Plumbagin induces ferroptosis in colon cancer cells by regulating p53-related SLC7A11 expression.

Objective: This study examined the mechanism through which plumbagin induces ferroptosis of colon cancer cells. Methods: CCK-8 assay was performed to examine the viability of colon cancer cells (SW480 and HCT116 cells) after they were treated with 0-, 5-, 10-, 15- and 20-µmol/L plumbagin. Colony formation assay and Transwell assay were used to examine the effects of 15-µmol/L plumbagin on the proliferation, invasive ability. The ferroptosis of SW480 and HCT116 cells and the expression of p-p53, p53 and SLC7A11 were analysed. The effects of blocking necrosis, apoptosis and ferroptosis on the anti-cancer effects of plumbagin were examined. After p53 was silenced, the effects of plumbagin on proliferation, invasion, ferroptosis and SLC7A11 expression were assessed. A tumour-bearing nude mouse model was used to examine the effects of p53 silencing and/or plumbagin on tumour growth, ferroptosis and SLC7A11 expression. Results: Plumbagin inhibited the proliferation of SW480 and HCT116 cells and their invasive and colony-forming abilities. It increased Fe2+ levels but significantly decreased GSH and GPX4 levels. When ferroptosis was inhibited, the effects of plumbagin on colon cancer cells were significantly alleviated. Plumbagin promoted the expression and phosphorylation of p53 and inhibited the mRNA and protein levels of SLC7A11. Silencing of p53 counteracted the effects of plumbagin on the ferroptosis and biological behaviour of SW480 and HCT116 cells. In mouse models of colon cancer, silencing of p53 attenuated the tumour-suppressing effects of plumbagin as well as its inhibitory effects on the protein level of SLC7A11 and restored the expression of GSH and GPX4. Conclusion: Plumbagin promotes ferroptosis and inhibits cell proliferation and invasion by decreasing the protein expression of SLC7A11 through p53.

Boosting degradation and defluorination efficiencies of PFBS in a vacuum-ultraviolet/S(â £) process with iodide involvement.

Perfluorobutane sulfonate (PFBS) is considered to be a promising alternative of perfluorooctane sulfonates (PFOS), while it is also hazardous. The UV/S (Ⅳ) system has been confirmed to be effective for PFOS removal from water, while it is inefficient for PFBS decomposition. A hybrid vacuum-ultraviolet (VUV)/S (Ⅳ)/KI process was investigated for the degradation of PFBS in aqueous solution. With KI involvement, the degradation rate of PFBS was boosted from 1.8802 µg h-1 up to 3.5818 µg h-1 in the VUV/S (Ⅳ) process. Alkaline conditions significantly increased the degradation efficiency of PFBS, which can be explained that S (Ⅳ) was dominated by SO32- rather than HSO3- and H2SO3 in alkaline conditions. Cl-, HCO3-, NO3-, NO2-, and HA would inhibit the performance of the VUV/S (Ⅳ)/KI process via various reactions. In addition, the toxicity of PFBS was significantly reduced by the VUV/S (Ⅳ)/KI process. Even in actual waters, the VUV/S (Ⅳ)/KI process also presented a satisfying performance in the degradation of PFBS.

Co-infection analysis of bacterial and viral respiratory pathogens from clinically healthy swine in Eastern China.

Porcine respiratory disease complex (PRDC) is one of the most challenging health concerns for pig production worldwide. The aim of the present study was to determine the prevalence of pathogens associated with PRDC, including porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) and bacterial agents, such as Streptococcus suis, Haemophilus parasuis and Actinobacillus pleuropneumoniae, in clinically healthy pigs in Eastern China. Molecular detection revealed positive single-pathogen detection rates of 59.9%, 27.2%, 52.3%, 33.2% and 0.4% for PCV2, PRRSV, S. suis, H. parasuis and A. pleuropneumoniae, respectively. Co-infection with more than one pathogen was frequently detected in these samples, with PCV2/S. suis, H. parasuis and PCV2/H. parasuis mixed infection rates of 35.4%, 33.2% and 21.6%, respectively, and PCV2/S. suis/H. parasuis and PRRSV/PCV2/S. suis co-infection rates of 21.6% and 6.2%, respectively. These results suggest that mixed infections are prevalent among PRDC cases in swine, which may pose a greater threat to the health of herds compared with single-pathogen infections.

