Identification and characterization of Laodelphax striatellus (Insecta: Hemiptera: Delphacidae) neutral sphingomyelinase.

The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus, a direct sap-sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase-enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively. LsSMase was an Mg2+ or Mn2+ dependent enzyme, but different concentration of each were needed. The activity of LsSMase was significantly stimulated by Ethylene glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA), whereas it was inhibited by ethylenediaminetetraacetic acid. Millimolar concentrations of Zn2+ completely inhibited LsSMase. The reducing agents dithiothreitol and ß-mercaptoethanol varied in their effects on activity. Phospholipids were not found to stimulate LsSMase.

Nutrient assessment of olive leaf residues processed by solid-state fermentation as an innovative feedstuff additive.

AIMS: Olive leaf residue feedstuff additives were prepared by solid-state fermentation (SSF), and its feeding effects on broiler chickens were examined. METHODS AND RESULTS: The fermentation's nutrient value, that is, protein enrichment, cellulase activity, tannic acid degradation and amino acid enhancement, was determined. The effect of different strains, including molds (Aspergillus niger, Aspergillus oryzae and Trichoderma viride) and yeasts (Candida utilis, Candida tropicalis and Geotrichum candidum), and the fermentation time on the nutrient values of the feedstuff additives was investigated. The experimental results showed that the optimal parameters for best performance were A. niger and C. utilis in a 1 : 1 ratio (v/v) in co-culture fermentation for 5 days. Under these conditions, the total content of amino acids in the fermented olive leaf residues increased by 22·0% in comparison with that in the raw leaf residues. Both Glutamic acid and Aspartic acid contents were increased by more than 25·4%. Broiler chickens fed with different amounts of feedstuff additives were assessed. The results demonstrated that the chicken weight gains increased by 120%, and normal serum biochemical parameters were improved significantly after 10% of the feedstuff additives were supplemented to the daily chicken feed for 28 days. CONCLUSIONS: The co-culture combination of A. niger and C. utilis with SSF for olive leaf residue had the best nutrient values. The addition of 10% fermented olive leaf residue facilitated the chicken growth and development. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reveals that olive leaf residues fermented by SSF exhibited considerable potential as feed additives for feeding poultry.

Content and distribution of different phosphorus forms in broiler chickens.

To understand the background value of phosphorus in chickens, the quantitative distribution of different phosphorus forms, including total phosphorus (TP), free phosphate (FP) and phospholipid (PL), in viscera, blood and bones of broiler chickens was investigated. Results showed that phosphorus contents exhibited significant differences in different parts of chickens. TP content of breast and thigh meat was over 5.0 g/kg, in which most of the phosphorus was in the form of water-soluble phosphates. TP content in viscera was higher than that in meat, and spleen was observed to contain the highest amount of phosphorus (10.0 g/kg). In all tested organs, FP and PL contents in liver were the highest, ranging between 1207-1989 and 81-369 mg/kg respectively. TP content in chicken bone was in the range of 52,716-136,643 mg/kg, and FP content in the bone was relatively lower than that in chicken meat.

Effect of lentivirus-mediated RNA interference of APC-Cdh1 expression on spinal cord injury in rats.

This study investigated cadherin-1 (Cdh1) expression in the sensorimotor cortex of rats after spinal cord injury (SCI). The repairing effect of Cdh1 was evaluated by silencing its expression with lentivirus-mediated RNAi. Twenty male Sprague-Dawley (SD) rats were randomly divided into a normal group and an operation group. Rats of the operation group were given SCI by the Allen method (T10-T11). Cdh1 expression in the sensorimotor cortex was examined by quantitative real-time polymerase chain reaction (PCR) and Western blot analysis. Thirty male SD rats were divided into a sham-operation (SO) group, a lentivirus vector (LV) group, and a recombinant lentivirus (RL) group. Rat behavior was evaluated using the Basso-Beattie-Bresnahan (BBB) test every week. Ten days after injection, Cdh1 expression was examined by quantitative real-time PCR and Western blot. Six weeks after injury, animals were injected with biotinylated dextran amine-Texas Red (BDA-TR), and then at 8 weeks, spinal cords were removed and sectioned in serial order. The expression of Cdh1 mRNA was significantly higher in the operation than in the normal group (P < 0.05). The expression of Cdh1 mRNA was lower in the RL than in the SO or LV groups at 10 days after injection (P < 0.05). In addition, the BBB score was higher for the RL than for the SO or LV groups at 6 weeks after injury (P < 0.05). A novel population of BDA-labeled axons was observed extending past the lesion in the RL group, which was rarely observed in the SO and LV groups. These results suggest that the anaphase-promoting complex-Cdh1 may play an important role in inhibiting axonal growth.

