Anti-inflammatory properties and characterization of water extracts obtained from Callicarpa kwangtungensis Chun using in vitro and in vivo rat models.

Callicarpa kwangtungensis Chun (CK) is a common remedy exhibits anti-inflammatory properties and has been used in Chinese herbal formulations, such as KangGongYan tablets. It is the main component of KangGongYan tablets, which has been used to treat chronic cervicitis caused by damp heat, red and white bands, cervical erosion, and bleeding. However, the anti-inflammatory effects of CK water extract remains unknown. This study assessed the anti-inflammatory effects of CK in vivo and in vitro, characterized its main components in the serum of rats and verified the anti-inflammatory effects of serum containing CK. Nitric oxide (NO), tumour necrosis factor α (TNF-α) and interleukin-6 (IL-6) release by RAW264.7 cells was examined by ELISA and Griess reagents. Inflammation-related protein expression in LPS-stimulated RAW264.7 cells was measured by western blotting. Furthermore, rat model of foot swelling induced by λ-carrageenan and a collagen-induced arthritis (CIA) rat model were used to explore the anti-inflammatory effects of CK. The components of CK were characterized by LC-MS, and the effects of CK-containing serum on proinflammatory factors levels and the expression of inflammation-related proteins were examined by ELISA, Griess reagents and Western blotting. CK suppressed IL-6, TNF-α, and NO production, and iNOS protein expression in LPS-stimulated RAW264.7 cells. Mechanistic studies showed that CK inhibited the phosphorylation of ERK, P38 and JNK in the MAPK signaling pathway, promoted the expression of IκBα in the NF-κB signaling pathway, and subsequently inhibited the expression of iNOS, thereby exerting anti-inflammatory effects. Moreover, CK reduced the swelling rates with λ-carrageenan induced foot swelling, and reduced the arthritis score and incidence in the collagen-induced arthritis (CIA) rat model. A total of 68 compounds in CK water extract and 31 components in rat serum after intragastric administration of CK were characterized. Serum pharmacological analysis showed that CK-containing serum suppressed iNOS protein expression and NO, TNF-α, and IL-6 release. CK may be an anti-inflammatory agent with therapeutic potential for acute and chronic inflammatory diseases, especially inflammatory diseases associated with MAPK activation.

Sinomenine Hydrochloride Protects IgA Nephropathy Through Regulating Cell Growth and Apoptosis of T and B Lymphocytes.

Purpose: Sinomenine hydrochloride (SH) is used to treat chronic inflammatory diseases such as rheumatoid arthritis and may also be efficacious against Immunoglobulin A nephropathy (IgAN). However, no trial has investigated the molecular mechanism of SH on IgAN. Therefore, this study aims to investigate the effect and mechanism of SH on IgAN. Methods: The pathological changes and IgA and C3 depositions in the kidney of an IgAN rat model were detected by periodic acid-Schiff (PAS) and direct immunofluorescence staining. After extracting T and B cells using immunomagnetic beads, we assessed their purity, cell cycle phase, and apoptosis stage through flow cytometry. Furthermore, we quantified cell cycle-related and apoptosis-associated proteins by Western blotting. Results: SH reduced IgA and C3 depositions in stage 4 IgAN, thereby decreasing inflammatory cellular infiltration and mesangial injury in an IgAN model induced using heteroproteins. Furthermore, SH arrested the cell cycle of lymphocytes T and B from the spleen of IgAN rats. Regarding the mechanism, our results demonstrated that SH regulated the Cyclin D1 and Cyclin E1 protein levels for arresting the cell cycle and it also regulated Bax and Bcl-2 protein levels, thus increasing Cleaved caspase-3 protein levels in Jurkat T and Ramos B cells. Conclusion: SH exerts a dual regulation on the cell cycle and apoptosis of T and B cells by controlling cell cycle-related and apoptosis-associated proteins; it also reduces inflammatory cellular infiltration and mesangial proliferation. These are the major mechanisms of SH in IgAN.

