Small molecule deoxynyboquinone triggers alkylation and ubiquitination of Keap1 at Cys489 on Kelch domain for Nrf2 activation and inflammatory therapy.

Activation of nuclear factor erythroid 2-related factor 2 (Nrf2) by Kelch-like ECH-associated protein 1 (Keap1) alkylation plays a central role in anti-inflammatory therapy. However, activators of Nrf2 through alkylation of Keap1-Kelch domain have not been identified. Deoxynyboquinone (DNQ) is a natural small molecule discovered from marine actinomycetes. The current study was designed to investigate the anti-inflammatory effects and molecular mechanisms of DNQ via alkylation of Keap1. DNQ exhibited significant anti-inflammatory properties both in vitro and in vivo. The pharmacophore responsible for the anti-inflammatory properties of DNQ was determined to be the α, ß-unsaturated amides moieties by a chemical reaction between DNQ and N-acetylcysteine. DNQ exerted anti-inflammatory effects through activation of Nrf2/ARE pathway. Keap1 was demonstrated to be the direct target of DNQ and bound with DNQ through conjugate addition reaction involving alkylation. The specific alkylation site of DNQ on Keap1 for Nrf2 activation was elucidated with a synthesized probe in conjunction with liquid chromatography-tandem mass spectrometry. DNQ triggered the ubiquitination and subsequent degradation of Keap1 by alkylation of the cysteine residue 489 (Cys489) on Keap1-Kelch domain, ultimately enabling the activation of Nrf2. Our findings revealed that DNQ exhibited potent anti-inflammatory capacity through α, ß-unsaturated amides moieties active group which specifically activated Nrf2 signal pathway via alkylation/ubiquitination of Keap1-Kelch domain, suggesting the potential values of targeting Cys489 on Keap1-Kelch domain by DNQ-like small molecules in inflammatory therapies.

Clinical and genomic features in patients with second primary glioblastoma following first primary renal cell carcinoma.

PURPOSE: To explore the potential pathogenesis and clinical features of second primary glioblastoma (spGBM) following first primary renal cell carcinoma (fpRCC). METHODS: Patients with spGBM after fpRCC were enrolled from our institution and the SEER dataset. Sanger sequencing, whole genome sequencing, and immunehistochemistry were used to detect molecular biomarkers. RESULTS: Four and 122 cases from our institution and the SEER dataset, respectively, were collected with an overall median age of 69 years at spGBM diagnosis following fpRCC. The median interval time between fpRCC and spGBM was 50.7 months and 4 years, for the four and 122 cases respectively. The median overall survival time was 11.2 and 6.0 months for the two datasets. In addition, spGBM patients of younger age (< 75 years) or shorter interval time (< 1 year) had favorable prognosis (p = 0.081 and 0.05, respectively). Moreover, the spGBM cases were molecularly classified as TERT only paired with TP53 mutation, PIK3CA mutation, EGFR alteration, low tumor mutation burden, and stable microsatellite status. CONCLUSIONS: This is the first study to investigate the pathogenesis and clinical features of spGBM following spRCC. We found that spGBMs are old-age related, highly malignant, and have short survival time. Moreover, they might be misdiagnosed and treated as brain metastases from RCC. Thus, the incidence of spGBMs after fpRCC is underestimated. Further studies are needed to investigate the underlying molecular mechanisms and clinical biomarkers for the development of spGBM following fpRCC.

Preoperative blood testing for glioblastoma, brain metastases, and primary central nervous system lymphoma differentiation.

Background: Differentiating glioblastoma (GBM), brain metastases, and primary central nervous system lymphoma (PCNSL) in clinical practice is difficult. This study aimed to evaluate the diagnostic value of routine blood biomarkers in patients with GBM, brain metastases, and PCNSL and find a preoperative differential diagnostic tool for these tumors. Methods: The perioperative medical records of 70 GBM, 41 PCNSL, and 81 brain metastases patients and their preoperative blood test results were compared and analyzed, and a diagnostic model to differentiate among them established. Results: Patient age, plateletcrit, international normalized ratio (INR), and thrombin time (TT) were independently associated with differential diagnosis by multinomial logistic regression. Compared with GBM patients, brain metastases patients were significantly older (OR =1.055, 95% CI: 1.016-1.094, P=0.005) and had lower plateletcrit levels (OR =0.008, 95% CI: 0.004-0.017, P=0.027). In addition, patients with GBM had lower INR and higher TT than patients with the other two tumor types. A diagnostic model including these parameters, had an accuracy of 88.2% and 76.1% for brain metastases and GBM, respectively. Conclusions: Preoperative plateletcrit, INR, and TT may be used as inexpensive blood diagnostic biomarkers for differentiating brain metastases from other intracranial malignant tumors.

