Complete genome sequence of a novel mitovirus isolated from the fungus Fusarium oxysporum f. sp. ginseng causing ginseng root rot.

A novel mitovirus, tentatively designated as "Fusarium oxysporum mitovirus 2" (FoMV2), was isolated from the pathogenic Fusarium oxysporum f. sp. ginseng strain 0414 infecting Panax ginseng. The complete genome of FoMV2 is 2388 nt in length with a GC content of 30.57%. It contains a large open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) of 713 amino acids with a molecular weight of 83.05 kDa. The sequence identity between FoMV2 and Botrytis cinerea mitovirus 8 and Fusarium verticillioides mitovirus 1 was 87.94% and 77.85%, respectively. Phylogenetic analysis showed that FoMV2 belongs to the genus Unuamitovirus in the family Mitoviridae. To the best of our knowledge, this is the first report of an unuamitovirus isolated from F. oxysporum f. sp. ginseng causing ginseng root rot.

Tulsi (Ocimum sanctum) mediated Co nanoparticles with their anti-inflammatory, anti-cancer, and methyl orange dye adsorption properties.

Nanotechnology is an emerging technology that uses medicinal plants to extract nanoparticles for conventional applications. In the present investigation, the medical plant Tulsi (Ocimum sanctum) has used in the synthesis of cobalt (Co) nanoparticles in a cost-effective, feasible process. The efficiency of nanoparticles in removing methyl orange dye was evaluated by analyzing their applications in wastewater treatment. An analysis of the anti-inflammatory and anti-cancer properties of Tulsi-mediated Co nanoparticles was conducted to examine their medical application. Morphological analysis of Co nanoparticles showed that the synthesized nanoparticles were in crystal shape with a mean particle size of 110 nm. A batch adsorption study has shown that incubation periods of 5 h, pH 2, temperatures of 70 °C, and adsorbent dosage of 125 µg/mL are optimal for removing methyl orange dye from wastewater. To examine the anti-inflammatory properties of Tulsi-mediated Co nanoparticles, protein denaturation and nitric oxide scavenging assays were performed. The maximum anti-inflammatory response was recorded at a concentration of 250 µg/mL of Co nanoparticles. MTT assays against MDA-MB-231 human breast cancer cells were used to evaluate the anti-cancer properties of Co nanoparticles. This study investigates the economical extraction of Co nanoparticles from tulsi and its potential use in wastewater purification and biomedical applications.

Changes in the relationship between vapour pressure deficit and water use efficiency with the drought recovery time: A case study of the Yellow River Basin.

Drought is a major driver of interannual variability in the gross primary productivity (GPP) of global terrestrial ecosystems, and drought recovery time has been widely used to assess ecosystem responses to drought. However, the response of the carbon-water coupled cycle to drought, especially changes in the correlation between drought intensity and carbon-water coupling throughout the recovery time, remains unclear. In this study, the Yellow River Basin (YRB) located mostly in drylands was the study area. We assessed the correlation between the standardized water vapour pressure deficit (VPD) and the water use efficiency of ecosystems (WUEe) and water use efficiency of canopies (WUEc) every month with the drought recovery time of GPP. We found that the drought intensity in the middle reach of the YRB (MYRB) was greater and the drought recovery time was longer than those in the upper reach (UYRB) and lower reach (LYRB) during the period from 2003 to 2017. In terms of the correlation between drought intensity and carbon-water coupling, the greater the VPD was, the lower the WUEc. In addition, the correlation of WUEc with VPD was higher than that of WUEe in most areas of the YRB, especially in the LYRB. On the watershed level, the correlation between the two types of WUE and VPD increased gradually with the recovery time, while the correlation between WUEc and VPD (mostly negative) changed more than the correlation between WUEe and VPD (mostly positive). Therefore, the response of WUEc to meteorological drought should be given more attention, especially during the middle and late stages of drought, since it exhibited an opposite signal compared to that of WUEe during drought recovery.

Effects of Lysine on the Interfacial Bonding of Epoxy Resin Cross-Linked Soy-Based Wood Adhesive.

