C-type lectin 12B/4E of black rockfish (Sebastes schlegelii) macrophages as pattern recognition receptors in the antibacterial mechanism of exploration.

As lower vertebrates, fish have both innate and adaptive immune systems, but the role of the adaptive immune system is limited, and the innate immune system plays an important role in the resistance to pathogen infection. C-type lectins (CLRs) are one of the major pattern recognition receptors (PRRs) of the innate immune system. CLRs can combine with pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs) to trigger NF-κB signaling pathway and exert immune efficacy. In this study, Ssclec12b and Ssclec4e of the C-type lectins, were found to be significantly up-regulated in the transcripts of Sebastes schlegelii macrophages stimulated by bacteria. The identification, expression and function of these lectins were studied. In addition, the recombinant proteins of the above two CLRs were obtained by prokaryotic expression. We found that rSsCLEC12B and rSsCLEC4E could bind to a variety of bacteria in a Ca2+-dependent manner, and promoted the agglutination of bacteria and blood cells. rSsCLEC12B and rSsCLEC4E assisted macrophages to recognize PAMPs and activate the NF-κB signaling pathway, thereby promoting the expression of inflammatory factors (TNF-α, IL-1ß, IL-6, IL-8) and regulating the early immune inflammation of macrophages. These results suggested that SsCLEC12B and SsCLEC4E could serve as PRRs in S. schlegelii macrophages to recognize pathogens and participate in the host antimicrobial immune process, and provided a valuable reference for the study of CLRs involved in fish innate immunity.

Genome-wide identification, characterization, molecular evolution and expression profiling analysis of scavenger receptors in black rockfish (Sebastes schlegelii).

The scavenger receptors (SRs) gene family is considered as the membrane-associated pattern recognition receptors that plays important roles in the immune responses of organisms. However, there is currently limited research on the systematic identification of the SRs gene family in teleost and their role in the innate immunity of S. schegelii. In this study, we identified and annotated 15 SRs genes in S. schegelii. Through phylogenetic analysis, analysis of conserved domains, gene structure, and motif composition, we found that SRs gene family within different classes were relatively conserved. Additionally, we used qRT-PCR to analyze the expression patterns of SRs genes in immune-related tissues from healthy and Acinetobacter johnsonii-infected S. schegelii. The results showed that SRs genes exhibited different tissue expression patterns and the expression of SRs genes significantly changed after A. johnsonii infection. These results provided a valuable basis for further understanding of the functions of SRs in the innate immune response of S. schegelii.

Coulomb blockade and Coulomb staircases in CoBi nanoislands on SrTiO3(001).

We successfully fabricated two-dimensional metallic CoBi nanoislands on SrTiO3(001) substrate by molecular beam epitaxy, and systematically investigated their electronic structures by scanning tunneling microscopy and spectroscopyin situat 4.2 K. Coulomb blockade and Coulomb staircases with discrete and well-separated levels are observed for the individual nanoisland, which is attributed to single-electron tunneling via two tunnel junction barriers. They are in excellent agreement with the simulations based on orthodox theory. Furthermore, we demonstrated that the Coulomb blockade becomes weaker with increasing temperature and almost disappears at ∼22 K in our variable temperature experiment, and its full-width at half-maximum of dI/dVpeaks with temperature is ∼6 mV. Our results provide a new platform for designing single-electron transistors that have potential applications in future microelectronics.

Molecular characterization, expression and immune functional analysis of cystatin 10 in turbot.

BACKGROUND: Cystatin is a protease inhibitor that also regulates genes expression linked to inflammation and plays a role in defense and regulation. METHODS AND RESULTS: Cystatin 10 (Smcys10) was cloned from Scophthalmus maximus and encodes a 145 amino acid polypeptide. The results of qRT-PCR showed that Smcys10 exhibited tissue-specific expression patterns, and its expression was significantly higher in the skin than in other tissues. The expression level of Smcys10 was significantly different in the skin, gill, head kidney, spleen and macrophages after Vibrio anguillarum infection, indicating that Smcys10 may play an important role in resistance to V. anguillarum infection. The recombinant Smcys10 protein showed binding and agglutinating activity in a Ca2+-dependent manner against bacteria. rSmcys10 treatment upregulated the expression of IL-10, TNF-α and TGF-ß in macrophages of turbot and hindered the release of lactate dehydrogenase (LDH) from macrophages after V. anguillarum infection, which confirmed that rSmcys10 reduced the damage to macrophages by V. anguillarum. The NF-κB pathway was suppressed by Smcys10, as demonstrated by dual-luciferase analysis. CONCLUSIONS: These results indicated that Smcys10 is involved in the host antibacterial immune response.

