An ultra-sensitive CRISPR-Cas12a and aptamer-based biosensor utilizing Entropy-driven catalytic DNA networks for precise detection of DNA Methyltransferase 1.

DNA Methyltransferase 1 (DNMT1) serves as a crucial biomarker associated with various diseases and is essential for evaluating DNA methylation levels, diagnosing diseases, and evaluating prognosis. As a result, a convenient, quantitative, and sensitive assay for detecting DNMT1 is in high demand. However, current techniques for DNMT1 detection struggle to balance accuracy, low cost, and high sensitivity, limiting their clinical usefulness. To address this challenge, we have developed a DNMT1 detection method (CAED), which combines aptamer-specific recognition with a highly programmable Entropy-driven catalysis DNA network and is further integrated with the CRISPR-Cas12a system. This innovative approach achieves a detection limit as low as 90.9 fmol/L. To demonstrate the clinical applicability and significance of our CAED method, we successfully measured DNMT1 levels in 10 plasma samples 10 cervical tissue samples. These results underscore the potential of our method as an accurate, affordable, and ultra-sensitive tool for evaluating DNMT1 levels. This innovative method offers a potent means for assessing DNMT1 levels and significantly advances disease diagnosis and health risk prediction. Plus, it establishes an innovative design framework for CRISPR-Cas12a-based biosensors, tailored explicitly for enzyme content quantification.

Pathways and contributions of sulfate reducing-bacteria to arsenic cycling in landfills.

Sulfate-reducing bacteria (SRB) are generally found in sanitary landfills and play a role in sulfur (S) and metal/metalloid geochemical cycling. In this study, we investigated the influence of SRB on arsenic (As) metabolic pathways in refuse-derived cultures. The results indicated that SRB promote As(III) methylation and are beneficial for controlling As levels. Heterotrophic and autotrophic SRB showed significant differences during As cycling. In heterotrophic SRB cultures, the As methylation rate increased with As(III) concentration in the medium and reached a peak (85.1%) in cultures containing 25 mg L-1 As(III). Moreover, 4.0-12.6% of SO42- was reduced to S2-, which then reacted with As(III) to form realgar (AsS). In contrast, autotrophic SRB oxidized As(III) to less toxic As(V) under anaerobic conditions. Heterotrophic arsM-harboring SRB, such as Desulfosporosinus, Desulfocurvibacter, and Desulfotomaculum, express As-related genes and are considered key genera for As methylation in landfills. Thiobacillus are the main autotrophic SRB in landfills and can derive energy by oxidizing sulfur compounds and metal(loid)s. These results suggest that different types of SRB drive As methylation, redox reaction, and mineral formation in landfills. These study findings have implications for the management of As pollutants in landfills and other contaminated environments.

Green synthesis of Fe3O4@ceramsite from sludge improving anaerobic digestion performance of waste activated sludge.

Anaerobic digestion (AD) is a promising technique for waste management, which can achieve sludge stabilization and energy recovery. This study successfully prepared Fe3O4@ceramsite from WAS and applied it as an additive in sludge digestion, aiming to improve the conversion of organics to biomethane efficiency. Results showed that after adding the Fe3O4@ceramsite, the methane production was enhanced by 34.7% compared with the control group (88.0 ± 0.1 mL/g VS). Further mechanisms investigation revealed that Fe3O4@ceramsite enhanced digesta stability by strong buffering capacity, improved sludge conductivity, and promoted Fe (III) reduction. Moreover, Fe3O4@ceramsite has a larger surface area and better porous structure, which also facilitated AD performance. Microbial community analysis showed that some functional anaerobes related to AD such as Spirochaeta and Smithella were enriched with Fe3O4@ceramsite treatment. Potential syntrophic metabolisms between syntrophic bacteria (Syntrophomonas, associated with DIET) and methanogens were also detected in the Fe3O4@ceramsite treatment AD system.

Distribution and survival of pathogens from different waste components and bioaerosol traceability analysis in household garbage room.

