Interferon-γ in the tumor microenvironment promotes the expression of B7H4 in colorectal cancer cells, thereby inhibiting cytotoxic T cells.

The bioactivity of interferon-γ (IFN-γ) in cancer cells in the tumor microenvironment (TME) is not well understood in the current immunotherapy era. We found that IFN-γ has an immunosuppressive effect on colorectal cancer (CRC) cells. The tumor volume in immunocompetent mice was significantly increased after subcutaneous implantation of murine CRC cells followed by IFN-γ stimulation, and RNA sequencing showed high expression of B7 homologous protein 4 (B7H4) in these tumors. B7H4 promotes CRC cell growth by inhibiting the release of granzyme B (GzmB) from CD8+ T cells and accelerating apoptosis in CD8+ T cells. Furthermore, interferon regulatory factor 1 (IRF1), which binds to the B7H4 promoter, is positively associated with IFN-γ stimulation-induced expression of B7H4. The clinical outcome of patients with CRC was negatively related to the high expression of B7H4 in cancer cells or low expression of CD8 in the microenvironment. Therefore, B7H4 is a biomarker of poor prognosis in CRC patients, and interference with the IFN-γ/IRF1/B7H4 axis might be a novel immunotherapeutic method to restore the cytotoxic killing of CRC cells.

Strengthened Interficial Adhesive Fracture Energy by Young's Modulus Matching Degree Strategy in Carbon-Based HTM Free MAPbI3 Perovskite Solar Cell with Enhanced Mechanical Compatibility.

Carbon-based hole transport layer-free perovskite solar cells (PSCs) based on methylammonium lead triiodide (MAPbI3 ) have become one of the research focus due to low cost, easy preparation, and good optoelectronic properties. However, instability of perovskite under vacancy defects and stress-strain makes it difficult to achieve high-efficiency and stable power output. Here, a soft-structured long-chain 2D pentanamine iodide (abbreviated as "PI") is used to improve perovskite quality and interfacial mechanical compatibility. PI containing CH3 (CH2 )4 NH3 + and I- ions not only passivate defects at grain boundaries, but also effectively alleviate residual stress during high temperature annealing via decreasing Young's modulus of perovskite film. Most importantly, PI effectively increases matching degree of Young's modulus between MAPbI3 (47.1 GPa) and carbon (6.7 GPa), and strengthens adhesive fracture energy (Gc ) between perovskite and carbon, which is helpful for outward release of nascent interfacial stress generated under service conditions. Consequently, photoelectric conversion efficiency (PCE) of optimal device is enhanced from 10.85% to 13.76% and operational stability is also significantly improved. 83.1% output is maintained after aging for 720 h at room temperature and 25-60% relative humidity (RH). This strategy of regulation from chemistry and physics provides a strategy for efficient and stable carbon-based PSCs.

Fate of glyphosate in lakes with varying trophic levels and its modification by root exudates of submerged macrophytes.

Accelerated eutrophication in lakes reduces the number of submerged macrophytes and alters the residues of glyphosate and its degradation products. However, the effects of submerged macrophytes on the fate of glyphosate remain unclear. We investigated eight lakes with varying trophic levels along the middle and lower reaches of the Yangtze River in China, of which five lakes contained either glyphosate or aminomethylphosphate (AMPA). Glyphosate and AMPA residues were significantly positively correlated with the trophic levels of lakes (P < 0.01). In lakes, glyphosate is degraded through the AMPA and sarcosine pathways. Eight shared glyphosate-degrading enzymes and genes were observed in different lake sediments, corresponding to 44 degrading microorganisms. Glyphosate concentrations in sediments were significantly higher in lakes with lower abundances of soxA (sarcosine oxidase) and soxB (sarcosine oxidase) (P < 0.05). In the presence of submerged macrophytes, oxalic and malonic acids secreted by the roots of submerged macrophytes increased the abundance of glyphosate-degrading microorganisms containing soxA or soxB (P < 0.05). These results revealed that a decrease in the number of submerged macrophytes in eutrophic lakes may inhibit glyphosate degradation via the sarcosine pathway, leading to a decrease in glyphosate degradation and an increase in glyphosate residues.

Investigation of phytoplankton community structure and formation mechanism: a case study of Lake Longhu in Jinjiang.