Continuous dynamic measurement of frequency scanning interferometry based on motion phase synchronization compensation and calibration.

We present a continuous dynamic frequency scanning interferometry (DFSI) measurement method based on motion phase synchronization compensation and calibration. By introducing heterodyne interferometry (HI) synchronization measurement and frequency scanning interferometry (FSI) motion phase compensation, dynamic continuous measurement is achieved and effectively suppresses the distance error introduced by the Doppler effect (DE). Based on this, the influence of the initial optical frequency deviation (OFD) of the tunable laser and the OFD of the HI laser on the dynamic absolute distance measurement (DADM) is analyzed; the relationships between the error of DADM with the variation of the OFD and the target motion parameters are investigated; and the residual DE introduced by the OFD is shown as the fundamental cause of the degradation of the accuracy of DFSI. We propose an online optical frequency measurement method based on HI combined with H13C14N gas absorption cells to resolve this problem. High-precision motion phase compensation is achieved by calibrating the optical frequency (fixed frequency) of the measured HI laser and the initial frequency of the tunable laser online during measurement and then performing motion phase calibration. To verify the effectiveness of our method, an optical frequency calibration experiment, a continuous DADM experiment, and a precision evaluation experiment were conducted, and a highly accurate continuous DADM was achieved.

Real-time high-precision FMCW laser range extraction method based on a hardware multiplier array.

Frequency-modulated continuous wave (FMCW) laser interferometry is an ideal large-scale absolute distance measurement method. It has advantages of high precision and noncooperative target measurement capability, with no blind spot for ranging. To meet the requirements of high-precision, high-speed 3D topography measurement technologies, a faster measurement speed of FMCW LiDAR at each measurement point is required. To solve the shortcomings of the existing technology, a real-time high-precision hardware solution method (including but not limited to FPGA and GPU) for lidar beat frequency signals is provided here based on hardware multiplier arrays to reduce lidar beat frequency signal processing time and to save energy and resource consumption during processing. A high-speed FPGA architecture was also designed for the frequency-modulated continuous wave lidar range extraction algorithm. The whole algorithm was designed and implemented in real time based on the principle of full-pipelines and parallelism. The results show that the processing speed of the FPGA system is faster than that of current top-performing software implementations.

Dynamic nonlinearity errors in laser Doppler vibrometer measurements induced by environmental vibration and error correction.

Laser Doppler vibrometers (LDVs) are widely used for vibration testing in various fields. Nonlinearity errors are the key factor affecting the measurement accuracy of LDVs. The conventional Heydemann method cannot correct nonlinearity errors produced by noisy environments. Thus, we establish a novel model to describe dynamic nonlinearity errors produced in noisy environments and propose a compensation method to mitigate signal distortion. The performance of the proposed method is assessed by performing both simulations and experiments. The results of experiments carried out in a noisy environment indicate that the proposed method suppresses the nonlinearity to 30 nm compared to 737 nm using the conventional Heydemann correction. The proposed method can improve the accuracy of LDV measurements in industrial environments.

Associations between ATG16L1 gene polymorphism and antineutrophil cytoplasmic antibody-associated vasculitis in the Chinese Guangxi population: A case-control study.