[Structure and Distribution Characteristics of Bacterial Community in Boqing River Water].

The structure and distribution of bacterial communities should be influenced by urban and industrial activities. The Boqing River, which flows through towns and a copper tailing reservoir, is an important tributary of Xiaolangdi Reservoir in South Shanxi. In order to clarify the structure and distribution characteristics of the bacterial community in the Boqing River, we collected water samples along the Boqing River. The diversity characteristics of bacterial communities were analyzed, and their relationships with environmental factors were also explored. The results showed that the abundance and diversity of the bacterial community were higher in the downstream than that in the upstream of the river. Both of the parameters first decreased and then increased along the river. The lowest and highest bacterial abundance and diversity existed in the copper tailing reservoir and the site adjacent to the Xiaolangdi Reservoir, respectively. Proteobacteria, Actinobacteriota, Bacteroidota, and Firmicutes were dominant taxa at the bacterial phylum level in the river, and Acinetobacter, Limnohabitans, Pseudoarthrobacter, and Flavobacterium were dominant taxa at the genus level. Acinetobacter had the highest relative abundance in urban water of the river, which was significantly positively correlated with TC. Flavobacterium was significantly correlated with As. Considering its co-occurrence with As, we speculated that As might contribute to the spread of pathogenic bacteria in the study area. The results of this study were of great importance for the assessment of aquatic health under a complex environment.

[Effects of Different Vegetation Types on Soil Microbial Functional Genes and Enzyme Activities in Reclaimed Coal Mine].

Long-term coal mining has seriously damaged soil structures and ecological environments. Reclaimed vegetation could effectively improve the ecological environment. There have been many studies on soil physical-chemical characteristics and microbial community structure, but there are few on soil functional microorganisms in mining areas. In this study, soil physical and chemical properties, soil enzyme activities, and the abundance of functional genes in carbon, nitrogen, phosphorus, and sulfur cycling were determined in five reclaimed lands of different vegetation types (Platycladus orientalis, Picea asperata, Pinus tabuliformis, Pinus sylvestris, and Sabina chinensis) in the Jinhuagong mining area, Datong City, Shanxi province. The results showed that the restoration types produced significant effects on soil physical and chemical properties, soil enzyme activities, and the abundance of 75 functional genes. The highest contents of total carbon, total nitrogen, and total sulfur were in the S. chinensis site. The activities of dehydrogenase and urease were the highest in P. asperata, and the lowest alkaline protease activity was in P. asperata. The alkaline protease activity was the highest in P. sylvestrist. The abundance of functional genes in the P. asperata forest was the highest, but the diversity index was significantly lower than that in other plots, which might be due to the fact that the total abundance of ureC, acsA, and mct in P. asperata accounted for more than 52%. Soil urease was significantly correlated with the abundance of six functional genes. The ß diversity of the functional microbial community was significantly different among the different vegetation types, indicating that functional genes could better explain the effects of vegetation types on microbial function. The highest and lowest integrated fertility indexes were found in the P. sylvestris and P. orientalis plots, respectively. In conclusion, soil functional genes are sensitive indicators signifying the effects of different reclamation types on soil microorganisms, and P. asperata and P. sylvestris are more suitable for reclamation in this mining area.

Virulence and biofilm inhibition of 3-methoxycinnamic acid against Agrobacterium tumefaciens.