In order to explore the species composition, spatial distribution and relationship between the phytoplankton community and environmental factors in Lake Longhu, the phytoplankton community structures and environmental factors were investigated in July 2020. Clustering analysis (CA) and analysis of similarities (ANOSIM) were used to identify differences in phytoplankton community composition. Generalized additive model (GAM) and variance partitioning analysis (VPA) were further analyzed the contribution of spatial distribution and environmental factors in phytoplankton community composition. The critical environmental factors influencing phytoplankton community were identified using redundancy analysis (RDA). The results showed that a total of 68 species of phytoplankton were found in 7 phyla in Lake Longhu. Phytoplankton density ranged from 4.43 × 105 to 2.89 × 106 ind./L, with the average density of 2.56 × 106 ind./L; the biomass ranged from 0.58-71.28 mg/L, with the average biomass of 29.38 mg/L. Chlorophyta, Bacillariophyta and Cyanophyta contributed more to the total density, while Chlorophyta and Cryptophyta contributed more to the total biomass. The CA and ANOSIM analysis indicated that there were obvious differences in the spatial distribution of phytoplankton communities. The GAM and VPA analysis demonstrated that the phytoplankton community had obvious distance attenuation effect, and environmental factors had spatial autocorrelation phenomenon, which significantly affected the phytoplankton community construction. There were significant distance attenuation effects and spatial autocorrelation of environmental factors that together drove the composition and distribution of phytoplankton community structure. In addition, pH, water temperature, nitrate nitrogen, nitrite nitrogen and chemical oxygen demand were the main environmental factors affecting the composition of phytoplankton species in Lake Longhu.

Photodegradation of glyphosate in water and stimulation of by-products on algae growth.

Glyphosate is the most widely used herbicide in global agricultural cultivation. However, little is known about the environmental risks associated with its migration and transformation. We conducted light irradiation experiments to study the dynamics and mechanism of photodegradation of glyphosate in ditches, ponds and lakes, and evaluated the effect of glyphosate photodegradation on algae growth through algae culture experiments. Our results showed that glyphosate in ditches, ponds and lakes could undergo photochemical degradation under sunlight irradiation with the production of phosphate, and the photodegradation rate of glyphosate in ditches could reach 86% after 96 h under sunlight irradiation. Hydroxyl radicals (•OH) was the main reactive oxygen species (ROS) for glyphosate photodegradation, and its steady-state concentrations in ditches, ponds and lakes were 6.22 × 10-17, 4.73 × 10-17, and 4.90 × 10-17 M. The fluorescence emission-excitation matrix (EEM) and other technologies further indicated that the humus components in dissolved organic matter (DOM) and nitrite were the main photosensitive substances producing •OH. In addition, the phosphate generated by glyphosate photodegradation could greatly promote the growth of Microcystis aeruginosa, thereby increasing the risk of eutrophication. Thus, glyphosate should be scientifically and reasonably applied to avoid environmental risks.

New insights into the mechanism of phosphate release during particulate organic matter photodegradation based on optical and molecular signatures.

Phosphate release from particulate organic matter (POM) dominates phosphorus (P) cycling in aquatic ecosystems. However, the mechanisms underlying P release from POM remain poorly understood because of complex fractionation and analytical challenges. In this study, the release of dissolved inorganic phosphate (DIP) during POM photodegradation was assessed using excitation-emission matrix (EEM) fluorescence spectroscopy and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). POM in suspension was significantly photodegraded under light irradiation, concomitantly with the production and release of DIP in the aqueous solution. Chemical sequential extraction revealed that organic phosphorus (OP) in POM participated in photochemical reactions. Moreover, FT-ICR MS analysis revealed that the average molecular weight of P-containing formulas decreased from 374.2 to 340.1 Da. Formulas containing P with a lower oxidation degree and unsaturation were preferentially photodegraded, generating oxygen-enriched and saturated formula compounds, such as protein- and carbohydrate-like P-containing formulas, benefiting further utilization of P by organisms. Reactive oxygen species played an important role in the photodegradation of POM, and excited triplet state chromophoric dissolved organic matter (3CDOM*) was mainly responsible for POM photodegradation. These results provide new insights into the P biogeochemical cycle and POM photodegradation in aquatic ecosystems.