BACKGROUND: Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is an autoimmune disease often accompanied by rapidly progressive renal failure, and the genetic background is still unknown. Our study was performed to test whether autophagy-related 16 like 1 (ATG16L1) rs4663402 and rs4663396 single nucleotide polymorphisms (SNPs) were associated with AAV in the Chinese Guangxi population. METHODS: One hundred seventy seven unrelated AAV patients and 216 healthy controls were included in this case-control study. Multiplex polymerase chain reaction combined with high-throughput sequencing was used for typing, and SNPStats and SHEsis were used for association analysis, pairwise linkage disequilibrium, and haplotype analysis. RESULTS: rs4663402 and rs4663396 were in Hardy-Weinberg equilibrium in AAV and control groups. The frequencies of rs4663402 AA, AT, and TT genotypes were 82.5%, 16.9%, and 0.6%, respectively, in patients with AAV, and 83.5%, 16.2%, and 0.5%, respectively, in controls. The frequencies of rs4663396 CC, CT, and TT genotypes were 63.8%, 33.9%, and 2.3%, respectively, in patients with AAV, and 69.2%, 26.6%, and 4.2%, respectively, in controls. Haplotype analysis revealed two SNPs in a single haplotype block (D' = 1.0). Our logistic regression adjusted for sex and age showed no association between rs4663402 and rs4663396 and the risk for AAV in genetic models (p > 0.05). However, ATG16L1 rs4663396 CC and CT + TT genotypes exhibited statistically significant differences in the incidence of arthralgia (p = 0.03). CONCLUSIONS: Our results indicated that ATG16L1 rs4663402 and rs4663396 polymorphisms were not associated with AAV in the Chinese Guangxi population. ATG16L1 rs4663396 CT + TT genotype may be associated with arthralgia.

Silencing of tumor-suppressive NR_023387 in renal cell carcinoma via promoter hypermethylation and HNF4A deficiency.

Dysregulation of the epigenetic status of long noncoding RNAs (lncRNAs) has been linked to diverse human diseases including human cancers. However, the landscape of the whole-genome methylation profile of lncRNAs and the precise roles of these lncRNAs remain elusive in renal cell carcinoma (RCC). We first examined lncRNA expression profiles in RCC tissues and corresponding adjacent normal tissues (NTs) to identify the lncRNA signature of RCC, then lncRNA Promoter Microarray was performed to depict the whole-genome methylation profile of lncRNAs in RCC. Combined analysis of the lncRNAs expression profiles and lncRNAs Promoter Microarray identified a series of downregulated lncRNAs with hypermethylated promoter regions, including NR_023387. Quantitative real-time polymerase chain reaction (RT-PCR) implied that NR_023387 was significantly downregulated in RCC tissues and cell lines, and lower expression of NR_023387 was correlated with shorter overall survival. Methylation-specific PCR, MassARRAY, and demethylation drug treatment indicated that hypermethylation in the NR_023387 promoter contributed to its silencing in RCC. Besides, HNF4A regulated the expression of NR_023387 via transcriptional activation. Functional experiments demonstrated NR_023387 exerted tumor-suppressive roles in RCC via suppressing the proliferation, migration, invasion, tumor growth, and metastasis of RCC. Furthermore, we identified MGP as a putative downstream molecule of NR_023387, which promoted the epithelial-mesenchymal transition of RCC cells. Our study provides the first whole-genome lncRNA methylation profile in RCC. Our combined analysis identifies a tumor-suppressive and prognosis-related lncRNA NR_023387, which is silenced in RCC via promoter hypermethylation and HNF4A deficiency, and may exert its tumor-suppressive roles by downregulating the oncogenic MGP.

Investigation of the Mn dopant-enhanced photoluminescence performance of lead-free Cs2AgInCl6 double perovskite crystals.

The lead-free double perovskite Cs2AgInCl6 is a potential candidate for LEDs, the photoluminescence performance of which is reinforced greatly by Mn doping. Here, we analyzed the geometric, electronic and photoluminescence properties of Mn-doped Cs2AgInCl6 by means of first-principle calculations. We found that in the interior of Cs2AgInCl6, the Mn dopant formed defect complexes by substituting an Ag atom and generating an Ag vacancy (MnAgVAg) owing to the charge balance and the weak distortion of the metal octahedra. The MnAgVAg defect introduced two defect bands in the forbidden gap, which was contributed predominantly by the 3d orbitals of the Mn2+ ions. The electron transition of the Mn2+ ions from the first excited state to the ground state, i.e., from 4T1 to 6A1 states, gives rise to the PL spectrum that is lower than the bandgap. Therefore, we show that the Mn dopant indeed reinforces the PL performance of Cs2AgInCl6 greatly and is beneficial for its use as an LED material.

