Attenuated Salmonella typhimurium L forms suppress tumor growth and promote apoptosis in murine ovarian tumors.

To study the effects of attenuated Salmonella typhimurium L forms on the in vivo tumorigenicity and apoptosis of murine epithelial ovarian cancer cells, as well as the related mechanisms. Attenuated Salmonella typhimurium VNP20009 was induced into bacterial L forms by using antibiotic ceftriaxone. CCK-8 cell proliferation assay showed that attenuated S. typhimurium L forms can inhibit the proliferation of murine ovarian epithelial cancer ID8 cells. Attenuated ST L forms can induce apoptosis and inhibit invasion ability of epithelial ovarian cancer cells in vitro. TUNEL assay showed that attenuated ST L forms can induce apoptosis of ID8 cells in murine ovarian tumors. Meanwhile, attenuated ST L forms inhibit tumor growth in murine ovarian tumors. The tumorigenicity-related proteins of xenograft tumors detected by immunohistochemistry and fluorescence quantitative RT-PCR assays showed that attenuated ST L forms can reduce the expression of proteins that promote tumor growth and metastasis, such as Lgals9 and MMP9. This study confirmed that attenuated ST L forms can suppress tumor growth and promote apoptosis in murine ovarian tumors. Attenuated ST L forms may serve as a novel biological agent for bacterial-mediated tumor therapy in epithelial ovarian cancer.

LncRNA HOTAIR down-expression inhibits the invasion and tumorigenicity of epithelial ovarian cancer cells by suppressing TGF-ß1 and ZEB1.

BACKGROUND: Epithelial ovarian cancer (EOC) is a pathological type with a higher mortality rate among gynecological cancers today. Long-chain noncoding RNAs (lncRNAs) can regulate the transcription and expression of cellular genes. However, the downstream molecules regulated by lncRNA HOTAIR have not been well studied. The effects of downregulated lncRNA HOTAIR on EOC invasiveness and tumorigenicity in nude mice, along with TGF- ß1 and ZEB1 in epithelial ovarian cancer cells, need to be investigated in further research. RESULTS: RT-qPCR was used to detect lncRNA HOTAIR and TGF-ß1 and ZEB1 mRNA expression in EOC SKOV3 cells. The expression of lncRNA HOTAIR in SKOV3 cells transfected with the recombinant shHOTAIR interference plasmid was significantly lower than that of the negative control. Compared with the negative control, the matrix gel invasion ability of shHOTAIR SKOV3 cells in vitro and their tumorigenicity in nude mice were significantly reduced. Moreover, compared with the control, the expression of ZEB1 protein in shHOTAIR-SKOV3 xenograft tumors was significantly reduced. Downregulation of lncRNA HOTAIR expression significantly reduced TGF-ß1 and ZEB1 mRNA expression, but increased the expression of E-cadherin mRNA. In summary, downregulated lncRNA HOTAIR in EOC SKOV3 cells transfected with shHOTAIR can inhibit TGF-ß1, reduce ZEB1, increase E-cadherin, and significantly reduce the invasiveness and tumorigenicity of ovarian epithelial cancer SKOV3 cells. CONCLUSIONS: These results suggest that the lncRNA HOTAIR may be an effective target for the treatment of human EOC.

Photochemical synthesis of group 10 metal nanoclusters for electrocatalysis.

Four group 10 metal nanoclusters, Ni10(4-MePhS)20, Ni11(PhS)22, Pd9(PhS)18 and Pd10(PhS)20 were synthesized from disulfides based on a photochemical reduction-oxidation cascade process, which proceeds via a different mechanism to that of the conventional two-step reduction process. The as-obtained nanoclusters possess oxidative resistance and structural robustness under different conditions. Their atomically precise structures are determined to be nickel or palladium rings in which the metal atoms are bridged by Ar-S groups. Their catalytic performance in oxygen reduction reaction was compared, and the ring size-dependent catalytic activity of the group 10 metal nanoclusters was revealed. This work provides an efficient route to atomically precise and structurally stable group 10 metal nanoclusters, and sheds light on their further applications in electrocatalysis.

Hydrothermal Pretreatment of KOH for the Preparation of PAC and Its Adsorption on TC.