Identification and biochemical characterization of Laodelphax striatellus neutral ceramidase.

Ceramidases are a group of enzymes that catalyse hydrolysis of ceramides to generate fatty acid and sphingosine. In this study, we report the cloning and characterization of the rice small brown planthopper Laodelphax striatellus neutral ceramidase (nCDase), LsnCer. LsnCer was identified by sequencing the transcriptome of L. striatellus and is a protein of 717 amino acids with a predicted molecular weight of 79.3 kDa. Similarly to other known nCDases, the optimum pH for LsnCer is 8.0 and the optimum temperature is 37 °C for its in vitro activity. LsnCer activity is inhibited by Zn(2+) significantly and Fe(2+) slightly. LsnCer has broad substrate specificity with a preference for ceramides with a medium acyl-chain or a monounsaturated long acyl-chain. Infection with rice strip virus (RSV) or treatment with insecticides significantly increased LsnCer mRNA expression and its enzymatic activity in L. striatellus. These results suggest that LsnCer is a bona fide nCDase that may have a role in adaption of L. striatellus to environmental stresses.

Core-shell structure disclosed in self-assembled Cu-Ag nanoalloy particles.

Core-shell segregation of copper and silver in self-assembled, free nanoparticles is established by means of photoelectron spectroscopy in a wide range of relative Cu-Ag concentrations. These conclusions are based on the analysis of the photon-energy-dependent changes of the Cu 3d and Ag 4d photoelectron spectra. The nanoparticles are formed from mixed Cu-Ag atomic vapor created by magnetron sputtering of a bimetallic sample in a gas-aggregation cluster source. Even at similar Cu and Ag fractions in the primary vapor the surface of the nanoparticles is dominated by silver. Only at low Ag concentration copper appears on the surface of nanoparticles. For the latter case, a threefold decrease in the Ag 4d spin-orbit splitting has been detected. The specific component distribution and electronic structure changes are discussed in connection with the earlier results on Cu-Ag macroscopic and surface alloys.

Survey of sweetpotato viruses in China.

Ten sweetpotato viruses were surveyed in 3 major sweetpotato planting region (covering 11 provinces) in China from 2006 to 2010 to understand the distribution of sweetpotato viral diseases. Nine out of the 10 viruses were found in every major planting region. The most frequently detected virus in the Northern and the Yangtze River region was SPMSV. In the Southern region, SPVG was the most frequently detected virus. Compared to the results of the survey done in 1989, the incidences of all the viral diseases increased.

Comprehensive analysis of the immune and prognostic implication of MASP1 in stomach adenocarcinoma.

