Generation of a control induced pluripotent stem cell line (SDUCHi001-A) from a healthy male donor.

An induced pluripotent stem cell (iPSC) line (SDUCHi001-A) was established using peripheral blood mononuclear cells (PBMCs) from a healthy 6 years old boy. Reprogramming of the PBMCs was achieved through non-integrating delivery of OCT4, SOX2, KFL4, BCL-XL, and c-MYC. The iPSC line expressed pluripotency markers, had a normal karyotype and trilineage differentiation potential.

Biomass 3D Printing: Principles, Materials, Post-Processing and Applications.

Under the background of green and low-carbon era, efficiently utilization of renewable biomass materials is one of the important choices to promote ecologically sustainable development. Accordingly, 3D printing is an advanced manufacturing technology with low energy consumption, high efficiency, and easy customization. Biomass 3D printing technology has attracted more and more attentions recently in materials area. This paper mainly reviewed six common 3D printing technologies for biomass additive manufacturing, including Fused Filament Fabrication (FFF), Direct Ink Writing (DIW), Stereo Lithography Appearance (SLA), Selective Laser Sintering (SLS), Laminated Object Manufacturing (LOM) and Liquid Deposition Molding (LDM). A systematic summary and detailed discussion were conducted on the printing principles, common materials, technical progress, post-processing and related applications of typical biomass 3D printing technologies. Expanding the availability of biomass resources, enriching the printing technology and promoting its application was proposed to be the main developing directions of biomass 3D printing in the future. It is believed that the combination of abundant biomass feedstocks and advanced 3D printing technology will provide a green, low-carbon and efficient way for the sustainable development of materials manufacturing industry.

A hybrid coded-aperture and Compton camera based on cerium-doped Gd3Al2Ga3O12 scintillators coupled with multi-pixel photon counter arrays.

In this study, we propose a hybrid coded-aperture and Compton camera based on cerium-doped Gd3Al2Ga3O12 (GAGG:Ce) scintillator arrays coupled with Multi-Pixel Photon Counter (MPPC) arrays. The sensitive detector of the gamma camera consists of a single GAGG:Ce crystal coupled with a single-chip MPPC unit module. An impedance bridge circuit and a 64-channel data acquisition system were employed to record the code-aperture events and Compton coincidence events. After the calibration of position and energy, the total energy resolution for 662 keV gamma-rays from 137Cs was 6.6%. The hybrid camera had the characteristics of mechanical collimation and electronic collimation at the same time. In the code aperture mode, the reconstructed images were obtained by direct deconvolution and maximum likelihood expectation maximization (MLEM) methods. In the Compton imaging mode, the energy-dependent method was applied to order the sequence of Compton scatter events. The simple back-projection algorithm and list-mode MLEM algorithm were adopted for image reconstruction. Practical performances demonstrated that the angular resolutions in two modes were measured as 5.2° and 11.4°, respectively. In addition, the hybrid camera had a desirable imaging capability in a wide energy range (32 keV-2.6 MeV) and a wide field of view (∼210° in the horizontal direction). As for the sensitivity, the camera had a commercially available sensitivity level of localizing a 137Cs point source, producing ∼0.026 µSv/h in 5 min. Furthermore, the function of distinction for different radiation sources was preliminarily realized.

A new radionuclide identification method for low-count energy spectra with multiple radionuclides.

Radionuclide identification is to recognize the radionuclides in the environment by analyzing the energy spectrum. Rapid and accurate identification is important for nuclear security. Current radionuclide identification methods based on traditional peak search require background subtraction. As a result, they have difficulties to deal with complex situations in practical applications such as low-count energy spectrum and mixed nuclides. In this paper, we propose a new radionuclide identification method with a feature enhancer and a one-dimensional neural network. The training dataset in this method is from simulated data generated by Geant4. By preprocessing the input energy spectrum data through the feature enhancer and extracting the nonlinear information through the neural network, this approach performs well on experimental energy spectra even at low count. The method also shows a high recognition accuracy and little misjudgments when dealing with mixed radionuclides spectra. Due to its good performance in identifying mixed nuclides and low-count spectra, the method has been deployed in portable instrument for radionuclide identification in real-time measurement.

