Weakly ionized gold nanoparticles amplify immunoassays for ultrasensitive point-of-care sensors.

Gold nanoparticle-based lateral flow immunoassays (AuNP LFIAs) are widely used point-of-care (POC) sensors for in vitro diagnostics. However, the sensitivity limitation of conventional AuNP LFIAs impedes the detection of trace biomarkers. Several studies have explored the size and shape factors of AuNPs and derivative nanohybrids, showing limited improvements or enhanced sensitivity at the cost of convenience and affordability. Here, we investigated surface chemistry on the sensitivity of AuNP LFIAs. By modifying surface ligands, a surface chemistry strategy involving weakly ionized AuNPs enables ultrasensitive naked-eye LFIAs (~100-fold enhanced sensitivity). We demonstrated how this surface chemistry-amplified immunoassay approach modulates nanointerfacial bindings to promote antibody adsorption and higher activity of adsorbed antibodies. This surface chemistry design eliminates complex nanosynthesis, auxiliary devices, or additional reagents while efficiently improving sensitivity with advantages: simplified fabrication process, excellent reproducibility and reliability, and ultrasensitivity toward various biomarkers. The surface chemistry using weakly ionized AuNPs represents a versatile approach for sensitizing POC sensors.

Analysis of genetic diversity by the SLAF-seq among the farmed Onychostoma macrolepis populations.

OBJECTIVE: The objective of this study was to examine the genetic diversity within and between farmed populations of Onychostoma macrolepis, and to establish a foundation for enhancing the genetic resources of breeding groups through the introduction of new individuals and crossbreeding. A total of 49 individuals were subjected to sequencing using Specific-Locus Amplified Fragment Sequencing (SLAF-seq), one of the restriction site-associated DNA sequencing technologies. The single nucleotide polymorphisms(SNPs)were identified to conduct the analyzation of phylogeny population structure, principal component and genetic diversity. RESULTS: A total of 853,067 SNPs were identified. The results of the phylogenetic analysis revealed that each sample was genetically clustered into three distinct groups: ZhenPing (ZP), LanGao parents (LG), and their progeny population (LG-F1). Each population was observed to be clustered together. Analysis of population genetic diversity revealed that the observed heterozygosity (Ho) ranged from 0.200 to 0.230, the expected heterozygosity (He) ranged from 0.280 to 0.282, and the polymorphic information content (PIC) ranged from 0.228 to 0.230. These results indicate that the genetic diversity of the population is low and the signs of long-term interbreeding are obvious, but there are differences between the populations, and the genetic diversity of the population can be improved by hybridization in different regions.

A Versatile Continuum Gripping Robot with a Concealable Gripper.

Continuum robots with their inherent compliance provide the potential for crossing narrow unstructured workspace and safely grasping various objects. However, the display gripper increases the size of the robots, and therefore, it tends to get stuck in constrained environments. This paper proposes a versatile continuum grasping robot (CGR) with a concealable gripper. The CGR can capture large objects with respect to the robot's scale using the continuum manipulator and can grasp various objects using the end concealable gripper especially in narrow and unstructured workspaces. To perform the cooperative operation of the concealable gripper and the continuum manipulator, a global kinematic model based on screw theory and a motion planning approach referred to as "multi-node synergy method" for the CGR are presented. The simulation and experimental results show that objects of different shapes and sizes can be captured by the same CGR even in complex and narrow environments. Finally, in the future, the CGR is expected to serve for satellite capture in harsh space environments such as high vacuum, strong radiation, and extreme temperatures.

Time-Varying Functional Connectivity of Rat Brain during Bipedal Walking on Unexpected Terrain.

The cerebral cortex plays an important role in human and other animal adaptation to unpredictable terrain changes, but little was known about the functional network among the cortical areas during this process. To address the question, we trained 6 rats with blocked vision to walk bipedally on a treadmill with a random uneven area. Whole-brain electroencephalography signals were recorded by 32-channel implanted electrodes. Afterward, we scan the signals from all rats using time windows and quantify the functional connectivity within each window using the phase-lag index. Finally, machine learning algorithms were used to verify the possibility of dynamic network analysis in detecting the locomotion state of rats. We found that the functional connectivity level was higher in the preparation phase compared to the walking phase. In addition, the cortex pays more attention to the control of hind limbs with higher requirements for muscle activity. The level of functional connectivity was lower where the terrain ahead can be predicted. Functional connectivity bursts after the rat accidentally made contact with uneven terrain, while in subsequent movement, it was significantly lower than normal walking. In addition, the classification results show that using the phase-lag index of multiple gait phases as a feature can effectively detect the locomotion states of rat during walking. These results highlight the role of the cortex in the adaptation of animals to unexpected terrain and may help advance motor control studies and the design of neuroprostheses.