OBJECTIVE: Stomach adenocarcinoma (STAD) is the major cancer worldwide with high morbidity and mortality rate. Late diagnosis and limited treatment options of STAD lead to disease progression, spread, and metastasis. Therefore, finding a new biomarker to diagnosis and treatment is very important for STAD in clinical practice. MATERIALS AND METHODS: The clinical data, transcriptome data and CCLE data were downloaded from TCGA database and CCLE database, respectively. TIMER website, TISIDB website and CIBERSORT methodology were used to analyse immune infiltration. R software and R package were used to analyse gene difference expression, determine co-expression genes, conduct gene enrichment analyses, construct a prognostic signature and establish nomogram. RESULTS: MASP1 was decreased in STAD compared with normal tissue at the mRNA level (p < 0.001). The enrichment analysis showed that mismatch repair (MMR) was related to the MASP1 gene. Up-regulation of MAPS1 expression was positively associated with dendritic cells (p < 0.01), neutrophils (p < 0.05), macrophages (p < 0.001), CD4+ T cells (p < 0.001) and B cells (p < 0.05). A four-gene prognostic signature was determined based on MASP1-related immunomodulators. The prognostic signature was an independent prognostic predictor in STAD. Finally, we established a nomogram to forecast survival and the nomogram has a good prediction accuracy. CONCLUSIONS: In STAD, MASP1 is closely related to immunity. MASP1 has the potential to positively regulate the abundance of immune cells. The MASP1-related prognosis signature and nomogram can accurately predict the survival of patients with STAD. Therefore, MASP1 is likely to be a diagnosis and promising immunotherapy target spot in STAD clinical practice.

Combined inhibitory effects of celecoxib and fluvastatin on the growth of human hepatocellular carcinoma xenografts in nude mice.

This study was designed to investigate the in vivo growth inhibitory effects of celecoxib, a cyclo-oxygenase-2 inhibitor, and fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on the hepatocellular carcinoma (HCC) cell line, BEL-7402. Athymic nude mice implanted with BEL-7402 cells were given celecoxib and fluvastatin, either alone or in combination, and the effect of treatment on tumour growth was evaluated after 6 weeks. The combination of celecoxib and fluvastatin enhanced inhibition of tumour growth, induction of apoptosis, inhibition of tumour cell proliferation, and inhibition of tumour angiogenesis compared with either treatment alone. The combination of celecoxib and fluvastatin also increased levels of the cyclin-dependent kinase inhibitor p21(Waf1/Cip1), decreased levels of p-Akt, myeloid cell leukaemia-1 (Mcl-1) and survivin protein, but had no effect on Akt protein levels in tumours. These results suggest that celecoxib combined with fluvastatin would be more efficacious for the treatment of HCC than either treatment alone and this combination of therapy warrants further research.

Keratinocytes play a role in the immunity to Herpes simplex virus type 2 infection.

Herpes simplex virus type 2 (HSV-2) infection is the most common cause of genital ulcerative disease in the developed world. Keratinocytes are the primary cells involved in clinical lesions caused by HSV-2. In our study, we intensively examined cytokine expression in the HSV-2-infected keratinocytes. We observed upregulation of a series of cytokines including early-induced antiviral cytokines as interferons α, ß (IFN-α, ß), tumor necrosis factor α (TNF-α), colony stimulating factors (CSFs) as G-CSF, GM-CSF, interleukin 3 (IL-3), growth factors (EGF, KGF, and IGF-ß1), defensins, selectins, leukocyte function-associated antigens (LFAs,) and toll-like receptors (TLR-2, 3, 4, and 9). More importantly, we found that HSV-2-infected keratinocytes stimulated the proliferation of lymphocytes in co-cultivation system. These data suggest that keratinocytes participate in the immune response to HSV-2 infection in two ways. They secrete inflammatory cytokines to resist the HSV-2 infection directly and recruit the immune cells to eliminate the primary infection indirectly and enhance the adaptive immunity to prevent subsequent infections.

MiR-155 affects renal carcinoma cell proliferation, invasion and apoptosis through regulating GSK-3ß/ß-catenin signaling pathway.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-155 affects renal carcinoma cell proliferation, invasion and apoptosis through regulating GSK-3ß/ß-catenin signaling pathway, by R.-J. Wei, C.-H. Zhang, W.-Z. Yang, published in Eur Rev Med Pharmacol Sci 2017; 21(22): 5034-5041-DOI: 10.26355/eurrev_201711_13813-PMID: 29228417" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/13813.

Suppression of connexin 43 expression by miR-106a promotes melanoma cell proliferation.