AIMS: In the current study the anti-virulence and anti-biofilm activities of the cinnamic acid derivative, 3-methoxycinnamic acid, was investigated against Agrobacterium tumefaciens. METHODS AND RESULTS: Based on the disc diffusion test and ß-galactosidase activity assay, 3-methoxycinnamic acid was shown to interfere with the quorum sensing (QS) system of A. tumefaciens. Crystal violet staining assay, phenol-sulfuric acid method, Bradford protein assay and confocal laser scanning microscopy (CLSM) revealed that the biofilm formation of A. tumefaciens was inhibited after the treatment of 3-methoxycinnamic acid. Employing high-performance liquid chromatography (HPLC) analysis of culture supernatant revealed that the production of 3-oxo-octanoylhomoserine lactone (3-oxo-C8-HSL) decreased concentration-dependently after treatment with 3-methoxycinnamic acid. Swimming and chemotaxis assays also indicated that 3-methoxycinnamic acid had a good effect on reducing the motility and chemotaxis of A. tumefaciens. In addition, the RT-qPCR, molecular docking and simulations further demonstrated that 3-methoxycinnamic acid could competitively inhibit the binding of 3-oxo-C8-HSL to TraR and down-regulate virulence-related genes. CONCLUSIONS: 3-Methoxycinnamic acid is proved to have good anti-virulence and anti-biofilm activities against A. tumefaciens. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study that investigates the anti-virulence and anti-biofilm activities of 3-methoxycinnamic acid against A. tumefaciens. With its potential QS-related virulence and biofilm inhibitory activities, 3-methoxycinnamic acid is expected to be developed as a potent pesticide or adjuvant for the prevention and treatment of crown gall caused by A. tumefaciens.

Effects of Chinese medicine herbal residues on antibiotic resistance genes and the bacterial community in chicken manure composting.

The use of livestock manure is an important way for antibiotic resistance genes (ARGs) to enter the environment, and composting is an effective method for removing ARGs from livestock manure. In this study, different volume ratios of Chinese medicinal herbal residues (CMHRs) were added to laboratory-scale chicken manure composting to evaluate their effects, if any, on the behavior of ARGs, mobile genetic elements (MGEs), and the bacterial community. At the end of the composting period, the composition of the microbial community changed. Firmicutes decreased and Bacteroidetes increased. The most striking effect was that the relative abundance of the 21 ARGs and 5 MGEs detected decreased by varying degrees in the different treatments (except for sulI and intI1). The removal rate of the ARGs increased with the increased addition of CMHRs. The correlations between transferase genes (tnpA and tnpA-02) and ARGs were significant (p < 0.05); therefore, transposons play an important role in the horizontal gene transfer of ARGs in chicken manure. The results imply that CMHRs would be an effective bulking agent for the removal of ARGs from chicken manure composting.

Effectiveness of methylprednisolone therapy in patients with a high-risk common type of COVID-19 pneumonia: a retrospective cohort study.

The optimal timing of glucocorticoid treatment for coronavirus disease 2019 (COVID-19) pneumonia is uncertain. We evaluated the clinical outcomes of methylprednisolone therapy (MPT) for patients with a high-risk common type (HRCT) COVID-19 pneumonia. We conducted a multicenter retrospective cohort study in Northeast China. A comparison was performed between the standard treatment (SDT) group and the SDT + MPT group to determine the efficacy of methylprednisolone in treating HRCT COVID-19 pneumonia. We collected the medical records of 403 patients with HRCT COVID-19 pneumonia (127 in the SDT + MPT group and 276 in the SDT group). None of the patients had received mechanical ventilation or died. Furthermore, there were no side effects associated with MPT. Patients in the SDT + MPT group treated with methylprednisolone received an intravenous injection for a median interval of five days (interquartile range of 3 to 7 days). The trends in lymphocyte count, C-reactive protein, interleukin 6, lactic acid dehydrogenase, respiratory rate, SpO2, PaO2, D-dimer and body temperature were similar between the SDT + MPT and SDT groups. The results for the SDT + MPT group seemed to improve faster than those for the SDT group; however, the results were not statistically significant. Clinical outcomes revealed that the average hospitalized days and the rate of progression to severe type COVID-19 pneumonia in both the SDT + MPT group and the SDT group were 14.56 ± 0.57 days versus 16.55 ± 0.3 days (p = 0.0009) and 21.26% (27/127) versus 32.4% (89/276) (p = 0.0247), respectively. The 16-day nucleic acid negative rate was higher in the SDT + MPT group than in the SDT group, 81.73% (104/127) versus 65.27% (180/276) (p = 0.0006). MPT effectively prevents patients with HRCT COVID-19 pneumonia from progressing to the severe stage.

