Multi-Granularity Context Network for Efficient Video Semantic Segmentation.

Current video semantic segmentation tasks involve two main challenges: how to take full advantage of multi-frame context information, and how to improve computational efficiency. To tackle the two challenges simultaneously, we present a novel Multi-Granularity Context Network (MGCNet) by aggregating context information at multiple granularities in a more effective and efficient way. Our method first converts image features into semantic prototypes, and then conducts a non-local operation to aggregate the per-frame and short-term contexts jointly. An additional long-term context module is introduced to capture the video-level semantic information during training. By aggregating both local and global semantic information, a strong feature representation is obtained. The proposed pixel-to-prototype non-local operation requires less computational cost than traditional non-local ones, and is video-friendly since it reuses the semantic prototypes of previous frames. Moreover, we propose an uncertainty-aware and structural knowledge distillation strategy to boost the performance of our method. Experiments on Cityscapes and CamVid datasets with multiple backbones demonstrate that the proposed MGCNet outperforms other state-of-the-art methods with high speed and low latency.

Photo-electrocatalytic based removal of acetaminophen: Application of visible light driven heterojunction based BiVO4/BiOI photoanode.

The presence of organic micro-pollutants (OMPs) in wastewater treatment effluents is becoming a major threat to the water safety for aquatic and human health. Photo-electrocatalytic based advanced oxidation process (AOP) is one of the emerging and effective techniques to degrade OMPs through oxidative mechanism. This study investigated the application of heterojunction based BiVO4/BiOI photoanode for acetaminophen (40 µg L-1) removal in demineralized water. Photoanodes were fabricated by electrodeposition of BiVO4 and BiOI photocatalytic layers. Optical (UV-vis diffusive reflectance spectroscopy), structural (XRD, SEM, EDX) and opto-electronic (IPCE) characterization confirmed the successful formation of heterojunction for enhanced charge separation efficiency. The heterojunction photoanode showed incident photon to current conversion efficiency of 16% (λmax = 390 nm) at an external voltage of 1 V under AM 1.5 standard illumination. The application of the BiVO4/BiOI photoanode in the removal of acetaminophen at 1 V (external bias) vs Ag/AgCl under simulated sunlight showed 87% removal efficiency within the first 120 min compared to 66% removal efficiency of the BiVO4 photoanode. Similarly, combining BiVO4 and BiOI exhibited 57% increase in first order removal rate coefficient compared to BiVO4. The photoanodes also showed moderate stability and reusability by showing 26% decrease in overall degradation efficiency after three cycles of each 5 h experiment. The results obtained in this study can be considered as a stepping stone towards the effective removal of acetaminophen as an OMP present in wastewater.

Engineering inducible biomolecular assemblies for genome imaging and manipulation in living cells.

Genome architecture and organization play critical roles in cell life. However, it remains largely unknown how genomic loci are dynamically coordinated to regulate gene expression and determine cell fate at the single cell level. We have developed an inducible system which allows Simultaneous Imaging and Manipulation of genomic loci by Biomolecular Assemblies (SIMBA) in living cells. In SIMBA, the human heterochromatin protein 1α (HP1α) is fused to mCherry and FRB, which can be induced to form biomolecular assemblies (BAs) with FKBP-scFv, guided to specific genomic loci by a nuclease-defective Cas9 (dCas9) or a transcriptional factor (TF) carrying tandem repeats of SunTag. The induced BAs can not only enhance the imaging signals at target genomic loci using a single sgRNA, either at repetitive or non-repetitive sequences, but also recruit epigenetic modulators such as histone methyltransferase SUV39H1 to locally repress transcription. As such, SIMBA can be applied to simultaneously visualize and manipulate, in principle, any genomic locus with controllable timing in living cells.

Roles of EP Receptors in the Regulation of Fluid Balance and Blood Pressure.

Prostaglandin E2 (PGE2) is an important prostanoid expressing throughout the kidney and cardiovascular system. Despite the diverse effects on fluid metabolism and blood pressure, PGE2 is implicated in sustaining volume and hemodynamics homeostasis. PGE2 works through four distinct E-prostanoid (EP) receptors which are G protein-coupled receptors. To date, pharmacological specific antagonists and agonists of all four subtypes of EP receptors and genetic targeting knockout mice for each subtype have helped in uncoupling the diverse functions of PGE2 and discriminating the respective characteristics of each receptor. In this review, we summarized the functions of individual EP receptor subtypes in the renal and blood vessels and the molecular mechanism of PGE2-induced fluid metabolism and blood pressure homeostasis.

