Employing CRISPR-Cas9 to Generate CD133 Synthetic Lethal Melanoma Stem Cells

Malignant melanoma is a lethal skin cancer containing melanoma-initiating cells (MIC) implicated in tumorigenesis, invasion, and drug resistance, and is characterized by the elevated expression of stem cell markers, including CD133. The siRNA knockdown of CD133 enhances apoptosis induced by the MEK inhibitor trametinib in melanoma cells. This study investigates the underlying mechanisms of CD133’s anti-apoptotic activity in patient-derived BAKP and POT cells, harboring difficult-to-treat NRAS^Q61K and NRAS^Q61R drivers, after CRISPR-Cas9 CD133 knockout or Dox-inducible expression of CD133. MACS-sorted CD133(+) BAKP cells were conditionally reprogrammed to derive BAKR cells with sustained CD133 expression and MIC features. Compared to BAKP, CD133(+) BAKR exhibit increased cell survival and reduced apoptosis in response to trametinib or the chemotherapeutic dacarbazine (DTIC). CRISPR-Cas9-mediated CD133 knockout in BAKR cells (BAKR-KO) re-sensitized cells to trametinib. CD133 knockout in BAKP and POT cells increased trametinib-induced apoptosis by reducing anti-apoptotic BCL-xL, p-AKT, and p-BAD and increasing pro-apoptotic BAX. Conversely, Dox-induced CD133 expression diminished apoptosis in both trametinib-treated cell lines, coincident with elevated p-AKT, p-BAD, BCL-2, and BCL-xL and decreased activation of BAX and caspases-3 and -9. AKT1/2 siRNA knockdown or inhibition of BCL-2 family members with navitoclax (ABT-263) in BAKP-KO cells further enhanced caspase-mediated apoptotic PARP cleavage. CD133 may therefore activate a survival pathway where (1) increased AKT phosphorylation and activation induces (2) BAD phosphorylation and inactivation, (3) decreases BAX activation, and (4) reduces caspases-3 and -9 activity and caspase-mediated PARP cleavage, leading to apoptosis suppression and drug resistance in melanoma. Targeting nodes of the CD133, AKT, or BCL-2 survival pathways with trametinib highlights the potential for combination therapies for NRAS-mutant melanoma stem cells for the development of more effective treatments for patients with high-risk melanoma.     

基于蓝牙技术的语音接入系统设计与实现

文中充分发挥互连网和无线接入的优势,引入蓝牙作为语音无线传输,成功架构和实现了基于蓝牙技术的语音接入系统.该系统能够使得用户在离计算机10米的范围内轻松实现网络语音聊天、接收实时消息、收听音乐等.基于CSR(Cambridge Silicon Radio)公司的 Bluecore2-Audio蓝牙芯片和BlueLab平台设计和实现了手持终端.在对手持终端硬件组成进行分析的基础上,详细介绍了手持终端和PC端驱动软件的实现方法.     

Combined Treatment with Cryptocaryone and Ultraviolet C Promotes Antiproliferation and Apoptosis of Oral Cancer Cells

Cryptocaryone (CPC) was previously reported as preferential for killing natural products in oral cancer cells. However, its radiosensitizing potential combined with ultraviolet C (UVC) cell killing of oral cancer cells remains unclear. This study evaluates the combined anti-proliferation effect and clarifies the mechanism of combined UVC/CPC effects on oral cancer cells. UVC/CPC shows higher anti-proliferation than individual and control treatments in a low cytotoxic environment on normal oral cells. Mechanistically, combined UVC/CPC generates high levels of reactive oxygen species and induces mitochondrial dysfunction by generating mitochondrial superoxide, increasing mitochondrial mass and causing the potential destruction of the mitochondrial membrane compared to individual treatments. Moreover, combined UVC/CPC causes higher G2/M arrest and triggers apoptosis, with greater evidence of cell cycle disturbance, annexin V, pancaspase, caspases 3/7 expression or activity in oral cancer cells than individual treatments. Western blotting further indicates that UVC/CPC induces overexpression for cleaved types of poly (ADP-ribose) polymerase and caspase 3 more than individual treatments. Additionally, UVC/CPC highly induces γH2AX and 8-hydroxy-2’-deoxyguanosine adducts as DNA damage in oral cancer cells. Taken together, CPC shows a radiosensitizing anti-proliferation effect on UVC irradiated oral cancer cells with combined effects through oxidative stress, apoptosis and DNA damage.     

