Inhibition of GLUD1 mediated by LASP1 and SYVN1 contributes to hepatitis B virus X protein-induced hepatocarcinogenesis.

Glutamate dehydrogenase 1 (GLUD1) is implicated in oncogenesis. However, little is known about the relationship between GLUD1 and hepatocellular carcinoma (HCC). In the present study, we demonstrated that the expression levels of GLUD1 significantly decreased in tumors, which was relevant to the poor prognosis of HCC. Functionally, GLUD1 silencing enhanced the growth and migration of HCC cells. Mechanistically, the upregulation of interleukin-32 through AKT activation contributes to GLUD1 silencing-facilitated hepatocarcinogenesis. The interaction between GLUD1 and AKT, as well as α-ketoglutarate regulated by GLUD1, can suppress AKT activation. In addition, LIM and SH3 protein 1 (LASP1) interacts with GLUD1 and induces GLUD1 degradation via the ubiquitin-proteasome pathway, which relies on the E3 ubiquitin ligase synoviolin (SYVN1), whose interaction with GLUD1 is enhanced by LASP1. In hepatitis B virus (HBV)-related HCC, the HBV X protein (HBX) can suppress GLUD1 with the participation of LASP1 and SYVN1. Collectively, our data suggest that GLUD1 silencing is significantly associated with HCC development, and LASP1 and SYVN1 mediate the inhibition of GLUD1 in HCC, especially in HBV-related tumors.

Hepatitis B virus core protein stabilizes RANGAP1 to upregulate KDM2A and facilitate hepatocarcinogenesis.

PURPOSE: As a vital component of the hepatitis B virus (HBV) nucleocapsid, HBV core protein (HBC) contributes to hepatocarcinogenesis. Here, we aimed to assess the effects of RANGAP1 and KDM2A on tumorigenesis induced by HBC. METHODS: Co-immunoprecipitation (Co-IP) combined with mass spectrometry were utilized to identify the proteins with the capacity to interact with HBC. The gene and protein levels of RANGAP1 and KDM2A in hepatocellular carcinoma (HCC) and HBV-positive HCC tissues were evaluated using different cohorts. The roles of RANGAP1 and KDM2A in HCC cells mediated by HBC were investigated in vitro and in vivo. Co-IP and western blot were used to estimate the interaction of HBC with RANGAP1 and KDM2A and assess RANGAP1 stabilization regulated by HBC. RESULTS: We discovered that HBC could interact with RANGAP1 and KDM2A, the levels of which were markedly elevated in HCC tissues. Relying on RANGAP1 and KDM2A, HBC facilitated HCC cell growth and migration. The increased stabilization of RANGAP1 mediated by HBC was relevant to the disruption of the interaction between RANGAP1 and an E3 ligase SYVN1. RANGAP1 interacted with KDM2A, and it further promoted KDM2A stabilization by disturbing the interaction between KDM2A and SYVN1. HBC enhanced the interaction of KDM2A with RANGAP1 and upregulated the expression of KDM2A via RANGAP1 in HCC cells. CONCLUSIONS: These findings demonstrate a novel mechanism by which HBC facilitates hepatocarcinogenesis. RANGAP1 and KDM2A could act as potential molecular targets for treating HBV-associated malignancy.

Soluble egg antigen of Schistosoma japonicum induces pyroptosis in hepatic stellate cells by modulating ROS production.