Monocytes engineered with iSNAP inhibit human B-lymphoma progression.

Monocytes are important regulators for the maintenance of homeostasis in innate and adaptive immune system and have been reported to play important role in cancer progression. CD47-SIRPα recognition is a coinhibitory immune signal to inhibit phagocytosis in monocytes and macrophages and has been well-known as the "Don't eat me" signal. By using an approach of integrated sensing and activating proteins (iSNAPs), we have rewired the CD47-SIRPα axis to create iSNAP-M which activates pathways in engineered human monocytes (iSNAP-MC). The mRNA expression levels of the monocyte/macrophage markers CD11b, CD14, and CD31 are upregulated in iSNAP-monocytes (iSNAP-MC). With PMA induction, the iSNAP-MC-derived macrophages (iSNAP-MΦ) showed upregelation in CD86 and CD80, but not CD206. TNFα expression and secretion were also increased in iSNAP-MΦ. Furthermore, the injection of iSNAP-MC into mice bearing human B-lymphoma tumors led to the suppression of tumor progression. Therefore, the engineered monocytes, via blockage of coinhibitory immune signals by rewiring CD47-SIRPα axis, can be applied to suppress target tumors for cancer immunotherapy.

Immobilization of soil Cd by sulfhydryl grafted palygorskite in wheat-rice rotation mode: A field-scale investigation.

The safe utilization of heavy metal contaminated farmland has attracted extensive attention of the whole society, and there is an urgent need to develop novel high-efficiency amendments. To clarify the actual remediation effect and potential for large-scale application of sulfhydryl grafted palygorskite (SGP) in Cd polluted soil in wheat-rice rotation mode, a field-scale experiment was conducted. SGP at the dosages of 0.5 g/kg-2.0 g/kg could reduce gain Cd contents by 27.15-59.05% and 16.16-79.47% for wheat and rice, respectively. The maximal decreases of soil available Cd figured out by DTPA extraction in wheat and rice season were 58.18% and 33.67%, respectively. The immobilization ratio for Cd was much more than that of trace elements, including Fe, Mn, Cu, and Zn, Ni. SGP showed an effective immobilization rate for soil Cd under the interference of many elements in the soil, pointing to the targeting and selectivity of its high-efficiency immobilization. It had no lifting effect on soil pH but decreased zeta potentials of soil particles. The sorption of Cd2+ on SGP amended soil could be fitted by the second-order kinetic model and Langmuir isotherm, and the changes of thermodynamic parameters showed SGP strengthened the fixation. SGP made the biological accumulation coefficient and transfer factor of rice grain drop dramatically but had no noticeable effect on these parameters of winter wheat, indicating different botanical responses. SGP as a novel immobilization amendment may provide an efficient and sustainable solution for the remediation of contaminated soil in wheat-rice rotation mode in field-scale.

Maintaining homeostasis of mitochondria and endoplasmic reticulum with NSC228155 alleviates cisplatin-induced acute kidney injury.

Acute kidney injury (AKI) is a common complication of hospitalization with high mortality. Approximately 30% of patients receiving cisplatin, the first-line chemotherapy treatment, develop AKI. NSC228155 is a novel compound with potential anti-cancer and anti-bacterial effects. Its therapeutic efficacy in other diseases is unclear. In the present study, we investigated the effect of NSC228155 on cisplatin-induced AKI. The mice were consecutively treated with 2.5 mg/kg of NSC228155 for five days and injected with cisplatin (22 mg/kg) via intraperitoneal injection on day three. NSC228155 strikingly improved the renal function by decreasing the serum creatinine by 52.6% in the cisplatin-induced AKI mice model. Pathologically, NSC228155 profoundly alleviated the tubular damage in Periodic Acid-Schiff staining, and significantly reduced the expression of tubular injury markers and apoptosis in the cisplatin-injured mice kidneys. NSC228155 effectively restored the mitochondrial homeostasis by decreasing damaged mitochondria, activating signals for mitochondrial dynamics and recycling, and corrected mitochondrial dysfunction in ATP production and oxidative stress in the cisplatin model. Transcriptomics and metabolomics analysis on the mice renal cortex suggested that NSC228155 profoundly corrected energy metabolism, especially citrate cycle-related pathways, oxidative stress, and endoplasmic reticulum (ER) stress in the cisplatin-induced AKI kidneys. NSC228155 effectively inhibited ER stress induced by cisplatin or tunicamycin in mice kidneys and HK-2 cells. Co-treatment of NSC228155 with 4-phenylbutyrate or MnTBAP showed a similar therapeutic effect in AKI as the inhibitors or NSC228155 alone did, and corrected the mitochondrial dysfunction and ER stress, respectively, indicating the crosstalk between ER and mitochondria played essential roles in the therapeutic effect of NSC228155 in AKI. Together, these results consistently demonstrated that NSC228155 alleviated cisplatin-induced AKI by restoring the homeostasis in mitochondria and ER, suggesting a therapeutic potential and perhaps a novel strategy for drug discovery.