Fat mass- and obesity-associated (FTO) gene promoted myoblast differentiation through the focal adhesion pathway in chicken.

The action of FTO on myoblasts proliferation and differentiation and molecular mechanism underlying it were investigated by transfecting with FTO lentiviral overexpression vector and gene expression profile sequencing. Compared with the control group, myoblasts with FTO transfection was significantly enhanced proliferation; the expression of MYOG and MYOD mRNA was significantly increased. In cells transfected with FTO, 129 differentially expressed genes were determined compared with control group, with 104 up-regulated and 25 down-regulated genes. Twelve pathways (Phagosome, Focal adhesion, Adrenergic signaling in cardiomyocytes, Endocytosis, Cardiac muscle contraction, Toll-like receptor, Ribosome, Tight junction, Regulation of actin cytoskeleton, Cytokine-cytokine receptor interaction, Adrenergic signaling in cardiomyocytes and MAPK) were significantly enriched. Eight genes known to be directly or indirectly related to skeletal muscle development (LAMA5, SPP1, CAV3, RASGRF1, FAK, PDGFB, PDGFRα, and RAC2) were enriched in the focal adhesion and expressed differentially. Altogether, these data suggested that FTO stimulated differentiation of myoblasts through regulation of eight genes enriched in the focal adhesion.

Novel compact high-speed passive gamma-ray assay equipment for uranium enrichment measurement of fuel rods.

Uranium enrichment measurement is an essential quality inspection for fuel rods before delivery to users. Generally, compared with active neutron assay (ANA) equipment, passive gamma-ray assay (PGA) equipment is more economical and safer. However, the current PGA equipment based on photomultipliers is too slow (1 m/min) to meet the growing needs in China. Recently, we have developed a set of compact high-speed PGA equipment including four detection modules (128 units in total), a 128-channel data acquisition system (DAS), a power supply, special software, and an automatic loading and unloading mechanism. The detection unit is based on silicon photomultipliers in virtue of its compact size and good performance. The DAS processes signals of all units in parallel into a sequence of data packets carrying the energy information and the corresponding unit ID. The software integrates the data packets into a fluctuating count curve in a time-delay superposition method and identifies possible abnormal pellets. After calibrations, our equipment can locate abnormal pellets accurately at a speed of 6 m/min. In addition, it can directly measure the enrichment of fresh pellets not in secular equilibrium without waiting for two months. So far, the equipment has been successfully run for one year on the assembly line of China North Nuclear Fuel Co. and shows good potential to replace the traditional ANA equipment.

SSeCKS promoted lipopolysaccharide-sensitized astrocytes migration via increasing ß-1,4-galactosyltransferase-I activity.

Astrocytes migration is essential in the formation of the glial scar during the injury response process of the central nervous system (CNS) especially during inflammation. Integrin ß1 is part of the extracellular matrix receptors in the CNS and it has been reported that integrin ß-deficient astrocytes randomly migrate into wounds. Previous studies have found that ß-1,4 Galactosyltransferase-I (ß-1,4-GalT-I) enhanced the ß-1,4-galactosylation of integrin ß1. Src-suppressed C kinase substrate (SSeCKS) is an inflammatory response protein which functionally interacts with ß-1,4 Galactosyltransferase-I (ß-1,4-GalT-I). In this study we aim to investigate the role of SSeCKS and ß-1,4-GalT-I in the migration of astrocytes during lipopolysaccharide (LPS)-induced inflammation. Coimmunoprecipitation and immunofluorescence assays have demonstrated that SSeCKS and ß-1,4-GalT-I were significantly enhanced in LPS-treated astrocytes and their interactions may occur in the Trans-Golgi Network. Lectin blot showed that the knockdown of ß-1,4-GalT-I could inhibit the ß-1,4-galactosylation of glycoproteins including integrin ß1 with and without LPS, and that SSeCKS knockdown inhibits the ß-1,4-galactosylation of glycoproteins including integrin ß1 only in LPS-induced astrocytes. Additionally, wound healing assays indicated that ß-1,4-GalT-I knockdown could inhibit astrocytes migration with and without LPS but SSeCKS inhibited cell migration only when LPS was present. Therefore our findings suggest that SSeCKS affects astrocytes migration by regulating the ß-1,4-galactosylation of glycoproteins including integrin ß1, via ß-1,4-GalT-I expression in LPS-sensitized astrocytes.