Household garbage rooms release abundant bioaerosols and are an important source of pathogens; however, information on the distribution and survival patterns of pathogens in different waste components is limited. In this study, a culture method and 16S rRNA high-throughput sequencing were used to determine bacterial communities, culturable pathogens, and human bacterial pathogens (HBPs). The results showed that abundant culturable bacteria were detected in all waste types, and a large number of S. aureus was detected on the surface of recyclable wastes, whereas S. aureus, total coliforms, Salmonella, Enterococcus, and hemolytic bacteria were detected in food waste and other waste. The activities of these detected pathogenic bacteria decreased after 24 h of storage but re-activated within one week. Factors affecting the emergence of pathogens varied with different waste components. Sequencing results showed that Pseudomonas, Acinetobacter, and Burkholderia were abundant in the waste samples, whereas Achromobacter, Exiguobacteriums, Bordetella, and Corynebacterium were the primary pathogens in the bioaerosol and wall attachment. The results of traceability analysis showed that bioaerosol microbes were mainly derived from raw kitchen waste (5.98%) and plastic and paper contaminated with food waste (19.93%) in garbage rooms. In addition, bioaerosols were the main source of microflora in the wall attachment, which possessed high HBP diversity and required more attention. These findings will help in understanding the microbial hazards in different waste components and provide guidance for the control and risk reduction of bioaerosols during waste management and recycling.

Evaluation of pathogen spread risk from excavated landfill.

Landfill is a huge pathogen reservoir and needs special attention. Herein, the distribution and spread risk of pathogen were assessed in excavated landfill scenario. The results show that landfill excavation will greatly increase the risk of environmental microbial contamination. The highest total concentration of culturable bacteria among landfill refuse, topsoil and plant leaves was found to be as high as 1010 CFU g-1. Total coliforms, Hemolytic bacteria, Staphylococcus aureus, Salmonella, Enterococci, and Fecal coliforms were detected in the landfill surrounding environment. Notably, pathogens were more likely to adhere to plant leaves, making it an important source of secondary pathogens. The culturable bacteria concentration in the air samples differed with the landfill zone with different operation status, and the highest culturable bacteria concentration was found in the excavated area of the landfill (3.3 × 104 CFU m-3), which was the main source of bioaerosol release. The distribution of bioaerosols in the downwind outside of the landfill showed a tendency of increasing and then decreasing, and the highest concentration of bioaerosols outside of the landfill (6.56 × 104 CFU m-3) was significantly higher than that in the excavated area of the landfill. The risk of respiratory inhalation was the main pathway leading to infection, whereas the HQin (population inhalation hazardous quotient) at 500 m downwind the excavation landfill was still higher than 1, indicating that the neighboring residents were exposed to airborne microbial pollutants. The results of the study provide evidence for bioaerosols control protective measures taken to reduce health risk from the excavated landfill.

Source analysis of chlorine in municipal solid waste under waste classification: a case study of Hangzhou, China.

Inorganic chlorine is susceptible to water and soil salinization due to its non-degradability and high mobility. To clarify the environmental risks associated with the active inorganic chlorine in municipal solid waste (MSW), the specific characteristics and contributions of inorganic chlorine in different MSW categories were investigated in this study. MSW samples were collected from eight representative waste classification residential areas in Hangzhou, China. It was found that the inorganic chlorine content in different MSW categories varied significantly (0-113 mg/g). Perishable waste, paper, and plastic were found to be the main sources of inorganic chlorine in MSW. A four-category classification system was used to quantify the contribution of inorganic chlorine from each waste category. It was found that the misclassification of inorganic chlorine contributions from perishable waste and other waste accounted for 51.96% and 48.04%, respectively. However, when correctly classified into the four-category system, their contributions were reduced to 67.14% and 30.65%, respectively. Therefore, MSW classification showed a significant reduction in the overall contribution of inorganic chlorine. The misclassification reduces the contribution of inorganic chlorine to 48.04%, while correct classification increases the reduction to 69.35%.

Enhanced removal of toluene in heterogeneous aquifers through injecting encapsulated ozone micro-nano bubble water.