Engineering CAR T cells for enhanced efficacy and safety.

Despite its success in treating hematologic malignancies, chimeric antigen receptor (CAR) T cell therapy faces two major challenges which hinder its broader applications: the limited effectiveness against solid tumors and the nonspecific toxicities. To address these concerns, researchers have used synthetic biology approaches to develop optimization strategies. In this review, we discuss recent improvements on the CAR and other non-CAR molecules aimed to enhance CAR T cell efficacy and safety. We also highlight the development of different types of inducible CAR T cells that can be controlled by environmental cues and/or external stimuli. These advancements are bringing CAR T therapy one step closer to safer and wider applications, especially for solid tumors.

Control of the activity of CAR-T cells within tumours via focused ultrasound.

Focused ultrasound can deliver energy safely and non-invasively into tissues at depths of centimetres. Here we show that the genetics and cellular functions of chimeric antigen receptor T cells (CAR-T cells) within tumours can be reversibly controlled by the heat generated by short pulses of focused ultrasound via a CAR cassette under the control of a promoter for the heat-shock protein. In mice with subcutaneous tumours, locally injected T cells with the inducible CAR and activated via focused ultrasound guided by magnetic resonance imaging mitigated on-target off-tumour activity and enhanced the suppression of tumour growth, compared with the performance of non-inducible CAR-T cells. Acoustogenetic control of the activation of engineered T cells may facilitate the design of safer cell therapies.

Reversible Patterning Cross-Linked, Humidity-Responsive Polymer Films with Programmatically and Accurately Controlled Deformation.

A series of novel humidity-responsive and photosensitive polymer films (PCA-PAA-PEG) are prepared. These films can be patterning cross-linked by the photodimerization of coumarin pendant groups. The humidity-induced deformation can be well controlled by the pattern because of the different modulus and hydrophilicity between cross-linked and un-cross-linked segments. In addition, the pattern can be erased and the deformation direction can be changed programmatically by the de-cross-linking-re-cross-linking approach due to the reversible photodimerization of coumarin groups. The cross-linking degree also affects the humidity responsiveness of the film. The deformation of the gradient patterning cross-linked film can be more accurately controlled. Moreover, the length and width ratio (L/Ws/Wh) of the un-cross-linked segment to the cross-linked segment affects the deformation of the films as well. When L/Ws/Wh is 5/2/1 or 5/3/1, the deformation is controllable, and when L/Ws/Wh is 5/1/1 or 5/4/1, the deformation is random at the initial stage, but the whole film will bend along the short axis in the end.

Mechanogenetics for cellular engineering and cancer immunotherapy.

Recent synthetic biology advancements have shown that cells can be engineered to respond to external stimuli such as chemical compounds and light, which significantly improves the specificity and controllability of CAR T therapy. However, the lack of both spatiotemporal and depth control is still the main issue in the clinic of CAR T treatment. At the same time, mechanogenetics, capable of penetrating deep tissues with high spatiotemporal precision, is rapidly evolving and advancing to reveal its potential for cancer immunotherapy. In the past few years, researchers have demonstrated the precise and remote control of engineered cells with mechanical perturbation originated from ultrasound, which may become a new solution to circumvent the limitations of CAR T therapy in the future. This review will discuss mechanobiology and the state-of art designs of controllable CAR T cells. A specific focus of this review will be on the mechanical control of CAR T therapy.

Engineering light-controllable CAR T cells for cancer immunotherapy.

T cells engineered to express chimeric antigen receptors (CARs) can recognize and engage with target cancer cells with redirected specificity for cancer immunotherapy. However, there is a lack of ideal CARs for solid tumor antigens, which may lead to severe adverse effects. Here, we developed a light-inducible nuclear translocation and dimerization (LINTAD) system for gene regulation to control CAR T activation. We first demonstrated light-controllable gene expression and functional modulation in human embryonic kidney 293T and Jurkat T cell lines. We then improved the LINTAD system to achieve optimal efficiency in primary human T cells. The results showed that pulsed light stimulations can activate LINTAD CAR T cells with strong cytotoxicity against target cancer cells, both in vitro and in vivo. Therefore, our LINTAD system can serve as an efficient tool to noninvasively control gene activation and activate inducible CAR T cells for precision cancer immunotherapy.