[The expression and role of MIF, NF-κB and IL-1ß in nasal polyps].

OBJECTIVE: To investigate the expression of MIF, NF-κB p65 and IL-1ß in the tissue of nasal polyps and normal inferior turbinate, to analyze their relevance, and to explore their role in nasal polyps. METHOD: The infiltrating results of EOS and others inflammatory cells in 48 cases diagnosed as nasal polyps (nasal polyps group) were detected by HE staining, and the expression of MIF, NF-κB p65 and IL-1ß were investigated by immunohistochemistry. Twenty-one patients who were performed septoplasty orthotics were included as the control group; the VAS and Lund-Kennedy score were used to evaluate the degree of nasal polyps in patients and the correlation analysis was conducted between the disease severity and the expression levels of this three factors. RESULT: (1) The infiltrating results of EOS and the expression level of MIF, NF-κB p65, IL-1ß in nasal polyps group are obviously higher than these in the control group (P < 0.05); Spearman correlation analysis shows that MIF, NF-κb p65 and IL-1ß are positively correlated with each other (r = 0.74, 0.66, 0.60, P < 0.05); the nuclear activation rate of NF-κB p65 is positively correlated with MIF, IL-1ß (r = 0.67, 0.63, P < 0.05); the infiltration degree of EOS is positively correlated with MIF, IL-1ß (r = 0.49, 0.55, P < 0.05), but has no correlation with the NF-κB p65 and its nuclear activation rate. (2) The VAS grade of the nasal polyps group is 8.24 ± 1.72 and the nasal endoscopic examination grade is 8.63 ± 3.81. Spearman correlation analysis shows that the VAS grade is positively correlated with the level of MIF (r = 0.71, P < 0.05), but had no correlation with NF-κB p65, its nuclear activation rate and IL-1ß. The nasal endoscopic examination grade is positively correlated with MIF and the nuclear activation rate of NF-κB p65 (r = 0.79, 0.73, P < 0.05), but has no correlation with the level of NF-κB p65 and IL-1ß (P > 0.05). CONCLUSION: MIF, NF-κB p65 and IL-1ß may promote the development of the nasal polyps, and there may exist the IL-1ß--NF-κB--MIF approach in nasal polyps; MIF and NF-κB may participate in maintaining physiological function of inferior turbinate and have relations with the lightest sustained inflammation of inferior turbinate. The MIF and NF-κB p65 nuclear activation rate can be used as a standard of the nasal polyp severity and the judgement prognosis.

A functional module-based exploration between inflammation and cancer in esophagus.

Inflammation contributing to the underlying progression of diverse human cancers has been generally appreciated, however, explorations into the molecular links between inflammation and cancer in esophagus are still at its early stage. In our study, we presented a functional module-based approach, in combination with multiple data resource (gene expression, protein-protein interactions (PPI), transcriptional and post-transcriptional regulations) to decipher the underlying links. Via mapping differentially expressed disease genes, functional disease modules were identified. As indicated, those common genes and interactions tended to play important roles in linking inflammation and cancer. Based on crosstalk analysis, we demonstrated that, although most disease genes were not shared by both kinds of modules, they might act through participating in the same or similar functions to complete the molecular links. Additionally, we applied pivot analysis to extract significant regulators for per significant crosstalk module pair. As shown, pivot regulators might manipulate vital parts of the module subnetworks, and then work together to bridge inflammation and cancer in esophagus. Collectively, based on our functional module analysis, we demonstrated that shared genes or interactions, significant crosstalk modules, and those significant pivot regulators were served as different functional parts underlying the molecular links between inflammation and cancer in esophagus.

The role of Cyclin G1 in cellular proliferation and apoptosis of human epithelial ovarian cancer.