OBJECTIVE: Connexin 43 (Cx43), a vital gap junction protein is reported to be involved in melanoma progression. The aim of the study is to investigate the regulatory role of Cx43 in melanoma. MATERIALS AND METHODS: Western blot assay was used to detect the protein expression of Cx43 in melanoma cells and the human epidermal melanocytes (HEMn). MTT cell proliferation and cell colony formation assays were used to assess cell proliferation. Bioinformatics prediction, luciferase reporter assay, Western blot and qRT-PCR assays were applied to demonstrate that Cx43 was a direct target of miR-106a in melanoma cells. RESULTS: Connexin 43 (Cx43) was lower expressed in melanoma cells compared with human epidermal melanocytes (HEMn). Cx43 overexpression significantly inhibited melanoma cell proliferation and colony formation ability in vitro. However, knockdown of Cx43 had opposite effects on cell proliferation and colony formation. Bioinformatics prediction and luciferase reporter assays demonstrated that miR-106a targeted the 3' untranslated region (3'UTR) of Cx43 and regulated its mRNA and protein expression levels in melanoma cells. MiR-106a was upregulated in melanoma cells, and its overexpression attenuated the effects caused by upregulating Cx43 expression. CONCLUSIONS: Thus, our results indicated that Cx43 was downregulated in melanoma cells and may be a potential target of melanoma treatment.

LncRNA HOTAIR inhibited osteogenic differentiation of BMSCs by regulating Wnt/ß-catenin pathway.

OBJECTIVE: This study aims to investigate whether HOX transcript antisense RNA (HOTAIR) can participate in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the Wnt/ß-catenin pathway, thereby participating in the pathogenesis of osteoporosis. PATIENTS AND METHODS: We detected the expression level of HOTAIR in 60 osteoporosis patients and 60 normal controls by Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Meanwhile, BMSCs derived from human or rats were subjected to determination of HOTAIR level. Subsequently, the effects of HOTAIR on osteogenic differentiation were evaluated by the activity of Alkaline Phosphatase (ALP), Alizarin Red S (ARS) staining, ALP staining and osteogenic-specific gene expression. The expression level of proteins related to the Wnt/ß-catenin was determined by Western blot, and ALP activity was detected by ALP activity determination kit and alizarin red staining after knockdown or overexpression of HOTAIR, as well as the treatment of DKK1 or the Wnt pathway antagonist. Finally, osteoporosis model in rats was established by ovariectomy (OVX). We examined protein levels of HOTAIR, ß-catenin, CyclinD, C-myc, and Runx2 in rat bone tissues. Bone morphology was observed in each group as well. RESULTS: The serum and BMSCs levels of HOTAIR in patients with osteoporosis were remarkably higher than that in normal people. Inhibition of HOTAIR induced increased ALP activity increased osteogenic marker genes and enhanced number of calcified nodules in BMSCs. However, the overexpression of HOTAIR exhibited the opposite effects. HOTAIR inhibited the expression level of Wnt/ß-catenin pathway-related protein. Also, Wnt pathway antagonist DKK1 partially reversed the regulatory effects of HOTAIR on Wnt/ß-catenin. DKK1 treatment markedly reduced the promotive effect of HOTAIR knockdown on ALP activity, ALP content and calcification ability of BMSCs. DKK1 administration in rats undergoing OVX showed worse bone morphology relative to controls. Protein levels of HOTAIR, ß-catenin, CyclinD, C-myc and Runx2 remarkably downregulated in OVX rats administrated with DKK1. CONCLUSIONS: HOTAIR inhibits osteoblast differentiation of rat BMSCs. The underlying mechanism of which may be related to the mediation of Wnt/ß-catenin pathway.

Analysis of related factors of orolingual angioedema after rt-PA intravenous thrombolytic therapy.