BACKGROUND: Inflammation-induced dysfunction of hepatic stellate cells (HSCs) is involved in schistosomiasis-associated liver fibrosis, and soluble egg antigen (SEA) is a crucial pathogen-associated molecular pattern associated with liver injury in schistosomiasis. In addition, numerous studies have shown that caspase-1-mediated pyroptosis participates in the development of multiple inflammation-related diseases. However, whether pyroptotic cell death of HSCs is involved in SEA-mediated liver damage is not well understood. METHODS: Primary cultured HSCs and Schistosoma japonicum-infected mouse liver tissue were analysed for histological changes and caspase-1 activation, and the role of pyroptosis in the mechanisms underlying SEA-induced HSC death was investigated. Accumulation of reactive oxygen species (ROS) in infected livers and SEA-stimulated HSCs was measured by flow cytometry and immunofluorescence. RESULTS: Caspase-1 activity was elevated in both liver tissues and HSCs of S. japonicum-infected mice. Furthermore, SEA stimulation increased the proportion of pyroptotic HSCs, as shown by lactate dehydrogenase (LDH) release assays and by flow cytometric analysis of propidium iodide (PI) and caspase-1 double staining in cells. In addition, ROS generation was elevated in infected liver tissues and SEA-stimulated HSCs, and ROS inhibition downregulated SEA-induced caspase-1 activation and pyroptosis in HSCs. CONCLUSIONS: Our present study demonstrates that pyroptotic cell death in HSCs induced by SEA via ROS-mediated caspase-1 activation may serve as a significant mechanism to initiate the inflammatory response and thereby exacerbate liver injury during S. japonicum infection.

Ag/Pyridine Co-Mediated Oxidative Arylthiocyanation of Activated Alkenes.

An efficient Ag/pyridine co-mediated oxidative arylthiocyanation of activated alkenes via radical addition/cyclization cascade process was developed. This reaction could be carried out under mild conditions to provide biologically interesting 3-alkylthiocyanato-2-oxindoles in good to excellent yields. Mechanistic studies suggested a unique NCS• radical addition path and clarified the dual roles of catalytic pyridine as base and crucial ligand to accelerate the oxidation of Ag(I) to Ag(II), which is likely oxidant responsible for the formation of NCS• radical. These mechanistic results may impact the design and refinement of other radical based reactions proceeding through catalytic oxidations mediated by Ag(I)-pyridine/persulfate. The chemical versatility of thiocyanate moiety was also highlighted via SCN-tailoring chemistry in post-synthetic transformation for new S-C(sp³/sp²/sp), S-P, and S-S bonds constructions. The protocol provides an easy access to many important bioisosteres in medicinal chemistry and an array of sulfur-containing 2-oxindoles that are difficult to prepare by other approaches.

Chitinophaga salinisoli sp. nov., isolated from saline soil.

A novel aerobic bacterium, designated strain LAM9153T, was isolated from a saline soil sample collected from Lingxian County, Shandong Province, China. Cells of strain LAM9153T were observed to be Gram-stain negative, non-motile, non-spore-forming and rod-shaped. The new isolate grew optimally at 30-35 °C, pH 7.0 and 0.5% of NaCl concentration (w/v). According to the phylogenetic analysis based on the 16S rRNA gene sequence, strain LAM9153T shares high similarity with Chitinophaga terrae Gsoil 238T (96.9%) and Chitinophaga niabensis JS 13-10T (95.9%), forming a subcluster with C. terrae Gsoil 238T, Chitinophaga cymbidii R156-2T, C. niabensis JS 13-10T and Chitinophaga soli Gsoil 219T in the phylogenetic tree. The major cellular fatty acids (> 10%) were identified as iso-C15:0, iso-C17:0 3-OH and summed features 3 (C16:1 ω6c and/or C16:1 ω7c). The predominant respiratory quinone was identified as menaquinone MK-7. The polar lipids consisted of phosphatidylethanolamine, aminophospholipid, three unidentified aminolipids and five unidentified lipids. The genomic DNA G+C content was determined to be 53.2 ± 1.6 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic data, strain LAM9153T is concluded to represent a novel species of the genus Chitinophaga, for which the name Chitinophaga salinisoli sp. nov. is proposed. The type strain is LAM9153T (= ACCC 19960T = JCM 30847T).

Arenimonas alkanexedens sp. nov., isolated from a frozen soil sample.