Quorum sensing inhibition of hordenine analogs on Pseudomonas aeruginosa and Serratia marcescens.

Quorum sensing (QS) plays an essential role in virulence factor production, biofilm formation, and antimicrobial resistance. As a potent QS inhibitor, hordenine can inhibit both QS and biofilm formation in Pseudomonas aeruginosa and Serratia marcescens. In this work, we tested the QS inhibitory potential of 27 hordenine analogs against QS and biofilm formation in P. aeruginosa and S. marcescens. Among the tested analogs, seven (12, 28, 27, 26, 2, 23, and 7) exhibited strong QS inhibitory activity against P. aeruginosa, five of which (12, 28, 27, 26, and 2) showed better inhibitory activity than hordenine. In addition, seven analogs (28, 12, 23, 7, 26, 2, and 27) exhibited better biofilm inhibition against P. aeruginosa than hordenine. Four analogs (7, 28, 2, and 12) showed QS inhibitory activity against S. marcescens, two of which (7 and 28) demonstrated better inhibitory activity than hordenine. Furthermore, analog 7 showed similar biofilm inhibition against S. marcescens as hordenine. Structure-activity relationship (SAR) analysis indicated that the inhibitory activities of the analogs were related to four factors, i.e., carbon chain length, presence or absence of an α,ß-C[bond, double bond]C bond, amino group with/without lipophilic group, such as methyl group, and hydroxyl group in benzene ring.

A positive feedback loop between Periostin and TGFß1 induces and maintains the stemness of hepatocellular carcinoma cells via AP-2α activation.

BACKGROUND: Liver cancer stem cells (LCSCs) play key roles in the metastasis, recurrence, and chemotherapeutic resistance of hepatocellular carcinoma (HCC). Our previous research showed that the POSTN gene is closely related to the malignant progression and poor prognosis of HCC. This study aimed to elucidate the role of POSTN in generating LCSCs and maintaining their stemness as well as the underlying mechanisms. METHODS: Human HCC tissues and matched adjacent normal tissues were obtained from 110 patients. Immunohistochemistry, western blotting (WB), and RT-PCR were performed to detect the expression of POSTN and stemness factors. The roles of transforming growth factor (TGF)-ß1 and AP-2α in the POSTN-induced stemness transformation of HCC cells were explored in vitro and in vivo using LCSCs obtained by CD133+ cell sorting. RESULTS: The high expression of POSTN was correlated with the expression of various stemness factors, particularly CD133, in our HCC patient cohort and in TCGA and ICGC datasets. Knockdown of POSTN expression decreased the abilities of HCC cell lines to form tumours in xenograft mouse models. Knockdown of POSTN expression also suppressed cell viability and clone formation, invasion, and sphere formation abilities in vitro. Knockdown of AP-2α attenuated the generation of CD133+ LCSCs and their malignant behaviours, indicating that AP-2α was a critical factor that mediated the POSTN-induced stemness transformation and maintenance of HCC cells. The role of AP-2α was verified by using a specific αvß3 antagonist, cilengitide, in vitro and in vivo. Activation of POSTN could release TGFß1 from the extracellular matrix and initiated POSTN/TGFß1 positive feedback signalling. Furthermore, we found that the combined use of cilengitide and lenvatinib suppressed the growth of HCC cells with high POSTN expression more effectively than the use of lenvatinib alone in the patient-derived xenograft (PDX) mouse model. CONCLUSIONS: The POSTN/TGFß1 positive feedback pathway regulates the expression of stemness factors and the malignant progression of HCC cells by regulating the transcriptional activation of AP-2α. This pathway may serve as a new target for targeted gene therapy in HCC.