The environment has been heavily contaminated with tetracycline (TC) due to its excessive use; however, activated carbon possessing well-developed pores can effectively adsorb TC. This study synthesized pinecone-derived activated carbon (PAC) with high specific surface area (1744.659 cm2/g, 1688.427 cm2/g) and high adsorption properties (840.62 mg/g, 827.33 mg/g) via hydrothermal pretreatment methods utilizing pinecones as precursors. The results showed that PAC treated with 6% KOH solution had excellent adsorption properties. It is found that the adsorption process accords with the PSO model, and a large amount of C=C in PAC provides the carrier for π-πEDA interaction. The results of characterization and the isothermal model show that TC plays a key role in the adsorption process of PAC. It is concluded that the adsorption process of TC on PAC prepared by hydrothermal pretreatment is mainly pore filling and π-πEDA interaction, which makes it a promising adsorbent for TC adsorption.

Comparative Study on the Pyrolysis Behaviors of Pine Cone and Pretreated Pine Cone by Using TGA-FTIR and Pyrolysis-GC/MS.

Pine cone (PC) is a potential biomass energy source and is rich in nonstructural substances (NSS). To understand the impact of these NSS on the pyrolysis behavior and its products, in this study, phenol alcohol extraction was used for the separation of NSS from PC (the PC after separation of NSS was labeled as A-PC), and then thermogravimetric analysis -Fourier transform infrared and PY-gas chromatography/MS detection techniques were used to conduct a systematic comparison of the thermal degradation behaviors and kinetics parameters of PC and A-PC. Results showed that the N content of PC was higher than that of other biomass, and the activation energies of PC and A-PC generally decreased at first and then increased as the conversion rate increased. Furthermore, the activation energy of PC decreased with conversion rates in the range of 0.25-0.30, while A-PC lagged significantly behind PC. On the other hand, the maximum absorption peak of CH4 for PC was higher than that of A-PC, and the maximum absorption peak of CO2 was quite the opposite. After extracting NSS from the PC, its activation energy was significantly increased.

Technical and Benefit Evaluation of Fruit-Wood Waste Gasification Heating Coproduction of an Activated Carbon System.

Biomass gasification polygeneration technology can well address both the economic and environmental issues that impeded the development of biomass gasification technology. To further improve the utilization efficiency of biomass, preactivation of gasified carbon is realized in the gasification reactor. The aim of this study is to adopt a new gasification reactor and an environmental protection combustion chamber to obtain high value-added activated carbon with clean heating. In this paper, an experimental study on the fruit-wood waste gasification heating coproduction of an activated carbon system was carried out. The results show that the yield of gasified carbon is 20.22%, the specific surface area of gasified carbon reaches 590 m2/g, the yield of activated carbon is 10.37%, and the gas yield is 1.9 Nm3/kg. The gasification efficiency of the system is 57.83%, the energy that is transferred to the activated carbon is 18.72%, and the percentage of fixed carbon is 24.3%. Compared with the biomass particle, coal, and natural gas heating projects, the environmental protection benefits of the project are significant, and the negative emission of CO2 is realized. Compared with the heating benefit of coal and natural gas, the economic benefit of this project is more significant.

Features and Commercial Performance of a System of Biomass Gasification for Simultaneous Clean Heating and Activated Carbon Production.

Biomass is a renewable and clean energy. Moreover, clean heating plays a vital role in solving issues related to the heating source structures in northern China. This paper reports on our novel technology: a system of biomass (mainly fruitwood waste, referred to in short as FWW) gasification for simultaneous clean heating and fruitwood activated carbon (FAC) production. In particular, we will discuss the features of our gasification system and product characteristics, as well as energy efficiency, environmental benefits, and economic benefits. The results showed that the energy conversion from FWW gasification was as follows: 48.10% hot gas, 49.08% fruitwood gasified carbon (FGC), and 2.82% energy loss. The NO x emissions of this system were about 126 mg/Nm3. The iodine adsorption values of the derived FGC and FAC were about 550 and 1000 mg/g, respectively. The system of gasification consumed 36 t of FWW per day, obtained 10 t of FGC, and produced 5 t of FAC. The emissions of CO2 were neutral during the operation, and the clean heating area was 4100 m2/d in Chengde, Hebei, China, with the payback period under one heating season. These results show that the system is practical, economical, energy-saving, and environmentally friendly.

Upgrading of bio-oil via solar pyrolysis of the biomass pretreated with aqueous phase bio-oil washing, solar drying, and solar torrefaction.