Efficient degradation of diclofenac sodium by periodate activation using Fe/Cu bimetallic modified sewage sludge biochar/UV system.

Iron/copper bimetallic nanoparticles based sludge biochar (Fe/Cu-SBC) was prepared by using a modified co-precipitation route. The Fe/Cu-SBC system prepared was subsequently applied to activate periodate (IO4-) to degrade diclofenac sodium (DCF) by using UV light at room temperature (25 °C). The physicochemical properties of both SBC and Fe/Cu-SBC such as morphology, physical properties, crystal structures and functional groups were examined. The type and number of surface functional groups were found to be increased and the catalytic performance was improved by the modification of Fe/Cu bimetallic nanoparticles. The influence of various parameters to evaluate the catalytic efficiency such as periodate (PI) concentration, dosage of catalysts, UV power, initial pH and coexisting anions were investigated. Under the optimized conditions (pH 6.9, UV-power 60 W, PI concentration of 5 mM and 0.1 g Fe/Cu-SBC), it was observed that 99.7% of DCF was degraded with a pseudo-first-order kinetics reaction constant 9.39 × 10-2 min-1. The radical scavenging experiments showed that IO3 radicals were the predominantly reactive oxidants in the Fe/Cu-SBC/UV system. Therefore, this investigation provides a feasible alternative for the degradation of PPCPs in wastewater.

Review on plant uptake of PFOS and PFOA for environmental cleanup: potential and implications.

Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) have gained increasing concern due to their persistent characteristics, wide distribution, biotoxicity, and bioaccumulative properties. The current remediation technologies for PFOA and PFOS are primarily focused on physical and chemical techniques. Phytoremediation has provided promising alternatives to traditional cleanup technologies due to their low operational costs, low maintenance requirements, end-use value, and aesthetic nature. In this review, uptake, translocation, and toxic effects of PFOS and PFOA are summarized and discussed. Several potential hyperaccumulators of PFOS and PFOA are provided according to the existing data. Biomass, chlorophyll, soluble protein, enzyme activities, oxidative stress, and other variables are assessed for potential indicator of PFOS/PFOA biotoxicity. The various studies on multiple scales are compared for identifying the threshold values. Several important implications and recommendations for future research are proposed at the end. This review provides an overview of current studies on plant uptake of PFOS and PFOA from the perspective of phytoremediation.

Human Mesenchymal Stem Cell-Derived Exosomal microRNA-143 Promotes Apoptosis and Suppresses Cell Growth in Pancreatic Cancer via Target Gene Regulation.

