Caspase recruitment domain 6 protects against hepatic ischemia/reperfusion injury by suppressing ASK1.

BACKGROUND & AIMS: The hepatic injury caused by ischemia/reperfusion (I/R) insult is predominantly determined by the complex interplay of sterile inflammation and liver cell death. Caspase recruitment domain family member 6 (CARD6) was initially shown to play important roles in NF-κB activation. In our preliminary studies, CARD6 downregulation was closely related to hepatic I/R injury in liver transplantation patients and mouse models. Thus, we hypothesized that CARD6 protects against hepatic I/R injury and investigated the underlying molecular mechanisms. METHODS: A partial hepatic I/R operation was performed in hepatocyte-specific Card6 knockout mice (HKO), Card6 transgenic mice with CARD6 overexpression specifically in hepatocytes (HTG), and the corresponding control mice. Hepatic histology, serum aminotransferases, inflammatory cytokines/chemokines, cell death, and inflammatory signaling were examined to assess liver damage. The molecular mechanisms of CARD6 function were explored in vivo and in vitro. RESULTS: Liver injury was alleviated in Card6-HTG mice compared with control mice as shown by decreased cell death, lower serum aminotransferase levels, and reduced inflammation and infiltration, whereas Card6-HKO mice had the opposite phenotype. Mechanistically, phosphorylation of ASK1 and its downstream effectors JNK and p38 were increased in the livers of Card6-HKO mice but repressed in those of Card6-HTG mice. Furthermore, ASK1 knockdown normalized the effect of CARD6 deficiency on the activation of NF-κB, JNK and p38, while ASK1 overexpression abrogated the suppressive effect of CARD6. CARD6 was also shown to interact with ASK1. Mutant CARD6 that lacked the ability to interact with ASK1 could not inhibit ASK1 and failed to protect against hepatic I/R injury. CONCLUSIONS: CARD6 is a novel protective factor against hepatic I/R injury that suppresses inflammation and liver cell death by inhibiting the ASK1 signaling pathway. LAY SUMMARY: The protein CARD6 plays an important role during the process of liver blood flow restriction (ischemia) and restoration (reperfusion). By suppressing the activity of ASK1, CARD6 can protect against hepatocyte injury. Targeting CARD6 is a potential strategy for prevention and treatment of ischemia/reperfusion injury.

Dusp14 protects against hepatic ischaemia-reperfusion injury via Tak1 suppression.

BACKGROUND & AIMS: Hepatic ischaemia-reperfusion (I/R) injury is characterised by severe inflammation and extensive cell death. Multiple signalling pathways, including NF-κB and mitogen-activated protein kinase (MAPK)/c-Jun NH2-terminal kinase (JNK), have important roles in this process. Identifying the unknown critical regulators of these signalling pathways could provide potential targets for therapeutic application. Dual-specificity phosphatase 14 (DUSP14) acts as a negative regulator of NF-κB signalling. However, its function in hepatic I/R injury is unknown. METHODS: Hepatocyte-specific Dusp14 knockout (HKO) and transgenic (TG) mice were subjected to hepatic I/R surgery to examine Dusp14 function in vivo. Primary hepatocytes isolated from Dusp14-HKO and Dusp14-TG mice were cultured and subjected to hypoxia/reoxygenation insult in vitro. Inflammatory cytokine production was measured using quantitative reverse transcription PCR and ELISA. Liver damage was analysed using histopathology. Co-immunoprecipitation and pull-down assays followed by Western blot were performed to detect Dusp14 and transforming growth factor (Tgf)-ß-activated kinase 1 (Tak1) interactions. RESULTS: Dusp14 was significantly downregulated in liver tissues from liver transplantation patients and mice subjected to hepatic I/R surgery. Dusp14-HKO and Dusp14-TG mouse models demonstrated that Dusp14 reduced cell death, ameliorated inflammation, and promoted hepatocyte proliferation and/or regeneration. Dusp14 also suppressed NF-κB and MAPK signalling via a physical interaction with Tak1, leading to its subsequent inhibition. Tak1 inhibition by 5Z-7-ox abolished Dusp14 function in vivo, indicating that TAK1 is required for Dusp14 function in hepatic I/R injury. Finally, mutant Dusp14 lost the ability to bind Tak1 and failed to protect against hepatic I/R injury. CONCLUSIONS: Dusp14 is a protective factor in hepatic I/R injury, and the Dusp14-Tak1-Jnk1/2 regulatory axis is important for the pathogenesis of hepatic I/R injury. Modulation of this axis could be a novel therapeutic strategy to prevent or interfere with this pathological process. LAY SUMMARY: Reductions in the level of the protein Dusp14 are closely associated with liver damage caused by inadequate blood supply followed by restoration of blood flow to the liver. Dusp14 protects against liver damage by suppressing the activity of Tak1. Targeting Dusp14 could be a strategy for prevention and treatment of this disease.