Cyclin G1 plays an essential role in the development of human carcinoma. Here, we characterized the clinical significance of Cyclin G1 and investigated its role in cellular proliferation and apoptosis of epithelial ovarian cancer (EOC). Western blot was used to evaluate the expression of Cyclin G1 in nine fresh EOC tissues and three fresh normal ovarian tissues. Immunohistochemistry analysis was performed on formalin-fixed paraffin-embedded section of 119 cases of EOCs. Using cell counting kit (CCK)-8 and colony formation assays, we analyzed the effect of Cyclin G1 in cellular proliferation of EOC. Besides, the immunofluorescence and flow cytometry analysis was performed to study the role of Cyclin G1 in cellular apoptosis of EOC. We found Cyclin G1 was up-regulated in EOC tissues compared with the normal ovary tissues. Cyclin G1 expression in EOC was closely correlated with differentiation grade (P = 0.009) and malignant tumor cells in ascites (P = 0.009). The Kaplan-Meier curve showed that higher expression of Cyclin G1 was associated with significantly shorter survival in EOC patients. Multivariate analysis suggested Cyclin G1 expression was an independent prognostic factor for overall survival. CCK-8 and colony formation assays revealed that depletion of Cyclin G1 inhibited the proliferation and clone formation. Combined immunofluorescence and flow cytometry analysis showed that silencing of Cyclin G1 with shRNA could promote apoptosis of ovarian cancer cells. Additionally, the result of immunoprecipitation test showed Cyclin G1 interacted with CDK2 in EOC cells. In summary, our findings suggest that Cyclin G1 may be involved in the prognosis of EOC patients and be a useful therapeutic target for EOC.

Global gene expression distribution in non-cancerous complex diseases.

For gene expression in non-cancerous complex diseases, we systemically evaluated the sensitivities of biological discoveries to violation of the common normalization assumption. Our results indicated that gene expression may be widely up-regulated in digestive system and musculoskeletal diseases. However, global signal intensities showed little difference in other four disease types.

Early mitotic inhibitor-1, an anaphase-promoting complex/cyclosome inhibitor, can control tumor cell proliferation in hepatocellular carcinoma: correlation with Skp2 stability and degradation of p27(Kip1).

Early mitotic inhibitor-1 (Emi1) is a key cell-cycle regulator that promotes S-phase and M-phase entry by inhibiting anaphase-promoting complex/cyclosome (APC/C) activity. Immunohistochemical analysis was performed in 114 human hepatocellular carcinoma (HCC) samples, and the data were correlated with clinicopathologic features. Univariate and multivariate survival analyses were performed to determine the prognostic significance of the proteins. Expression of Emi1 correlated directly with the stage of HCC. More importantly, high expression of Emi1 was associated with a poor outcome. Western blot analysis showed that Emi1 was highly expressed in HCC compared with the adjacent noncancerous tissue. In vitro, after the release of HCC cell lines from serum starvation, the expression of Emi1 APC/C substrates (cyclins A, B) and Skp2 was up-regulated, whereas p27(Kip1) was down-regulated. In addition, we used small interfering RNA to knock out Emi1 expression and observed its effects on HCC growth in vitro to determine whether loss of Emi1 could inhibit cell proliferation by blocking S-phase and mitotic entry. Western blot analyses indicated that deletion of Emi1 was positively correlated with APC/C substrates (cyclins A, B) and Skp2 but was negatively correlated with p27(Kip1). Emi1 inhibits APC/C activity, whereas Skp2 degradation is mediated by APC/C, and degradation of Skp2 can stabilize p27(kip1). These results suggested that Emi1 participates in HCC cell proliferation and that progression is controlled by APC/C inhibition, which stabilized Skp2 and enabled p27(kip1) degradation. These findings provide a potential therapeutic strategy for HCC.

Overexpression of forkhead box J2 can decrease the migration of breast cancer cells.