Design of a Fuel Explosion-Based Chameleon-Like Soft Robot Aided by the Comprehensive Dynamic Model.

Soft robotics have advantages over the traditional rigid ones to achieve the bending motion but face with challenges to realize the rapid and long-distance linear motion due to the lack of a suitable actuation system. In this paper, a new explosion-based soft robot is proposed to generate the axial fast extension by the explosion pressure. To support and predict the performance of this explosion-based soft robot, a novel dynamic model is developed by considering the change of working fluid (molecular numbers) and some unavoidable and influential factors in the combustion process. Then, based on the physical prototype, a set of experiments is conducted to test the performance of the explosion-based soft robot in performing the axial extensions, as well as to validate the model proposed in this article. It is found that the novel explosion-based soft robot can achieve rapid axial extension by the developed explosion-based actuation system. The explosion-based soft robot can achieve 41-mm displacement at a fuel mass of 180 mg. In addition, the proposed dynamic model can be validated with an average error of 1.5%. The proposed approach in this study provides a promising solution for future high-power density explosion-based soft robots.

Primary pulmonary anaplastic large cell lymphoma: case report and literature review.

Primary anaplastic large cell lymphoma (ALCL) is a rare pulmonary malignancy. Due to its nonspecific clinical and radiologic manifestations, the disease presents a great challenge to pulmonologists. Appropriate invasive biopsy and immunohistochemistry are important for its diagnosis. Here, we report an ALCL case of a 27-year-old Chinese woman who presented to our hospital complaining of coughing for 10+ days and breath holding for 4-5 days after the event. Positive signs on physical examination were dull percussion sounds and decreased right lung breath sounds. Chest CT scans revealed central carcinoma and atelectasis of the right lung, pleural effusion, and lung mass. Pathology consultation showed a right main bronchial ALCL that involved the parabronchial lymph nodes but not the bronchial tangent. The patient discontinued treatment after right pneumonectomy and died two months later. Postoperative lung biopsy showed anaplastic tumor cells with large and multiple nuclei. The ALCL was characterized by the expression of T cell antigens, CD30 and ALK, as indicated by immunohistochemistry. We also reviewed the atypical cases of ALCL that were previously published. The results indicated that primary pulmonary ALCL is an extremely rare and easily misdiagnosed disease with non-specific clinical and imaging manifestations. Its diagnosis is based on biopsy and immunohistochemistry, and its prognosis is poor.

Autofluorescence bronchoscope diagnosis for lung nodules and masses.

OBJECTIVE: This study set out to enhance the positive detection rate of lung tumors, diagnosis of nodular lesions and improve the accuracy of lung cancer classification by evaluating the clinical value of autofluorescence bronchoscopy (AFB). METHODS: Forceps biopsy, brush biopsy, needle aspiration, and washing techniques were performed alone and in combination with AFB among 38 subjects who were analyzed cytologically and histologically. RESULTS: Our results showed that 33 out of 38 patients were diagnosed with lung cancer, with an overall positive diagnostic rate of 86.8% when the combined methods and AFB was performed; the positive diagnostic rates obtained by forceps, needle aspiration, brush biopsy and washing methods were 68.4%, 84.2%, 55.3% and 36.8%, respectively. Compared to the overall positive rate, the positive diagnostic rates detected by forceps and needle aspiration were not statistically significant (P>0.05). However, the positive rates gained by the brush biopsy and washing method were statistically significant (P<0.01). In addition, we compared the 38 cases evaluated with AFB and 43 cases evaluated with "traditional" white light bronchoscopy (WLB), using the same methods. CONCLUSIONS: AFB and WLB markedly improved the positive diagnosis rate when combined with forceps and needle aspiration. The overall positive diagnostic rate of lung cancer scanned by WLB was increased from 60.5% to 86.8% (P<0.01) by using the combined methods of forceps, brush biopsy, needle aspiration and washing. Moreover, the accuracy of lung cancer classification combined with cytology and cell immunohistochemistry was improved. These results showed that a variety of inspection techniques and diagnostic technologies effectively play a complementary role in the diagnosis and classification of lung cancer.