An AND-Gated Drug and Photoactivatable Cre-loxP System for Spatiotemporal Control in Cell-Based Therapeutics.

While engineered chimeric antigen receptor (CAR) T cells have shown promise in detecting and eradicating cancer cells within patients, it remains difficult to identify a set of truly cancer-specific CAR-targeting cell surface antigens to prevent potentially fatal on-target off-tumor toxicity against other healthy tissues within the body. To help address this issue, we present a novel tamoxifen-gated photoactivatable split-Cre recombinase optogenetic system, called TamPA-Cre, that features high spatiotemporal control to limit CAR T cell activity to the tumor site. We created and optimized a novel genetic AND gate switch by integrating the features of tamoxifen-dependent nuclear localization and blue-light-inducible heterodimerization of Magnet protein domains (nMag, pMag) into split Cre recombinase. By fusing the cytosol-localizing mutant estrogen receptor ligand binding domain (ERT2) to the N-terminal half of split Cre(2-59aa)-nMag, the TamPA-Cre protein ERT2-CreN-nMag is physically separated from its nuclear-localized binding partner, NLS-pMag-CreC(60-343aa). Without tamoxifen to drive nuclear localization of ERT2-CreN-nMag, the typically high background of the photoactivation system was significantly suppressed. Upon blue light stimulation following tamoxifen treatment, the TamPA-Cre system exhibits sensitivity to low intensity, short durations of blue light exposure to induce robust Cre-loxP recombination efficiency. We finally demonstrate that this TamPA-Cre system can be applied to specifically control localized CAR expression and subsequently T cell activation. As such, we posit that CAR T cell activity can be confined to a solid tumor site by applying an external stimulus, with high precision of control in both space and time, such as light.

[Effect of Quercetin on Apoptosis of Platelets and Its Mechanism].

OBJECTIVE: To investigate the effects of quercetin on the apoptosis of platelets and to analyze the intrinsic mechanism. METHODS: Firstly, the effects of quecetin on the apoptosis of platelets was detected by flow cytometry. Secondly, Western blot was used to detect the expression of apoptosis-related proteins in the platelets treated with quercetin for 2 and 4 day. RESULTS: By flow cytometry, it was found that the apoptosis of platelets in the quercetin-treated group (2, 4 and 8 µmol/L) was inhibited, the apoptosis rate of platelets in 2, 4 and 8 µmol/L quercetin group was 3.12%±0.32%, 2.89%±0.15% and 2.31%±0.28%, respectively, which were signigicantly lover than that in control group (P＜0.01). With the increase of quecetin concentration, the proportion ratio of platelets significantly decreased in a concentration-dependent manner（r=-0.9985）. Similar results were observed on the 4th day. Western blot showed that the treatment with quercetin (2, 4 and 8 µmol/L) promoted the expression of anti-apoptotic protein BCL-2, inhibited the expression of pro-apoptotic protein BAX, resulting in a significant increase in the ratio of BCL-2/BAX (P＜0.01), thereby inhibiting the apoptosis of platelets. Similar results were observed on the 4th day. CONCLUSION: Quercetin can inhibit platelet apoptosis by increasing the ratio of apoptosis-related protein BCL-2/BAX in a concentration-dependent manner.

Hydrogen-bonding-induced bathochromic effect of Si-coumarin and its use in monitoring adipogenic differentiation.

Introduction of a heteroatom into a fluorophore was carried out for coumarin through a replacement of its bridging oxygen atom with a silicon atom. The maximum-emission wavelength of Si-coumarin (SiC B) bathochromically shifted from 426 nm in cyclohexane to 626 nm in water. The adipogenic differentiation processes in mesenchymal stem cells were monitored using SiC B.

Biophysical basis underlying dynamic Lck activation visualized by ZapLck FRET biosensor.

Lck plays crucial roles in TCR signaling. We developed a new and sensitive FRET biosensor (ZapLck) to visualize Lck kinase activity with high spatiotemporal resolutions in live cells. ZapLck revealed that 62% of Lck signal was preactivated in T-cells. In Lck-deficient JCam T-cells, Lck preactivation was abolished, which can be restored to 51% by reconstitution with wild-type Lck (LckWT) but not a putatively inactive mutant LckY394F. LckWT also showed a stronger basal Lck-Lck interaction and a slower diffusion rate than LckY394F. Interestingly, aggregation of TCR receptors by antibodies in JCam cells led to a strong activation of reconstituted LckY394F similar to LckWT. Both activated LckY394F and LckWT diffused more slowly and displayed increased Lck-Lck interaction at a similar level. Therefore, these results suggest that a phosphorylatable Y394 is necessary for the basal-level interaction and preactivation of LckWT, while antibody-induced TCR aggregation can trigger the full activation of LckY394F.