Resin-based iron-manganese binary oxide for phosphate selective removal.

Adsorption technology can effectively remove phosphorus from water and realize phosphorus recovery. Hence, it is used to curb the eutrophication of water and alleviate the crisis caused by the shortage of phosphorus resources. Resin has been attracting increasing interest as an ideal adsorption material; however, its practical application is greatly affected by environmental factors. To solve the competitive adsorption and pore blockage caused by humic acid and coexisting ions during the removal of phosphorus by ion-exchange resin, this study has developed an iron-manganese oxide-modified resin composite adsorbent (Fe/Mn-402) based on the nanoconfinement theory. The structural characterization results of XRD, FT-IR, SEM, and XPS showed that the iron-manganese binary oxide was successfully loaded on the skeleton of the strongly alkaline anion resin and showed good stability under both neutral and alkaline conditions. The batch adsorption experiments showed that the maximum adsorption capacity of Fe/Mn-402 for phosphorus can reach up to 50.97 mg g-1 under the optimal raw material ratio (Fe:Mn = 1:1). In addition, Fe/Mn-402 shows good selectivity for phosphorus removal. Fe/Mn-402 can maintain good adsorption performance for phosphate even under high concentrations of SO42-, HCO3-, and humic acid. The regenerated Fe/Mn-402 can be recycled without any obvious change in its treatment capacity. Hence, it is suitable for stable, long-term usage. In general, this work puts forward a new idea for the development of phosphorus-removal adsorbents for the treatment of wastewater containing coexisting ions and HA.

Comparative study for removal of phosphorus from aqueous solution by natural and activated bentonite.

The novelty of the current article is to investigate the adsorption potential of the Egyptian natural and activated bentonite (Na-bentonite) to inorganic and organic phosphorus (IP, OP) in aqueous media. The natural bentonite (NB) was activated to Na-bentonite (Na-B) by a new facile method within 2 h. NB and Na-B were also characterized using XRD, XRF, BET ESM, and FT-IR. The batch experiment has been employed to select the ideal conditions for the removal of inorganic and organic phosphorus (IP, OP) from aqueous solutions. The findings clearly showed that the Na-bentonite is enriched with sodium in the form of Na-montmorillonite with a higher specific area 138.51 m2/g than the value for the natural bentonite 74.21 m2/g. The batch experiment showed maximum absorption for both IP and OP adsorbents occurred at an equilibrium pH = 6, contact time of 40 to 50 min, 40 °C temperature, and a dose rate of 2 mg/L and 1 mg/L, respectively. The equilibrium data displayed better adjustment to Langmuir than the Freundlich, Temkin, and Dubinin-Radushkevich isotherms and adsorption kinetics followed the pseudo-second-order-type kinetic, and the parameters of thermodynamics reveal that adsorption occurs spontaneously and exothermic nature. Na-bentonite proved to be more efficient in removing target material than natural bentonite. The spent bentonites were easily regenerated by chemical methods.

Spatial optimization of ecological ditches for non-point source pollutants under urban growth scenarios.

Non-point source (NPS) pollution is regarded as the major threat to water quality worldwide, and ecological ditches (EDs) are considered an important and widely used method to collect and move NPS pollutants from fields to downstream water bodies. However, few studies have been conducted to optimize the spatial locations of EDs, particularly when the watershed experiences urbanization and rapid land-use changes. As land-use patterns change the spatial distribution of NPS loads, this study used a cellular automata-Markov method to simulate future land-use changes in a typical agricultural watershed. Three scenarios are included as follows: historical trend, rapid urbanization, and ecological protection scenarios. The spatial distributions of particulate phosphorus loads were simulated using the revised universal soil loss equation and sediment transport distribution model. The results suggested that the total particulate phosphorus (TP) load in the Zhuxi watershed decreased by 10,555.2 kg from 2000 to 2020, primarily because the quality and quantity of forests in Zhuxi County improved over the last 20 years. The TP load in Zhuxi watershed would be 2588.49, 2639.15, and 2553.32 kg in 2040 in historical trend, rapid urbanization, and ecological protection scenarios, respectively, compared with 2308.1 kg in 2020. This indicated that urban expansion increases the TP load, and the faster the expansion rate, the more the TP load. Consequently, the optimal locations of EDs were determined based on the intercepted loads and the period during which they existed during land-use changes. The results suggested that rapid urbanization would consequently reduce the space available for building EDs and also increase the cost of building EDs to control the NPS pollution in the watershed.