Nestin+ Peyer's patch resident MSCs enhance healing of inflammatory bowel disease through IL-22-mediated intestinal epithelial repair.

Inflammatory bowel disease (IBD) is a chronic condition characterized by gastrointestinal tract inflammation and still lacks satisfactory treatments. Mesenchymal stromal cells (MSCs) show promising potential for treating IBD, but their therapeutic efficacy varies depending on the tissue of origin. We aim to investigate whether intestine Peyer's patch (PP)-derived MSCs have superior immunomodulatory effects on T cells and better therapeutic effects on IBD compared with bone marrow-derived MSCs. We isolated PPs-derived Nestin+ MSCs (MSCsPP ) and bone marrow-derived Nestin+ MSCs (MSCsBM ) from Nestin-GFP transgenic mice to explore their curative effects on murine IBD model. Moreover, we tested the effects of IL-22 knockdown and IL-22 overexpression on the therapeutic efficacy of MSCsPP and MSCsBM in murine IBD, respectively. We demonstrated that Nestin+ cells derived from murine PPs exhibit MSC-like biological characteristics. Compared with MSCsBM , MSCsPP possess enhanced immunoregulatory ability to suppress T cell proliferation and inflammatory cytokine production. Moreover, we observed that MSCsPP exhibited greater therapeutic efficacy than MSCsBM in murine IBD models. Interestingly, IL-22, which was highly expressed in MSCsPP , could alleviate the severity of the intestinal inflammation, while knockdown IL-22 of MSCsPP remarkably weakened the therapeutic effects. More importantly, IL-22 overexpressing MSCsBM could significantly improve the symptoms of murine IBD models. This study systemically demonstrated that murine MSCsPP have a prominent advantage in murine IBD treatment, partly through IL-22.

Measurement of density profile and fluctuations using a multi-channel terahertz solid-state interferometer system on Keda Torus eXperiment (KTX).

A five-chord interferometer based on terahertz solid state sources has been successfully installed on the Keda Torus eXperiment (KTX), a reversed field pinch machine. The optical design has been carefully optimized for the uniform distribution of beam light to fully use the limited power source (∼2 mW). By setting the telescopic mirror unit, the beam waist is located in the center of the vacuum vessel and its diameter is in the range of the Rayleigh length. The beam width across the plasma area is improved to ∼20 mm to minimize crosstalk and beam energy loss. After careful beam alignment, the phase noise for each channel can reach 0.004π. The radial profiles of electron density on the KTX are inverted, and density fluctuation associated with instabilities is shown based on the forward-scattering signals.

Long-time measurements of line-integrated plasma electron density using a two-color homodyne optical fiber interferometer.

A two-color homodyne Mach-Zehnder (M-Z) optical fiber interferometer with wavelengths of 1.55 and 1.31 µm was developed for long-time measurement of line-integrated plasma electron density. A novel phase difference demodulation algorithm based on a single 3 × 3 optical coupler was implemented in a two-color optical fiber interferometer scheme for the first time. Our laboratory tests showed that this new optical fiber interferometer could determine the phase shift due to the low-frequency ambient vibration and could maintain high phase resolution measurement. The resolution of the new interferometer was less than 0.04 rad in 1000 s, corresponding to a line-averaged electron density of less than 1.0 × 1019 m-2. Actual plasma discharge experiments performed on KTX-CTI, which is a new compact torus injector (CTI) constructed at the Keda Torus eXperiment (KTX), showed that this interferometer has excellent several-second stability.

