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To overcome the structural complexity and improve the bioconversion efficiency of Pennisetum purpureum into bioethanol or/and biomethane, the effects of ensiling pretreatment, NaOH pretreatment, and their combination on digestion performance and mass flow were comparatively investigated. The coproduction of bioethanol and biomethane showed that 65.2 g of ethanol and 102.6 g of methane could be obtained from 1 kg of untreated Pennisetum purpureum, and pretreatment had significant impacts on the production; however, there is no significant difference between the results of NaOH pretreatment and ensiling-NaOH pretreatment in terms of production improvement. Among them, 1 kg of ensiling-NaOH treated Pennisetum purpureum could yield 269.4 g of ethanol and 144.5 g of methane, with a respective increase of 313.2% and 40.8% compared to that from the untreated sample; this corresponded to the final energy production of 14.5 MJ, with the energy conversion efficiency of 46.8%. In addition, for the ensiling-NaOH treated Pennisetum purpureum, the energy recovery from coproduction (process III) was 98.9% higher than that from enzymatic hydrolysis and fermentation only (process I) and 53.6% higher than that from anaerobic digestion only (process II). This indicated that coproduction of bioethanol and biomethane from Pennisetum purpureum after ensiling and NaOH pretreatment is an effective method to improve its conversion efficiency and energy output., We investigated the effects of pretreatment on the coproduction (ethanol and methane) performance and material flow from Pennisetum purpureum.

The effects of steam and sulfuric acid pretreatment on anaerobic digestion (AD) of whiskey byproducts (including draff, thin and thick stillage) were investigated in order to improve the digestion performance under both mesophilic and thermophilic temperatures. The results of biomethane potential assays suggested that thermophilic AD facilitated the release of free ammonia (ca. 1200 mg/L) from byproducts, resulting in strong ammonia inhibition and volatile fatty acid accumulation. In contrast, no free ammonia inhibition (ca. 700 mg/L) was observed under mesophilic AD; the methane yield from mesophilic AD was between 375.3 ± 13.6 mL/g volatile solid (VS; acid-treated sample) and 389.1 ± 8.5 mL/g VS (untreated sample). Although acid pretreatment (2% acid under 135 °C for 15 min) did not improve the methane yield from mesophilic AD, it reduced the digestion time by 14.3% compared to that of the untreated sample. Microbial community analysis showed that irrelevant of pretreatment, hydrogenotrophic methanogens of Methanobrevibacter (28.9%–49.8% in abundance) and Methanoculleus (26.0%–55.9% in abundance) were the dominant archaeal genus under mesophilic AD. In comparison, hydrogenotrophic Methanothermobacter (over 97% in abundance) were dominant in thermophilic AD. This study could be exploited to aid in decarbonizing the whiskey industry by optimizing the biogas process in a circular economy system.

This study investigated the effect of particle size on the performance of anaerobic digestion of Hybrid Pennisetum. Hybrid Pennisetum was ground and sieved to provide different particle sizes between 0.180 and 1.000 mm. Characterization of the different particle sizes suggested that the composition of ground Hybrid Pennisetum altered-carbohydrate content decreased and lignin content increased with a decrease in particle size. The highest specific methane yield was 291.9 ± 4.7 mL CH4·g−1 VS at a particle size of 0.250–0.380 mm and this value plateaued as the particle size was reduced to below 0.250 mm. Excessive size reduction did not improve the methane yield but did result in a reduction of digestion time by 28.6–35.7%. The net energy output from the process was calculated at 300 kWh/t VS. Therefore, grinding was proved to enhance the anaerobic fermentation efficiency and energy output of Hybrid Pennisetum.

In this study, the links between performance and microbial communities were investigated with municipal solid waste (MSW) at two feedstock ratios and eight organic loading rates (OLRs). The co-digestion systems were stably operated at OLRs of 2.0–6.0 g VS/(L·d). The performance of the co-digestion system varied with the feedstock ratio. Compared with the 50:50 (hybrid Pennisetum:MSW) system, the 75:50 system, GM31, obtained increases of 1.93–17.68% and 0.29–23.29% for the specific biogas and methane yields, respectively. Whereas a shift in bacterial and methanogen communities occurred as the operating conditions changed, particularly with OLR variations. The genera Saccharofermentans, Prevotella, Clostridium, Syntrophomonas, and Proteiniphilum became the dominant bacteria for the conversion of carbohydrates and nitrogen compounds as the OLR increased. Meanwhile, a shift from acetoclastic to hydrogenotrophic or multifunctional methanogens was observed.