Influence of glyphosate and its metabolite aminomethylphosphonic acid on aquatic plants in different ecological niches.

Glyphosate and its metabolite aminomethylphosphonic acid (AMPA) draw great concern due to their potential threat to aquatic ecosystems. The individual and combined effects of glyphosate and AMPA on aquatic plants in different ecological niches need to be explored. This study aimed to investigate the ecotoxicity of glyphosate and AMPA on the emergent macrophyte Acorus calamus, phytoplankton Chlorella vulgaris, and submerged macrophyte Vallisneria natans after their exposure to glyphosate and AMPA alone and to their mixture. Medium and low concentrations of glyphosate (≤ 0.5 mg L-1) significantly inhibited the growth of V. natans and promoted the growth of C. vulgaris (P < 0.05) but had no significant effect on the growth of A. calamus (P > 0.05). AMPA (≤ 5.0 mg L-1) did not significantly influence the relative growth rate (except C. vulgaris) or malonaldehyde levels but significantly altered the expression levels of chlorophyll-related genes and superoxide dismutase [Cu-Zn] genes in the aquatic plants examined. AMPA mainly affected the oxidative phosphorylation pathway in V. natans and not those in other two plants, indicating that V. natans was more sensitive to AMPA-induced oxidative damage. Moreover, antagonistic effects on plant growth were observed when plants were exposed to low concentrations of glyphosate + AMPA (≤ 0.1 + 0.1 mg L-1). When the concentration of glyphosate + AMPA reached 0.5 + 0.5 and 5.0 + 5.0 mg L-1, the growth of the submerged macrophyte was additively or synergistically inhibited, but the growth of the emergent macrophyte and phytoplankton was antagonistically inhibited. Our results indicated that both the individual and combined effects of glyphosate and AMPA might alter the vertical structure of shallow lakes and accelerate the conversion of shallow lakes from grass-based to algal-based lakes.

Novel ecological ditch system for nutrient removal from farmland drainage in plain area: Performance and mechanism.

The loading of nitrogen (N) and phosphorus (P) from agricultural drainage as the non-point sources is a worldwide environmental issue for aquatic ecosystem. However, how to remove these nutrients effectively from agricultural drainage remains a big challenge with increasing cemented ditches for better management. Here, we designed a novel ecological ditch system which integrated an earth ditch and a cemented ditch with iron-loaded biochar in the Chengdu Plain to reduce the loss of N and P from farmland. After a two-year monitoring, the removal efficiency of total N and total P reached 24.9% and 36.1% by the earth ditch and 30.7% and 57.8% by the integrated ditch system, respectively. The water quality was evidently improved after passing through the ditch system with the marked decrease in the concentrations of N and P. Dissolved organic N, nitrate, and particulate P became the dominant fractions of N and P loss. Rainfall soon after fertilization increased the concentrations of N and P in the ditch system and markedly affected their removal efficiency. The iron-loaded biochar effectively removed N and P from the drainage, especially at the high concentrations, which was mainly attributed to its high adsorption of the dissolved N and P fractions and the interception of the particulate nutrients. Our results indicate that the designed ecological ditch system has a high potential for alleviating agricultural non-point source pollution in the plain area and can be extended to other lowland agricultural ecosystems.

The effect of iron oxide types on the photochemical transformation of organic phosphorus in water.

Iron oxides play an important role in the transport and transformation of organic phosphorus in aquatic environments. However, the effect of different types of iron oxide on the environmental fate of organic phosphorus has remained unclear. In this study, the photodegradation of the organic phosphorus compound adenosine triphosphate (ATP) via the activity of crystalline (goethite) and amorphous (ferrihydrite) iron oxides was investigated. It was found that ATP was photodegraded by goethite, resulting in the release of dissolved inorganic phosphate under simulated sunlight irradiation. The concentration of ATP on goethite decreased by 75% after 6 h of simulated sunlight irradiation, while the concentration of ATP on ferrihydrite decreased by only 22%. ATR-FTIR spectroscopy revealed that the intensity of the peaks for the P-O and PO stretching vibrations in the goethite-ATP complex decreased significantly more after simulated sunlight irradiation than did those for the ferrihydrite treatment. Combined with the higher TOC/TOC0 values for the goethite treatment, the results indicate that a more vigorous photochemical reaction took place in the presence of goethite than with ferrihydrite. Reactive oxygen species analysis also showed that hydroxyl and superoxide anion radicals were generated when goethite was exposed to simulated sunlight irradiation, while ferrihydrite did not exhibit this ability. Overall, this study highlights that the type of iron oxide is an important factor in the transformation of organic phosphorus in aquatic environments.