Soy protein isolate (SPI) is an attractive natural material for preparing wood adhesives that has found broad application. However, poor mechanical properties and unfavorable water resistance of wood composites with SPI adhesive bonds limit its more extensive utilization. The combination of lysine (Lys) with a small molecular structure as a curing agent for modified soy-based wood adhesive allows Lys to penetrate wood pores easily and can result in better mechanical strength of soy protein-based composites, leading to the formation of strong chemical bonds between the amino acid and wood interface. Scanning electron microscopy (SEM) results showed that the degree of penetration of the S/G/L-9% adhesive into the wood was significantly increased, the voids, such as ducts of wood at the bonding interface, were filled, and the interfacial bonding ability of the plywood was enhanced. Compared with the pure SPI adhesive, the corresponding wood breakage rate was boosted to 84%. The wet shear strength of the modified SPI adhesive was 0.64 MPa. When Lys and glycerol epoxy resin (GER) were added, the wet shear strength of plywood prepared by the S/G/L-9% adhesive reached 1.22 MPa, which increased by 29.8% compared with only GER (0.94 MPa). Furthermore, the resultant SPI adhesive displayed excellent thermostability. Water resistance of S/G/L-9% adhesive was further enhanced with respect to pure SPI and S/GER adhesives through curing with 9% Lys. In addition, this work provides a new and feasible strategy for the development and application of manufacturing low-cost, and renewable biobased adhesives with excellent mechanical properties, a promising alternative to traditional formaldehyde-free adhesives in the wood industry.

Umbilical artery ultrasound haemodynamics combined with serum adiponectin levels can aid in predicting adverse pregnancy outcomes in patients with severe pre-eclampsia.

Severe pre-eclampsia is a leading cause of maternal and perinatal morbidity and mortality. This retrospective study explored pregnancy outcome predictive values of umbilical artery Doppler with serum adiponectin in severe pre-eclampsia. Fasting elbow venous blood was collected from 118 severe pre-eclampsia patients [maternal systolic pressure ≥ 160 mmHg and/or diastolic pressure ≥ 110 mmHg + minimal proteinuria, 56; mild hypertension + heavy proteinuria (≥2 g/24 h or random urinary protein ≥ 2+), 42; no proteinuria but new-onset hypertension + diseases of heart/lung/liver/kidney/other organs or abnormalities in blood/digestive/nervous systems, placental foetus involved, 20] and 90 controls (18.5-24.9 kg/m2) in the first morning of admission. Serum adiponectin and resistance/pulsatility indexes were separately measured and correlatively analysed by Pearson's coefficient analysis. Adverse outcomes included maternal primary postpartum haemorrhage and placental abruption, neonatal asphyxia, low birth weight, foetal distress, foetal growth restriction. In severe pre-eclampsia, serum adiponectin (downregulated) was negatively-correlated with resistance/pulsatility indexes (upregulated). The area under the curve of umbilical artery Doppler with serum adiponectin for predicting adverse outcomes of severe pre-eclampsia was 0.6545 (specificity 60.27%, sensitivity 60.00%). In conclusion, umbilical artery Doppler with serum adiponectin predicts adverse pregnancy outcomes in severe pre-eclampsia.Impact statementWhat is already known on this subject? Sad levels were lowered in sPE patients. UA ultrasound hemodynamic parameters can predict adverse pregnancy outcomes.What do the results of this study add? Our study revealed that ultrasonic hemodynamic indexes of UA combined with Sad levels had better efficacy in predicting pregnancy outcomes in patients with sPE, and our study is expected to improve the accuracy of clinical prediction of adverse outcomes in sPE patients.What are the implications of these findings for clinical practice and/or further research? Through the combined detection of multiple indicators of the foetus in the mother, our study expects to be able to monitor and predict the growth of the foetus in the mother more accurately in clinical practice, avoid excessive intervention or untimely intervention, and reduce the incidence of perinatal adverse pregnancy outcomes.

Thermal and flame-retardant properties of multilayered composites prepared through novel multilayering approach.