This study investigated whether bioaugmentation can improve the anaerobic digestion (AD) performance of food waste (FW), as well as the effects of addition dosage and frequency on the bioaugmentation’s performance and economic feasibility. The findings demonstrated that all the bioaugmented digesters, regardless of dosage and frequency, performed more effectively in biogas production than the non-bioaugmentation control. Furthermore, relatively higher dosages or frequencies increased AD performance. Introducing 0.25 g L−1 d−1 of bioaugmentation seed every three days increased OLR and volumetric biogas production 8-fold and 12-fold, respectively, compared to the non-bioaugmentation control. Whole-genome sequencing analysis showed that bioaugmentation enhanced the population of the acetoclastic Methanothrix (belong to the order Methanosarcinales). Moreover, high abundance of Methanothrix (exceeding 80%) contributed to a better AD performance. Economic analysis of an up-scale biogas plant suggested that an appropriate bioaugmentation process increased income, thus increasing the profit to 3696 CNY d−1 if treated at 21 t FW.

Microalgae have been considered as one of the most promising biomass resources for the generation of biodiesel, whereas algal residues (AR) after lipid extraction need to be utilized to enhance its economy and sustainability. This study proposed an integrated biochemical method for microalgae Chlorella sp. processing to achieve a high energy yield (12.33 KJ g−1VS). The microalgae were first pretreated with mixed enzymes for lipid extraction. After that, the AR obtained from the lipid extraction of microalgae was co-digested with energy grass (GR) Pennisetum hybrid for methane production. Results indicated that the lipid yield from microalgae was enhanced by 54.45% with mixed enzymes pretreatment of cellulase, xynalase and pectinase compared to that without enzymatic pretreatment. The maximal methane yield (207.35 ± 15.66 mLCH4 g−1VS) of AR and GR co-digestion was obtained with the AR and GR VS ratio at 1:3. Energy yields evaluation based on the biodiesel and methane yields indicated that the energy yield for this two-step combined method was 169% higher than that for biodiesel alone. Thus, the proposed integrated approach was verified to be applicable for microalgae processing for energy production.

Cell wall disruption is an essential downstream processing step for improving the efficiency of lipid extraction from microalgae. Enzyme-assisted extraction of lipid from microalga Scenedesmus sp. with cellulase, xylanase and pectinase, using various parameters, such as enzyme concentration, temperature, pH and incubation time, was optimized by central composite design (CCD) coupled with response surface methodology (RSM). Both the lipid extraction from microalgae and the fatty acid methyl esters (FAMEs) production under optimal conditions showed a ∼twofold in the yields compared to the control group with no enzymatic treatment. SEM images, FTIR measurement, XPS and HPLC analysis showed that the enzymatic pretreatment caused significant alterations in the cell wall structure of microalgae. And the disruption of microalgal cell walls was primarily attributed to the breakage of β-glucosidic linkages in cellulose and hemicellulose. The study showed a promising approach can lead to an improvement in the lipid extraction yield from microalgae and further provide valuable information for the use of enzymes in microalgal processes.

This study aims to investigate the performance of bioaugmentation with an acid-tolerant methanogenic culture to recover deteriorated anaerobic digestion caused by organic overloading. The function of bioaugmentation was evaluated in terms of substance metabolism, microbial community structure, and gene function. Our findings demonstrate that routine bioaugmentation effectively recovered the failing digester by degrading accumulated volatile fatty acids and increasing pH. In contrast, a non-bioaugmentation reactor (control) did not recover by itself, while abiotic augmentation restored the digestion performance temporarily but the digester failed again at an organic loading rate of 1.5 g L−1 d−1. Using whole genome pyrosequencing analysis, we found that after bioaugmentation, the populations of Methanothrix (acetoclastic methanogens) and Methanolinea (hydrogenotrophic methanogens) increased significantly, which may be the main contributors for the positive effect on methane production. On the genic level, bioaugmentation may enhance the function of genes involved in cell motility, signal transduction mechanisms for methanogens, and energy production and conversion for bacteria.