BACKGROUND: This study aimed to explore the regulatory mechanism of hsa-miR-143-3p and lncRNA RP11-363N22.3-functioning upstream of KRAS-in exosomes derived from human mesenchymal stem cells (hMSCs) in pancreatic cancer. METHODS: Western blotting and quantitative PCR were used to determine gene expression. In vitro, cell proliferation, apoptosis, and cell cycle and invasion were evaluated using CCK-8 assay, flow cytometry, and transwell assays, respectively. In vivo, the effect of hsa-miR143-3p was investigated using a tumorigenesis test in nude mice. The association between hsa-miR-143-3p and lncRNA RP11-363N22.3 was investigated using the dual-luciferase assay. RESULTS: hsa-miR-143-3p expression significantly increased in hMSC exosomes than in those in human pancreatic cancer cell line (CFPAC-1) exosomes. In vitro, compared to the MOCK (CFPAC-1 only) group, cell proliferation and invasion were inhibited and apoptosis was induced in the inhibitor NC (CFPAC-1 + MSC-hsa-miR-3p inhibitor NC) group, while these changes were reversed in the inhibitor (CFPAC-1 + MSC-hsa-miR-3p inhibitor) group. The expression of lncRNA RP11-363N22.3 and genes related to miR-143 significantly decreased in the inhibitor NC group compared to the MOCK group, and increased in the inhibitor group compared to inhibitor NC group. A targeted combinatorial effect was observed between lncRNA RP11-363N22.3 and hsa-miR-143-3p. In vivo, the tumor volume of the mimics (CFPAC-1 + MSC-hsa-miR-143-3p mimics) group was smaller than that of the mimics NC (CFPAC-1 + MSC-hsa-miR-143-3p mimics NC) and MOCK groups. H&E staining showed that there were no obvious pathological changes in MOCK and mimic NC groups, while cell necrosis was seen in some regions in mimic groups. CONCLUSION: hsa-miR-143-3p may promote apoptosis and suppress cell growth and invasion in pancreatic cancer.

Molecular epidemiology and genetic evolution of canine parvovirus in East China, during 2018-2020.

Canine parvovirus type 2 (CPV-2) emerged in the late 1970s, which caused high rates of morbidity and mortality in dogs. In last decade, five genetic variants (CPV-2a, CPV-2b, CPV-2c, New CPV-2a, and New CPV-2b) were frequently reported in the dog population, and replaced the original CPV-2, rising widespread concerns. However, little is known about their recent genetic diversity and evolution. The aim of this study was to analyze the characteristics of the CPV-2 strains collected in East China from 2018 to 2020. The 57 CPV-2 strains were isolated from rectal swab samples (n=140). They belong to three different genotypes, based on VP2 protein amino acid sequence. The results revealed a high prevalence of CPV-2c (77.19%) compared to the New CPV-2a (5.26%) and New CPV-2b (17.54%) strains. Further analysis showed that nucleotide homology of the VP2 gene among the 57 CPV strains was 98.9%~100%, and the homology with 24 reference strains from different countries and regions was 98.1%~100%. The phylogenetic tree of VP2 gene sequence showed that 44 CPV-2c strains were distantly related to CPV-2, CPV-2a, CPV-2b, New CPV-2a, New CPV-2b and European/American CPV-2c strains, and were closely related to Asian CPV-2c strains. The results showed that these Asian CPV-2c strains had become the dominant strain, which renewed the knowledge of CPV-2 molecular epidemiology in East China.

MiRNA-340-5p mediates the functional and infiltrative promotion of tumor-infiltrating CD8+ T lymphocytes in human diffuse large B cell lymphoma.

BACKGROUND: CD8+ tumor-infiltrating T lymphocytes (T-TILs) in the tumor microenvironment (TME) play an important role in tumor development, and miRNAs regulate tumor cell interactions with the microenvironment. T-TIL-based tumor immunotherapy provides a promising treatment strategy in diffuse large B-cell lymphoma (DLBCL). MiRNAs tend to be attractive targets for novel antitumor interventions. METHODS: Weighted gene coexpression network analysis (WGCNA), CIBERSORT analysis and Cox regression analysis were used to identify CD8+ T-TIL-related miRNAs. RT-PCR, western blotting, immunohistochemistry (IHC), in situ hybridization (ISH), luciferase reporter assay, coimmunoprecipitation and ubiquitination analyses were used to detect miRNA, mRNA and protein expression and their combination. The viability and function of CD8+ T cells after stimulation were evaluated by enzyme-linked immunosorbent assay (ELISA), cytotoxicity assay, functional avidity assessment, flow cytometry and Cell Counting Kit-8 (CCK-8) assay. DLBCL cell lines, primary cells and a murine xenograft model established with A20 cell injection were used as in vitro and in vivo experimental models. RESULTS: MiR-340-5p was positively correlated with CD8+ T-TILs in DLBCL patients, and KMT5A was a direct target gene of miR-340-5p. CD8+ T-cell function was significantly enhanced by miR-340-5p mimics both in vitro and in vivo, which was reversed by KMT5A overexpression. We demonstrated that COP1/CD73 was involved in the downstream mechanism of the miR-340-5p/KMT5A axis involving ubiquitination. In vivo, we validated an improved CD8+ T-TIL infiltration rate and tumor suppression with miR-340-5p treatment. Furthermore, miR-340-5p directly regulated the biological activity of DLBCL cells without CD8+ T-cell participation. CONCLUSIONS: MiR-340-5p promoted CD8+ T-TIL infiltration and antitumor function by regulating KMT5A and COP1 and further activating CD73 ubiquitination. MiR-340-5p is potentially a novel target for DLBCL immunotherapy.