[Soil Fungal Community Structure and Functional Diversity in a Copper Tailing Dam and Its Surrounding Areas].

Soil fungal communities are reconstructed under heavy metal stress. This study was conducted to explore the structural and functional diversity of soil fungal communities under different land-use patterns, namely grassland and farmland, in 15-year-old or 10-year-old poplar plantations (Populus deltoides cv. 'zhonghuahongye') near the Shibahe copper tailing dam and the surrounding area, located in Yuanqu County, Shanxi. The results indicated that the abundance and diversity of fungal communities were the highest in the 15-year-old poplar plantation and lowest in the grassland on the tailing dam. The dominant fungal groups in the study area were Ascomycota, Basidiomycota, and Mortierellomycota. The Ascomycota members were the most abundant at all four sampling sites, reaching 92.92% of the fungal community in the grassland on the tailing dam. The highest functional diversity of carbon utilization was found in the soil fungal communities of the 15-year-old poplar plantations; moreover, the diversity in the grassland on the tailing dam was significantly higher than that in the farmland and in the 10-year-old poplar plantation. Meanwhile, we also observed the highest glucoside carbon source utilization efficiency in the grassland. Notably, the correlation analysis revealed that Ascomycota was significantly correlated with the utilization efficiency of the glucoside carbon source in our study area. Moreover, the soil total nitrogen content, but not the soil heavy metal content, was significantly correlated with the abundance of the soil fungal community. There were significant correlations between the cadmium and arsenic contents and the soil fungal community diversity indices. Our results indicated that the soil fungal communities were more influenced by soil fertility and land use than by the soil heavy metal content. This study underlies the application of soil fungal communities in soil remediation of heavy metal-contaminated areas.

Quorum sensing inhibition and tobramycin acceleration in Chromobacterium violaceum by two natural cinnamic acid derivatives.

Chromobacterium violaceum, one free-living Gram-negative bacterium, is abundantly presented in tropics and sub-tropics soil and aquatic environment; it is also an opportunistic human pathogen. Here, two cinnamic acid derivatives, i.e., 4-dimethylaminocinnamic acid (DCA) and 4-methoxycinnamic acid (MCA), were identified as potential quorum sensing (QS) and biofilm inhibitors in C. violaceum ATCC12472. Both DCA (100 µg/mL) and MCA (200 µg/mL) inhibited the levels of N-decanoyl-homoserine lactone (C10-HSL) and reduced the production of certain virulence factors in C. violaceum, including violacein, hemolysin, and chitinase. Metabolomics analysis indicated that QS-related metabolites, such as ethanolamine and L-methionine, were down-regulated after treatment with DCA and MCA. Quantitative real-time polymerase chain reaction (qRT-PCR) demonstrated that DCA and MCA markedly suppressed the expression of two QS-related genes (cviI and cviR). In addition, DCA and MCA also inhibited biofilm formation and enhanced the susceptibility of biofilms to tobramycin, which was evidenced by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Our results indicated that DCA and MCA can serve as QS-based agent for controlling pathogens.Key Points • DCA and MCA inhibited QS and biofilm formation in C. violaceum.• The combination of DCA or MCA and tobramycin removed the preformed biofilm of C. violaceum. • DCA or MCA inhibited virulence factors and expressions of cviI and cviR of C. violaceum.• DCA or MCA are potential antibiotic accelerants for treating C. violaceum infection.

[Effect of Co-composting of Chicken Manure with Chinese Medicinal Herbal Residues on Antibiotic Resistance Genes].