Optical fiber interferometer application for high electron density measurements on compact torus plasmas.

An optical fiber Mach-Zehnder interferometer at a wavelength of 1.55 µm has been developed for measurements of high electron density on compact torus (CT) plasmas with a high time resolution of 0.1 µs and high phase resolution of 6.4 × 10-4 rad. To improve density measurement accuracy, the phase noise of the interferometer has been investigated in detail and optimized. In the bench test, the interferometer was calibrated using a piezoelectric ceramic actuator with known stroke. Initial results on CT plasma show that the optical fiber interferometer provides reliable density measurements at two spatial locations and the bulk velocity of plasma can be determined by the method of time of flight.

An automatic beam alignment system based on relative reference points for Thomson scattering diagnosis system.

An automatic beam alignment system based on relative reference points is developed for the Thomson scattering system on Keda Torus eXperiment. Two critical apertures around the vacuum vessel are designed to shield stray light, and a probe beam is required to go through the centers of these two apertures, which are the reference points for alignment. Since these two apertures are coated with light absorbing materials, three fibers with glowing tips are employed to indicate the centers of two apertures. CMOS cameras are used to monitor beam deviations. The misalignment correction is achieved by driving piezomotor mirror mounts via a program developed with LabVIEW, which includes the image acquisition and processing module and the deviation correction module. As a result, this system can correct beam misalignment in less than 20 s and suppress the long-term drift of laser pointing in ±10 µrad. Also, this system has the capability to correct up to about 2.3 mm of camera shift with our experiment condition.

LincK contributes to breast tumorigenesis by promoting proliferation and epithelial-to-mesenchymal transition.

BACKGROUND: Increasing evidence has demonstrated that mesenchymal stem cells (MSCs) play a role in the construction of tumor microenvironments. Co-culture between tumor cells and MSCs provides an easy and useful platform for mimicking tumor microenvironments and identifying the important members involved in tumor progress. The long non-coding RNAs (lncRNAs) have been shown to regulate different tumorigenic processes. In this study, we aimed to examine functional lncRNA deregulations associated with breast cancer malignancy instigated by MSC-MCF-7 co-culture. METHODS: The microarrays were used to profile the expression changes of lncRNAs in MCF-7 cells during epithelial-mesenchymal transition (EMT) induced by co-culture with MSCs. We found that an intergenic lncRNA KB-1732A1.1 (termed LincK, partly overlapped with GASL1) was significantly elevated. To investigate the biological function of LincK, the expression of EMT markers, cell migration, invasion, proliferation, and colony formation were evaluated in vitro and xenograft assay in nude mice were performed in vivo. Furthermore, we detected LincK expression in clinical samples using RNAscope® technology and verified aberrant expression of LincK in breast cancer data sets from The Cancer Genome Atlas (TCGA) by bioinformatic analysis. The underlying mechanisms of LincK were investigated using mRNA microarray analyses, Western blot, RNA pull down, and RNA immunoprecipitation. RESULTS: LincK induced an EMT progress in breast cancer cells (BCC) MCF-7, MDA-MB-453, and MDA-MB-231. The depletion of LincK decreased the growth, migration, and invasion in BCC, whereas the overexpression of LincK exerted the opposite effects. Moreover, knockdown of LincK repressed tumorigenesis, and ectopic expression of LincK promoted tumor growth in MCF-7 xenograft model. LincK ablation in MDA-MB-231 cells dramatically impaired lung metastasis when incubated intravenously into nude mice. Further, LincK was frequently elevated in breast cancer compared with normal breast tissue in clinical samples. Mechanistically, LincK may share common miRNA response elements with PBK and ZEB1 and regulate the effects of miR-200 s. CONCLUSION: LincK plays a significant role in regulating EMT and tumor growth and could be a potential therapeutic target in breast cancer.

A parametric method for correcting polluted plasma current signal and its application on Keda Torus eXperiment.