This study investigated the co-digestion performance of Pennisetum hybrid and cow manure via batch mode tests to optimize the feedstock ratio. On the basis of the batch test results, the long-term operational performance of the co-digestion system at different ratios was assessed at mesophilic temperature and organic loading rates (OLRs) of 2.0–5.5 g of volatile solids (VS) L–1 day–1 via semi-continuous mode. Batch tests showed that the maximum specific methane yield of 170.35 ± 11.61 mL/g of VS was obtained at a Pennisetum hybrid/cow manure ratio of 100:0, with the specific methane yield decreasing with an increased proportion of cow manure. In the semi-continuous experiments, the mono-digestion of Pennisetum hybrid failed at the OLR of 2.0 g of VS L–1 day–1. This failure was accompanied by the accumulation of volatile fatty acids (VFAs). The stable operational OLR increased by 1.0–1.5 times with the addition of cow manure. The volumetric biogas yield increased with the augmentation of the OLR from 2.0 t...

The acid-tolerant methanogenic propionate degradation culture was acclimated in a propionate-fed semi-continuous bioreactor by daily adjusting the digestate pH. The performance of propionate fermentation, the respond of microbial community structure to the acidic environment, and the microbial network for propionate degradation in the acid-tolerant culture was investigated. The results demonstrated that after long term of acclimation to low pH, the digester could produce methane from propionate at pH 4.8–5.5 with 0.3–0.4 L g−1 propionic acid (HPr) d−1 of the volatile solids (VS) methane production. The predominant methanogens shifted from acetoclastic methanogens (∼87%) to hydrogenotrophic methanogens (∼67%) in the bioreactor with the dropping pH, indicating that hydrogenotrophic methanogens were more acid-tolerant than acetoclastic methanogens. Smithella (∼11%), Syntrophobacter (∼7%) and Pelotomaculum (∼3%) were the main propionate oxidizers in the acid-tolerant propionate-utilizing culture. Methanothrix dominant acetoclastic methanogens, while Methanolinea and Methanospirillum were the major H2 scavengers to support Syntrophobacter and Pelotomaculum syntrophic propionate degradation.

Intercropping improves yield and is broadly adopted worldwide; however, the risk and regulatory mechanism of nitrate leaching in intercropping compared with monocropping remain elusive. A fixed plot field experiment was conducted with three cropping patterns (maize monocropping, potato monocropping, and maize–potato intercropping) under four (nitrogen) N levels in Kunming, China during 2014–2017. Based on crop N uptake and soil nitrate transformation, the underlying mechanism of potential nitrate-N (NO3−-N) leaching (NO3−-N accumulation below the root layer) and N regulation was explored under intercropping and monocropping. The results showed that the area-scaled potential NO3−-N leaching under intercropping was mitigated by 3.4–37.4%, with a mean of 15.8% relative to the weighted mean of maize- and potato-monocropping. Intercropping-related mitigation of potential NO3−-N leaching was enhanced with increasing N level. Intercropping increased the ammonia-oxidizing archaea population and soil nitrification potential but had no significant impact on the abundance of ammonia-oxidizing bacteria and narG. Redundancy analysis and structural equation modeling indicated that soil water played a crucial role in regulating the intercropping-related mitigation of potential NO3−-N leaching, while the improvement in N uptake by intercropped maize reduced the potential NO3−-N leaching at low and medium N levels. The results highlight the intercropping advantage over monocropping in mitigating the risk of NO3−-N leaching. This study provides insights into the environmental impacts of diversified cropping systems and regulating mechanisms.

The methanogenic propionate degradation consortia were enriched in a propionate-fed semi-continuous bioreactor. The microbial community shift with depth, the microbial network and its correlation with metabolic pathway were also investigated. The results demonstrated that the maximum organic loading rate (OLR) of the reactor was 2.5 g propionic acid (HPr) L−1 d−1 with approximately 1.20 LL−1 d−1 of volumetric methane production (VMP). The organisms in the enrichment were spanning 36 bacterial phyla and 7 archaeal orders. Syntrophobacter, the main Hpr oxidizer in the digester, dominated bacteria with relative abundance changing from 63% to 37% with depth. The predominant methanogens shift from hydrogenotrophic Methanoculleus (∼60%) at the upper liquid layer to acetoclastic Methanothrix (∼51%) at the lower sediment layer in the bioreactor. These methanogens syntrophically support Syntrophobacter by degrading HPr catabolism by-products (H2 and acetate). Other bacteria could scavenge anabolic products (carbohydrate and protein) presumably derived from detrital biomass produced by the HPr-degrading community.