In this study, three types of biomass were first pretreated with an aqueous phase bio-oil instead of traditional acid washing. Then, the washed samples were pretreated with drying (100 â„ƒ, 30 min) and torrefaction (250 â„ƒ, 30 min) using a parabolic-trough solar receiver system. The subsequent pyrolysis was performed at 550 â„ƒ for 10 min using a parabolic-dish solar receiver system. Results showed that the solar energy can effectively ensure the temperature required for biomass drying, torrefaction, and pyrolysis, having thus a potential to replace the conventional electric heating or fossil fuel heating. Such a strategy combines the advantages of the independent pretreatments, i.e., leaching out of metallic species and reduction of oxygen content. Consequently, the high heating value of bio-oil increased remarkably, the generation of acids was strongly inhibited, whereas the formation of phenols and anhydrosugars was promoted. Therefore, the method proposed herein is promising for upgrading of biomass and bio-oil.

Study on antibiotic susceptibility of Salmonella typhimurium L forms to the third and forth generation cephalosporins.

Salmonella typhimurium is a pathogenic gram-negative bacterium, which is found primarily in the intestinal lumen. It often causes diarrhea in infants and young children and leads to food poisoning. Drug resistance of Salmonella typhimurium presented serious complications in clinical patients. In this study, we investigated the antibiotic susceptibility of Salmonella typhimurium standard strain L forms to the third and forth generation cephalosporins, in order to control and eliminate Salmonella typhimurium L forms in infection treatment. Salmonella typhimurium L forms were induced by ß-lactam antibiotic cefazolin in the culture medium of bacterial L forms. The antibiotic susceptibility of Salmonella typhimurium L forms was analyzed by K-B drug susceptibility testing. The change trend of drug susceptibility and resistance of Salmonella typhimurium L forms was obtained in accordance with USA clinical and laboratory standards institute (CLSI) evaluation data and statistical analysis. Drug resistance of Salmonella typhimurium L forms showed little increasing trend compared with their parent bacteria. The L form inhibition zone was smaller than in the parent bacteria. However, the drug susceptibility of L forms of Salmonella typhimurium to the third and forth generation cephalosporins remained sensitive.The antibiotic susceptibility of Salmonella typhimurium L forms to the third and forth generation cephalosporins remains sensitive, and the combined use of multi-antibiotics is a convenient and effective method to reduce Salmonella typhimurium L forms occurrence.

Enhancement of the production of bio-aromatics from renewable lignin by combined approach of torrefaction deoxygenation pretreatment and shape selective catalytic fast pyrolysis using metal modified zeolites.

In this work, combined approach of torrefaction deoxygenation pretreatment (TDP) and shape selective catalytic fast pyrolysis (SS-CFP) using bifunctional catalyst (metal modified HZSM-5) were employed to improve the yield of bio-BTX derived from the renewable starting material of lignin. Results showed that after TDP, the oxygen element could be removed effectively. The oxygen removal efficiency reached its maximum value of 22.27% at 300 °C, resulting in markedly decrease of unnecessary oxygenates in bio-oil. Compared to parent HZSM-5, all metal modified HZSM-5 (Ga/HZSM-5, Zn/HZSM-5, and Ga-Zn/HZSM-5) promoted the formation of bio-BTX. Zn/HZSM-5 showed the highest selective yield of bio-BTX because of the enhancement deoxygenation reaction of oxygenates and the aromatization reaction of olefins. The combined approach of TDP and SS-CFP remarkably improved the selective yield of bio-BTX, reaching the maximum value of 65.19%, which was much higher than that from single approach of TDP (33.84%) and SS-CFP (47.36%).

miR-200c overexpression inhibits the invasion and tumorigenicity of epithelial ovarian cancer cells by suppressing lncRNA HOTAIR in mice.