Three-dimensional printing - Assisted planning for complete and safe resection of retroperitoneal tumor.

BACKGROUND: Surgery is usually recommended to treat retroperitoneal tumors. However, complete surgical resections often remain challenging. OBJECTIVE: To assess the assistant role of three-dimensional (3D) imaging and printing model in retroperitoneal tumor resection, as well as compare the difference between 3D printing and computed tomography (CT) in preoperative planning and confidence building. METHODS: We admitted a patient with retroperitoneal mass (13.0×6.4×14.8 cm) adjacent to important abdominal blood vessels whose surgery was thought to be difficult. 3D printing and CT was arranged. A novel questionnaire and scoring system consisting of surgery difficulty and safety were designed to compare doctors understanding and confidence for surgery based on 3D printing and CT. Twenty-four doctors completed the scoring table based on CT and then 3D imaging, respectively. Paired t-test was applied for statistics analysis. RESULTS: Preoperative evaluation based on 3D printing indicated that the tumor could be removed completely. The operation lasted 120 minutes to successfully remove the tumor and the estimated blood loss was less than 100 ml. Scores based on 3D printing is significantly higher than CT in difficulty and safety of surgery (p < 0.001). Interestingly, the junior doctors seem to benefit more from 3D printing than the senior doctors. CONCLUSIONS: 3D imaging and printing model provides greater help for preoperative planning and confidence building than using CT in resection of retroperitoneal tumor, especially for the junior doctors.

Gamma-Ray Irradiation to Accelerate Crystallization of Mesoporous Zeolites.

Gamma-ray (γ-ray) irradiation was introduced into zeolite synthesis. The crystallization process of zeolite NaA, NaY, Silicalite-1, and ZSM-5 were greatly accelerated. The crystallization time of NaA zeolite was significantly decreased to 18 h under γ-ray irradiation at 20 °C, while more than 102 h was needed for the conventional process. Unexpectedly, more mesopores were created during this process, and thus the adsorption capacity of CO2 increased by 6-fold compared to the NaA prepared without γ-ray irradiation. Solid experimental evidence and density function theory (DFT) calculations demonstrated that hydroxyl free radicals (OH*) generated by γ-rays accelerated the crystallization of zeolite NaA. Besides NaA, mesoporous ZSM-5 with MFI topology was also successfully synthesized under γ-ray irradiation, which possessed excellent catalytic performance for methanol conversion, suggesting the universality of this new synthetic strategy for various zeolites.

Regulation of glucose metabolism by p62/SQSTM1 through HIF1α.

The signaling adaptor sequestosome 1 (SQSTM1)/p62 is frequently overexpressed in tumors and plays an important role in the regulation of tumorigenesis. Although great progress has been made, biological roles of p62 and relevant molecular mechanisms responsible for its pro-tumor activity remain largely unknown. Here, we show that p62 knockdown reduces cell growth and the expression of glycolytic genes in a manner that depends on HIF1α activity in renal cancer cells. Knockdown of p62 decreases HIF1α levels and transcriptional activity by regulating mTORC1 activity and NF-κB nuclear translocation. Furthermore, p62 interacts directly with the von Hippel-Lindau (VHL) E3 ligase complex to modulate the stability of HIF1α. Mechanistically, p62 binds to the VHL complex and competes with HIF1α. Expression of p62 inhibits the interaction of DCNL1 (also known as DCUN1D1) with CUL2 and attenuates the neddylation of CUL2, and thus downregulates the VHL E3 ligase complex activity. Functionally, HIF1α expression is required for p62-induced glucose uptake, lactate production and soft agar colony growth. Taken together, our findings demonstrate that p62 is a crucial positive regulator of HIF1α, which is a facilitating factor in p62-enhanced tumorigenesis.

miR-490-5p suppresses tumour growth in renal cell carcinoma through targeting PIK3CA.