The effect of variable sodium concentrations on the anaerobic digestion performance of Sargassum sp. was investigated, and the variation in bacterial and archaeal communities was analyzed by high-throughput gene sequencing. Results showed that the maximum methane yield of 290.41 ± 34.21 mL of CH4 g–1 of volatile solids was obtained at a sodium concentration of 4.42 g L–1, increased by 38.1% compared to the control group. Meanwhile, higher volatile fatty acids production was observed at the sodium concentration of 4.42 g L–1. The inhibitory concentration values of 10%, 50%, and 90% occurred at the sodium concentrations of 6.3, 11.3, and 18.7 g L–1, respectively. The hydrolytic, acidogenic, and acetogenic bacteria and the hydrogenotrophic methanogens could normally metabolize at sodium concentrations of 2.46–24.08 g L–1, whereas the acetoclastic methanogens were severely inhibited at sodium concentrations greater than 20.15 g L–1. The microbial community structure showed that Paludibacter and Fibrobacter pl...

BACKGROUND Propionate accumulation may lead to digester failure. This study aimed to investigate the effect of ammonia, a metabolic product of protein, on propionate degradation. The shift of microbial community was also investigated. RESULTS Propionate accumulated over the experimental period in the reactor with a total ammonia nitrogen (TAN) concentration of 2.5 g N L−1, as a result the digester failed after 4 hydraulic retention times (HRT) at an organic load rate (OLR) of 0.8 g propionic acid (HPr) L−1 d−1. The average HPr degradation rate was below 54% during the fourth HRT, while >97% of the degraded HPr was converted to methane. The reactor without ammonia stress did not experience HPr accumulation and OLR was increased stepwise to 1.2 g L−1 d−1 at the 8th HRT. The average HPr degradation rate and methane recovery rate of this reactor in the last HRT was 99% and 74%, respectively. According to the shifts of microbial community, acetoclastic methanogen was more vulnerable to ammonia than hydrogenotrophic methanogen CONCLUSION TAN concentration of 2.5 g N L−1 inhibited propionate degradation more severely than methanogenesis. The loss of abundance of Clostridiaceae and Syntrophobacter might be the main reason for the poor performance under ammonia stress. © 2017 Society of Chemical Industry

BACKGROUND The main obstacle associated with methane production in anaerobic digestion processes from marine macroalgae is inhibited by the sea salts. This study aims to investigate the anaerobic digestion performance of marine macroalgae by acclimating inoculum to varying salinity, and to analyze the influence of salinity on microbial community by high-throughput gene sequencing. RESULTS The relationship between specific methane yield and salinity followed the third-order polynomial equation: y = 271.533 + 6.449X − 0.261X2 + 0.002X3. The acclimating inoculum produced methane efficiently at 35 g L−1 with a specific methane yield of 220.88 mL CH4 g−1 VS. Methanogenesis was considerably impeded at salinity greater than 55 g L−1. The bacterial phyla Bacteroidetes, Firmicutes, and Proteobacteria and the Methanobacterium, Methanosaeta, and Methanosarcina genera in archaea were predominant at different salinities. Hydrogenotrophic methanogens such as Methanobacterium can tolerate salinity up to 85 g L−1, whereas acetoclastic methanogens, Methanosaeta and Methanosarcina were severely inhibited at salinity greater than 65 g L−1. CONCLUSION The acclimating inoculum can be used as anaerobic microbial resources for efficient methane production from marine macroalgae under seawater conditions of 35 g L−1. The hydrolytic, acidogenic, acetogenic bacteria and hydrogenotrophic methanogens normally metabolized at different salinities, whereas acetoclastic methanogens were severely inhibited at high saline conditions. © 2017 Society of Chemical Industry

The biomass of grass-based Pennisetum hybrid commonly use for a biogas production via anaerobic digestion. However, it is necessary to determine a method to optimize the plant harvest time for high biogas production. Moreover, ensiling of biomass in the presence of diverse microbes may offer a solution to improve biogas production. In this study, whole plant of Pennisetum biomass (including stems and leaves) was collected at different harvesting time (plant heights of 70, 100, 150 cm), and then comparatively assessed for further ensiling and biogas production. Compared to leaves, stems exhibited a significant linear relationship (R2 = 0.99) with whole plants in terms of ensiling quality (i.e. pH and NH3-N). Microbial analysis further revealed that Lactobacillus was the dominant bacterial genus during ensiling of stems and whole plants, with the highest relative abundance of 50.08% obtained at the height of 100 cm. Ensiling of biomass at a height of 100 cm achieved the best digestion performance, with the methane yields of 316 ± 20 mL/g VS for leaves, 361 ± 43 mL/g VS for stems, and 356 ± 28 mL/g VS for whole plants. A harvesting time at the plant height of 100 cm was the optimal from the silage quality and anaerobic digestion performance.