Changes of microRNAs expression profiles from red swamp crayfish (Procambarus clarkia) hemolymph exosomes in response to WSSV infection.

MicroRNAs (miRNAs) as short noncoding RNAs play important regulatory roles in diverse biological processes by degrading the target mRNAs, and could be delivered by exosomes. WSSV is a highly pathogenic and prevalent virus, and has brought high mortality of P. clarkia. Till present, no studies focus on the miRNAs changes in exosomes during WSSV infection. To understand the different virulence of WSSV on miRNAs expression in P. clarkia hemolymph exosome, the deep sequencing was performed to compare the small RNA libraries from the hemolymph exosome of P. clarkia individuals with or without WSSV infections. From the TEM observations, NTA and Western Blot analysis, the extracted exosomes were well identified with classic characteristics. The 209 conserved miRNAs and 250 novel miRNAs were identified from the small RNA libraries. In response to WSSV infection, there were about 98 miRNAs significantly up-regulated and 59 miRNAs significantly down-regulated. The target genes prediction, GO and KEGG enrichment analysis revealed that some target genes of P. clarkia miRNAs were grouped mainly into the categories of biological regulation, immune system process, signal pathway and other more functions. This is the first report of comprehensive identification of P. clarkia hemolymph exosome miRNAs being differentially regulated in response to WSSV infection. These results will help to understand the hemolymph exosome miRNAs response to different virulence WSSV infection.

Involvement of a short-type peptidoglycan recognition protein (PGRP) from Chinese giant salamanders Andrias davidianus in the immune response against bacterial infection.

PGRPs (Peptidoglycan recognition proteins) could recognize peptidoglycan and play vital roles in innate immunity among different animals. Till present, the functions of PGRP have been studied in various animals, but few reports have studied the amphibian PGRPs. In the current research, a short type PGRP was identified from Chinese giant salamander and its involvement in the innate immunity was studied. The ORF of AdPGRP-SC2 cDNA was 573 bp, which encoded 190 amino acids, and contained a PGRP and an amidase_2 domain. The qPCR analysis revealed that AdPGRP-SC2 mRNA transcripts expressed in different tissues, with the highest expression level in muscle, intestine and spleen. Results of immune challenges with peptidoglycan (PGN) demonstrated that expression patterns of AdPGRP-SC2 were significantly up-regulated in erythrocyte and spleen at the early injection stage. The recombinant AdPGRP-SC2 protein was successfully produced and purified, and it could show binding affinity to different bacteria. In the presence of Zn2+, the rAdPGRP-SC2 could exhibit a broad PAMPs binding activities, strongly agglutinate bacteria and exhibit amidase enzyme activity. Collectively, these data indicate AdPGRP-SC2 could act as PRR to recognize the invading microorganisms and as the antimicrobial effectors during the innate immune response of A. davidianus.

Tissue distribution and elimination features of florfenicol in hybrid sturgeon cultured in cool water.

In order to understand pharmacokinetics in vivo and tissue residues of florfenicol in Hybrid sturgeon cultured in cold water, changes in concentration of florfenicol in plasma, muscle, liver and kidney of sturgeon after administration were measured by high performance liquid chromatography mass spectrometry. The results show that after 15.0 mg/kg dose administration, the concentration change in plasma and liver, kidney and muscle of sturgeon are suitable for the one compartment open model. Florfenicol pharmacokinetic parameters in plasma, liver, kidney and sturgeon muscle showed that the peak concentration (Cmax ) were 549.7, 551.8, 958.2 and 405.4 µg/kg, respectively. The peak time (Tmax ) was 17.5, 14.2, 14.0 and 20.8 hr, respectively. The elimination half-life (T1/2 ) was 75.1, 99.8, 68.8 and 75.5 hr, respectively. Area under the curve were 70,048.9, 63,850.6, 109,579.9 and 53,437.3 µg/kg hr, respectively. Mean residence time was 100.6, 96.4, 96.5 and 104.6 hr, respectively. The absorption speed and elimination of florfenicol in cold water cultured sturgeon is slow, and the peak concentration is low, but the florfenicol was widely distributed in the body of sturgeon. The results provide a theoretical basis for formulation of medication and withdrawal period of florfenicol used in sturgeon cultured in cold water.