BACKGROUND INFORMATION: Dysregulated micro-RNAs have been reported in many human cancers, including renal cell carcinoma. Recent studies indicated that miR-490 is involved in tumour development and progression. However, the expression profile and function in renal cell carcinoma remains unknown. RESULTS: Herein, we showed that miR-490-5p was down-regulated in renal cell carcinoma tissues and cells compared with the adjacent normal tissues and normal cells. We also provided evidence that miR-490-5p acts as a tumour suppressor in renal carcinoma in a variety of in vitro and in vivo assays. Mechanistically, miR-490-5p was verified to directly bind to 3' UTR of the PIK3CA mRNA and reduce the expression of PIK3CA at both mRNA and protein levels, which further inhibits phosphatidylinositol 3-kinase/Akt signalling pathway. We further showed that knockdown of PIK3CA can block the growth inhibitory effect of miR-490-5p, and over-expression of PIK3CA can reverse the inhibitory effect of miR-490-5p on renal cancer cell tumourigenicity. CONCLUSIONS: Taken together, our results indicated for the first time that miR-490-5p functions as a tumour suppressor in renal carcinoma by targeting PIK3CA. SIGNIFICANCE: Our findings suggest that miR-490-5p may be a potential gene therapy target for the treatment of renal cell carcinoma.

Laparoscopic cholecystectomy under spinal anaesthesia vs. general anaesthesia: a meta-analysis of randomized controlled trials.

BACKGROUND: Laparoscopic Cholecystectomy (LC) is conventionally performed under general anaesthesia (GA), but there are multiple studies which have found spinal anaesthesia (SA) as a safe alternative. This meta-analysis was performed after adding many recent randomized controlled trials (RCTs) to clarify this issue. METHODS: Relevant articles published in English were identified by searching PubMed, Embase, Web of Knowledge, and the Cochrane Controlled Trial Register from January 1, 2000 to December 1, 2014. Reference lists of the retrieved articles were reviewed to identify additional articles. Primary outcomes (postoperative pain scores) and secondary outcomes (operating time (OT) and postoperative complications) were pooled. Quantitative variables were calculated using the weighted mean difference (WMD), and qualitative variables were pooled using odds ratios (OR). RESULTS: Seven appropriate RCTs were identified from 912 published articles. Seven hundred and twelve patients were treated, 352 in SA group and 360 in GA group. LC under SA was superior to LC under GA in postoperative pain within 12 h (visual analogue score (VAS) in 2-4 h, WMD = -1.61, P = 0.000; VAS in 6-8 h, WMD = -1.277, P = 0.015) and postoperative complications (postoperative nausea and vomiting (PONV) WMD = 0.427, P = 0.001; Overall Morbidity WMD = 0.691, P = 0.027). The GA group was superior to SA group in postoperative urinary retention (WMD = 4.273, P = 0.022). There were no significant differences in operating time (WMD = 0.184, P = 0.141) between two groups. CONCLUSIONS: SA as the sole anaesthesia technique is feasible, safe for elective LC.

Towards a dynamic clamp for neurochemical modalities.

The classic dynamic clamp technique uses a real-time electrical interface between living cells and neural simulations in order to investigate hypotheses about neural function and structure. One of the acknowledged drawbacks of that technique is the limited control of the cells' chemical microenvironment. In this manuscript, we use a novel combination of nanosensor and microfluidic technology and microfluidic and neural simulations to add sensing and control of chemical concentrations to the dynamic clamp technique. Specifically, we use a microfluidic lab-on-a-chip to generate distinct chemical concentration gradients (ions or neuromodulators), to register the concentrations with embedded nanosensors and use the processed signals as an input to simulations of a neural cell. The ultimate goal of this project is to close the loop and provide sensor signals to the microfluidic lab-on-a-chip to mimic the interaction of the simulated cell with other cells in its chemical environment.

Cisplatin-induced epigenetic activation of miR-34a sensitizes bladder cancer cells to chemotherapy.