We have developed a parametric method for eliminating the background component of the plasma current, which is measured by a Rogowski coil and polluted by the toroidal magnetic field in the vacuum vessel of the Keda Torus eXperiment (KTX) reversed field pinch (RFP) device. The method considers the toroidal magnetic field windings, the KTX vacuum chamber, and the Rogowski coil as a linear time-invariant system; in this case, a constant frequency response function characterizes the system. Using this response function, the current component caused by pollution from the toroidal magnetic field can be predicted exactly for an arbitrary input current to the toroidal magnetic field windings. Compared with the traditional proportional compensation method, the proposed method has great flexibility and universality and it is potentially applicable to cases in which the toroidal field current signal changes over time with plasma feedback signals. Furthermore, the method can be applied to other similarly affected signals, such as magnetic field signals. As an example, we have corrected the poloidal and toroidal magnetic field signals better to reveal the true physical processes for the RFP state.

Diagnostic capacity of electron cyclotron emission imaging system with continuous large observation area on EAST tokamak.

Electron cyclotron emission imaging on EAST provides direct measurements of the 2-D electron temperature dynamics in a continuous large observation area with high temporal and spatial resolution. Besides the normal MHD investigation, a system with a view field large enough to cover the core plasma region has been applied to extract more plasma information, such as the plasma center location, the deposition location of auxiliary heating, and the core toroidal rotation speed. These results solely based on electron cyclotron emission imaging (ECEI) data are consistent with the results of the equilibrium fitting (EFIT), numerical code, and other diagnostics, which indicate the powerful diagnostic capacity of this ECEI system.

Electromagnetic diagnostic system for the Keda Torus eXperiment.

A system for electromagnetic measurements was designed and installed on the Keda Torus eXperiment (KTX) reversed field pinch device last year. Although the unique double-C structure of the KTX, which allows the machine to be opened easily without disassembling the poloidal field windings, makes the convenient replacement and modification of the internal inductive coils possible, it can present difficulties in the design of flux coils and magnetic probes at the two vertical gaps. Moreover, the KTX has a composite shell consisting of a 6 mm stainless steel vacuum chamber and a 1.5 mm copper shell, which results in limited space for the installation of saddle sensors. Therefore, the double-C structure and composite shell should be considered, especially during the design and installation of the electromagnetic diagnostic system (EDS). The inner surface of the vacuum vessel includes two types of probes. One type is for the measurement of the global plasma parameters, and the other type is for studying the local behavior of the plasma and operating the new saddle coils. In addition, the probes on the outer surface of the composite shell are used for measurements of eddy currents. Finally, saddle sensors for radial field measurements for feedback control were installed between the conducting shell and the vacuum vessel. The entire system includes approximately 1100 magnetic probes, 14 flux coils, 4×26×2 saddle sensors, and 16 Rogowski coils. Considering the large number of probes and limited space available in the vacuum vessel, the miniaturization of the probes and optimization of the probe distribution are necessary. In addition, accurate calibration and careful mounting of the probes are also required. The frequency response of the designed magnetic probes is up to 200 kHz, and the resolution is 1 G. The EDS, being spherical and of high precision, is one of the most basic and effective diagnostic tools of the KTX and meets the demands imposed by requirements on basic machine operating information and future studies.

Identification of Differential Genes Expression Profiles and Pathways of Bone Marrow Mesenchymal Stem Cells of Adolescent Idiopathic Scoliosis Patients by Microarray and Integrated Gene Network Analysis.