Characterization of galectin-1 from Chinese giant salamanders Andrias davidianus and its involvements during immune response.

Galectins are considered as a multifunctional protein which play essential roles in cell adhesion and apoptosis, inflammation, tumor progression and immune response. In spite of extensive studies of galectin importance in immune system among different animals, few studies have been devoted to their functions in amphibian. In the present study, we characterized one proto type of galectin (named AdGal1) from Chinese giant salamander Andrias davidianus and studied its function in immune response. AdGal1 cDNA possesses an open reading frame of 598 bp, which encodes a putative galectin of 134 amino acids containing one carbohydrate recognition domains (CRDs). The constitutive expression of mRNA transcripts was detected in a wide range of tissues, with the highest expression in kidney. Immune challenges with Aeromonas hydrophila and Chinese giant salamander iridovirus (GSIV), the transcript level of AdGal1 in kidney was significantly upregulated. The mature protein of AdGal1 was successfully expressed and purified in Escherichia coli BL21 (DE3). The recombinant AdGal1 (rAdGal1) could show bind activity to different Gram negative and Gram positive bacteria. It could also strongly agglutinate different kinds of bacteria at different concentrations. Collectively, these data from the present study indicate that AdGal1 is a vital pattern recognition receptor to recognize different microbes in the innate immune system of Andrias davidianus.

RNA-Seq analysis and gene discovery of Andrias davidianus using Illumina short read sequencing.

The Chinese giant salamander, Andrias davidianus, is an important species in the course of evolution; however, there is insufficient genomic data in public databases for understanding its immunologic mechanisms. High-throughput transcriptome sequencing is necessary to generate an enormous number of transcript sequences from A. davidianus for gene discovery. In this study, we generated more than 40 million reads from samples of spleen and skin tissue using the Illumina paired-end sequencing technology. De novo assembly yielded 87,297 transcripts with a mean length of 734 base pairs (bp). Based on the sequence similarities, searching with known proteins, 38,916 genes were identified. Gene enrichment analysis determined that 981 transcripts were assigned to the immune system. Tissue-specific expression analysis indicated that 443 of transcripts were specifically expressed in the spleen and skin. Among these transcripts, 147 transcripts were found to be involved in immune responses and inflammatory reactions, such as fucolectin, ß-defensins and lymphotoxin beta. Eight tissue-specific genes were selected for validation using real time reverse transcription quantitative PCR (qRT-PCR). The results showed that these genes were significantly more expressed in spleen and skin than in other tissues, suggesting that these genes have vital roles in the immune response. This work provides a comprehensive genomic sequence resource for A. davidianus and lays the foundation for future research on the immunologic and disease resistance mechanisms of A. davidianus and other amphibians.

Literature-based analysis on relationship of symptoms, drugs and therapies in treatment of intestinal diseases.