BACKGROUND: Accumulating evidence suggests a tumor suppressive role for miR-34a in human carcinogenesis. However, its precise biological role remains largely elusive. This study aimed to reveal the association of the miR-34a expression and its modulation of sensitivity to cisplatin in muscle-invasive bladder cancer (MIBC). METHODS: miR-34a expression in MIBC cell lines and patient tissues was investigated using qPCR. The methylation analysis of miR-34a promoter region was performed by MassARRAY. Synthetic short single or double stranded RNA oligonucleotides and lentiviral vector were used to regulate miR-34a expression in MIBC cells to investigate its function in vitro and in vivo. RESULTS: miR-34a expression was frequently decreased in MIBC tissues and cell lines through promoter hypermethylation while it was epigenetically increased in MIBC cells following cisplatin treatment. Increased miR-34a expression significantly sensitized MIBC cells to cisplatin and inhibited the tumorigenicity and proliferation of cancer cells in vitro and in vivo. Furthermore, we identified CD44 as being targeted by miR-34a in MIBC cells following cisplatin treatment, and increased CD44 expression could efficiently reverse the effect of miR-34a on MIBC cell proliferation, colongenic potential and chemosensitivity. CONCLUSIONS: Cisplatin-based chemotherapy induced demethylation of miR-34a promoter and increased miR-34a expression, which in turn sensitized MIBC cells to cisplatin and decreased the tumorigenicity and proliferation of cancer cells that by reducing the production of CD44.

miRNA-34a suppresses cell proliferation and metastasis by targeting CD44 in human renal carcinoma cells.

PURPOSE: We investigated the potential functions of miR-34a in CD44 transcriptional complexes in renal cell carcinoma. MATERIALS AND METHODS: We detected miR-34a expression by quantitative real-time polymerase chain reaction. Oligonucleotides were used to over express miR-34a. Cell proliferation and xenograft assays, colony formation and flow cytometry were done to examine effects on cancer cell proliferation in vitro and in vivo. Luciferase assay was performed to verify the precise target of miR-34a. RESULTS: Promoter methylation contributed to miR-34a loss in the ACHN, 786-O and SN12PM6 renal carcinoma cell lines. Ectopic over expression of miR-34a restrained cell growth, tube formation and migration/invasion, and significantly suppressed the growth of renal carcinoma xenografts and metastasis in nude mice. Dual luciferase assay revealed that CD44 was a direct target of miR-34a in renal cancer cells and CD44 knockdown by RNAi in renal cancer cells suppressed tumor progression. In contrast, CD44 ectopic expression partially reversed the antitumor effects of miR-34a in renal cancer cells. CONCLUSIONS: Our findings indicate that miR-34a targets CD44 in renal cancer cells and suppresses renal cancer cell growth, tube formation and metastasis in vitro and in vivo. Thus, miR-34a may be a potential molecular target for novel therapeutic strategies for clear cell renal carcinoma.

Fibulin-1 is epigenetically down-regulated and related with bladder cancer recurrence.

BACKGROUND: Bladder cancer is one of the most common cancers worldwide. Fibulin-1, a multi-functional extracellular matrix protein, has been demonstrated to be involved in many kinds of cancers, while its function in bladder cancer remains unclear. So here we investigated the expression and function of fibulin-1 in Bladder cancer. METHODS: We used real-time PCR, Western blot analysis and immunohistochemistry to determine the expression of fibulin-1 in Bladder cancer cells and patient tissues respectively. Methylation-specific PCR and quantitative sequencing were used to examine the methylation status of FBLN1 gene promoter. Eukaryotic expression plasmid and lentiviral vector were used to overexpress fibulin-1 in Bladder cancer cells 5637, HT-1376 to investigate its function in vitro and in vivo. RESULTS: We identified that fibulin-1 was significantly down-regulated in bladder cancer, and its dysregulation was associated with non-muscle-invasive bladder cancer (NMIBC) grade and recurrence. The promoter region of FBLN1 was generally methylated in bladder cancer cell lines and tissues, further investigation in patient tissues showed that the methylation status was associated with the fibulin-1 expression. Overexpression of fibulin-1 significantly suppressed tumor growth, induced tumor cell apoptosis, decreased cell motility, and inhibited angiogenesis in cultured bladder cancer cells and xenograft tumor in nude mice. CONCLUSIONS: Altogether, our results indicated that fibulin-1 expression is associated with NMIBC grade and recurrence, it is epigenetically down-regulated and functions as a tumor suppressor gene and angiogenesis inhibitor in bladder cancer.