A novel facultatively anaerobic bacterium, designated strain LAM-WHM-D11T, was isolated from a frozen soil sample of China. Cells of strain LAM-WHM-D11T were observed to be Gram-stain negative, non-motile and rod-shaped. Colonies were yellowish, and circular with convex shape. Strain LAM-WHM-D11T was found to be able to grow at 4-40 °C (optimum 15 °C), pH 7.5-2.0 (optimum 9.5) and 0-2.5% NaCl (w/v) (optimum 1.5%). The 16S rRNA gene sequence similarity analysis showed that strain LAM-WHM-D11T is closely related to Arenimonas metalli CF5-1T (98.0%), Arenimonas aquaticum NA-09T (97.9%), Arenimonas donghaensis HO3-R19T (95.6%) and Arenimonas aestuarii S2-21T (95.3%). The DNA-DNA hybridization values between the isolate and A. metalli CGMCC 1.10787T, A. aquaticum KACC 14663T, A. donghaensis KACC 11381T were 41.0 ± 1.7, 44.7 ± 1.4 and 42.8 ± 1.2%, respectively. The genomic DNA G+C content was found to be 66.5 mol% as determined by the T m method. The major cellular fatty acids were identified as iso-C15:0 and iso-C16:0. The major isoprenoid quinone was identified as ubiquinone 8 (Q-8). The major polar lipids were found to be diphosphatidyglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, two phospholipids and five unidentified lipids. Based on the phylogenetic, phenotypic and chemotaxonomic characteristics, strain LAM-WHM-D11T is concluded to represent a novel species within the genus Arenimonas, for which the name Arenimonas alkanexedens sp. nov. is proposed. The type strain is LAM-WHM-D11T (ACCC 19750T = JCM 30464T).

MHC II-, but not MHC II+, hepatic Stellate cells contribute to liver fibrosis of mice in infection with Schistosoma japonicum.

Hepatic stellate cells (HSCs) are considered as the main effector cells in vitamin A metabolism and liver fibrosis, as well as in hepatic immune regulation. Recently, researches have revealed that HSCs have plasticity and heterogeneity, which depend on their lobular location and whether liver is normal or injured. This research aimed to explore the biological characteristics and heterogeneity of HSCs in mice with Schistosoma japonicum (S. japonicum) infection, and determine the subpopulation of HSCs in pathogenesis of hepatic fibrosis caused by S. japonicum infection. Results revealed that HSCs significantly increased the expressions of MHC II and fibrogenic genes after S. japonicum infection, and could be classified into MHC II+ HSCs and MHC II- HSCs subsets. Both two HSCs populations suppressed the proliferation of activated CD4+T cells, whereas only MHC II- HSCs displayed a myofibroblast-like phenotype. In response to IFN-γ, HSCs up-regulated the expressions of MHC II and CIITA, while down-regulated the expression of fibrogenic gene Col1. In addition, praziquantel treatment decreased the expressions of fibrogenic genes in MHC II- HSCs. These results confirmed that HSCs from S. japonicum-infected mice have heterogeneity. The MHC II- α-SMA+ HSCs were major subsets of HSCs contributing to liver fibrosis and could be considered as a potential target of praziquantel anti-fibrosis treatment.

Cobalt-Catalyzed Peroxidation of 2-Oxindoles with Hydroperoxides.

A highly efficient and facile cobalt-catalyzed C-H activation and peroxidation of 2-oxindoles was reported, which provides a new pathway for the synthesis of biologically active 3-peroxy-2-oxindoles from readily available starting materials in excellent chemical yields. The resulting products could be further transformed into various substituted 3-peroxyoxindoles in good to excellent yields. The developed method has been successfully applied to the synthesis of the natural product (±)-N-[2-(3-hydroxy-2-oxo-2,3-dihydro-1H-indol-3-yl)ethyl]acetamide.

Paenibacillus salinicaeni sp. nov., isolated from saline silt sample.