In this study, batch experiments were conducted to compare the effect of temperature and substrate source on microbial communities in the anaerobic digestion of kitchen waste. The results showed that the microbial communities of anaerobic digestion were not sensitive to varied sources of waste, but shifted with the change in operating temperatures. In the reactors operated at mesophilic conditions, Levilinea, Syntrophomonas, Methanothrix, and Methanosphaerula, etc. were the dominant microbes during the process. While in thermophilic reactors, Levilinea, Ornatilinea, Methanosphaerula and Methanomassiliicoccus, etc. prevailed. Meanwhile, an enrichment in Coprothermobacter, Defluviitoga, Defluviitalea, Tepidimicrobium, Lutispora and Fonticella were observed as the temperature changed from mesophilic to thermophilic, suggesting these genera could be selectively enriched at thermophilic conditions. The results provided fundamental understanding of the microbiology that could support the scale up of food waste anaerobic digestion.

The effect of additives on the silage quality, microbial community, and anaerobic digestion performance of Pennisetum purpureum with high moisture content was studied. The sample treated with a mixed additive had best silage quality with the lowest pH and highest lactic acid/acetic acid ratio. Different additives influenced the dominant desirable bacteria. Correspondingly, Enterobacter was the dominant bacterial genus for sample with non-fermentative additives, whereas for the samples with fermentative or mixed additives, both Enterobacter and Lactobacillus had high relative abundance. The parameters of NH3-N, hemicellulose and lactic acid were positively correlated with the specific methane yield, while the lignin content was inversely correlated with the specific methane yield. The higher specific methane yield of 293.81 ± 0.15–334.69 ± 22.75 mL/g VS was obtained for samples treated with fermentative additive. Therefore, the mixed additive and fermentative additive are recommended for the silage of material with high-moisture content to improve the silage quality and methane yield.

To investigate whether bioaugmentation could improve the digestion performance of high C/N ratio feedstock without co-digestion with nitrogen-rich substrate, different forms of enriched methanogenic culture were introduced to the continuous feed digesters. The performance efficiency of bioaugmentation on digestion improvement was compared. The effect of bioaugmentation on microbial community composition was revealed as well. Results demonstrated that routine bioaugmentation with liquid culture (containing the microbes and the medium remains) showed the best performance, with the organic loading rate (OLR), methane percentage, volumetric methane production (VMP) and volatile solid methane production (VSMP) higher at 1.0 g L−1 d−1, 24%, 0.22 L L−1 d−1 and 0.23 L g−1 VS d−1 respectively, compared to the non-bioaugmentation control. Whole genome pyrosequencing analysis suggested that consecutive microbial consortium addition could reconstruct the methanogens community by increasing the populations of acetoclastic methanogens Methanothrix, which could accelerate the degradation of acetate and methane production.

Benzothiadiazole (BTH) is an artificial inducer of systemic acquired resistance. Due to rice being an important food crop and model plant, we investigated its response to BTH using label-free proteomics technology coupled with bioinformatics. Protein expression levels were verified using the multi-reaction monitoring mode and semi-quantitative RT-PCR. BTH treatment can up- or down-regulate many proteins produced by the rice host at all four periods, with the numbers of proteins being 6/24, 9/10, 14/10, and 8/20, respectively. Compared with mock treatments (phosphate buffered saline with 0.1 % dimethylsulfoxide and 0.5 % Tween-20), some proteins related to plant resistance were only detected after BTH treatments, such as ascorbate peroxidase (POD) 3, chitinase A, thioredoxin-dependent POD 2, beta-1,3-glucanase 2, POD superfamily protein, major facilitator superfamily (MFS) protein, copper/zinc-superoxide dismutase (SOD) 1, pathogenesis-related protein (PR) 1. Other proteins showing up-regulation after BHT treatment included PR-5, glyceraldehyde-3-phosphate dehydrogenase C, plasma-membrane associated cation-binding protein 1, and oxidoreductase family proteins. These results indicated that BTH was involved with inducing rice resistance. Some up-regulated proteins were also involved in other metabolic processes. The activity and expression level of POD, phenylalanine ammonia-lyase (PAL), and SOD, lipoxygenase (LOX), beta-1,3-glucanases, and chitinases were determined using the enzyme activity assay and semi-quantitative RT-PCR. These results indicated that BTH can enhance the activity of beta-1,3-glucanases, LOX, PAL, and POD. BTH can also induce up-regulation of the copper/zinc-SOD, ascorbate POD, glutathione POD 1, Chitinase, and LOX1 genes.