Thermal and flame-retardant properties of traditional composites have limitations that are not satisfied for the various applications. Multilayered materials have great potential to improve material properties. The present paper focused on designing new multilayering approach to fabricate flame retardant multilayered materials with a very basic instrument and several simple steps. Montmorillonite nanoparticles filled maleic anhydride grafted polypropylene composites were prepared by the melt-blending method, and the multilayered composites with polypropylene alternating multilayers were fabricated by the quadruple-layering approach. The multilayer structure was characterized by the scanning electron microscopy/energy dispersive spectrometer. The influence of layer structure on the thermal stability, thermal conductivity and flame-retardant properties was investigated by the comparison with the conventional composites. Multilayered composites showed enhanced flame-retardant properties with lower peak heat release rate and better char formation compared to conventional composites with the same mass fraction of montmorillonite. Multilayered composites had higher mass fraction of montmorillonite in filled layers and no fillers in other layers, which caused the unequal distribution of montmorillonite, resulting in changes of thermal and flame-retardant properties of the materials especially in the perpendicular direction to the film surface. This study demonstrates a unique multilayering approach that has potential to be used in variety applications such as food and medical packaging.

Multilayered Nanocomposites Prepared through Quadruple-Layering Approach towards Enhanced Mechanical Performance.

Multilayered materials are widely studied due to their special structures and great properties, such as their mechanical ones. In this paper a novel and effective technique, a quadruple-layering approach, was used to fabricate multilayered materials. This approach increases the number of layers rapidly via simple operations. Materials with 4, 16, and 64 layers with alternating layers of polypropylene and nanocomposites were fabricated using this approach, and their film morphology and mechanical properties were studied. The influence of the number of layers on the mechanical properties of the materials and the relationship between the mechanical properties of each material were investigated. The results illustrated that the tensile modulus and strength were enhanced and elongation at the break increased when the layer numbers of the multilayered materials increased. However, this approach has a defect in that as the layer number increases, the layer thickness was not uniform, thus restricting the improvement of properties. This may need to be further studied in future work.

Lead contamination alters enzyme activities and microbial composition in the rhizosphere soil of the hyperaccumulator Pogonatherum crinitum.

Pogonatherum crinitum is a promising lead (Pb) hyperaccumulator; however, the effects of Pb contamination on P. crinitum rhizosphere soil enzymatic activities and microbial composition remain largely unexplored. Thus, an indoor experiment was conducted by cultivating P. crinitum seedlings and exposing them to four Pb concentrations (0, 1,000, 2000 and 3000 mg/kg Pb). Protease, urease, acid phosphatase and invertase activities were determined using standard methods while soil bacterial composition was determined by 16 S rDNA sequencing. The results showed that rhizosphere soil acid phosphatase activity significantly increased with increasing Pb concentration, while urease activity was significantly greater in rhizosphere soil contaminated with 1000 and 2000 mg/kg than in the control. There was a clear shift in bacterial composition during phytoremediation by P. crinitum. Compared to the control, Bacteroidetes was more abundant in all Pb-contaminated soils, Actinobacteria was more abundant in 1000 mg/kg Pb-treated soil, and Firmicutes was more abundant in 3000 mg/kg Pb-treated soil. Positive correlations were observed between dominant bacterial phyla and soil enzyme activities. Metabolic pathways, such as ABC transporter, quinine reductase, and ATP-binding protein were significantly increased in rhizosphere soil bacteria with Pb contamination. In conclusion, Pb contamination differentially influenced the activities of rhizosphere soil enzymes, specifically increasing acid phosphatase and urease activities, and alters the dominance of soil bacteria through up-regulation of genes related to some metabolic pathways. The strong correlations between dominant bacterial phyla and enzymatic activities suggest synergetic effects on the growth of P. crinitum during Pb contamination.

Bamboo grid versus polyvinyl chloride as packing material in cooling tower: Energy efficiency and environmental impact assessment.