Epithelial ovarian cancer (EOC) is a common ovarian cancer in gynecological cancers today. It has been found that microRNAs and long-chain noncoding RNA (lncRNA) regulate the gene transcriptional expression in cells. However, it is not well understood that the upstream and downstream regulatory molecules of lncRNA HOX antisense intergenic RNA (HOTAIR). The effects of miR-200c overexpression on the invasion and nude mouse tumorigenicity, as well as lncRNA HOTAIR and snail expression of EOC SKOV3 cells, should be further explored. The expression of miR-200c and lncRNA HOTAIR was detected by reverse transcription PCR (RT-PCR) in EOC SKOV3 cells. The whole miR-200c gene fragment was cloned into a lentiviral plasmid vector. The miR-200c expression in transducted SKOV3 cells with reconstructed miR-200c lentivirus was significantly higher than the negative control (P < .01). The lentivirus-miR-200c-SKOV3 cells show that the invasion ability was significantly decreased compared with the negative control (P < .01). The nude mouse tumorigenicity was significantly decreased compared with that of the control group (P < .01). The snail protein expression in lentivirus-miR-200c-SKOV3 xenograft tumor was significantly decreased compared with the negative control lentivirus-SKOV3 group (P < .05). The miR-200c overexpression significantly decreased the expressions of lncRNA HOTAIR and snail, but increased E-cadherin expression in the lentivirus-miR-200c transducted SKOV3 cells of xenograft tumor, compared with the negative control (P < .05). The miR-200c overexpression in SKOV3 cells with transducted lentivirus-miR-200c can inhibit lncRNA HOTAIR expression, decrease snail, increase E-cadherin and significantly reduce the invasion and tumorigenicity of EOC SKOV3 cells. These results suggest that the miR-200c and lncRNA HOTAIR could be effective therapeutic targets for human epithelial ovarian cancer treatment.

New sight on the lignin torrefaction pretreatment: Relevance between the evolution of chemical structure and the properties of torrefied gaseous, liquid, and solid products.

In order to reveal the deoxygenation mechanism of lignin torrefaction, the relevance between evolution of chemical structure of torrefied lignin and the properties of torrefied gaseous, liquid, and solid products was established in this study. Results showed that the contents of oxygen element, ßO4 linkages, oxygen-containing functional groups (aliphatic OH, aliphatic COOH, aromatic OCH3) in lignin decreased with the increase of the torrefaction temperature from 210 to 300 °C. The oxygen removal efficiency of lignin torrefaction reached the maximum value of 25.53% at 300 °C. The removed oxygen in the torrefied lignin was transferred into the torrefied gaseous product (e.g. CO2, H2O, and CO) and torrefied liquid product (e.g. G-type and P-type phenols, acids). Among the torrefied gaseous products, CO2 was the dominant oxygen carrier, followed by CO and H2O. Among the torrefied liquid products, G-type phenols were the dominant oxygen carrier, followed by P-type phenols and acids.

Investigation of the relevance between biomass pyrolysis polygeneration and washing pretreatment under different severities: Water, dilute acid solution and aqueous phase bio-oil.

Washing pretreatments of rice straw were performed using three different solutions, namely water, dilute hydrochloric acid solution (HCl solution, pH = 2.9), and aqueous phase bio-oil (APBO, pH = 2.9). The raw and pretreated samples were pyrolyzed at 550 °C in a fixed bed reactor. Results showed that among the three pretreatments, washing with APBO had the highest removal efficiency of alkali metal and alkaline earth metals (AAEMs). Among the pyrolysis products, bio-oil from APBO washed sample had the highest mass, energy, and carbon yields, lowest water content of 36.9%, highest HHV of 17.2 MJ/kg, and highest relative content of anhydrosugars of 31.2%. Its biochar had the lowest ash content of 27.3% and highest specific surface area of 98.6 m2/g, and its non-condensable gases had the highest HHV of 11.9 MJ/m3. Therefore, APBO washing was effective in improving the quality of biomass and its subsequent pyrolysis products.

[Effects of Targeting lncRNA HOTAIR on the Invasion and Nude Mouse Tumorigenicity of Epithelial Ovarian Cancer Cells].