Organic contaminants have a tendency to accumulate in low-permeability aquifers, making their removal challenging and creating a bottleneck in groundwater remediation efforts. The use of ozone micro-nano bubbles, due to their smaller size compared to traditional macrobubbles, shows potential for efficient penetration into the low-permeability aquifer and effective oxidization of contaminants. This study conducted batch experiments, column studies, and 2D tank experiments to systematically investigate the remediation efficiency of toluene in a heterogeneous aquifer using ozonated water (OW), ozone micro-bubble water (OMBW), and encapsulated ozone micro-nano bubble water (EOMBW) with rhamnolipid. Experimental results showed that rhamnolipid effectively increased the densities and reduced the sizes of micro-nano bubbles, leading to improved ozone preservation and enhanced toluene degradation. Nanobubbles exhibited higher mobility compared to microbubbles in porous media, while rhamnolipid increased the density of penetrated nanobubbles by 9.6 times. EOMBW demonstrated superior efficiency in oxidizing toluene in low-permeability aquifers, and a numerical model was developed to successfully simulate the ozone and toluene concentration. The model revealed that the increased oxidation rate by EOMBW was attributed to the preservation of ozone in micro-nano bubbles and the enhanced toluene oxidation rate. These findings contribute significantly to the application of EOMBW in heterogeneous aquifer remediation.

Transformation behavior of heavy metal during Co-thermal treatment of hazardous waste incineration fly ash and slag/electroplating sludge.

In this study, the behavior of heavy metal transformation during the co-thermal treatment of hazardous waste incineration fly ash (HWIFA) and Fe-containing hazardous waste (including hazardous waste incineration bottom slag (HWIBS) and electroplating sludge (ES)) was investigated. The findings demonstrated that such a treatment effectively reduced the static leaching toxicity of Cr and Pb. Moreover, when the treatment temperature exceeded 1000 °C, the co-thermal treated sample exhibited low concentrations of dynamically leached Cr, Pb, and Zn, indicating that these heavy metals were successful detoxified. Thermodynamic analyses and phase transformation results suggested that the formation of spinel and the gradual disappearance of chromium dioxide in the presence of Fe-containing hazardous wastes contributed to the solidification of chromium. Additionally, the efficient detoxification of Pb and Zn was attributed to their volatilization and entry into the liquid phase during the co-thermal treatment process. Therefore, this study sets an excellent example of the co-thermal treatment of hazardous wastes and the control of heavy metal pollution during the treatment process.

Sulfate reduction behavior in response to landfill dynamic pressure changes.

During the long-term stabilization process of landfills, the pressure field undergoes constant changes. This study constructed dynamic pressure changes scenarios of high-pressure differentials (0.6 MPa) and low-pressure differentials (0.2 MPa) in the landfill pressure field at 25 °C and 50 °C, and investigated the sulfate reduction behavior in response to landfill dynamic pressure changes. The results showed that the pressurization or depressurization of high-pressure differentials caused more significant differences in sulfate reduction behavior than that of low-pressure differentials. The lowest hydrogen sulfide (H2S) release peak concentration under pressurization was only 29.67% of that under initial pressure condition; under depressurization, the highest peak concentration of H2S was up to 21,828 mg m-3, posing a serious risk of H2S pollution. Microbial community and correlation analysis showed that pressure had a negative impact on the sulfate-reducing bacteria (SRB) community, and the SRB community adjusted its structure to adapt to pressure changes. Specific SRBs were further enriched with pressure changes. Differential H2S release behavior under pressure changes in the 25 °C pressure environments were mediated by Desulfofarcimen (ASV343) and Desulfosporosinus (ASV1336), while Candidatus Desulforudis (ASV24) and Desulfohalotomaculum (ASV94) played a key role at 50 °C. This study is helpful in the formulation of control strategies for the source of odor pollution in landfills.

Enhanced biomethane production from waste activated sludge anaerobic digestion by ceramsite and amended Fe2O3 ceramsite.