Livestock manure is an important pathway by which antibiotic resistance genes (ARGs) enter the environment. To reduce the occurrence of antibiotic resistance genes in manures, we studied the variations of ARGs and mobile gene elements (MGEs) during the 46-day co-composting of chicken manure with Chinese medicinal herbal residues. The abundance of 100 ARGs and 5 MGEs were measured by Real-Time Quantitative PCR with 0, 3, 7, 14, 28, and 46 d co-composting. We detected 21 ARGs, 2 integrase genes (intI1 and intI2), and 3 transposase genes (tnpA-01, tnpA-02, and tnpA-03). The abundance of 5 MGEs significantly declined with co-composting time, particularly tnpA-01 and tnpA-02, which were reduced by two orders of magnitude. The abundance of aacA/aphD and aadE were significantly reduced (P<0.05) in aminoglycoside resistance genes. In ß-lactam resistance genes, the strongest relationships were demonstrated between blaOXA1 and compost days (P=0.016), and the removal rate was 78.63%. The average removal rate was 90.39% for amide resistance genes, which decreased significantly with composting time. The removal ratios were different among tetracycline resistance genes. For example, the removal ratios of tetG and tetR were 99.77% and 31.72%, respectively. The highest removal rate of qnrD was 99.89%. The removal rate of sulⅢ was as high as 99.88%, while sulⅠ showed an increasing trend. Correlations between ARGs and MGEs were significant correlation for tnpA-01 and ARGs (P<0.05). The trend of ARGs with composting time indicates that the composting of Chinese medicine residues and chicken manure can significantly reduce the abundance of ARGs, thus reducing the risk of ARGs being distributed via livestock manure application.

[Effect of the Sampling Scale and Number on the Heterogeneity of Soil Respiration in a Mixed Broadleaf-conifer Forest].

By upscaling the observed results at the plot scale, the carbon efflux from soils in a region can be estimated. Therefore, it is very important to investigate the spatial relations of soil respiration (Rs) and its environment and to evaluate the effect of the sampling scale and number on the accuracy of Rs measurement at the spatial scale. Based on field observation data for a mixed broadleaf-conifer forest in the Pangquangou Nature Reserve of the Shanxi Province, two analysis methods, that is, traditional statistics and geostatistics, were used to analyze the influence of the soil water content (Ws), soil temperature (T10), litter mass (Lw), litter moisture content (Lm), soil total carbon (C), total nitrogen (N), and ratio of C/N and sulfur (S) on the Rs heterogeneity at 4, 2, and 1 m sampling scales. The results show no significant differences between the average Rs values for the three sampling scales, but the degree of variation of Rs, which was evaluated based on the coefficient of determination, increases with increasing sampling scales, ranging from 16% to 22%. At the 4 m sampling interval, the correlations between Rs and Ws, Lw, C, and C/N are highly significant (P<0.01) and significant for N (P <0.05). At the 2 m sampling interval, Rs shows a highly negative significant correlation with T10 (P<0.01) and insignificant correlations with the other factors. At the 1 m sampling interval, significant relations between Rs and all other factors were not observed. With the decrease of the sampling interval scale, the spatial autocorrelation of Rs decreases gradually, ranging from high to weak autocorrelations.This indicates that the role the structural factors play decreases with the decrease of the sampling scale, but that of the random factors increases gradually. At the same confidence level for a certain sampling number, the estimated error in Rs decreases with decreasing sampling scale. The analysis of the effect of the sampling number at different sampling scales on the accuracy of Rs shows that the error of Rs at both the 2 and 1 m sampling scales is approximately±12% at the 95% confidence interval and±16% at the 4 m sampling scale. At the 90% confidence interval, the error of Rs at both the 2 and 1 m sampling scales is less than ±10%; at the 4 m scale, it is ±13%. Our results provide insights into how to arrange the sampling sites at the plot scale to measure the seasonal Rs.

Rosiglitazone Inhibits Activation of Hepatic Stellate Cells via Up-Regulating Micro-RNA-124-3p to Alleviate Hepatic Fibrosis.