Highly expressed of SERPINA3 indicated poor prognosis and involved in immune suppression in glioma.

INTRODUCTION: The prognosis of patients with glioma is dismal. It has been reported that Serpin peptidase inhibitor clade A member 3 (SERPINA3) is associated with the mobility and invasion of tumor cells. Our study was designed to explore the value of SERPINA3 messenger RNA (mRNA) expression in the biological process, prognosis, and immune significance in glioma. METHODS: We analyzed the biological functions of SERPINA3 through data from the Chinese Glioma Genome Atlas databases. Differentially expressed genes and enrichment analysis were performed and correlations between SERPINA3 expression and immune cell infiltration were analyzed. Further, we validated the expression and the survival prediction role of SERPINA3 by using tissue microarrays and RNAscope in situ hybridization in 321 gliomas. The correlations between the expression and clinical-pathological parameters as well as other biomarkers were examined. RESULTS: Univariate and multivariate regression both indicated that the level of SERPINA3 transcript represented an independent prognostic factor. High levels of SERPINA3 correlated with poor survival in patients with glioma. Expression of SERPINA3 mRNA was observed positively correlated with MCM6, IGFBP2, and FKBP10. Enrichment analysis showed SERPINA3 mainly enriched in immune-related terms and signaling pathways including MAPK, TNF, P53, PI3K-Akt, nuclear factor-κB. Immune infiltration analysis further declare the SERPINA3 expression negatively correlated with levels of Macrophages M1, native CD4+ T cell, monocytes, and Mast cell activated. And overexpression of SERPINA3 correlated with low CD4+ T cell infiltration in glioma tissues. CONCLUSIONS: SERPINA3 may play a key role in the biological process of glioma cells especially in immune suppression activities. SERPINA3 may serve as an independent survival prediction factor in glioma patients.

Seawater silicate fertilizer facilitated nitrogen removal via diatom proliferation.

Dissolved inorganic nitrogen (DIN) enrichment accompanied by silicate deficiency in seawater can promote dinoflagellate growth over diatom growth and induce further negative ecological consequences. Here, we propose an easily exercisable method for silicate fertilization as a remedy of eutrophication. In the laboratory, rice husk ash (RHA) released silicate and phosphate in an atomic ratio range of 38-113 without a significant influence on DIN. During incubations of silicate-limited waters, low-dose fertilization increased the diatom/dinoflagellate ratio by 1-5 times. With the high-dose fertilizer addition, DIN, with an initial concentration of 7.63 ±â€¯0.95 µmol l-1, was exhausted in three days, and the diatom abundance increased by 19 times on the 5th day. The silicate fertilization method presented here can be applied independently in eutrophicated waters for dinoflagellate suppression and dissolved nitrogen removal; this method could also work as a supplementary measure to existing nutrient (N, P) reduction and biomanipulation efforts.

Forkhead box P3 gene silencing inhibits the expression of chemokines and chemokine receptors associated with cell growth, migration, and apoptosis in hepatocellular carcinoma cells.

The aberrant expression of forkhead box P3 (FOXP3) leads to the formation of malignant tumors. FOXP3 expression levels are also elevated in hepatocellular carcinoma (HCC). The aim of the present study was to investigate the effects of FOXP3 silencing on cell proliferation, migration, apoptosis and chemokine/chemokine receptor expression in the MHCC-97H HCC cell line. Three FOXP3 short hairpin (sh)RNA constructs were designed: Sh-FOXP3-1-pGreenPuro, sh-FOXP3-2-pGreenPuro, and sh-FOXP3-3-pGreenPuro. MHCC-97H cells were transfected with shRNA-FOXP3, and the mRNA and protein expression levels of C-X-C motif chemokine (CXC) ligand 12 (CXCL12), CXCL11, CXC receptor 4 (CXCR4) and CXCR7 were measured. Cell Counting Kit-8, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labeling and Transwell assays were used to evaluate cell proliferation, apoptosis and migration, respectively. Of the three FOXP3 lentivirus carriers constructed, sh-FOXP3-1 significantly reduced FOXP3 expression levels and was chosen for further experiments. sh-FOXP3-1 inhibited cell proliferation, promoted apoptosis and inhibited cell migration compared with the negative control. The mRNA and protein expression levels of CXCL12, CXCL11, CXCR4 and CXCR7 were decreased significantly in response to FOXP3 silencing. FOXP3 silencing may therefore inhibit cell growth, induce apoptosis and inhibit migration in HCC cells, possibly by impairing the chemokine/chemokine receptor axes.