As an abundant and fast-growing biomass, bamboo can be used as construction materials owing to its desirable physical and mechanical properties, environmentally friendly features, and alternative to replace toxic and hazardous wastes in industrial processing. In this study, grid material made from bamboo (termed 'bamboo grid') was developed and compared to commercially used polyvinyl chloride (PVC) as packing material in cooling towers; PVC packing has drawbacks such as fouling, deposit buildup, low durability, and is harmful to environments. The cooling capacity, energy efficiency and environmental impact of bamboo grid packing were evaluated via life cycle assessment (LCA), particularly the cumulative energy demand (CED) and the Building for Environmental and Economic Sustainability (BEES). Although the thermal performance of the PVC packing was found higher than that of the bamboo grid packing, the bamboo grid packing showed improved resistance characteristic, recording a total saving of 529.2 tons of standard coal during a six-month field test in a real thermal power generation plant. LCA results revealed that the utilization of bamboo-grid packing to replace PVC packing in cooling towers reduced total CED from 3420 MJ to 561 MJ per functional unit, achieving 6 times reduction. A desirable reduction ranging from 1.5 to 10.5 times was also recorded for the BEES indices. This LCA comparison analysis confirmed the improvement of energy efficiency and reduction of environmental impact by using the bamboo grid to replace PVC as packing material in cooling towers. The major environmental impact (BEES) indices (e.g., the total Global warming potential, Acidification, Eutrophication and Smog) were reduced by 1.5-10.5 times via the use of bamboo grid. The results demonstrate that bamboo grid packing is a good alternative to replace existing grid packing materials such as concrete and PVC that are harmful to human health and environments.

Production of magnetic sodium alginate polyelectrolyte nanospheres for lead ions removal from wastewater.

Polyelectrolyte composite nanospheres are relatively new adsorbents which have attracted much attention for their efficient pollutant removal and reuse performance. A novel polyelectrolyte nanosphere with magnetic function (SA@AM) was synthesized via the electrostatic reaction between the polyanionic sodium alginate (SA) and the surface of a prepared terminal amino-based magnetic nanoparticles (AMs). SA@AM showed a size of 15-22 nm with 6.85 emu·g-1 of magnetization value, exhibiting a high adsorption capacity on Pb(II) ions representing a common heavy metal pollutant, with a maximum adsorption capacity of 105.8 mg g-1. The Langmuir isotherm adsorption fits the adsorption curve, indicating uniform adsorption of Pb(II) on the SA@AM surfaces. Repeated adsorption desorption experiments showed that the removal ratio of Pb(II) by SA@AM was more than 76%, illustrating improved regeneration performance. These results provide useful information for the production of bio-based green magnetic nano scale adsorption materials for environmental remediation applications.

Strength and size of phosphorus-rich patches determine the foraging strategy of Neyraudia reynaudiana.

BACKGROUND: Under natural conditions, soil nutrients are heterogeneously distributed, and plants have developed adaptation strategies to efficiently forage patchily distributed nutrient. Most previous studies examined either patch strength or patch size separately and focused mainly on root morphological plasticity (increased root proliferation in nutrient-rich patch), thus the effects of both patch strength and size on morphological and physiological plasticity are not well understood. In this study, we examined the foraging strategy of Neyraudia reynaudiana (Kunth) Keng ex Hithc, a pioneer grass colonizing degraded sites, with respect to patch strength and size in heterogeneously distributed phosphorus (P), and how foraging patchily distributed P affects total plant biomass production. Plants were grown in sand-culture pots divided into ½, », 1/6 compartments and full size and supplied with 0 + 0/30, 0 + 7.5/30 and 7.5 + 0/30 mg P/kg dry soil as KH2PO4 or 0 + 15/15, 0 + 18.5/ 18.5, 7.5 + 15/15 mg kg - 1 in the homogenous treatment. The first amount was the P concentration in the central region, and that the second amount was the P concentration in the outer parts of the pot. RESULTS: After 3 months of growth under experimental conditions, significantly (p < 0.05) high root elongation, root surface area, root volume and average root diameter was observed in large patches with high patch strength. Roots absorbed significantly more P in P-replete than P-deficient patches. Whole plant biomass production was significantly higher in larger patches with high patch strength than small patches and homogeneous P distribution. CONCLUSION: The result demonstrates that root morphological and physiological plasticity are important adaptive strategies for foraging patchily distributed P and the former is largely determined by patch strength and size. The results also establish that foraging patchily distributed P resulted in increased total plant biomass production compared to homogeneous P distribution.

Case report: Analysis of BRCA1 and BRCA2 gene mutations in a hereditary ovarian cancer family.