Alkaline pretreatment with NaOH was used to improve methane yield from Pennisetum Hybrid. The pretreatments were carried out with different NaOH solutions (2–8% w/w) at three temperatures (35, 55 and 121 °C) for different periods of time (24, 24 and 1 h). All treated and untreated Pennisetum Hybrid were digested under mesophilic conditions (37 °C) to biogas, significant effects of the pretreatments on the yield of methane were observed. Results showed the modified Gompertz equation was reliable (determination coefficients (R2) greater than 0.96) to describe the kinetic behavior of anaerobic digestion of Pennisetum Hybrid. The best result, obtained by the treatment at 35 °C 2% NaOH for 24 h, resulted in the methane yield of 301.7 mL/g VS, corresponding to 21.0% improvement in the methane yield. Compositional, SEM, XRD and FTIR analysis confirmed that lignin removal, structural modification and cellulose crystalline variation were responsible for the improvement.

The silage quality and anaerobic digestion performance of Pennisetum purpereum with molasses-processed wastewater addition were assessed. The silage samples with molasses-processed wastewater addition obtained higher lactic acid concentration of 15.18-23.38mg/g FM, lower pH value of 3.96-4.45 and lower NH3-N content of 0.47-0.64mg/g FM. No obvious difference was observed in the dominant phyla and genus, but the relative abundance of Lactobacillus reached up to 77.39%, increased by 84% compared to the silage samples without molasses-processed wastewater addition. Combined the silage quality and bacterial community, the decreased in pH value of fresh material caused by molasses-processed wastewater addition was the main reason for improving the silage quality. Meanwhile, the increased in COD concentration is beneficial for improving the specific methane yield and the maximum specific methane yield of 259±5.75mL/g VS was obtained with adding 20g/kg molasses-alcoholic wastewater. Molasses-processed wastewater is an alternative additive for silage.

Sine oculis homeobox homolog 1 (SIX1) protein is a member of the homeobox transcription factor family. Overexpression of SIX1 contributes to cancer progression and is associated with adverse outcomes in various cancer types including breast, ovarian, uterine cervical and liver. To investigate the clinicopathological significance of SIX1 protein expression in gastric adenocarcinomas (GAC), localization of the SIX1 protein was determined in MKN-1, a gastric cancer cell line, using immunofluorescence (IF) staining; SIX1 mRNA level was detected in fresh tissues of GAC and normal gastric mucosa using quantitative real-time polymerase chain reaction (qRT-PCR); and SIX1 protein expression was assessed in 163 GAC, 35 gastric dysplasia and 26 normal gastric mucosa using immunohistochemical (IHC) staining. Correlations between SIX1 protein expression and pathological parameters of GAC were analyzed using Chi-square tests, differences in survival curves were analyzed using log-rank tests, and multivariate survival analysis was performed using the Cox proportional hazards regression model. SIX1 protein showed a mainly cytoplasmic staining pattern in GAC using IF and IHC staining. The positive SIX1 protein expression rate was 80.4% in GAC, which was significantly higher than in either gastric dysplasia (45.7%) or normal gastric mucosa (26.9%) (P

Runt-related transcription factor 3 (RUNX3) has recently been reported to be a possible predictor of sensitivity of cancer cells for photodynamic therapy (PDT), a promising therapeutic modality for keloids. In this study, we aimed to elucidate the implications of RUNX3 for keloid pathogenesis and sensitivity to pheophorbide a-based PDT (Pa-PDT). RUNX3 and proliferating cell nuclear antigen (PCNA) expression were examined in 6 normal skin samples and 32 keloid tissue samples by immunohistochemistry. We found that RUNX3 expression was detected more often in keloid tissues than in dermis of normal skin. In keloid tissues, RUNX3 expression was significantly increased in patients presenting with symptoms of pain or pruritus, and was also significantly related to PCNA expression. The therapeutic effect of Pa-PDT was comparatively investigated in keloid fibroblasts (KFs) with and without RUNX3 expression. Significant differences were found after Pa-PDT between KFs with and without RUNX3 expression in cell viability, proliferative ability, type I collagen expression, generation of reactive oxygen species (ROS), and apoptotic cell death. In addition, RUNX3 expression was significantly decreased after Pa-PDT in KFs, and KFs with downregulation of RUNX3 showed significantly increased cell viability after Pa-PDT. Pa-PDT may be a potential therapeutic modality for keloids, and RUNX3, as a possible contributor to keloid pathogenesis, may improve sensitivity to Pa-PDT in KFs.