Desumoylating Isopeptidase 2 (DESI2) Inhibits Proliferation and Promotes Apoptosis of Pancreatic Cancer Cells through Regulating PI3K/AKT/mTOR Signaling Pathway.

This study aimed to investigate the effects of desumoylating isopeptidase 2 (DESI2) on tumor cell proliferation, apoptosis and invasion of pancreatic cancer, and to assess the signaling pathway involved. Overexpression and silence of DESI2 were designed and the experiments were divided into 5 groups: a normal control group, an interference control group (shRNA-NC); an interference group (sh-DESI2); an overexpression control group (NC), an overexpression group (DESI2). Quantitative real time polymerase chain reaction (qRT-PCR) was used to screen the appropriate interference sequence. The silencing and overexpression of DESI2 were confirmed by qRT-PCR and western blotting. Cell cycling, apoptosis, invasion, and the expression of phosphatidylinositol-3-kinase (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) pathway and caspase 3 were measured. Overexpression and silence of DESI2 were successfully designed in two pancreatic cancer cells, and the interference effect of sh-DESI2-3 showed the best silencing effects. The biological activities of DESI2 were detected in both ASPC-1 and PANCE-1 cells. Our results showed that cell proliferation was significantly increased in the sh-DESI2 group, while decreased in DESI2 group compared with the control group in both cell lines. In ASPC-1 cells, the events in G1 phase decreased and in S phase increased obviously in the sh-DESI2 group, compared with control group. An opposite result was found when DESI2 was overexpressed. In PANCE-1 cells, the events in G2 phase were higher in the sh-DESI2 group, while in the DESI2 group was significantly lower than that in control group. In ASPC-1 and PANCE-1 cells, sh-DESI2 group showed decreased apoptosis, increased cell invasion and increased expression of AKT, p-Akt, PI3K, p-PI3K, p-mTOR and mTOR and decreased caspase 3 expression compared with the control group, while overexpression of DESI2 leaded to increased apoptosis, decreased cell invasion and reduced expression of AKT, p-Akt, PI3K, p-PI3K, p-mTOR and mTOR and increased expression of caspase 3. DESI2 regulates the proliferation and apoptosis of pancreatic cancer cells through PI3K/AKT/mTOR signaling pathway.

[Shovel-shaped electrode transurethral plasmakinetic enucleation versus plasmakinetic resection of the prostate in the treatment of benign prostatic hyperplasia].

OBJECTIVE: To compare the safety and effectiveness of shovel-shaped electrode transurethral plasmakinetic enucleation of the prostate (PKEP) with those of plasmakinetic resection of the prostate (PKRP) in the treatment of benign prostatic hyperplasia (BPH). METHODS: We retrospectively analyzed the clinical data about 78 BPH patients received in Shanghai Ninth People's Hospital from June 2016 to January 2017, 39 treated by shovel-shaped electrode PKEP and the other 39 by PKRP. We observed the patients for 6 months postoperatively and compared the effects and safety of the two surgical strategies. RESULTS: No statistically significant difference was observed between the PKEP and PKRP groups in the operation time (ï¼»69.3 ± 8.8ï¼½ vs ï¼»72.2 ± 7.9ï¼½ min, P = 0.126), but the former, as compared with the latter, showed a markedly less postoperative loss of hemoglobin (ï¼»3.9 ± 2.8ï¼½ vs ï¼»13.9 ± 5.2ï¼½ g/L, P <0.001) and shorter bladder irrigation time (ï¼»12.5 ± 1.2ï¼½ vs ï¼»43.4 ± 2.8ï¼½ h, P <0.001), catheterization time (ï¼»64.0 ± 4.5ï¼½ vs ï¼»84.8 ± 3.0ï¼½ h, P <0.001) and hospital stay (ï¼»3.1 ± 0.3ï¼½ vs ï¼»5.5 ± 0.4ï¼½ d, P <0.001). There were no statistically significant differences between the PKEP and PKRP groups in the postoperative maximum urinary flow rate (Qmax) (ï¼»21.62 ± 1.07ï¼½ vs ï¼»21.03 ± 0.96ï¼½ ml/s, P = 0.12), International Prostate Symptoms Score (IPSS) (5.85 ± 0.90 vs 6.03 ± 0.81, P = 0.279), quality of life score (QoL) (2.0 ± 0.73 vs 2.28 ± 0.72, P = 0.09), postvoid residual urine volume (PVR) (ï¼»19.59 ± 6.01ï¼½ vs ï¼»20.21 ± 5.16ï¼½ ml, P = 0.629), or the incidence rates of urinary incontinence (2.56% ï¼»1/39ï¼½ vs 7.69% ï¼»3/39ï¼½, P >0.05) and other postoperative complications. CONCLUSIONS: Both PKEP and PKRP are effective methods for the treatment of BPH, but PKEP is worthier of clinical recommendation for a better safety profile, more thorough removal of the prostate tissue, less blood loss, shorter hospital stay, and better improved quality of life of the patient.

