Knockdown of MAGEA6 Activates AMP-Activated Protein Kinase (AMPK) Signaling to Inhibit Human Renal Cell Carcinoma Cells

Background/Aims: Melanoma antigen A6 (MAGEA6) is a cancer-specific ubiquitin ligase of AMP-activated protein kinase (AMPK). The current study tested MAGEA6 expression and potential function in renal cell carcinoma (RCC). Methods: MAGEA6 and AMPK expression in human RCC tissues and RCC cells were tested by Western blotting assay and qRT-PCR assay. shRNA method was applied to knockdown MAGEA6 in human RCC cells. Cell survival and proliferation were tested by MTT assay and BrdU ELISA assay, respectively. Cell apoptosis was tested by the TUNEL assay and single strand DNA ELISA assay. The 786-O xenograft in nude mouse model was established to test RCC cell growth in vivo. Results: MAGEA6 is specifically expressed in RCC tissues as well as in the established (786-O and A498) and primary human RCC cells. MAGEA6 expression is correlated with AMPKα1 downregulation in RCC tissues and cells. It is not detected in normal renal tissues nor in the HK-2 renal epithelial cells. MAGEA6 knockdown by targeted-shRNA induced AMPK stabilization and activation, which led to mTOR complex 1 (mTORC1) in-activation and RCC cell death/apoptosis. AMPK inhibition, by AMPKα1 shRNA or the dominant negative AMPKα1 (T172A), almost reversed MAGEA6 knockdown-induced RCC cell apoptosis. Conversely, expression of the constitutive-active AMPKα1 (T172D) mimicked the actions by MAGEA6 shRNA. In vivo, MAGEA6 shRNA-bearing 786-O tumors grew significantly slower in nude mice than the control tumors. AMPKα1 stabilization and activation as well as mTORC1 in-activation were detected in MAGEA6 shRNA tumor tissues. Conclusion: MAGEA6 knockdown inhibits human RCC cells via activating AMPK signaling

The protective effects of Lipoxin A(4) during the early phase of severe acute pancreatitis in rats

Objective. Our aim was to investigate the protective effects of a Lipoxin A(4) analogue (LXA(4)) in the early phase of acute pancreatitis in rats. Materials and methods. Severe acute pancreatitis (SAP) was induced by injection of 5% sodium taurocholate into the pancreatic duct. Rats with SAP were treated with LXA(4) (0.1 mg/kg), 10 min after the 5% sodium taurocholate injection, after which LXA(4) was administrated every 8 hours, three times (LXA(4) group). The sham group was only given the vehicle after operation. Plasma amylase activity, serum levels of interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were measured at 4, 12, and 24 h after induction of SAP. The pancreatic index and histopathologic observations were evaluated and the expression of intercellular adhesion molecule-1 (ICAM-1) and NF-kappa B p65 in the pancreas, and the expression of ICAM-1 in the lungs were detected by immunohistochemistry. Results. LXA(4) treated rats had lower serum levels of TNF-alpha, IL-1, and IL-6 at all time points measured (p < 0.05), but significantly differed in plasma amylase activity only at 24 h as compared with the SAP group. The pancreatic index and the scores of pancreatitic histopathologic evaluations were lower in the LXA(4) group as compared to the SAP group. Immunohistochemistry showed that LXA(4) attenuated the expression of ICAM-1 and NF-kappa B p65 in the pancreas, as well as the expression of ICAM-1 in the lungs in animals with pancreatitis (p < 0.05). Conclusions. We demonstrate that LXA(4) has protective effects in experimental SAP, which may be achieved by inhibiting the NF-kappa B signalling pathway, thereby reducing the production of proinflammatory cytokines

Efficient De Novo Biosynthesis of Heme by Membrane Engineering in <i>Escherichia coli</i>

Heme is of great significance in food nutrition and food coloring, and the successful launch of artificial meat has greatly improved the application of heme in meat products. The precursor of heme, 5-aminolevulinic acid (ALA), has a wide range of applications in the agricultural and medical fields, including in the treatment of corona virus disease 2019 (COVID-19). In this study, E. coli recombinants capable of heme production were developed by metabolic engineering and membrane engineering. Firstly, by optimizing the key genes of the heme synthesis pathway and the screening of hosts and plasmids, the recombinant strain EJM-pCD-AL produced 4.34 ± 0.02 mg/L heme. Then, the transport genes of heme precursors CysG, hemX and CyoE were knocked out, and the extracellular transport pathways of heme Dpp and Ccm were strengthened, obtaining the strain EJM-ΔCyoE-pCD-AL that produced 9.43 ± 0.03 mg/L heme. Finally, fed-batch fermentation was performed in a 3-L fermenter and reached 28.20 ± 0.77 mg/L heme and 303 ± 1.21 mg/L ALA. This study indicates that E. coli recombinant strains show a promising future in the field of heme and ALA production

Development of a Lightweight Single-Band Bathymetric LiDAR

Traditional bathymetry LiDAR (light detection and ranging) onboard manned and/or unmanned airborne systems cannot operate in the context of narrow rivers in urban areas with high buildings and in mountainous areas with high peaks. Therefore, this study presents a prototype of a lightweight bathymetry LiDAR onboard an unmanned shipborne vehicle (called &ldquo;GQ-Cor 19&rdquo;). The GQ-Cor 19 system primarily includes an emitting optical module, a receiving optical module, control module, detection module, high-speed A/D sampling module, and data processing system. Considering that the &ldquo;GQ-Cor 19&rdquo; is extremely close to the water surface, various new technical challenges are encountered, such as significant laser scattering energy from the surface of the water, which saturates signals received by the photomultiplier tube detector. Therefore, this study presents various new technical solutions, including (1) an improved Bresenham algorithm, (2) a small and lightweight receiving optical system with a split-field method, and (3) a data acquisition module with a high-speech A/D collector. Following a series of different experimental verifications, the results demonstrate that the new generation of single-band LiDAR onboard an unmanned shipborne vehicle can swiftly measure the underwater depth, and the maximum measurement depth is more than 25 m. The measurement accuracy is better than 30 cm and the weight is less than 12 kg