Variable selection from a feature representing protein sequences: a case of classification on bacterial type IV secreted effectors.

BACKGROUND: Classification of certain proteins with specific functions is momentous for biological research. Encoding approaches of protein sequences for feature extraction play an important role in protein classification. Many computational methods (namely classifiers) are used for classification on protein sequences according to various encoding approaches. Commonly, protein sequences keep certain labels corresponding to different categories of biological functions (e.g., bacterial type IV secreted effectors or not), which makes protein prediction a fantasy. As to protein prediction, a kernel set of protein sequences keeping certain labels certified by biological experiments should be existent in advance. However, it has been hardly ever seen in prevailing researches. Therefore, unsupervised learning rather than supervised learning (e.g. classification) should be considered. As to protein classification, various classifiers may help to evaluate the effectiveness of different encoding approaches. Besides, variable selection from an encoded feature representing protein sequences is an important issue that also needs to be considered. RESULTS: Focusing on the latter problem, we propose a new method for variable selection from an encoded feature representing protein sequences. Taking a benchmark dataset containing 1947 protein sequences as a case, experiments are made to identify bacterial type IV secreted effectors (T4SE) from protein sequences, which are composed of 399 T4SE and 1548 non-T4SE. Comparable and quantified results are obtained only using certain components of the encoded feature, i.e., position-specific scoring matix, and that indicates the effectiveness of our method. CONCLUSIONS: Certain variables other than an encoded feature they belong to do work for discrimination between different types of proteins. In addition, ensemble classifiers with an automatic assignment of different base classifiers do achieve a better classification result.

Photosynthesis of alfalfa (Medicago sativa) in response to landfill leachate contamination.

Thousands of unlined landfills and open dumpsites have put great threat on the security of soil and ground water due to leachate leakage. Alfalfa is believed potential as a phytoremediation plant for leachate contamination based on strong root system and the excellent capacity of removing various kinds of pollutants. A lab-scale investigation was conducted to figure out the sensitiveness of alfalfa photosynthesis in response to leachate contamination. The results demonstrated that both of the maximum photosynthetic efficiency (Fv/Fm) and net photosynthetic rate (Pn) were slightly inhibited in the high-dosage group. Based on statistical analysis, higher sensitivity of Pn to leaching parameters than Fv/Fm was observed. There were significant correlations between most of leaching parameters (pH, ammonium and COD) and Pn with correlation coefficients of 0.530, -0.580 and -0.578 (p < 0.01), respectively. Therefore, Pn is potential for acting as an effective indicator for staple leaching characteristics of leachate contaminated soils.

Contribution of eukaryotic-type serine/threonine kinase to stress response and virulence of Streptococcus suis.