SSThe coexistence and diversity of Candidatus methylomirabilis oxyfera-like and anammox bacteria in sediments of an urban eutrophic lake / SLa coexistencia y diversidad de bacterias Candidatus methylomirabilis oxyfera y anammox en sedimentos de un lago urbano eutrófico

Tangxun Lake is the largest urban lake in China, which is polluted by multiple wastewaters, and now is severely eutrophic. We detected diversity, abundance, and the coexistence of Candidatus Methylomirabilis oxyfera-like and anammox bacteria in different horizontal and vertical directions of the lake sediments through qPCR and clone library. Phylogenetic tree analysis showed that the Ca. Methylomirabilis oxyfera-like and anammox bacteria exhibited high diversity, and they belonged to group B—E and Ca. Brocadia genus, respectively. These two bacteria displayed higher diversity in polluted area than in other areas. Furthermore, they had great spatial variation of abundance both horizontally and vertically. The abundance of anammox bacteria was significantly higher than that of Ca. Methylomirabilis oxyfera-like bacteria. The stronger the human interference were, the higher abundances these two bacteria exhibited horizontally, whereas both their abundances and the ratio of anammox to Ca. Methylomirabilis oxyfera-like bacteria decreased with the increasing depth. Redundancy analysis indicated that nitrate was the most influential environmental factor to the abundance of these two bacteria. Ammonia, nitrite, total nitrogen, and organic matters were in positive correlation with the abundance of these two bacteria. Nitrate was slightly negatively correlated with the abundance of Ca. Methylomirabilis oxyfera-like bacteria, while it was positively correlated with that of anammox bacteria. Our results provided an insight into the effects of environmental factors such as ammonia, nitrite, and nitrate on the diversity and abundances of these two bacteria and theoretical basis for restoration of water.(AU)

Molecular insight into the release of phosphate from dissolved organic phosphorus photo-mineralization in shallow lakes based on FT-ICR MS analysis.

Dissolved organic phosphorus (DOP) is a key factor in the water eutrophication process because of its high potential bioavailability and inorganic phosphate (Pi) compensation ability through bio- and photo-mineralization. However, the research on the characterization and transformation of DOP is insufficient owing to their complex composition. This study investigates the release of dissolved Pi from DOP photo-mineralization in Lake Dong based on Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) analysis. The results showed that the photo-release of dissolved Pi is spatially heterogeneous in Lake Dong and is consistent with the distribution of DOP concentration. The FT-ICR MS results showed that the simulated irradiation decreased the relative abundance (RA) of the DOP molecular formulae with higher molecular weight (MW) and higher double bond equivalence values (DBE), while the RA of DOP molecular formulae with lower MW and lower DBE value increased or remained. Besides, the total RA of lipid-like formulae increased from 49.09% to 55.90%, while the oxy-aromatic-like formulae decreased from 50.91% to 44.10%, suggesting that simulated irradiation would influence the potential bioavailability of DOP. As the main photolysis medium during DOP photo-mineralization, the hydroxyl radicals (∙OH) are mainly derived from dissolved organic matter (DOM) compared to the nitrate (NO3-) and iron ion (Fe3+) in Lake Dong. These results are important in understanding the ability and mechanism of DOP photo-mineralization and provide suggestions for cycling phosphorus in eutrophic shallow lakes.

Organic carbon quantity and composition jointly modulate the differentiation of nitrate reduction pathways in sediments of the Chinese eutrophic lake, Lake Chaohu.