A novel facultatively anaerobic bacterium, designated strain LAM0A28(T), was isolated from a saline silt sample collected from the Chinese Sea of Death located in Suining city, Sichuan province, China. Cells of strain LAM0A28(T) were observed to be Gram-stain positive, motile, endospore-forming and straight-rod shaped. Strain LAM0A28(T) was found to be able to grow at 15-45 °C (optimum: 30-35 °C), pH 5.0-10.0 (optimum: 7.5) and 0-5 % NaCl (w/v) (optimum: 0.5 %). The 16S rRNA gene sequence similarity analysis showed that strain LAM0A28(T) is closely related to Paenibacillus jilunlii DSM 23019(T) (97.5 %) and Paenibacillus graminis DSM 15220(T) (97.2 %). The DNA-DNA hybridization values between the isolate and P. jilunlii DSM 23019(T), P. graminis DSM 15220(T) were 30.2 ± 1.6 % and 44.7 ± 2.1 %, respectively. The DNA G+C content was found to be 51.2 mol% as determined by the T m method. The major cellular fatty acids were identified as anteiso-C15:0, C16:0, iso-C16:0 and C14:0. The major isoprenoid quinone was identified as MK-7. The cell wall peptidoglycan was found to contain meso-diaminopimelic acid. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two aminophospholipids and six unidentified lipids. Based on the phylogenetic, phenotypic and chemotaxonomic characteristics, strain LAM0A28(T) is concluded to represent a novel species within the genus Paenibacillus, for which the name Paenibacillus salinicaeni sp. nov. is proposed. The type strain is LAM0A28(T) (=ACCC 00741(T) = JCM 30850(T)).

FeCl3-Mediated Radical Tandem Reactions of 3-Benzyl-2-oxindoles with Styrene Derivatives for the Stereoselective Synthesis of Spirocyclohexene Oxindoles.

A novel FeCl3-mediated reaction of 3-benzyl-2-oxindoles with styrene derivatives was developed. The reaction provided spirocyclohexene oxindoles in good yields and excellent diastereoselectivities via a tandem radical addition/cyclization process.

A metal-free yne-addition/1,4-aryl migration/decarboxylation cascade reaction of alkynoates with Csp(3)-H centers.

A metal-free cascade reaction of aryl alkynoates with five different types of radical precursors (R-H) through an yne-addition/1,4-aryl migration/decarboxylation process was reported, which allowed facile and convenient access to functionalized vinyl products with "R" and protons located at the identical carbon of the formed double bond.

Lentibacillus amyloliquefaciens sp. nov., a halophilic bacterium isolated from saline sediment sample.

A Gram-stain positive, non-motile, non-sporogenous, aerobic, rod-shaped and halophilic bacterium, designated LAM0015(T), was isolated from a saline sediment sample collected from Yantai City in China. The isolate was found to be able to grow at NaCl concentrations of 5-25 % (w/v) (optimum: 7-12 %), 15-45 °C (optimum: 35 °C) and pH 5.0-9.0 (optimum: 7.0). The major fatty acids were determined to be anteiso-C15:0 and anteiso-C17:0. The predominant respiratory quinone was identified as MK-7. The cell wall peptidoglycan was determined to contain meso-diaminopimelic acid. The polar lipids were found to be diphosphatidyglycerol, phosphatidylglycerol, five phospholipids and one glycolipid. The DNA G+C content was 43.1 mol% as determined by the T m method. Analysis of the 16S rRNA gene sequence indicated that the isolate belongs within the genus Lentibacillus and is closely related to Lentibacillus persicus DSM 22530(T), Lentibacillus salicampi JCM 11462(T) and Lentibacillus jeotgali JCM 15795(T) with 97.3, 96.7 and 96.4 % sequence similarity, respectively. The DNA-DNA hybridization value between LAM0015(T) and L. persicus DSM 22530(T) was 51.2 ± 1.4 %. Based on its phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0015(T) is concluded to represent a novel species of the genus Lentibacillus, for which the name Lentibacillus amyloliquefaciens sp. nov. is proposed. The type strain is LAM0015(T) (=ACCC 06401(T) = JCM 19838(T)).

Paenibacillus populi sp. nov., a novel bacterium isolated from the rhizosphere of Populus alba.