OBJECTIVE: Breast cancer susceptibility gene 1/2 (BRCA1/2) is the most important susceptibility gene associated with hereditary ovarian cancer (HOC). We aimed to screen BRAC1 and BRAC2 gene mutations in a member of a hereditary ovarian cancer family in China, and to analyze the structure and function of the mutant protein. METHODS: A typical HOC family was selected. Blood samples and pathological tissue samples were taken from the female members of the family. Blood samples from two patients with sporadic ovaries of the same pathological type were taken as a control group. After RNA extraction, PCR amplification was applied and the PCR products were directly sequenced and aligned, prediction and analysis of protein structure and molecular conformation that may be caused by BRCA1/2 mutation. RESULTS: The whole gene analysis of BRCA1 and BRCA2 in ovarian cancer patients in the family showed that there were 8 mutations in BRCA1 whole gene sequencing, including 3 nonsense mutations (2314C>T, 2543T>C, 4540T>C); two mutations have been recorded, which are associated with cervical cancer (2844C>T) and endometriosis (3345A>G); three newly discovered mutations (3780A>G, 5069A>G, 3326A>T). Among them, 3780A>G and 5069A>G caused amino acid changes, while 3326A>T mutation caused Arg mutation to stop codon. A total of 7 mutations were detected in BRCA2 whole-genome sequencing, including 5 non-significant mutations (3623A>G, 4034T>C, 4790A>G, 6740G>C, 7469A>G); one no-record mutation (1716T>A), and 1 recorded mutation (1342A>C), which was associated with breast cancer and ovarian cancer. BRCA1 (3326A>T) and BRCA2 (1342A>C) mutations were co-existing in patients (II1, II3, and II5) identified as serous adenocarcinoma grade II. Two cases of ovarian serous cystadenocarcinoma with no history of family tumors were normalized for BRCA1/2 gene sequencing. In the gene detection of III generation female, four females with BRCA2 (1342A>C) mutation were found, and one of them also carried the BRCA1 (3326A>T) mutation, who can be considered a high-risk group of HOC in this family. Online protein structure predictions revealed that BRCA1 (3326A>T) mutations mutated AGA at this site to TGA resulting in a translated Arg (arginine) mutation as a stop codon, while BRCA2 (1342A>C) mutated AAT at this site to CAT resulting in a translated Asn mutation to His. CONCLUSION: The BRCA1 (3326A>T) and BRCA2 (1342A>C) were detected in the HOC family, which may be the susceptibility gene of the family's HOC. The BRCA1/2 gene screening may be possible to obtain high-risk populations in this family.

Transcriptional Regulation Between the Two Global Regulators RovA and CRP in Yersinia pestis biovar Microtus.

Yersinia pestis is a dangerous bacterial pathogen that can cause plague. Both RovA and cyclic AMP receptor protein (cAMP-CRP) are required for regulating biofilm- and virulence-related genes in Y. pestis. In this study, the transcriptional regulation between RovA and cAMP-CRP were analyzed by using primer extension, quantitative RT-PCR, LacZ fusion, and electrophoretic mobility shift assay. The results indicated that RovA repressed crp transcription in an indirect manner, while that RovA had no regulatory action on cyaA at the transcriptional level. In addition, cAMP-CRP did not regulate the transcription of rovA. Taken together with our previous results, complex regulatory interactions of RovA, cAMP-CRP, and PhoP/PhoQ in Y. pestis were revealed, which would promote us gain deeper understanding about coordinative modulation of biofilm- and virulence-related regulator genes.

SIRT1 activation by pterostilbene attenuates the skeletal muscle oxidative stress injury and mitochondrial dysfunction induced by ischemia reperfusion injury.