Wastes recycling and reutilization technique could simultaneously fulfill waste control and energy recovery sustainably, which has attracted increasing attention. This work proposed a novel waste reuse technology utilizing ceramsite and amended Fe2O3-ceramsite made from waste activated sludge (WAS) as additives to promote the yield of methane from WAS anaerobic digestion (AD). Experimental results demonstrated that compared to the control (85.05 ± 0.2 mL CH4/g-VS), the cumulative methane yield was effectively enhanced by 14% and 40% when ceramsite and Fe2O3-ceramsite were added. Further investigation revealed that ceramsite, especially the Fe2O3-ceramsite, enriched the populations of key anaerobes involved in hydrolysis, acidification, and methanogenesis. Meanwhile, potential syntrophic metabolisms between syntrophic bacteria and methanogens were confirmed in the Fe2O3-ceramsite AD system. Mechanisms studies exhibited that ceramsite and Fe2O3-ceramsite reinforced intermediate processes for methane production. The favorable pore structure, enhanced Fe (III) reduction capacity and conductivity also contributed a lot to the AD process.

Microplastic release and sulfate reduction response in the early stage of a simulated landfill.

Landfills are essential facilities for treating and disposing municipal solid waste. They emit sulfur-containing odors and serve as an important sink for a new type of pollutant called microplastics (MPs). This study focused on the initial stage of anaerobic degradation to establish the relationship between the release of MPs and odor generation. Our findings show the rapid release of MPs into the leachate in the early stage of landfill and their predominant accumulation in the leachate sediment. The circulating leachate contained 1.45 times higher concentrations of MPs than the noncirculating leachate, with a peak concentration of 39 items·L-1. In addition, fragmentation of MPs occurred. The percentage of MPs with particle sizes of 2.5-5 mm decreased from 66.70 % to 22.32 %, while those measuring 0.1-0.5 mm increased by 33.12 %. A positive correlation was observed between MP release and sulfate reduction. Although leachate circulation increased the release of MPs, it also reduced the overall release time and total amount of MPs exported from the landfill. Compared with the initial landfill waste, the leachate operation mode, regardless of circulation, resulted in a 6.15-8.93-fold increase in MP release. These findings provide a valuable foundation for the simultaneous regulation of traditional pollutant odor and new pollutants (MPs) in landfills.

Sulfate reduction behavior in response to changing of pressure coupling with temperature inside landfill.

The behavior of sulfate reduction, which was the source of hydrogen sulfide (H2S) odor, was investigated under changing pressure and temperature conditions inside landfills. The results showed that the release of H2S and methyl mercaptan (MM) was significantly inhibited at 25 °C and 50 °C under pressure, and the highest H2S and MM concentrations released were only 0.82 %-1.30 % and 1.87 %-4.32 % of atmospheric pressure, respectively. Analysis of the microbial community structure and identification of sulfate-reducing bacteria (SRB) revealed that temperature significantly altered the microbial community in the landfill environment, while pressure inhibited some bacteria and induced the growth and reproduction of specific bacteria. Key SRB (Desulfosporosinus-ASV212, Desulfitibacter-ASV1744) mediated differentiated sulfate reduction behavior in the pressure-bearing environment at 25 °C, while key SRB (Dethiobacter-ASV177, Desulfitibacter-ASV2355 and ASV316) were involved at 50 °C. This study provides a theoretical basis for the formulation of landfill gas management and control strategies.

Insight into the impact of industrial waste co-disposal with MSW on groundwater contamination at the open solid waste dumping sites.