OBJECTIVE: To evaluate clinical articles published in the past 30 years using the Traditional Chinese Medicine (TCM) theory of the lung being connected with large intestine to treat intestinal diseases. We also analyzed the relationship between symptoms, drugs and therapies with data-excavating technology to aid management. METHODS: After retrieving relevant clinical articles, we set up a database, used Microsoft Structured Query Language Server 2005 Analysis Services as a data-excavating tool, and applied the association rule to study the relationship between the symptoms, drugs and therapies of intestinal diseases. RESULTS: The key symptoms of dyschesia, constipation, abdominal fullness, fatigue and pale tongue could be treated with Kuxingren (Semen Armeniacae Amarum), Huangqi (Radix Astragali Mongolici) and Gualou (Fructus et Semen Trichosanthis) to invigorate Qi and moisten the intestine. Among these agents, Kuxingren (Semen Armeniacae Amarum) was used most frequently. Clearing Fu-organs was the most prevalent therapy for abdominal fullness, dyschesia, constipation and red tongue. Clearing Fu-organs could be achieved with Kuxingren (Semen Armeniacae Amarum) and Gua-lou (Fructus et Semen Trichosanthis), whereas Qi could be invigorated using Huangqi (Radix Astragali Mongolici), Gancao (Radix Glycyrrhizae), Baizhu (Rhizoma Atractylodis Macrocephalae) and Kuxingren (Semen Armeniacae Amarum). Moistening the intestine was possible with Kuxingren (Semen Armeniacae Amarum), Huomaren (Fructus Cannabis) and Jiegeng (Radix Platycodi). Also, moistening the lungs was done with Kuxingren (Semen Armeniacae Amarum), ventilating the lungs with Kuxingren (Semen Armeniacae Amarum) and Gualou (Fructus et Semen Trichosanthis), and nourishing the lungs with Huangqi (Radix Astragali Mongolici), Gancao (Radix Glycyrrhizae) and Kuxingren (Semen Armeniacae Amarum). These data demonstrated that Kuxingren (Semen ArmeniacaeAmarum) was a key agent. CONCLUSION: Our analyses of the literature showed clear relationships between symptoms (constipation, dyschesia, abdominal fullness), drugs [Gualou (Fructus et Semen Trichosanthis), Kuxingren (Semen Armeniacae Amarum), Huangqi (Radix Astragali Mongolici)] and therapies (moistening the intestine, clearing Fu-organs, invigorating Qi, ventilating the lungs).

Molecular characterization, tissue distribution and functional analysis of macrophage migration inhibitory factor protein (MIF) in Chinese giant salamanders Andrias davidianus.

The macrophage migration inhibitory factor (MIF) produced in numerous cell types is a multi-functional cytokine mediating both innate and adaptive immune responses. To obtain a better understanding of the innate immune ability of Andrias davidianus and their defensive mechanisms, we identify the A. davidianus MIF (AdMIF) cDNA sequence from a skin cDNA library. The full-length cDNA of AdMIF was of 661bp, consisting of 134bp 5'-terminal UTR, 348bp open reading frame and 179bp 3'-terminal UTR. The deduced protein was composed of 115 amino acids, with an estimated molecular mass of 12.53kDa and a predicted pI of 6.07. AdMIF primary structure is as conserved as the other known sequences. Real time quantitative PCR (qRT-PCR) analysis indicated that AdMIF gene was ubiquitously expressed in selected tissues, with the highest level in liver, moderate level in spleen, intestine, stomach, and the lowest level in heart and skin. The cDNA fragment encoding mature peptide of AdMIF was recombined and expressed in Escherichia coli BL21 (DE3). By means of Ni(2+)-chelating chromatography, the recombinant protein of AdMIF (rAdMIF) was purified successfully. The rAdMIF protein was proved to have enzymatic redox and tautomerase activity in vitro. This study represents the first report for characterization of A. davidianus MIF, demonstrating the successful isolation of MIF from Chinese giant salamanders, and the purified rAdMIF protein is important to produce the monoclonal antibodies and provides a foundation for further investigation of the physiological function of AdMIF.

Analysis on clinically drug-used law for lung-intestine related diseases.

OBJECTIVE: To analyze the application laws of clinical drugs by descriptive statistical method with the clinical literatures guided by "lung being connected with large intestine" used as the object. METHODS: Retrieve publicly published clinical literatures about "lung being connected with large intestine" in recent 30 years, establish the database of clinical literature, and study on the clinically drug-used laws by descriptive statistical analytic method. RESULTS: The common high frequent drugs used for the lung-intestine related diseases were Gualou (Ructus Trichsanthis), Dahuang (Radix et Rhizoma), Kuxingren (Semen Armeniacae Amarum), Gancao (Radix Glycyrrhizae), Houpu (Cortex Magnoliae Officinalis), Zhishi (Fructus Aurantii Immaturus). For the different clinical syndrome types, in the commonly used drugs there were different characteristics, reflecting the principle of TCM syndrome differentiation treatment. CONCLUSION: The common high frequent drugs used for the lung-intestine related diseases are Gualou (Fructus Trichsanthis), Dahuang (Radix et Rhizoma), Kuxingren (Semen Armeniacae Amarum), Gancao (Radix Glycyrrhizae), Houpu (Cortex Magnoliae Officinalis), Zhishi (Fructus Aurantii Immaturus), which are commonly-used drugs for the lung-intestine related diseases, with differences for different syndrome types.