Streptococcus suis serotype 2 (SS2) is an important swine and human pathogen responsible for septicemia and meningitis. The bacterial homologues of eukaryotic-type serine/threonine kinases (ESTKs) have been reported to play critical roles in various cellular processes. To investigate the role of STK in SS2, an isogenic stk mutant strain (Δstk) and a complemented strain (CΔstk) were constructed. The Δstk showed a significant decrease in adherence to HEp-2 cells, compared with the wild-type strain, and a reduced survival ratio in whole blood. In addition, the Δstk exhibited a notable reduced tolerance of environmental stresses including high temperature, acidic pH, oxidative stress, and high osmolarity. More importantly, the Δstk was attenuated in both the CD1 mouse and piglet models of infection. The results of quantitative reverse transcription-PCR (qRT-PCR) analysis indicated that the expressions of a few genes involving in adherence, stress response and virulence were clearly decreased in the Δstk mutant strain. Our data suggest that SsSTK is required for virulence and stress response in SS2.

Characterization and proteome analysis of inosine 5-monophosphate dehydrogenase in epidemic Streptococcus suis serotype 2.

Streptococcus suis serotype 2 (SS2) is an important zoonotic pathogen that causes severe disease symptoms in pigs and humans. In the present study, we found one isogenic mutant lacking inosine 5-monophosphate dehydrogenase (IMPDH) ΔZY05719 was attenuated in pigs compared with the wild-type SS2 strain ZY05719. Comparative proteome analysis of the secreted proteins expression profiles between ZY05719 and ΔZY05719 allowed us to identify Triosephosphate isomerase (TPI) and glyceraldehyde phosphate dehydrogenase (GAPDH), which were down expressed in the absence of the IMPDH. Both of them are glycolytic enzymes participating in the glycolytic pathway. Compared with ZY05719, ΔZY05719 lost the ability of utilize mannose, which might relate to down expression of TPI and GAPDH. In addition, GAPDH is a well-known factor that involved in adhesion to host cells, and we demonstrated ability of adhesion to HEp-2 and PK15 by ΔZY05719 was significantly weakened, in contrast to ZY05719. The adhesion to host cells is the crucial step to cause infection for pathogen, and the reduction adhesion of ΔZY05719, to some extent illustrates the attenuated virulence of ΔZY05719.

Monoclonal antibodies against Nsp2 protein of the highly pathogenic porcine reproductive and respiratory syndrome virus.

Porcine reproductive and respiratory syndrome (PRRS) continues to be a serious threat, causing an economically significant impact on the swine industry worldwide. In this study, non-structural protein Nsp2 of porcine reproductive and respiratory syndrome virus (PRRSV) was expressed in Escherichia coli and purified by dialysis. An important monoclonal antibody (MAb 2H6) against Nsp2 protein was generated by fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from Nsp2 protein immunized mice. Then activity of the MAb was characterized by enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and indirect immunofluorescence assays. The results demonstrated that the MAb has a positive reaction to HP-PRRSV in IFA at 1:100 dilution and in Western blot analysis at 1:500 dilution, and no reaction with classic PRRSV. These indicated that this MAb against Nsp2 protein of PRRSV might be a good candidate for a specific diagnostic method and functional exploration of the Nsp2 protein.

Complete genome sequence of a recombinant porcine epidemic diarrhea virus strain from eastern china.

A field porcine epidemic diarrhea virus (PEDV) strain, JS2008, was isolated from stool samples of a piglet with acute diarrhea on a vaccinated farm in eastern China. We sequenced and analyzed the complete genome of strain JS2008, which will help increase our understanding of the molecular characteristics of the epidemic PEDV in China.

In vitro and in vivo isolation of a novel rearranged porcine circovirus type 2.

Here, we present the first report of a novel rearranged porcine circovirus type 2 (PCV2) strain named BIV, isolated from both in vitro and in vivo sources. The complete circular genome of BIV is 896 nucleotides in length. The data will help us to update current knowledge of the replication of PCV2 viruses in cell culture and of their molecular evolution, as well as their diagnosis.

A novel porcine circovirus-like agent P1 is associated with wasting syndromes in pigs.