Due to the lack of normalized management, industrial waste is often co-disposed at open solid waste dumping sites, which could aggravate the groundwater pollution. In this study, 5 practical open solid waste dumping sites dealing with municipal solid wastes (MSW) (2 of 5) and industrial wastes mixed with MSW (3 of 5) were chosen to investigate the effect of waste co-disposal on the groundwater contamination. The industrial waste was mainly from rubber production, leather production, machinery industry, pharmaceutical industry and plastic production. 3 to 6 groundwater wells were excavated from each dumping site and 148 indices were analyzed, including regular chemicals, heavy metals, biological pollutants, volatile organic compounds (VOCs), semi-volatile organic compounds (SVOCs) and pesticide residues. Nemerow index analysis showed that 5 indices were severely polluted in the groundwater from every industrial waste co-disposal landfill, while only 0 and 1 severely polluted index was found for the two MSW landfill, respectively. The principal component analysis (PCA) analysis indicated that 2 biological pollutant (plate-counting bacteria (TPB) and total coliforms (TCs)), 4 chemical pollutants (permanganate index, ammonia, S2- and petroleum) were closely connected with the disposal of industrial waste. Besides, co-disposal of industrial waste also brought in series of PAHs and dichloromethane, with di(2-ethylhexyl)phthalate exceeding the standard limit (10.5 mg L-1). Attention should be paid to TPB and TCs, whose maximal concentrations exceeded the standard limit by extraordinary 3200 and 1600 times, respectively. The distribution pattern of the pollutants showed that the biological pollutants at the downstream area, and chemical pollutants at the leakage points exhibited the highest concentration, which indicated the downstream area and seepage points should be specially concerned for the industry waste co-disposed dumping sites.

Microbial contamination risk of landfilled waste with different ages.

Landfills are reservoirs of antibiotic resistance genes (ARGs) and pathogens, and humans are exposed to these pollutants during extensive excavation of old landfills. However, the microbial contamination risk of landfilled waste with different ages has not been assessed. In this study, human bacterial pathogens (HBPs), ARGs, and virulence factors (VFs) were systematically determined using metagenomic analysis. Results showed that the abundance of HBPs, ARGs, and VFs increased with landfill age, the percentage of HBPs in refuse with deposit age of 10-12 years (Y10) was 23.75 ± 0.49%, which was higher than that in fresh refuse (Y0, 17.99 ± 0.14%) and refuse with deposit age of 5-6 years (Y5, 19.14 ± 0.15%), indicating that old refuse had higher microbial contamination risk than fresh refuse. Multidrug, macrolide, lincosamide, streptogramine, and tetracycline resistance genes were the primary ARGs, whereas lipooligosaccharides, type IV pili, and polar flagella were the dominant VFs in refuse. The HBPs showed a significant positive correlation with ARGs and VFs. Listeria monocytogenes, Salmonella enterica, Streptococcus pneumoniae, Acinetobacter baumannii, and Escherichia coli possibly possess both multiple ARGs and VFs and could be listed as high-risk HBPs in refuse. Mobile genetic elements, especially transposons, showed positive correlations with most ARGs and VFs, and they were identified as the primary factors accounting for the variations in ARGs and VFs. These findings will help understand the spread of ARGs and VFs in landfills and evaluate the potential risk of microbiological contamination in refuse of different landfill ages, thus providing guidance for preventing disease infection during landfill excavations.

TRIM28 recruits E2F1 to regulate CBX8-mediated cell proliferation and tumor metastasis of ovarian cancer.

Chromobox protein homolog 8 (CBX8) is a transcriptional suppressor participated in various cancers. However, the function and mechanism of CBX8 in the progression of ovarian cancer (OC) are unclear. In this study, we found that CBX8 was upregulated in OC tissues originating from GEPIA and TNM databases, OC patients' samples from hospital, and OC cell lines. Furthermore, CBX8 knockdown by short hairpin RNA (shRNA) technology markedly inhibited proliferation and invasion, induced migration, cell cycle arrest, and apoptosis in vitro. Mechanistically, CBX8 activated PI3K/AKT/mTOR signaling pathway to take effect. In addition, TRIM28 and E2F1 were enriched in OC tissues from the TNM database and OC patients' samples similar to the results of CBX8. Correlation analysis indicated positive correlations among TRIM28, E2F1, and CBX8. E2F1 was proved to bind to the promoter regions of CBX8 and TRIM28, while TRIM28 recruited E2F1 to increase the expression of CBX8 to further increase cell viability, proliferation, and invasion, and decrease migration, apoptosis, and cell cycle progression. Finally, CBX8 or TRIM28 knockdown repressed tumor growth and metastasis of OC in vivo. Therefore, our study showed that the promoting effect of CBX8 on tumor growth and metastasis of OC was participated in the PI3K/AKT/mTOR signaling, TRIM28 and E2F1. Our findings suggested that CBX8 could serve as a potential marker and therapeutic target for OC patients.