Selective suppression of in situ proliferation of scyphozoan polyps by biofouling.

An increase in marine artificial constructions has been proposed as a major cause of jellyfish blooms, because these constructions provide additional substrates for organisms at the benthic stage (polyps), which proliferate asexually and release a large amount of free-swimming medusae. These hard surfaces are normally covered by fouling communities, the components of which have the potential to impede the proliferation of polyps. In this study, we report an in situ experiment of polyp survival of four large scyphozoan species found in East Asian marginal seas that were exposed to biofouling, a universal phenomenon occurring on marine artificial constructions. Our results showed that the polyps of three species (Nemopilema nomurai, Cyanea nozaki, and Rhopilema esculentum) attached to the artificial surfaces were completely eliminated by biofouling within 7-8months, and only those of moon jellyfish (Aurelia sp.1) in the upper layers could multiply on both artificial materials and other organisms (e.g., ascidians and bryozoans). Fouling-associated competition and predation and suppressed asexual reproduction of podocysts were observed to contribute to the loss of polyps. This study shows that the natural distribution of polyps is defined by the biofouling community that colonizes the surfaces of artificial constructions. Consequently, the contribution of marine constructions to jellyfish bloom is limited only to the ability of the jellyfish species to reproduce asexually through budding and inhabit solid surfaces of fouling organisms in addition to inhabiting original artificial materials. We anticipate that fragile polyps will colonize and proliferate in harsh environments that are deleterious to biofouling, and we propose special attention to polyps in antifouling practices for excluding the possibility that they occupy the available ecological space.

Marininema mesophilum gen. nov., sp. nov., a thermoactinomycete isolated from deep sea sediment, and emended description of the family Thermoactinomycetaceae.

A novel filamentous bacterium, strain SCSIO 10219(T), was isolated from a sediment sample collected from the South China Sea (113° 3.752' E 18° 1.722' N) at a depth of 2105 m. Growth was observed at 25-35 °C (optimum 30 °C) and pH 5.0-8.0 (optimum pH 6.0-7.0). The organism formed yellow-white colonies with radial wrinkles. Aerial mycelium was not produced on any of the growth media tested. Phenotypic characterization and 16S rRNA gene sequence analysis indicated that strain SCSIO 10219(T) belongs to the family Thermoactinomycetaceae. The strain contained ll-diaminopimelic acid in the cell wall. The predominant menaquinone was MK-7. The phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine and five unknown phospholipids. Major fatty acids were anteiso-C(15:0) and iso-C(15:0). The DNA G+C content was 46.5 mol%. On the basis of chemotaxonomic properties and phylogenetic analysis based on 16S rRNA gene sequence data, it is proposed that this strain represents a novel species in a new genus, Marininema mesophilum gen. nov., sp. nov., in the family Thermoactinomycetaceae. The type strain of the type species is SCSIO 10219(T) ( = CCTCC AA 2011006(T) = DSM 45610(T)). In addition, we propose that the description of the family Thermoactinomycetaceae should be further emended based on the present study.

Identification and characterization of a novel gene, TrCCD1, and its possible function in hyphal growth and conidiospore development of Trichoderma reesei.

To investigate genes with essential functions during hyphal growth or sporulation in the asexual filamentous fungus Trichoderma reesei, we screened a collection of T-DNA insertion mutants and identified the genomic integration events. Two mutants with abnormal phenotypes, named as ccdO and ccdP, were found to have independent T-DNA insertions into a putative TrCCD1 gene locus, the product of which has significant homology to carotenoid cleavage dioxygenases (CCDs). Compared to the parental strain, both mutants tended to produce slow-growing hyphae and had a more than 50% reduction in colony growth rate. Simultaneously, the hyphae of the growing mutants formed wilting tip while the parental strain elongated straightly. To the effect of the TrCCD1 mutation on the conidiospore development, less spores were formed in the mutants than in the parental strain. In addition, disruption of TrCCD1 resulted in another phenotype characterized by a remarkable enhancement in the total carotenoid content. When the wild-type TrCCD1 gene was reintroduced into the ccd mutants, the abnormal phenotypes were rescued. These results suggest that TrCCD1 is involved in carotenoid metabolism and likely required for hyphal growth and conidiospore development in filamentous fungi T. reesei.