In Vitro Angiogenic Behavior of HUVECs on Biomimetic SF/SA Composite Scaffolds

The 60Fc and 70Fc SF/SA blend scaffolds were prepared to mimic the functions of the native ECM for skin regeneration.Human Umbilical Vein Endothelial Cells (HUVECs) were used to examine the cell cytotoxicity,adhesion,growth factors secretion and the gene expression of associated angiogenic factors.Cell proliferation,adhesion and live-dead analyses showed that HUVECs could better attach,grow,and proliferate on the 70Fc scaffolds compared with 60Fc scaffolds and unmodified controls.Furthermore,the 70Fc scaffolds showed higher levels of specific angiogenic proteins and genes expression as well.This study suggests that the involvement of higher composition of SF (about 70%) than that of SA on the blended scaffolds could be advantageous as it is more suitable to promote angiogenesis,which is potential for vascularization during skin repair.     

Application of neural networks to prediction of phase transport characteristics in high-pressure two-phase turbulent bubbly flows

The phase transport phenomenon of the high-pressure two-phase turbulent bubbly flow involves complicated interfacial interactions of the mass, momentum, and energy transfer processes between phases, revealing that an enormous effort is required in characterizing the liquid–gas flow behavior. Nonetheless, the instantaneous information of bubbly flow properties is often desired for many industrial applications. This investigation aims to demonstrate the successful use of neural networks in the real-time determination of two-phase flow properties at elevated pressures. Three back-propagation neural networks, trained with the simulation results of a comprehensive theoretical model, are established to predict the transport characteristics (specifically the distributions of void-fraction and axial liquid–gas velocities) of upward turbulent bubbly pipe flows at pressures covering 3.5–7.0 MPa. Comparisons of the predictions with the test target vectors indicate that the averaged root-mean-squared (RMS) error for each one of three back-propagation neural networks is within 4.59%. In addition, this study appraises the effects of different network parameters, including the number of hidden nodes, the type of transfer function, the number of training pairs, the learning rate-increasing ratio, the learning rate-decreasing ratio, and the momentum value, on the training quality of neural networks.     

Corrosion of Mg alloys EV31A,WE43B,and ZE41A in chloride- and sulfate-containing solutions saturated with magnesium hydroxide

This study studied corrosion in 0.1 M Na₂SO₄, 0.1 M NaCl, and 0.6 M NaCl, all saturated with Mg(OH)₂, using weight loss, hydrogen evolution, and electrochemical measurements. Corrosion was similar in all cases. Nevertheless, the corrosion rates were alloy-dependent, were somewhat lower in 0.1 M Na₂SO₄ than in 0.1 M NaCl, and increased with NaCl concentration. The corrosion damage morphology was similar for all solutions; the extent correlated with the corrosion rate. The corrosion rates evaluated by the electrochemical methods were lower than those evaluated from hydrogen evolution, consistent with the Mg corrosion mechanism involving the unipositive Mg⁺ ion.     

Assessment of LTE Wireless Accessing for Managing Traffic Flow of IoT Services

Mobile Networks and Applications - There is a tremendous growth in the traffic of the Internet of Things (IoT) devices. Some of the IoT application scenarios may prefer the wireless access...     

Improved design of dust test chamber for uniform distribution of dust sedimentation rate

Soiling problems are always encountered for equipment installed in outdoor environments, such as headlamps of automobiles, air conditioners, solar collectors, and so on. How to prevent soiling problems on this equipment is one of the challenges for the design of their external layouts. Thus, evaluations of the dust sedimentation quantities on the surfaces of such equipment are necessary. Outdoor testing is usually straightforward but it takes a long time to experience various environmental parameters, such as dust-laden air, wind speed, wind direction, and so on. Indoor tests, performed in a dust test chamber, are instruments for controlling various environmental parameters independently. The conventional design for a dust test chamber is aimed at providing for a test under extreme environmental conditions. The uniformity of dust sedimentation rate within the chamber is not rigorously controlled. Nevertheless, some applications such as the effect of dust sedimentation on the glazing of solar collector require uniform distribution of dustfall in the test chamber for the indoor (laboratory) test. An improved design for a conventional dust test chamber is proposed. Performance tests done with the remodeled dust test chamber based on the improved design show that the normalized standard deviation of the dustfall concentration can be controlled within 2.41% ± 1.29%.     

GaN-based Indium-tin-oxide light emitting diodes with nanostructured silicon upper contacts

GaN-based indium-tin-oxide (ITO) light emitting diodes (LEDs) with p-GaN, n⁺-short period superlattice (SPS) and nanostructured silicon contact layers were fabricated. It was found that surface of the ITO LED with nanostructured silicon layer was very rough. It was also found that 20 mA forward voltages measured from ITO LEDs with p-GaN, n⁺-SPS and nanostructured silicon contact layers were 6.01, 3.25 and 3.26 V, respectively. Compared with ITO LED with n⁺-SPS, it was found that output power of ITO LED with nanostructured silicon contact was 17% larger. Furthermore, it was found that ITO LED with nanostructured silicon contact was more reliable.