OBJECTIVE: Orolingual angioedema (OA) is a rare clinical complication with a potentially fatal risk that occurs after the intravenous application of alteplase (rt-PA) in patients with acute ischemic stroke. The purpose of this work is to investigate the related factors of OA in patients with acute ischemic stroke after the administration of intravenous thrombolytic therapy, to improve the predictive ability of OA during intravenous thrombolytic therapy, and to reduce the prevalence of complications. PATIENTS AND METHODS: We recruited 1223 cases of patients with acute ischemic stroke that were treated in the Department of Neurology No. 4 of the Tianjin Huanhu Hospital from June 2014 to April 2015. The clinical manifestations of rt-PA related OA were recorded, the clinical prevalence was counted, related factors of OA after intravenous thrombolytic therapy were analyzed, and the risk assessment of rt-PA related OA was conducted. RESULTS: 14 cases of patients developed OA, with a prevalence rate of 1.14%. Among them, 5 had a history of urticaria, 4 of drug allergy, and 3 of food allergy. Among the 14 cases of patients, 10 developed OA in the process of intravenous thrombolysis and 4 after intravenous thrombolysis, 12 showed lip edema, 9 showed extensive swelling of tongue, 3 showed swelling of lateral tongue, 3 were complicated by respiratory distress, 10 showed infarction in the middle cerebral artery territory, and 6 had previously been given oral ACEI drugs. CONCLUSIONS: Orolingual angioedema is a rare complication that occurs after rt-PA intravenous thrombolytic therapy; when serious, it may endanger a patient's life. If patients take an oral hypotension such as ACEI drugs before the onset of OA, they have a history of allergies, or the lesion is an infraction in the dominated area of the middle cerebral artery, the risk of OA after rt-PA intravenous thrombolytic therapy will be increased. The prevalence of OA should be monitored during the rt-PA intravenous thrombolytic therapy process; timely detection and early intervention should be conducted, which can avoid serious adverse consequences.

Photosensitizer-induced self-assembly of antigens as nanovaccines for cancer immunotherapy.

Herein, the photosensitizer indocyanine green (ICG) is used to induce the self-assembly of antigens to form nanovaccines. Under near-infrared (NIR) laser irradiation, reactive oxygen species can be generated by nanovaccines to disrupt the membranes of endo/lysosomes, which helps to release antigens into the cytosol efficiently, thereby enhancing antigen cross-presentation and anti-cancer immunity. To the best of our knowledge, this study represents the first example of ICG as a biocompatible adjuvant to improve cancer vaccine efficacy.

Bioinformatic analysis of prognostic value of ARAP3 in breast cancer and the associated signaling pathways.

OBJECTIVE: In this study, we tried to pool previous annotated genomic data to assess the association between ARAP3 expression and metastatic relapse (MR) risk in patients with breast cancer. Moreover, we also investigated the signaling pathways in which ARAP3 might be involved in breast cancer. MATERIALS AND METHODS: The raw microarray data (GDS5666) that compared gene transcriptional profiles of 4T1 derived lung-aggressive explant and primary tumor explant were reanalyzed to identify the dysregulated genes. ARAP3 mRNA expression, its association with MR-free survival and its co-upregulated genes in breast cancer, were studied by data mining in TCGA database and Breast Cancer Gene-Expression Miner Version 4.0 (bc-GenExMiner 4.0). RESULTS: ARAP3 is a significantly upregulated gene in the metastatic breast tumor cells. The ER- patients with high ARAP3 expression had significantly increased the risk of MR, regardless of the nodal status. Patients in ER-/Nm group with high ARAP3 expression had the highest risk of MR (HR: 1.25; 95%CI: 1.10-1.41, p<0.001). In patients with basal-like tumors, High ARAP3 level is associated with significantly worse MR-free survival (HR: 1.63, 95%CI: 1.22-2.19, p=0.001). ARAP3 might be closely related to the NOTCH4 and CDH5 signaling pathways in breast cancer. CONCLUSIONS: The expression level of ARAP3 might be a useful indicator of the metastatic likelihood of the basal-like breast tumors. ARAP3 might be involved in NOTCH4 and CDH5 related signaling pathways, but the underlying mechanism should be further studied.

MiR-155 affects renal carcinoma cell proliferation, invasion and apoptosis through regulating GSK-3ß/ß-catenin signaling pathway.