Promising valorisation method of chitin biomass by producing 5-hydroxymethylfurfural using microwave hydrothermal treatment.

Chitin biomass is the second largest biomass resource on Earth but under-utilized. In this study, pretreated shrimp shells were converted into value-added platform chemical 5-hydroxymethylfurfural (HMF) using microwave hydrothermal treatment. Under the combined pretreatment of acid decalcification at room temperature and microwave-assisted alkali deacetylation, the HMF yield could reach 1.8 wt%. The key process parameters, including the holding temperature, holding time, and pH value, were evaluated and optimised. The highest HMF yield of 6.5 wt% was obtained at 202.6°C at a holding time of 5.8 min and a pH value of 1.5. This result demonstrates the potential of synchronously treating waste and recycling it, thereby offering a highly promising valorisation strategy for chitin-biomass utilisation.

Co-electron donors driven medium-chain fatty acids production: Roles of electron donors, reaction kinetics and metabolic pathways.

Energy conversion of waste activated sludge alkaline fermentation liquor (WASAFL) to medium-chain fatty acids (MCFAs) is promising for sludge treatment and carbon recovery. However, the single electron donor (ED) fermentation for MCFAs production has irreparable defects. To resolve the respective shortcomings of single electron donor (ED) and improve the MCFAs production efficiency from WASAFL, a novel biotechnical process utilizing ethanol and lactate as co-EDs within different combination ratios were investigated. The results verified that MCFAs production was highest with ethanol to lactate ratio of 1:3 (6988.54 ± 208.18 mg COD/L), being 1.46 and 1.87 times of that with ethanol and lactate as single ED. The kinetic analysis results confirmed that ethanol to lactate ratio of 1:3 resulted in the highest MCFAs yield and formation rate. The microbial taxa results uncovered that the relative abundance of Sphaerochaeta and Haloimpatiens showed positive correlation with MCFAs production. The metabolic pathway analysis indicated that the ethanol oxidization, lactate oxidization, acrylate pathway, reverse ß oxidization and fatty acid biosynthesis pathway might take place in the WASAFL fermentation system, contributing to the WASAFL-to-MCFAs conversion.

Co-vitrification of hazardous waste incineration fly ash and hazardous waste sludge based on CaO-SiO2-Al2O3 system.

Based on the CaO-SiO2-Al2O3 system, the feasibility of co-vitrification of hazardous waste incineration fly ash (FA) and hazardous waste sludge (HWS) was verified. In the CaO-SiO2-Al2O3 ternary system diagram, the melting point of the system gradually decreases with an appropriate increase in SiO2 content when the CaO/Al2O3 ratio is determined to be approximately 1. The TG-DSC results revealed that the liquid phase generation temperature in the FA and HWS mixture system was significantly lower than those of FA and HWS individually owing to the different CaO, SiO2, and Al2O3 contents; this is consistent with the results of the theoretical melting characteristics analysis, which show that the melting characteristic temperatures can be reduced by controlling the CaO-SiO2-Al2O3 ratio in the system. The co-vitrification experimental results confirmed that a vitreous content above 92%, a loss ratio on acid dissolution less than 1.74%, and leaching toxicity of heavy metals lower than 0.15 mg/L could be obtained by adjusting the CaO, SiO2, and Al2O3 contents in the FA and HWS system to 20 wt%-32.5 wt%, 35 wt%-61 wt% and 14 wt%-32.5 wt%, respectively, and under a melting temperature of 1350 °C.

Effect of dissolved organic matter on selective oxidation of toluene by ozone micro-nano bubble water.