A novel aerobic bacterium, designated strain LAM0705(T), was isolated from the rhizosphere of Populus alba in the Peking University Third Hospital. Cells of strain LAM0705(T) were observed to be Gram-stain positive, motile, spore-forming and rod-shaped. The optimal temperature and pH for growth were found to be 30 °C and pH 7.5, respectively. Strain LAM0705(T) was found to be able to grow in the presence 0-5 % NaCl (w/v) (optimum 1.0 %). The major fatty acids of strain LAM0705(T) were identified as anteiso-C15:0, C16:0 and iso-C16:0. The dominant polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The cell wall peptidoglycan of strain LAM0705(T) was found to contain meso-diaminopimelic acid. The predominant menaquinone was identified as MK-7. The G+C content of genomic DNA was found to be 48 mol% when determined by the T m method. The 16S rRNA gene sequence similarity analysis indicated that strain LAM0705(T) is closely related to Paenibacillus agaridevorans DSM 1355(T) and Paenibacillus thailandensis KCTC 13043(T) with 97.8 and 96.1 % sequence similarity, respectively. The DNA-DNA hybridization value between strain LAM0705(T) and P. agaridevorans DSM 1355(T) was 47 ± 0.8 %. On the basis of its phenotypic, phylogenetic and chemotaxonomic characteristics, strain LAM0705(T) is concluded to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus populi sp. nov. is proposed. The type strain is LAM0705(T) (=ACCC 06427(T) = JCM 19843(T)).

Paenibacillus vini sp. nov., isolated from alcohol fermentation pit mud in Sichuan Province, China.

A novel facultatively anaerobic bacterial strain, designated LAM0504(T), was isolated from a pit mud of Luzhou flavour liquor alcohol fermentation in Sichuan Province, China. Cells of strain LAM0504(T) were observed to be Gram-stain negative, spore-forming, rod shaped and motile by means of peritrichous flagella. Strain LAM0504(T) was found to be able to grow at 20-48 °C (optimum: 30 °C), pH 5.0-9.0 (optimum: 7.0) and 0-3 % NaCl (w/v) (optimum: 1.0 %). The 16S rRNA gene sequence similarity analysis showed that strain LAM0504(T) was most closely related to Paenibacillus konsisdensis JCM 14798(T), Fontibacillus phaseoli LMG 27589(T) and Paenibacillus motobuensis JCM 12774(T), with 97.0, 96.8 and 96.7 % sequence similarity, respectively. The DNA-DNA hybridization value between strain LAM0504(T) and P. konsisdensis JCM 14798(T) was 53.3 ± 1.2 %. The genomic DNA G+C content of strain LAM0504(T) was 43.0 mol% as determined by the Tm method. The major fatty acids of strain LAM0504(T) were identified as anteiso-C15:0, C16:0 and iso-C15:0. The cell-wall peptidoglycan was found to contain meso-diaminopimelic acid. The predominant menaquinone was identified as MK-7. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids, two unidentified glycolipids and three unidentified lipids. On the basis of its physiological and phylogenetic characteristics, strain LAM0504(T) is concluded to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus vini sp. nov. is proposed. The type strain is LAM0504(T) (=ACCC 06420(T) = JCM 19842(T)).

[Isolation and identification of Toxoplasma gondii strains from cats in Xuzhou region].

OBJECTIVE: To isolate Toxoplasma gondii (T. gondii) strains from stray cats and explore their prevalence in Xuzhou City. METHODS: The sera of 41 stray cats were collected to detect the antibodies of T. gondii by using a commercial enzyme-linked immunosorbent (ELISA) kit. The tissues including the heart, brain and tongue from these cats were digested by acid pepsin solution and inoculated to Kunming mice. These suspicious isolates were subsequently identified by a specific PCR method. RESULTS: A total of 11 strains were isolated from 41 stray cats, which were confirmed by the PCR results. Moreover, 17 cats (41.5%) were found to be positive with the antibodies of T. gondii. CONCLUSION: A high prevalence of T. gondii infection was found in Xuzhou City, which indicates that the stray cats infected with T. gondii would be an important infection source that may infect humans and other animals in this area.