OBJECTIVE: Glycogen Synthase Kinase-3ß (GSK-3ß) negatively regulates Wnt/ß-catenin signaling pathway through degrading ß-catenin protein. It plays an inhibitory role in various tumors, while the influence in the pathogenesis of renal carcinoma has not been elucidated. MicroRNA-155 (MiR-155) was found to be upregulated in renal carcinoma tissue. Bioinformatics analysis revealed the complementary binding site between miR-155 and 3'-UTR of GSK-3ß. This study investigated the influence of miR-155 in regulating GSK-3ß expression, Wnt/ß-catenin signaling pathway activity, and renal carcinoma cell proliferation, invasion, and apoptosis. PATIENTS AND METHODS: The targeted regulatory relationship between miR-155 and GSK-3ß were tested by dual luciferase assay. Renal carcinoma tissue and benign renal tissue were collected to detect miR-155 and GSK-3ß expressions. MiR-155, GSK-3ß, and ß-catenin levels were compared between HK-2 and 786-O cells. Renal carcinoma 786-O cells were cultured in vitro and divided into four groups, including miR-NC, anti-miR-155, pIRES2-blank, and pIRES2-GSK-3ß groups. Cell apoptosis was evaluated by flow cytometry. Cell invasion was determined by transwell assay. Cell proliferation was assessed by EdU staining. RESULTS: MiR-155 targeted regulated GSK-3ß expression. MiR-155 and ß-catenin expressions were significantly increased, while GSK-3ß level was significantly declined in renal carcinoma tissue compared with benign renal tissue. MiR-155 and ß-catenin expressions were significantly elevated, whereas GSK-3ß level was significantly downregulated in 786-O cells compared with HK-2 cells. Anti-miR-155 or pIRES2-GSK-3ß transfection significantly up-regulated GSK-3ß expression, attenuated ß-catenin level, restrained cell proliferation and invasion, and enhanced cell apoptosis. CONCLUSIONS: MiR-155 promoted renal carcinoma pathogenesis. Inhibition of miR-155 increased GSK-3ß expression, attenuated Wnt/ß-catenin signaling pathway, weakened proliferation and invasion, and facilitated apoptosis in renal carcinoma cells.

Absorption mechanism of three curcumin constituents through in situ intestinal perfusion method.

This study aimed to investigate the absorption mechanism of three curcumin constituents in rat small intestines. Self-emulsification was used to solubilize the three curcumin constituents, and the rat in situ intestinal perfusion method was used to study factors on drug absorption, including drug mass concentration, absorption site, and the different types and concentrations of absorption inhibitors. Within the scope of experimental concentrations, three curcumin constituents were absorbed in rat small intestines through the active transport mechanism.

A positive crosstalk between CXCR4 and CXCR2 promotes gastric cancer metastasis.

The molecular mechanism underlying gastric cancer (GC) invasion and metastasis is still poorly understood. In this study, we tried to investigate the roles of CXCR4 and CXCR2 signalings in gastric cancer metastasis. A highly invasive gastric cancer cell model was established. Chemokines receptors were profiled to search for the accountable ones. Then the underlying molecular mechanism was investigated using both in vitro and in vivo techniques, and the clinical relevance of CXCR4 and CXCR2 expression was studied in gastric cancer samples. CXCR4 and CXCR2 were highly expressed in a high invasive gastric cancer cell model and in gastric cancer tissues. Overexpression of CXCR4 and CXCR2 was associated with more advanced tumor stage and poorer survival for GC patients. CXCR4 and CXCR2 expression strongly correlated with each other in the way that CXCR2 expression changed accordingly with the activity of CXCR4 signaling and CXCR4 expression also changed in agreement with CXCR2 activity. Further studies demonstrated CXCR4 and CXCR2 can both activated NF-κB and STAT3 signaling, while NF-κBp65 can then transcriptionally activate CXCR4 and STAT3 can activate CXCR2 expression. This crosstalk between CXCR4 and CXCR2 contributed to EMT, migration and invasion of gastric cancer. Finally, Co-inhibition of CXCR4 and CXCR2 is more effective in reducing gastric cancer metastasis. Our results demonstrated that CXCR4 and CXCR2 cross-activate each other to promote the metastasis of gastric cancer.