkers was evaluated for discriminating non-small-cell lung cancer (NSCLC) from healthy controls.Our study included a discovery phase to find multiple biomarkers, and an independent validation phase to confirm the applicability of the selected biomarkers. In the discovery phase, CTC level in blood and 5 mRNA biomarkers in saliva (i.e., CCNI, Epidermal growth factor receptor [EGFR], FGF19, FRS2, and GREB1) were measured for 140 NSCLC patients and 140 healthy controls, followed by developing a predictive model. Next, this panel of biomarkers was applied to another patient cohort consisted of 60 patients with NSCLC and 60 healthy controls in the validation phase.We found that our novel biomarker panel could differentiate patients with NSCLC from healthy controls with high sensitivity (92.1%) and high specificity (92.9%) in the discovery phase. In the validation phase, we achieved sensitivity of 88.3% and specificity of 90.0%.To our best knowledge, it is the first time that a combined use of CTC and salivary mRNA biomarkers were applied for noninvasive detection of NSCLC.

Transcatheter arterial chemoembolization (TACE) for lymph node metastases in patients with hepatocellular carcinoma.

OBJECTIVE: To evaluate the efficacy and safety of transcatheter arterial chemoembolization (TACE) in hepatocellular carcinoma (HCC) with regional lymph node metastases. METHODS: Forty-eight patients with HCC and regional lymph node metastases were enrolled in this study. The patients were allocated into two groups: Group A (28 patients) underwent TACE for both intrahepatic tumors and lymph node metastasis and Group B (20 patients) received TACE for intrahepatic tumors only. RESULTS: The patients were followed-up by contrast enhanced CT scan 6-8 weeks after TACE treatment. In Group A, seven and nine patients achieved complete and partial response for lymph node metastasis, respectively, with 1-year and 2-year overall survival rates of 60.7% and 35.7%, respectively. In contrast, none of the patients in Group B achieved a complete response, whereas four patients achieved a partial response. The 1-year and 2-year survival rates for the patients in Group B were 40% and 0%, respectively. The difference in survival between the two groups was statistically significant (P = 0.001). CONCLUSIONS: TACE is an effective treatment to regional lymph node metastasis in HCC without significant side effects and could provide survival benefits to the patients with advanced HCC.

Pure transcatheter arterial chemoembolization therapy for intrahepatic tumors causes a shrink in pulmonary metastases of hepatocellular carcinoma.

OBJECTIVE: Pulmonary metastasis of hepatocellular carcinoma (HCC) could be defined as advanced HCC and systematic treatment is the main therapeutic modality. However, local therapy of intrahepatic tumor, which is significantly associated with the prognosis of HCC, remains important for advanced HCC. METHODS: Twenty-six HCC patients with pulmonary metastasis underwent intrahepatic transcatheter arterial chemoembolization (TACE). We investigated the progression of lung metastastic tumors, overall survival and risk factors related to survival of these patients. RESULTS: Of the 26 patients who underwent TACE for one to four times, 10 patients achieved complete remission (CR) of intrahepatic tumors and among these 10 patients, 4 patients successfully received hepatic artery-venous shunt embolization combined with TACE. The lung metastasis lesions also achieved CR and the survival time was significantly longer than the other 22 patients. The lung metastastic lesions of the other 6 patients of intrahepatic tumors achieved stable disease (SD). Six patients acquired partial remission (PR) of intrahepatic tumors after TACE, while the lung metastastic lesions showed SD or progress disease (PD). Patients who showed CR and PR of intrahepatic tumors had longer survival time than patients with SD and PD. Portal vein tumor thrombus and size of the lung metastastic lesions were significant prognostic factors in these advanced HCC patients. CONCLUSIONS: With respect to HCC patients with lung metastasis, TACE was an effective and important therapeutic tool to control pulmonary metastatic tumor growth, and prolong the survival of advanced HCC patients, especially patients with hepatic artery-venous shunt.

Ultrasensitive electrochemical immunosensor for carbohydrate antigen 72-4 based on dual signal amplification strategy of nanoporous gold and polyaniline-Au asymmetric multicomponent nanoparticles.

A sandwich electrochemical immunosensor is described for carbohydrate antigen 72-4 (CA72-4) based on a dual amplification strategy with nanoporous gold (NPG) film as the sensor platform and polyaniline-Au asymmetric multicomponent nanoparticles (PANi-Au AMNPs) as labels. In this study, the second anti-CA72-4 antibody (Ab2) adsorbed onto the Au of the PANi-Au AMNPs, which could be simply synthesized by interfacial reaction and have many characteristics of polyaniline and Au nanoparticle, such as well-controlled size, high conductivity, biocompatibility and catalysis. NPG film was used as electrode substrate material to fix a large number of antibodies, due to its unique properties: good biocompatibility, high conductivity, large surface area, and stability. The synergetic of NPG film and PANi-Au AMNPs could increase signal response, and significantly improve sensitivity of the immunosensor. The proposed immunosensor exhibited a wide linear range from 2 to 200 U/mL, with a detection limit of 0.10 U/mL CA72-4, good reproducibility, selectivity and stability. This new type of labels for immunosensors may provide many potential applications in the detection of carbohydrate antigen in immunoassays.

Ultrasound-assisted production of biodiesel from soybean oil using Brønsted acidic ionic liquid as catalyst.