BACKGROUND: The activation of hepatic stellate cells (HSCs) is involved in hepatic fibrogenesis and is regulated by the decreased expression of peroxisome proliferator-activated receptor γ (PPARγ). Rosiglitazone (RGZ) is a highly potent agonist of PPARγ. AIMS: To clarify molecular regulatory mechanism of RGZ in the activation of HSCs in hepatic fibrosis. METHODS: A mouse model of hepatic fibrosis was established by carbon tetrachloride with or without RGZ intervention. A vector carrying pcDNA-HOTAIR was constructed and injected into a mouse model. HSCs were isolated from liver tissue and activated by transforming growth factor-ß. The expression of miR-124-3p, HOTAIR, Col1A1, α-SMA, and PPARγ mRNAs was measured by quantitative real-time PCR. The level of PPARγ was measured by Western blotting. The interaction between HOTAIR and PPARγ was assessed by RNA immunoprecipitation (RIP) and RNA pull-down. The target gene of miR-124-3p was determined by luciferase reporter assay and RNA interference approaches. RESULTS: The expression of Col1A1 and α-SMA was reduced after RGZ intervention. Different expressions of HOTAIR and miR-124-3p were observed in liver tissue and HSCs. The luciferase reporter assay and RNA interference approaches indicated that miR-124-3p negatively regulated HOTAIR expression. RIP and RNA pull-down results revealed that PPARγ was interacted by HOTAIR. The therapeutic effect of RGZ on hepatic fibrosis was reversed by overexpression of HOTAIR. CONCLUSIONS: RGZ inhibits the activation of HSCs by up-regulating miR-124-3p. The silencing of HOTAIR by miR-124-3p in HSC activation provided the foundation to understand interactions of ncRNAs and potential treatment target in hepatic fibrosis.

Isocorydine suppresses doxorubicin-induced epithelial-mesenchymal transition via inhibition of ERK signaling pathways in hepatocellular carcinoma.

Doxorubicin (DOX) is a conventional and effective chemotherapeutic used in the treatment of hepatocellular carcinoma (HCC). However, doxorubicin administration may induce EMT, which results in the development of chemoresistance in HCC. Recent studies report that Isocorydine (ICD) selectively inhibits human cancer stem cells (CSCs), which have an important role in the development of chemoresistance. In this study, we observed that ICD co-administration enhanced DOX cytotoxicity in HCC cells, enabling the inhibition of DOX-induced epithelial-mesenchymal transition (EMT). Microarray data analysis revealed substantially decreased ERK signaling after ICD treatment. Additionally, we observed decreased IC50 for DOX upon ERK knockdown. Finally, we confirmed the enhanced efficacy of treatment with a combination of DOX and ICD in xenograft models. Collectively, the present study unveils the benefit of using DOX in combination with ICD for chemotherapy against HCC, revealing a novel potential anti-cancer strategy.

Aberrant SERPINE1 DNA methylation is involved in carboplatin induced epithelial-mesenchymal transition in epithelial ovarian cancer.

AIM: Resistance to platinum-based therapeutic agents is the major contributor to epithelial ovarian cancer (EOC) mortality. There is an urgent need to better understand the underlying mechanisms. Here we investigated the role of serpins in EOC chemoresistance and related mechanisms, and found that SERPINE1 played an important role in chemoresistance in A2780cp cells. MATERIALS AND METHODS: A2780cp and A2780s cells were used in our study. Microarray screening was used to identify the gene expression change under carboplatin treatment. A cell-counting kit-8 was used to detect the sensitivity of ovarian cancer cells after treatment. The expression of SERPINE1 was silenced by siRNA. The levels of SERPINE1 and epithelial-mesenchymal transition (EMT)-related proteins were confirmed by Western blot. MassArray EpiTYPER quantitative DNA methylation analysis was introduced to evaluate the methylation of the promoter of SERPINE1. RESULTS: Microarray data showed that SERPINE1 and SERPINE2 increased most dramatically under carboplatin treatment in A2780cp cells. Carboplatin treatment could significantly increase the expression of SERPINE1 and induce the EMT process, with decreased expression of E-cadherin and increased expression of Vimentin, Snail and Twist. Knockdown of SERPINE1, but not SERPINE2, in A2780cp cells could inhibit the EMT process. We also found that hypomethylation in the promoter of SERPINE1 might result in the increased expression of SERPINE1 and subsequent EMT process in A2780cp cells. CONCLUSION: Our study suggested that SERPINE1 may be a promising therapeutic target for chemoresistance.