Puppet resting behavior in the Tibetan antelope (Pantholops hodgsonii).

Rest contributes a large part of animals' daily life, and animals usually rest in two ways, standing or in recumbence. Small or medium sized ungulates bed to rest in most cases, and standing rest is very rare and hardly seen. Here we described a standing rest behavior of Tibetan antelopes (Pantholops hodgsonii) living on the Tibet Plateau which has not been reported before. We named the standing rest behavior Puppet behavior, since the antelope stand still for a certain time. Of the 304 individuals observed, 48.3% (98/203) of adult and sub-adult males expressed the Puppet behavior, whereas only 6.3% (6/96) of females did, indicating an obvious sexual difference. Puppet behavior occurred more frequently at noon and in the afternoon on sunny and cloudy days, meaning that daytime and weather were both influential factors. Puppet behavior was usually accompanied with rumination and sometimes ended with leg-shaking. Our results suggest that Puppet behavior may be an adaptive form of rest, which may serve a thermoregulatory and anti-predation function, and may be simpler and safer than recumbent rest.

Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface.

Monitoring enzymatic activities at the cell surface is challenging due to the poor efficiency of transport and membrane integration of fluorescence resonance energy transfer (FRET)-based biosensors. Therefore, we developed a hybrid biosensor with separate donor and acceptor that assemble in situ. The directed evolution and sequence-function analysis technologies were integrated to engineer a monobody variant (PEbody) that binds to R-phycoerythrin (R-PE) dye. PEbody was used for visualizing the dynamic formation/separation of intercellular junctions. We further fused PEbody with the enhanced CFP and an enzyme-specific peptide at the extracellular surface to create a hybrid FRET biosensor upon R-PE capture for monitoring membrane-type-1 matrix metalloproteinase (MT1-MMP) activities. This biosensor revealed asymmetric distribution of MT1-MMP activities, which were high and low at loose and stable cell-cell contacts, respectively. Therefore, directed evolution and rational design are promising tools to engineer molecular binders and hybrid FRET biosensors for monitoring molecular regulations at the surface of living cells.

Qidong hepatitis B virus infection cohort: a 25-year prospective study in high risk area of primary liver cancer.

Qidong hepatitis B virus (HBV) infection cohort (QBC) is a prospective community-based study designed to investigate causative factors of primary liver cancer (PLC) in Qidong, China, where both PLC and HBV infection are highly endemic. Residents aged 20-65 years, living in seven townships of Qidong, were surveyed using hepatitis B surface antigen (HBsAg) serum test and invited to participate in QBC from June 1991 to December 1991. A total of 852 and 786 participants were enrolled in HBsAg-positive and HBsAg-negative sub-cohorts in May 1992, respectively. All participants were actively followed up in person, received HBsAg, alanine aminotransferase (ALT), alpha-fetoprotein (AFP) tests and upper abdominal ultrasonic examination, and donated blood and urine samples once or twice a year. The total response rate was 99.6%, and the number of incident PLC was 201 till the end of February 2017. The ratio of incidence rates was 12.32 (95% confidence interval[CI]=7.16-21.21, P < 0.0001) in HBsAg-positive arm compared with HBsAg-negative arm. The relative risk of PLC was 13.25 (95% CI=6.67-26.33, P < 0.0001) and 28.05 (95% CI=13.87-56.73, P < 0.0001) in the HBsAg+/HBeAg- group and the HBsAg+/HBeAg+ group, respectively, as compared to the HBsAg-/HBeAg- group. A series of novel PLC-related mutations including A2159G, A2189C and G2203W at the C gene, A799G, A987G and T1055A at the P gene of HBV genome were identified by using samples from the cohort. The mutation in hepatitis B virus (HBV) basal core promoter region of HBV genome has an accumulative effect on the occurrence of PLC. In addition, the tripartite relationship of aflatoxin exposure, P53 mutation and PLC was also investigated. Dynamic prediction model for PLC risk by using its long-term follow-up information and serial blood samples for QBC was developed. This model is expected to improve the efficiency of PLC screening in HBV infection individuals.