The prognosis of breast cancer patients with metastases is generally poor, so it is essential to elucidate related molecules mechanisms. Forkhead Box J2 (FOXJ2) is a member of Forkhead Box transcription factors, many of which have been reported to participate in tumor migration and invasion. In this study, we showed the expression of FOXJ2 was higher in primary breast cancer tissues without lymph nodes metastases than those with, and there was statistical significance between the expression of FXOJ2 and the clinical factors. Hence, we identified a novel function of metastasis, which was not previously known for FOXJ2. Overexpression of FOXJ2 decreased the motility property of highly migrative MDA-MB-231 cells in vitro by wound healing assays and trans-well migration assays, and it was concurrent with the increased expression of epithelial marker E-cadherin and the decreased expression of mesenchymal marker vimentin by Western blot analysis, reverse transcription PCR analysis, and immunofluorescence analysis. Consistent with these observations, the repression of FOXJ2 in weakly metastatic MCF-7 cells remarkably promoted cellular motility. Our study demonstrates that FOXJ2 can inhibit the metastasis of human breast cancer by regulating the EMT key markers E-cadherin and vimentin.

Involvement of CtBP2 in LPS-induced microglial activation.

CtBP2 (C-terminal binding protein 2), which is widely expressed during developmental processes and differentiation, acts as a transcriptional repressor following recruitment to target promoters through repressors or other co-repressor proteins. In this study, we elucidated the dynamic expression changes and localization of CtBP2 in lipopolysaccharide (LPS)-induced neuroinflammatory processes in adult rats. CtBP2 expression was strongly induced in active glia cells (microglia and astrocytes) in inflamed spinal cord. In vitro studies indicated that the up-regulation of CtBP2 may be involved in the subsequent microglia activation following LPS exposure. And the knock-down of CtBP2 in microglia cell line HAPI by siRNA showed that CtBP2 increased the activation of microglia induced by LPS. Collectively, these results suggested CtBP2 may be important in host defense in microglia-mediated immune response. Understanding the cell signal pathway may provide a novel strategy against inflammatory and immune reaction in neuroinflammation in central nervous system.

ß-1,4-Galactosyltransferase I involved in Schwann cells proliferation and apoptosis induced by tumor necrosis factor-alpha via the activation of MAP kinases signal pathways.

ß-1,4-galactosyltransferase-I (ß-1,4-GalT-I) plays a critical role in the initiation and maintenance of peripheral nervous system inflammatory reaction. However, the exact function of ß-1,4-GalT-I in the regulation of SCs proliferation and apoptosis remains unclear. In this study, we found that low concentration of tumor necrosis factor-alpha (TNF-α) induced SCs proliferation, while high concentration of TNF-α induced SCs apoptosis. Meanwhile, the expressions of ß-1,4-GalT-I, TNFR1, and TNFR2 were changed following. When ß-1,4-GalT I overexpression, low concentration of TNF-α-induced SCs proliferation was partially repressed. Concurrently, the activity of ERK1/2 was decreased. While knocking down ß-1,4-GalT I expression, high concentration of TNF-α-induced SCs apoptosis was partially rescued. Consistent with this, the activity of P38 and JNK were decreased. We also found anti-TNFR2 antibody suppressed low concentration of TNF-α-induced SCs proliferation, while anti-TNFR1 antibody inhibited high concentration of TNF-α-induced SCs apoptosis. Thus, present data show that ß-1,4-GalT I may play an important role in SCs proliferation and apoptosis induced by TNF-α via different signal pathways and TNFR.

Changes in Pirh2 and p27kip1 expression following traumatic brain injury in adult rats.

Pirh2, a p53-induced ubiquitin-protein ligase, has been reported to promote ubiquitin-dependent degradation of p27(kip1), which plays an essential role in mammalian cell cycle regulation and neurogenesis in the developing central nervous system (CNS). However, their distributions and functions in the nervous system lesion and repair remain unclear. In this study, we observed that the up-regulated expression of Pirh2 was concomitant with decreased p27(kip1) level after traumatic brain injury by Western blot and immunohistochemistry. Immunofluorescence double-labeling revealed that Pirh2 was mainly co-expressed with GFAP and CD11b. Meanwhile, we also examined the expression profiles of proliferating cell nuclear antigen (PCNA) whose changes were correlated with the expression of Pirh2. In addition, Pirh2 colocalized with p27(kip1) and PCNA. Immunoprecipitation further showed that they interacted with each other in the pathophysiology process. In summary, our data indicated Pirh2 might be a negative regulator of p27(kip1) and associated with glial proliferation.