Twelve sampling sites from two basins of Lake Chaohu were studied seasonally from June 2020 to April 2021 in Hefei City (China) to better understand the effect of organic carbon (C) quantity and composition on nitrate (NO3--N) reduction pathways. Serious algal bloom in the west basin of Lake Chaohu (WLC) resulted in higher organic C accumulation and NO3--N deficiency in interstitial water compared to the east basin of Lake Chaohu (ELC), jointly leading to a high C/NO3--N ratio. This triggered dissimilatory nitrate reduction to ammonium (DNRA) over denitrification in terms of higher DNRA rate, nitrogen retaining index (NRI), and nrfA gene abundance mediating DNRA. Furthermore, high oxygen-alkyl C and abundance of functional genes mediating labile organic C decomposition and DNRA suggested that the alkyl carbon-oxygen bond was responsible for DNRA induction. Different bacterial community composition and diversity involved in C and nitrogen (N) metabolism in two basins indicated that bacteria in sediments of WLC were more active in NO3--N reduction. Spearman correlation analysis showed that the less represented genera, such as Thiobacillus and Clostridium, were positively correlated with both organic C and NO3--N reduction rates, respectively. Hence, organic C composition could affect NO3--N reduction function by shaping the specific bacterial community.

Current status of microbes involved in the degradation of pharmaceutical and personal care products (PPCPs) pollutants in the aquatic ecosystem.

Contamination of aquatic systems with pharmaceuticals, personal care products, steroid hormones, and agrochemicals has been an immense problem for the earth's ecosystem and health impacts. The environmental issues of well-known persistence pollutants, their metabolites, and other micro-pollutants in diverse aquatic systems around the world were collated and exposed in this review assessment. Waste Water Treatment Plant (WWTP) influents and effluents, as well as industrial, hospital, and residential effluents, include detectable concentrations of known and undiscovered persistence pollutants and metabolites. These components have been found in surface water, groundwater, drinking water, and natural water reservoirs receiving treated and untreated effluents. Several studies have found that these persistence pollutants, and also similar recalcitrant pollutants, are hazardous to a variety of non-targeted creatures in the environment. In human and animals, they can also have severe and persistent harmful consequences. Because these pollutants are harmful to aquatic organisms, microbial degradation of these persistence pollutants had the least efficiency. Fortunately, only a few wild and Genetically Modified (GMOs) microbial species have the ability to degrade these PPCPs contaminants. Hence, researchers have been studying the degradation competence of microbial communities in persistence pollutants of Pharmaceutical and Personal Care Products (PPCPs) and respective metabolites for decades, as well as possible degradation processes in various aquatic systems. As a result, this review provides comprehensive information about environmental issues and the degradation of PPCPs and their metabolites, as well as other micro-pollutants, in aquatic systems.

SSThe coexistence and diversity of Candidatus methylomirabilis oxyfera-like and anammox bacteria in sediments of an urban eutrophic lake.

Tangxun Lake is the largest urban lake in China, which is polluted by multiple wastewaters, and now is severely eutrophic. We detected diversity, abundance, and the coexistence of Candidatus Methylomirabilis oxyfera-like and anammox bacteria in different horizontal and vertical directions of the lake sediments through qPCR and clone library. Phylogenetic tree analysis showed that the Ca. Methylomirabilis oxyfera-like and anammox bacteria exhibited high diversity, and they belonged to group B-E and Ca. Brocadia genus, respectively. These two bacteria displayed higher diversity in polluted area than in other areas. Furthermore, they had great spatial variation of abundance both horizontally and vertically. The abundance of anammox bacteria was significantly higher than that of Ca. Methylomirabilis oxyfera-like bacteria. The stronger the human interference were, the higher abundances these two bacteria exhibited horizontally, whereas both their abundances and the ratio of anammox to Ca. Methylomirabilis oxyfera-like bacteria decreased with the increasing depth. Redundancy analysis indicated that nitrate was the most influential environmental factor to the abundance of these two bacteria. Ammonia, nitrite, total nitrogen, and organic matters were in positive correlation with the abundance of these two bacteria. Nitrate was slightly negatively correlated with the abundance of Ca. Methylomirabilis oxyfera-like bacteria, while it was positively correlated with that of anammox bacteria. Our results provided an insight into the effects of environmental factors such as ammonia, nitrite, and nitrate on the diversity and abundances of these two bacteria and theoretical basis for restoration of water.

PYY modulates the tumorigenesis and progression of colorectal cancer unveiled by proteomics.