A novel porcine pathogen tentatively named P1, which was obtained from the sera of the pigs exhibiting clinical signs of postweaning multisystemic wasting syndrome (PMWS) experimentally caused the classical clinic signs and pathologic lesions of the disease in pigs by direct in vivo injection with P1 DNA plasmids. Twenty colostrum-fed (CF) pigs that were free of PCV2 and P1 at 1 month of age were randomly designated equally to two groups. Group 1 pigs were each injected with 400 µg of the cloned P1 plasmid DNA into the superficial inguinal lymph nodes and Group 2 were injected with same amount of the empty pSK vector DNA and served as controls. Viremias were positively detected in 8 of 10 P1 infected pigs from 14-21 days post-inoculation (dpi). The 8 infected animals showed pallor of skin and diarrhea. Gross lesions in the pigs euthanized on 35 dpi were similarly characterized by encephalemia, haemorrhage of the bladder mucosa, haemorrhage of the superficial inguinal lymph nodes, lung atrophy and haemorrhage. Histopathological lesions were arteriectasis and telangiectasia of the cavitas subarachnoidealis, interstitial pneumonia, mild atrophy of the cardiac muscle cells, histiocytic hyperplasia of the follicles in the tonsils, and haemorrhage of the inguinal lymph nodes. P1 DNA and antigens were confirmed by PCR and immunohistochemistry in the tissues and organs of the infected pigs, including the pancreas, bladders, testicles/ovaries, brains, lungs and liver. There were no obvious clinical signs and pathological lesions in the control pigs. This study demonstrated that P1 infection is one of the important pathologic agents on pig farms.

In vitro intragenomic rearrangement of porcine circovirus type 2.

We report here for the first time the genome sequence of a rearranged porcine circovirus type 2 (PCV2) strain, CH-IVT1, isolated from PCV2-infected PK-15 cells. The complete circular genome of the CH-IVT1 is 605 nucleotides (nt) in length. The finding will help us to understand the molecular evolution of PCV2 and the relationship between PCV2 and PCV-associated diseases.

Complete genome sequence of the rearranged porcine circovirus type 2.

We first report here the genome sequences of 4 rearranged porcine circovirus type 2 strains, JSTZ, ZJQDH1, ZJQDH2, and JSHM, isolated from porcine sera in China. The complete circular genomes of these isolates are 578, 483, 574, and 772 nucleotides in length, respectively. They are predicted to be defective interfering particles of porcine circovirus type 2. The findings will help us to understand molecular evolution of porcine circovirus type 2 and the relationship between porcine circovirus type 2 and diseases.

Complete genome sequence of a highly prevalent porcine circovirus 2 isolated from piglet stool samples in China.

Porcine circovirus type 2 (PCV2) is the etiologic agent of porcine circovirus-associated disease. Here, we first report the complete genome sequence of PCV2 strain JSTZ, which was isolated from piglet stool samples and is highly prevalent in China. It will help in understanding the epidemiology and molecular characteristics of PCV2.

Development of a loop-mediated isothermal amplification method for rapid detection of porcine boca-like virus.

The porcine boca-like virus (Pbo-likeV) was recently discovered in Swedish pigs with post-weaning multisystemic wasting syndrome (PMWS). In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid, specific and sensitive detection of Pbo-likeV. A set of four primers specific for six regions of Pbo-likeV VP1/2 genes was designed with the online software. The reaction temperature and time were optimized to 65 °C and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change due to addition of fluorescent dye. The developed method was highly specific for detection of Pbo-likeV, and no cross-reaction was observed with other swine viruses, such as porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and classic swine fever virus (CSFV) found commonly in China. The lower detection limit of the LAMP assay was approximately 10 copies per reaction, and it was 100 times more sensitive than that of conventional PCR. Furthermore, the efficiency of LAMP for detection Pbo-likeV in clinical samples was comparable to PCR and sequencing. These results showed that the LAMP assay is a simple, rapid, sensitive and specific technique for detection of Pbo-likeV, and the procedure of LAMP does not rely on any special equipment. It has capacity for the detection of Pbo-likeV both in the laboratory and on farms.