The biomass yield and mesophilic anaerobic digestion performance of Pennisetum hybrid that were collected at different harvest periods and cutting frequencies were investigated. Results indicated that chemical composition, biomass yield, and specific methane yield of Pennisetum hybrid were significantly influenced by harvest period and cutting frequency. The contents of total solids, volatile solids, carbon, cellulose, lignin and starch increased with a prolonged harvest period, whereas the nitrogen and crude protein content decreased. Specific methane yields decreased considerably from 280 to 119 mL/g VS with a prolonged harvest period, whereas the biomass yield per hectare increased. A maximum dry matter yield of 41.63 t/ha was achieved when the Pennisetum hybrid was harvested in October. When the cutting frequencies increased from twice to three times per year, the specific methane yields increased as the biomass yield decreased. Because of the reverse trend of anaerobic digestion performance...

In this study, we sought to investigate the expression of microRNA (miR)-214 on the osteogenic differentiation of bone marrow‑derived mesenchymal stem cells (BMSCs) and explore the possible underlying mechanisms. We found that the overexpression of miR‑214 effectively promoted the adipocyte differentiation of BMSCs in vitro, reduced alkaline phosphatase (ALP) activity and the gene expression of collagen type I (Col I), osteocalcin (OCN) and osteopontin (OPN) in the BMSCs. We further found that the overexpression of miR‑214 suppressed the protein expression of fibroblast growth factor (FGF), phosphorylated c‑Jun N-terminal kinase (p-JNK) and phosphorylated p38 (p-p38) in the BMSCs. The downregulation of miR‑214 promoted the osteogenic differentiation of BMSCs, and increased ALP activity and Col I, OCN and OPN gene expression in the BMSCs. It also increased FGF p-JNK and p-p38 protein expression in the BMSCs. The use of JNK inhibitor (SP600125) enhanced the inhibitory effects of miR‑214 overexpression on osteogenic differentiation, ALP activity, and Col I, OCN and OPN gene expression in the BMSCs. Lastly, the use of p38 inhibitor (SB202190) also enhanced the inhibitory effects of miR‑214 overexpression on ALP activity, and Col I, OCN and OPN gene expression in the BMSCs. These results provide a mechanism responsible for the suppressive effects of miR‑214 on the osteogenic differentiation of BMSCs involving the inhibition of the JNK and p38 pathways.

Pretreatment methods play an important role in the improvement of biogas production from the anaerobic digestion of energy grass. In this study, conventional thermal and microwave methods were performed on raw material, namely, Pennisetum hybrid, to analyze the effect of pretreatment on anaerobic digestion by the calculation of performance parameters using Logistic function, modified Gompertz equation, and transference function. Results indicated that thermal pretreatment improved the biogas production of Pennisetum hybrid, whereas microwave method had an adverse effect on the performance. All the models fit the experimental data with R (2) 0.980, and the Reaction Curve presented the best agreement in the fitting process. Conventional thermal pretreatment showed an increasing effect on maximum production rate and total methane produced, with an improvement of around 7% and 8%, respectively. With regard to microwave pretreatment, maximum production rate and total methane produced decreased by 18% and 12%, respectively.

Accumulating evidence indicated that aberrantly expressed microRNAs play critical roles in the initiation and progression of human cancers. However, the underlying functions of miR-493 in human melanoma remains unknown. Here, our study found that miR-493 expression was downregulated in human melanoma tissues and cells. Overexpression of miR-493 suppressed cell proliferation and cell cycle in human melanoma cell line A375. IRS4 was defined as a target for downregulation by miR-493 and was confirmed by luciferase assay. We also found that knockdown of IRS4 counteracted the proliferation promotion by miR-493 inhibitor. In summary, these results demonstrated that miR-493 acts as a tumor suppressor and inhibits cell proliferation and cell cycle in human melanoma by directly targeting IRS4.