Biodiesel production from soybean oil with methanol was performed in the presence of a Brønsted acidic ionic liquid-based catalyst under ultrasound irradiation. The influences of various parameters on the transesterification reaction, including the amount of catalyst, the molar ratio of methanol to oil, the temperature and the ultrasound power, were investigated. The optimal conditions were: methanol/oil molar ratio of 9:1, 1.0 wt.% catalyst in oil, ultrasound power of 200 W, and reaction temperature of 60°C. Under these conditions, the conversion of triglycerides to fatty acid methyl esters was about 93.2% within the reaction time of 60 min.

Enzyme-modified nanoporous gold-based electrochemical biosensors.

On the basis of the unique physical and chemical properties of nanoporous gold (NPG), which was obtained simply by dealloying Ag from Au/Ag alloy, an attempt was made in the present study to develop NPG-based electrochemical biosensors. The NPG-modified glassy carbon electrode (NPG/GCE) exhibited high-electrocatalytic activity toward the oxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H(2)O(2)), which resulted in a remarkable decrease in the overpotential of NADH and H(2)O(2) electro-oxidation when compared with the gold sheet electrode. The high density of edge-plane-like defective sites and large specific surface area of NPG should be responsible for the electrocatalytic behavior. Such electrocatalytic behavior of the NPG/GCE permitted effective low-potential amperometric biosensing of ethanol or glucose via the incorporation of alcohol dehydrogenase (ADH) or glucose oxidase (GOD) within the three-dimensional matrix of NPG. The ADH- and GOD-modified NPG-based biosensors showed good analytical performance for biosensing ethanol and glucose due to the clean, reproducible and uniformly distributed microstructure of NPG. The stabilization effect of NPG on the incorporated enzymes also made the constructed biosensors very stable. After 1 month storage at 4 degrees C, the ADH- and GOD-based biosensors lost only 5.0% and 4.2% of the original current response. All these indicated that NPG was a promising electrode material for biosensors construction.

Immobilization of lignin peroxidase on nanoporous gold: enzymatic properties and in situ release of H2O2 by co-immobilized glucose oxidase.

Immobilization of enzymes on porous inorganic materials is very important for biocatalysis and biotransformation. In this paper, nanoporous gold (NPG) was used as a support for lignin peroxidase (LiP) immobilization. NPG with a pore size of 40-50 nm was prepared by dealloying Au/Ag alloy (50:50 wt%) for 17 h. By incubation with LiP aqueous solution, LiP was successfully immobilized on NPG. The optimal temperature of the immobilized LiP was ca. 40, 10 degrees C higher than that of free LiP. After 2h incubation at 45 degrees C, 55% of the initial activity of the immobilized LiP was still retained while the free LiP was completely deactivated. In addition, a high and sustainable LiP activity was achieved via in situ release of H(2)O(2) by a co-immobilized glucose oxidase. The present co-immobilization system was demonstrated to be very effective for LiP-mediated dye decolourization.

Effects of nonionic surfactant Triton X-100 on the laccase-catalyzed conversion of bisphenol A.

The laccase-catalyzed conversion of bisphenol A (BPA) in aqueous solutions was studied in the absence and presence of nonionic surfactant Triton X-100. It was found that the addition of Triton X-100 into the reaction system increased the conversion of BPA, especially near the critical micelle concentration of Triton X-100. Also it was found that the stability of laccase was greatly improved in the presence of TritonX-100. Studies on the endogenous fluorescence emission of laccase indicated that there existed an interaction between Triton X-100 and laccase, which was beneficial to folding and stabilizating of laccase. The binding of Triton X-100 to the laccase surface also mitigated the inactivation effect caused by the free radicals and polymerization products. Under otherwise identical conditions, a lower dosage of laccase was needed for the higher conversion of BPA in the presence of Triton X-100.

A novel nanoporous gold modified electrode for the selective determination of dopamine in the presence of ascorbic acid.

The electrochemical detection of dopamine (DA) at conventional solid electrodes was interfered by the coexisted ascorbic acid (AA). To circumvent this problem, many modified electrodes were tried. In this paper, an attempt was made to use nanoporous gold (NPG) as modifying materials. The NPGs with different pore sizes were prepared simply by dealloying Ag from Au/Ag alloy with concentrated nitric acid. The glassy carbon electrode (GCE) based modified electrode was fabricated by simply affixing the NPG film on the surface of a GCE. The electrochemical behaviors of AA and DA at the modified electrode were studied. The results indicated that the NPG/GCE exhibited substantial enhancement in electrochemical sensitivity for DA and AA due to its large surface area. Results also showed that the oxidation of AA at the electrode was a diffusion-controlled process, but for DA it was an adsorption-controlled process. This result, together with the different anodic peak potentials of the two species, made it possible the selective determination of DA in the presence of AA. Due to the interaction of the amino group of DA with the surface of nanoscale gold, DA could be accumulated on the NPG/GCE, while AA could not. This permitted the coexistence of large amount of AA. When differential pulse voltammetry (DPV) was used, a submicrolevel of DA could be detected in the millimolar level of AA with a detection limit of 17nM at a signal-to-noise ratio of 3. In a word, the modified electrode showed good sensitivity, selectivity and reproducibility.