Increased expression of transcription initiation factor IIB after rat traumatic brain injury.

The protein TFIIB is a general transcription initiation factor that plays a pivotal role in the preinitiation complex (PIC) and selects the transcription initiation site. However, its distribution and function in the central nervous system (CNS) remains unclear. In the present study, we mainly investigated the expression and cellular localization of TFIIB during traumatic brain injury (TBI). Western blot analysis revealed that TFIIB was present in normal rat brain cortex. It gradually increased, reached a peak at the 5th day after TBI, and then decreased. Importantly, more TFIIB was colocalized with astrocytes and microglia, which are largely proliferated. In addition, Western blot detection showed that the 5th day post injury was also the proliferation peak indicated by the elevated expression of PCNA. Importantly, injury-induced expression of TFIIB was colabelled by proliferating cell nuclear antigen (proliferating cells marker). These data suggested that TFIIB may be implicated in the proliferation of astrocytes and microglia and the recovery of neurological outcomes. But the inherent mechanisms remained unknown. Further studies are needed to confirm the exact role of TFIIB after brain injury.

The relationship between Src-suppressed C kinase substrate and ß-1,4 galactosyltransferase-I in the process of lipopolysaccharide-induced TNF-α secretion in rat primary astrocytes.

Src-suppressed C kinase substrate (SSeCKS), a protein kinase C substrate, is a major lipopolysaccharide (LPS) response protein. In addition, ß-1,4 Galactosyltransferase-I (ß-1,4-GalT-I) also plays an important role in the inflammation reactions of nervous system. It was reported that both SSeCKS and ß-1,4-GalT-I were involved in the LPS-induced tumor necrosis factor-alpha (TNF-α) expression in rat primary astrocytes. However, the functional interaction between SSeCKS and ß-1,4-GalT-I in the LPS-induced TNF-α secretion remains unclear. Therefore, in this study, using the inflammation model of astrocytes treated by LPS in vitro, we found that the changed expressions of SSeCKS and ß-1,4-GalT-I participated in LPS-induced TNF-α secretion through p38, JNK, and ERK signal transduction pathways in rat primary astrocytes. Knockdown by small-interfering RNAs (siRNAs) or overexpression of SSeCKS and ß-1,4-GalT-I could influence Mitogen-activated protein kinases (MAPKs) signaling pathways activation and TNF-α secretion. Besides, we confirmed that knockdown of SSeCKS could prevent the induction of ß-1,4-GalT-I in this process. Inversely, ß-1,4-GalT-I had no significant effect on SSeCKS expression in the same way. In summary, our data indicated that SSeCKS could regulate LPS-induced TNF-α secretion through ß-1,4-GalT-I in rat primary astrocytes.

ß-1,4-galactosyltransferase I promotes tumor necrosis factor-α autocrine via the activation of MAP kinase signal pathways in Schwann cells.

Recent studies have demonstrated that aberrant galactosylation is associated with some inflammation diseases. ß-1,4-Galactosyltransferase-I (ß-1,4-GalT I), which transferred galactose to the terminal N-acetylglucosamine of N- and O-linked glycans in a ß-1,4-linkage, was considered to be the major galactosyltransferase among the seven members of the subfamily responsible for ß4 galactosylation. To elucidate the expression and possible function of ß-1,4-GalT I in the peripheral nervous system (PNS) inflammatory diseases, we performed a tumor necrosis factor-alpha (TNF-α) autocrine inflammatory model in Schwann cells (SCs). In this study, we found that silencing of ß-1,4-GalT I suppressed TNF-α autocrine, while overexpression of ß-1,4-GalT I promoted TNF-α autocrine in TNF-α-treated SCs. Meanwhile, anti-TNFR1 antibody suppressed the expression of ß-1,4-GalT I, and TNF-α autocrine. ß-1,4-GalT I conferred its effect by promoting ERK, JNK, and P38 MAP kinase signal pathways activation in TNF-α-induced SCs. Thus, the present data shows that during SCs activation, ß-1,4-GalT I may play an important role in the release of inflammatory mediators.