[Effects of Different Vegetation Types and Reclamation Years on Soil Bacterial Community Structure in Reclaimed Mine Areas].

Effects of different vegetation types (Ulmus pumila, Larix gmelinii, Armeniaca vulgaris, Picea asperata and Robinia pseudoacacia) and reclamation years (15 and 20 years) on soil bacterial community structure in reclaimed Antaibao opencast mine areas were investigated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and clone sequencing. For 20-year reclaimed soils, the significantly highest and lowest bacterial diversity were found in U. pumila and A. vulgaris stand, respectively, whereas no significant differences were found between the other three vegetation types. Under 15-year plantations, soil bacterial diversity index of P. asperata was significantly higher than that of R. pseudoacacia. Soil bacterial diversity index significantly increased in R. pseudoacacia planted soils but decreased in P. asperata treatment with the increase of reclaimed years. No significant change of soil bacterial community structure was observed in the same reclamation years based on the similarity coefficient analysis, cluster analysis and principal component analysis (PCA). Pearson correlation analysis demonstrated that bacterial diversity index was significantly positively correlated with soil pH. Nitrospira, Sphingomonas, Arthrobacter, Brachybacterium, Rhizobium as well as Mesorhizobium, which play important roles in the nitrogen cycle, degradation of polycyclic aromatic hydrocarbons and other organic matter, were identified by clone sequencing of the DGGE bands. Our results indicated that U. pumila and P. asperata were conducive to the recovery of soil bacterial diversity. The most dominant bacterial community from reclaimed mine soil would be beneficial for restoring wasteland contaminated soil and improving soil fertility.

[Effects of Microbial Diversity on Soil Carbon Mineralization].

Soil microbial community plays an important role in ecosystem functions; however, little is known about the importance of microbial diversity to the ecosystems. In this study, serial dilution of soil suspension (10-1, 10-3, and 10-5) was performed and inoculated into the sterilized soils that form the broadleaf mixed forests in Pangquangou. The change in the carbon mineralization rate and the pattern of the carbon source utilization were studied by titration, Biolog Eco, and other experimental methods. The results show that after being incubated for six weeks, carbon mineralization rate, the cumulative amount of carbon mineralization, average well color development (AWCD), and diversity index (Shannon, McIntosh, and richness index) of D1 were significantly higher than those of the D5 treatment. The cumulative amount of carbon mineralization and AWCD was strongly and inversely correlated with richness. Principal component analysis and one-way ANOVA also indicated that the patterns of carbon source utilization of microbially diverse soil were different. Therefore, the loss of microbial diversity affects the carbon mineralization rate and the pattern of carbon source utilization, leading to functional changes in terrestrial ecosystems. In the management of forest soils, the effects of soil microbial diversity on ecosystem functions should be considered.

Study of the genus Gomphocerippus (Orthoptera, Acridoidea, Acrididae, Gomphocerinae) with a new species from northeast China.

A new species Gomphocerippus longipennis Li & Ren sp. nov. of the genus Gomphocerippus Roberts, 1941 is described and illustrated from the most northern region of China. The new species is allied to Gomphocerippus rufus (Linnaeus, 1758), but differs from the latter by length of lateral foveola 3.0 times width, tegmina partially surpassing the apex of hind femur, costal area of tegmina in male 1.5 times of subcostal area in width and media area of tegmina in male 1.2 times cubital area in width. The type specimens are deposited in the Jilin Provincial Key Laboratory of Animal Resource Conservation and Utilization, Northeast Normal University, Changchun, Jilin, China.