Despite decrease in mortality caused by colorectal cancer (CRC), there remains no effective therapeutic method for patients with CRC. We attempted to screen biomarkers with therapeutic values in CRC. Proteomic analysis was performed on tumor, tumor-adjacent, and normal tissues derived from five patients with colon adenocarcinoma (COAD) via label-free proteome profiling. Differentially expressed proteins (DEPs) were identified, and functional annotation was performed based on the gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. The effect of marker proteins on CRC was determined via cell function experiments and using tumor organoid models. The localization of the marker proteins was determined via immunofluorescence. A total of 126 DEPs were identified in COAD tissues compared with normal tissues, of which Peptide YY (PYY) overlapped among the tumor, adjacent, and normal groups. DEPs in the cancer group vs. normal group were enriched in the regulation of cell cycle checkpoint, developmental process, focal adhesion, and apoptosis-related pathways. The low expression of PYY in CRC tissues was verified via qRT-PCR, western blotting, and immunohistochemistry. Overexpression of PYY promoted apoptosis and inhibited the proliferation, migration, and invasion of HCT116 and HT29 cells. Furthermore, PYY was secreted by neurons and its supplementation suppressed tumor organoid growth in a dose-dependent manner. In conclusion, PYY exerted inhibitory action on CRC and could be a therapeutic target for CRC.

Directed-evolution of translation system for efficient unnatural amino acids incorporation and generalizable synthetic auxotroph construction.

Site-specific incorporation of unnatural amino acids (UAAs) with similar incorporation efficiency to that of natural amino acids (NAAs) and low background activity is extremely valuable for efficient synthesis of proteins with diverse new chemical functions and design of various synthetic auxotrophs. However, such efficient translation systems remain largely unknown in the literature. Here, we describe engineered chimeric phenylalanine systems that dramatically increase the yield of proteins bearing UAAs, through systematic engineering of the aminoacyl-tRNA synthetase and its respective cognate tRNA. These engineered synthetase/tRNA pairs allow single-site and multi-site incorporation of UAAs with efficiencies similar to those of NAAs and high fidelity. In addition, using the evolved chimeric phenylalanine system, we construct a series of E. coli strains whose growth is strictly dependent on exogenously supplied of UAAs. We further show that synthetic auxotrophic cells can grow robustly in living mice when UAAs are supplemented.

Tumor cell-derived SPON2 promotes M2-polarized tumor-associated macrophage infiltration and cancer progression by activating PYK2 in CRC.

BACKGROUND: Tumor-associated macrophages (TAMs) are key regulators of the complex interplay between cancer and the immune microenvironment. Tumor cell-derived spondin 2 (SPON2) is an extracellular matrix glycoprotein that has complicated roles in recruitment of macrophages and neutrophils during inflammation. Overexpression of SPON2 has been shown to promote tumor cell migration in colorectal cancer (CRC). However, the mechanism by which SPON2 regulates the accumulation of TAMs in the tumor microenvironment (TME) of CRC is unknown. METHODS: Immunohistochemistry was used to examine SPON2 expression in clinical CRC tissues. In vitro migration assays, transendothelial migration assays (iTEM), and cell adhesion assays were used to investigate the effects of SPON2 on monocyte/macrophage migration. Subcutaneous tumor formation and orthotopic implantation assays were performed in C57 BL/6 mice to confirm the effects of SPON2 on TAM infiltration in tumors. RESULTS: SPON2 expression is positively correlated with M2-TAM infiltration in clinical CRC tumors and poor prognosis of CRC patients. In addition, SPON2 promotes cytoskeletal remodeling and transendothelial migration of monocytes by activating integrin ß1/PYK2 axis. SPON2 may indirectly induce M2-polarization through upregulating cytokines including IL10, CCL2 and CSF1 expression in tumor cells. Blocking M2 polarization and Macrophage depletion inhibited the SPON2-induced tumors growth and invasion. Furthermore, blocking the SPON2/integrin ß1/PYK2 axis impairs the transendothelial migration of monocytes and cancer-promoting functions of TAMs in vivo. CONCLUSIONS: Our findings demonstrate that SPON2-driven M2-TAM infiltration plays an important role during CRC tumor growth and metastasis. SPON2 may be a valuable biomarker guiding the use of macrophage-targeting strategies and a potential therapeutic target in advanced CRC.