OBJECTIVE: To explore the effects of shRNA-mediated downregulating lncRNA HOTAIR on the invasion,nude mouse tumorigenicity and snail expression of epithelial ovarian cancer SKOV3 cells. METHODS: The expression of lncRNA HOTAIR was detected by RT-PCR in SKOV3 cells. The shRNA targeting the lncRNA HOTAIR gene was cloned into RNA interference plasmid. The construction shHOTAIR vector was transfected into ovarian cancer SKOV3 cells by lipofectamine 2000,and the stably transfected cells were isolated by G418 and single clone selection. The downregulating expression of lncRNA HOTAIR was detected by quantitative real time PCR(qRT-PCR). The characteristics of shHOTAIR transfected SKOV3 cells were analyzed from the assays of invasion and nude mouse tumorigenicity,as well as the expression of snail and E-cadherin mRNA detected by qRT-PCR,and snail detected by immunohistochemistry and Western blot methods in xenograft tumor,respectively. RESULTS: The lncRNA HOTAIR expression was proved by RT-PCR in SKOV3 cells. The lncRNA HOTAIR expression in shHOTAIR transfected SKOV3 cells was significantly lower than the scramble control (P<0.01). The shHOTAIR transfected SKOV3 cells show that the invasion ability was significantly decreased compared with the scramble control (P<0.01). The nude mouse tumorigenicity,including tumorigenicity mouse number and tumor volume,was significantly decreased compared with the control group (P<0.01). The snail protein expression detected by IHC and Western blot in shHOTAIR-SKOV3 xenograft tumor was significantly decreased compared with the control scramble- SKOV3 group (P<0.05). The lncRNA HOTAIR low expression resulted in increasing E-cadherin and decreasing snail expression detected by qRT-PCR in the shHOTAIR transfected SKOV3 cells of xenograft tumor,compared with the scramble control (P<0.05). CONCLUSION: Targeting lncRNA HOTAIR expression in SKOV3 cells with RNA interference can decrease snail,increase E-cadherin and significantly reduce the invasion and tumorigenicity of epithelial ovarian cancer SKOV3 cells. These results suggest that the lncRNA HOTAIR could be an effective therapeutic target for human epithelial ovarian caner treatment.

[Effect of cordycepin on apoptosis and autophagy of tongue cancer cells in vitro and the molecular mechanism].

OBJECTIVE: To study the effect of cordycepin on cell cycle, apoptosis and autophagy of human tongue cancer TCA-8113 cells and explore the mechanism of cordycepin for inhibiting the occurrence of tongue cancer. METHODS: CCK-8 method was used to assess the inhibitory effect of cordycepin on TCA-8113 cell proliferation in vitro. The cell cycle and cell apoptosis of TCA-8113 cells treated with different concentrations of cordycepin were analyzed using flow cytometry. The expressions of apoptosis-related genes caspase-3, caspase-9, Bcl-2, and Bax were examined using quantitative real-time PCR and Western blotting, and immunohistochemistry was used to detect the expressions of autophagy-related proteins LC-3ß, P62, p-mTOR, and AMPK. RESULTS: CCK-8 assay showed that cordycepin significantly inhibited the proliferation of TCA-8113 cells in a concentration-dependent manner with an IC50 of 3.548 mg/mL at 24 h and an IC50 of 1.185 mg/mL at 48 h. Flow cytometric analysis showed that cordycepin caused cell cycle arrest at S phase and dose-dependently increased the apoptotic rate of TCA-8113 cells. Treatment of the cells with cordycepin enhanced the expressions of Bax, caspase-3 and caspase-9 at both the mRNA and protein levels and inhibited the expression of the antiapoptotic gene Bcl-2. Immunohistochemistry demonstrated that cordycepin promoted the expression of LC-3ß and AMPK and inhibited the expression of P62 and p-mTOR. CONCLUSION: Cordycepin inhibits the proliferation and induces apoptosis of HCT-116 cells through the mitochondrial pathway and induces autophagy via the AMPK/mTOR pathway.

In-depth study of rice husk torrefaction: Characterization of solid, liquid and gaseous products, oxygen migration and energy yield.

Torrefaction is a promising method for biomass upgrading, and analysis of all products is the essential way to reveal torrefaction mechanism. In this study, torrefaction of rice husk was performed at 210-300 °C. Results showed that the fuel properties of solid products were greatly enhanced upon removal of oxygen. The gaseous products were mainly CO2 (52.9-73.8 vol%), followed by CO (26.3-39.2 vol%). The liquid product was mainly water and some tar, and the latter contained acids, furans, ketones, aldehydes, and phenols, among which the relative content of acids was the highest. Torrefaction temperature has obvious effects on the oxygen migration. Within the temperature range of 210-300 °C, 9.5-63.2% of oxygen in rice husk was migrated to the gaseous and liquid products. The H2O was the major contributor to deoxygenation, followed by CO2 and CO. Thus, formation of H2O, CO2, and CO during torrefaction is important as it achieves the purpose of intense deoxygenation.

[Knock-down of ZEB1 inhibits the proliferation, invasion and migration of gastric cancer cells].