The oxidation capacity of ozone micro-nano bubble water (OMBW) was always higher than ozonated water due to enhanced contact by bubble interface, while the effect of coexisted dissolved organic matter (DOM) on the oxidation efficiency was still unclear. In this paper, batch experiments were carried out to investigate the selective oxidation of toluene by both OMBW and ozonated water (OW) with coexisted DOM in water. Five types of background solutions were applied in this study, including humic acid solution, fulvic acid solution and three types of diluted landfill leachates at the same content of total organic carbon. Results showed that coexisted DOM had a greater inhibition effect on toluene oxidation rate by OMBW, and the oxidation rate of toluene by OMBW and OW became close. It was mainly caused by the decreased reaction rate between toluene and hydroxyl radical (kT-OH·) in OMBW after the introduction of DOM, which competed for the adsorption sites on micro-nano bubble interface. The fraction of ozone to oxidize toluene as well as kT-OH· was in positive correlations with SUVA254 and the content of humic acid-like substances, but negatively correlated with E2/E3, content of tryptophan-like proteins and content of fulvic acid-like substances. In addition, increasing the ozone dose was not effective in increasing the utilization rate of ozone in OMBW due to limited adsorption sites on micro-nano bubble interface. The paper was conductive to the application of ozone micro-nano bubble water in groundwater remediation with complex water matrices.

TEM1 up-regulates MMP-2 and promotes ECM remodeling for facilitating invasion and migration of uterine sarcoma.

OBJECTIVES: To explore the correlation between tumor endothelial marker 1 (TEM1) and matrix metalloproteinase 2 (MMP-2) in uterine sarcoma and their roles in the progression of uterine sarcoma. METHODS: Uterine leiomyosarcoma (uLMS, n = 25) and uterine leiomyoma (n = 25) specimens were collected from a total of 50 patients. Immunohistochemistry assay was conducted to determine the expression of TEM1, MMP-2 and MMP-9. TEM1 over expression (hTEM1) and low expression (shRNA-TEM1) MES-SA cell lines were established as in vitro uterine sarcoma models. MMP-2 mRNA, protein expression and enzymatic activity were verified using qPCR, Western blot and gelatin zymography respectively. MMP-2 expression was downregulated using MMP-2 siRNA in hTEM1 MES-SA cells to better study the role of MMP-2. The invasive and migratory capacities of hTEM1, shRNA-TEM1, and hTEM1 treated with MMP-2 siRNA MES-SA cells were determined using transwell assays. Extracellular matrix (ECM) remodeling mediated by TEM1 was examined using cell-ECM adhesion and fluorescent gelatin-ECM degradation assays. The immunofluorescence of F-actin was examined to analyze the formation of invadopodia. Subcutaneous and intraperitoneal xenografts were established to validate the role of TEM1 in promoting uterine sarcoma metastasis. RESULTS: TEM1 and MMP-2 were expressed in 92% (n = 23) and 88% (n = 22) of uterine leiomyosarcoma specimens, respectively. Both TEM1 and MMP-2 were highly expressed in 100% (n = 17) of high stage (III-IV) uterine leiomyosarcoma specimens. In addition, TEM1 expression was positively correlated with MMP-2 expression in uterine leiomyosarcoma. The successful establishment of in vitro uterine sarcoma models was confirmed with qPCR and Western blotting tests. TEM1 promoted the invasion and metastasis of uterine sarcoma in vivo and in vitro. MMP-2 expression and activity were up-regulated in hTEM1 cells but down-regulated in shRNA-TEM1 cells. Importantly, MMP-2 knockdown impaired the invasive and migratory capacity of hTEM1 cells. TEM1 promoted ECM remodeling by increasing cell-ECM adhesion and ECM degradation. TEM1 overexpression also induced the formation of invadopodia. CONCLUSION: TEM1 was co-expressed and positively correlated with MMP-2 in uterine leiomyosarcoma specimens. In addition, both TEM1 and MMP-2 were associated with tumor development. TEM1 promoted uterine sarcoma progression by regulating MMP-2 activity and ECM remodeling.