STUDY DESIGN: Microarray approach and integrated gene network analysis. OBJECTIVE: To explore the differential genetic expression profile, gene ontology terms, and Kyoto Encyclopedia of Genes and Genomes pathways in bone marrow mesenchymal stem cells (BM-MSCs) of idiopathic scoliosis (AIS) and non-AIS controls. SUMMARY OF BACKGROUND DATA: The pathogenesis of adolescent AIS and the accompanying generalized osteopenia remain unclear. Our previous study suggested increased proliferation ability and decreased osteogenic differentiation ability of BM-MSCs of AIS. Therefore, we hypothesized that MSCs may play a significant role in the etiology and pathogenesis of AIS. METHODS: In this study, microarray analysis was used to identify differentially expressed genes (DEGs) of BM-MSCs from AIS patients compared with those from healthy individuals. Comprehensive bioinformatics analyses were then used to enrich datasets for gene ontology and pathway. Based on the gene signal transduction network analysis of DEGs contained in significant pathways, 24 potential crucial genes were selected for validation by reverse transcription polymerase chain reaction. RESULTS: There are 1027 previously unrecognized DEGs in BM-MSCs from AIS patients. Pathway analysis revealed dysregulated mitogen-activated protein kinase (MAPK) signaling pathway, PI3K-Akt signaling pathway, calcium signaling pathway, peroxisome proliferator-activated receptor (PPAR) signaling pathway, ubiquitin-mediated proteolysis, and Notch signaling pathway, all of which have been reported to play an important role in regulating the osteogenic or adipogenic differentiation of MSCs. Furthermore, gene signal transduction networks analysis indicated that mitogen-activated protein kinase kinase 1 (MAP2K1), SMAD family member 3 (SMAD3), homeobox C6 (HOXC6), heat shock 70kDa protein 6 (HSPA6), general transcription factor IIi (GTF2I), CREB binding protein (CREBBP), phosphoinositide-3-kinase, regulatory subunit 2 (PIK3R2), and dual specificity phosphatase 2 (DUSP2) may play essential roles in AIS pathogenesis and accompanied osteopenia. CONCLUSION: This study reports the differential genes expression profiles of BM-MSCs from AIS patients and related potential pathways for the first time. These previously unrecognized genes and molecular pathways might play a significant role in not only the causal mechanism of osteopenia in AIS, but also the AIS initiation and development. The identification of these candidate genes provides novel insight into the underlying etiological mechanisms of AIS. LEVEL OF EVIDENCE: N/A.

Compact and lightweight support platform with electromagnetic disturbance elimination for interferometer on reversed field pinch Keda Torus eXperiment.

A compact and lightweight support platform has been used as a holder for the interferometer system on the Keda Torus eXperiment (KTX), which is a reversed field pinch device. The vibration caused by the interaction between the time-varying magnetic field and the induced current driven in the metal optical components has been measured and, following comparison with the mechanical vibration of the KTX device and the refraction effect of the ambient turbulent air flow, has been identified as the primary vibration source in this case. To eliminate this electromagnetic disturbance, nonmetallic epoxy resin has been selected as the material for the support platform and the commercially available metal optical mounts are replaced. Following these optimization steps and mechanical reinforcements, the stability of the interferometer platform has improved significantly. The phase shift caused by the vibration has been reduced to the level of background noise.

The eddy current probe array for Keda Torus eXperiment.

In a reversed field pinch device, the conductive shell is placed as close as possible to the plasma so as to balance the plasma during discharge. Plasma instabilities such as the resistive wall mode and certain tearing modes, which restrain the plasma high parameter operation, respond closely with conditions in the wall, in essence the eddy current present. Also, the effect of eddy currents induced by the external coils cannot be ignored when active control is applied to control instabilities. One diagnostic tool, an eddy current probe array, detects the eddy current in the composite shell. Magnetic probes measuring differences between the inner and outer magnetic fields enable estimates of the amplitude and angle of these eddy currents. Along with measurements of currents through the copper bolts connecting the poloidal shield copper shells, we can obtain the eddy currents over the entire shell. Magnetic field and eddy current resolutions approach 2 G and 6 A, respectively. Additionally, the vortex electric field can be obtained by eddy current probes. As the conductivity of the composite shell is high, the eddy current probe array is very sensitive to the electric field and has a resolution of 0.2 mV/cm. In a bench test experiment using a 1/4 vacuum vessel, measurements of the induced eddy currents are compared with simulation results based on a 3D electromagnetic model. The preliminary data of the eddy currents have been detected during discharges in a Keda Torus eXperiment device. The typical value of toroidal and poloidal eddy currents across the magnetic probe coverage rectangular area could reach 3.0 kA and 1.3 kA, respectively.