Objective To down-regulate the expression of zinc-finger E-box binding homeobox 1 (ZEB1) gene by shRNA, and investigate its effect on invasion, migration and proliferation, as well as the related gene expressions of lncRNA HOTAIR and E-cadherin in human gastric cancer BGC823 cells. Methods RNA interfering (RNAi) was used to knock down ZEB1 in gastric cancer BGC823 cells. The recombinant plasmid shZEB1 was constructed and transfected into the gastric cancer BGC823 cells by LipofectamineTM 2000, and the stably transfected cells were isolated by G418 selection and limited dilution. The expression of ZEB1 mRNA and protein was detected by real-time quantitative PCR and Western blot analysis. Cell proliferation was determined by MTT assay, and the invasion and migration abilities of BGC823 cells were monitored by TranswellTM invasion assay and wound healing assay, respectively. The expressions of lncRNA HOTAIR and E-cadherin mRNA were detected by real-time quantitative PCR. Results After ZEB1 expression was successfully down-regulated in BGC823 cells by siRNA, the proliferation, invasion and migration rates in shZEB1 transfection group were significantly lower than those in control group; meanwhile, the expression of lncRNA HOTAIR was reduced and E-cadherin expression was enhanced. Conclusion Knock-down of ZEB1 expression by RNA interference can decease lncRNA HOTAIR expression and restrain cell proliferation, invasion and migration in gastric cancer BGC823 cells.

An approach for upgrading biomass and pyrolysis product quality using a combination of aqueous phase bio-oil washing and torrefaction pretreatment.

Bio-oil undergoes phase separation because of poor stability. Practical application of aqueous phase bio-oil is challenging. In this study, a novel approach that combines aqueous phase bio-oil washing and torrefaction pretreatment was used to upgrade the biomass and pyrolysis product quality. The effects of individual and combined pretreatments on cotton stalk pyrolysis were studied using TG-FTIR and a fixed bed reactor. The results showed that the aqueous phase bio-oil washing pretreatment removed metals and resolved the two pyrolysis peaks in the DTG curve. Importantly, it increased the bio-oil yield and improved the pyrolysis product quality. For example, the water and acid content of bio-oil decreased significantly along with an increase in phenol formation, and the heating value of non-condensable gases improved, and these were more pronounced when combined with torrefaction pretreatment. Therefore, the combined pretreatment is a promising method, which would contribute to the development of polygeneration pyrolysis technology.

Combined pretreatment with torrefaction and washing using torrefaction liquid products to yield upgraded biomass and pyrolysis products.

This study presented an approach to upgrade biomass and pyrolysis products using a process based on torrefaction liquid washing combined with torrefaction pretreatment. The torrefaction of cotton stalk was first conducted at 250°C for 30min and then the resulting torrefaction liquid products were collected and reused to wash cottonstalk. The pyrolysis of the original and pretreated cotton stalk was performed at 500°C for 15min in a fixed-bed reactor. The results indicated that the combined pretreatment obviously reduced the metallic species in cotton stalk, decreased the water and acids contents while promoted phenols in bio-oil, declined the ash content in biochar, as well as improved the heating value of non-condensable gas. Overall, the combined pretreatment did not only allow to reuse the liquid products issued from torrefaction pretreatment but also improved the quality of biomass and the pyrolysis products, making it a novel promising pretreatment method.

Effect of pyrolysis temperature on the chemical oxidation stability of bamboo biochar.

Biochar produced by biomass pyrolysis has the advantage of carbon sequestration. However, some of the carbon atoms in biochar are not very stable. In this study, the effect of pyrolysis temperature on the chemical oxidation stability of bamboo biochar was investigated using the atomic ratios of H/C and O/C, Fourier transform infrared spectroscopy, and potassium dichromate (K2Cr2O7) oxidation spectrophotometric method. The results show that the carbon yield and ratios of H/C and O/C decreased from 71.72%, 0.71, and 0.32 to 38.48%, 0.22, and 0.06, respectively, as the temperature was increased from 300°C to 700°C. Moreover, the main oxygen-containing functional groups gradually decreased, while the degree of aromatization increased accordingly. The biochar showed a better stability at a higher pyrolysis temperature. The proportion of carbon loss, i.e., the amount of oxidized carbon with respect to the total carbon of the biochar, decreased from 16.52% to 6.69% with increasing temperature.