Ischemia reperfusion (IR) injury is harmful to skeletal muscles and causes mitochondrial oxidative stress. Pterostilbene (PTE), an analogue of resveratrol, has organic protective effects against oxidative stress. However, no studies have investigated whether PTE can protect against IR-related skeletal muscular injury. In this study, we sought to evaluate the protective effect of PTE against IR-related skeletal muscle injury and to determine the mechanisms in this process. Male Sprague-Dawley rats were pretreated with PTE for a week and then underwent limb IR surgery. The IR injury induced segmental necrosis and apoptosis, myofilament disintegration, thicker interstitial spaces, and inflammatory cell infiltration. Furthermore, mitochondrial respiratory chain activity in the muscular tissue was inhibited, methane dicarboxylic aldehyde concentration and myeloperoxidase activity were up-regulated, and superoxide dismutase was down-regulated after IR. However, these effects were significantly inhibited by PTE in a dose-dependent manner. The mechanism underlying IR injury is attributed to the down-regulation of silent information regulator 1 (SIRT1)-FOXO1/p53 pathway and the increase of the Bax/Bcl2 ratio, Cleaved poly ADP-ribose polymerase 1, Cleaved Caspase 3, which can be reversed with PTE. Furthermore, EX527, an SIRT1 inhibitor, counteracted the protective effects of PTE on IR-related muscle injury. In conclusion, PTE has protective properties against IR injury of the skeletal muscles. The mechanism of this protective effect depends on the activation of the SIRT1-FOXO1/p53 signaling pathway and the decrease of the apoptotic ratio in skeletal muscle cells.

The protective effects of Acanthus ilicifolius alkaloid A and its derivatives on pro- and anti-inflammatory cytokines in rats with hepatic fibrosis.

This study was designed to investigate the protective effects of Acanthus ilicifolius alkaloid A [4-hydroxy-2-benzoxazolone (HBOA)] and its acetylated derivatives including 4-acetoxy-2-benzoxazolone (TC-2) and 3-acetyl-4-acetoxy-2-benzoxazolone (TC-3) on carbon tetrachloride (CCl4 )-induced liver fibrosis in rats. Sprague-Dawley rats were given CCl4 twice per week for 8 weeks to induce liver fibrosis. Then, they were treated with HBOA, TC-2, and TC-3 daily for 4 weeks, respectively. The serum indicators including total protein (TP), albumin (Alb), globulin, hyaluronic acid (HA), and laminin (LN) were measured by commercial kits. The messenger ribonucleic acid expression of adiponectin, peroxisome proliferator-activated receptor-γ (PPAR-γ), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-ß1 (TGF-ß1 ) and Toll-like receptor 4 (TLR4 ) was determined by reverse-transcriptase -PCR. The proteins of adiponectin, TGF-ß1 , α-smooth muscle actin (α-SMA), and TLR4 were also detected by the immunohistochemical assay. The results showed that HBOA, TC-2, and TC-3 significantly attenuated the fibrotic degree induced by CCl4 as evidenced by higher levels of TP, Alb, adiponectin, and PPAR-γ, which in turn decreased the proliferation of hepatic stellate cells. Moreover, those drugs markedly decreased the levels of HA, LN, TNF-α, IL-6, TGF-ß1 , α-SMA, and TLR4 . Our study indicates that HBOA, TC-2, and TC-3 have beneficial effects against liver fibrosis, and the mechanisms may be related to the inhibition of inflammatory response.

High autophagy in the naked mole rat may play a significant role in maintaining good health.

BACKGROUND/AIMS: The maximum lifespan of the naked mole rat is over 28.3 years, which exceeds that of any other rodent species, suggesting that age-related changes in its body composition and functionality are either attenuated or delayed in this extraordinarily long-lived species. However, the mechanisms underlying the aging process in this species are poorly understood. In this study, we investigated whether long-lived naked mole rats display more autophagic activity than short-lived mice. METHODS: Hepatic stellate cells isolated from naked mole rats were treated with 50 nM rapamycin or 20 mM 3-methyladenine (3-MA) for 12 or 24 h. Expression of the autophagy marker proteins LC3-II and beclin 1 was measured with western blotting and immunohistochemistry. The induction of apoptosis was analyzed by flow cytometry. RESULTS: Our results demonstrate that one-day-old naked mole rats have higher levels of autophagy than one-day-old short-lived C57BL/6 mice, and that both adult naked mole rats (eight months old) and adult C57BL/6 mice (eight weeks old) have high basal levels of autophagy, which may be an important mechanism inhibiting aging and reducing the risk of age-related diseases. CONCLUSION: Here, we report that autophagy facilitated the survival of hepatic stellate cells from the naked mole rat, and that treatment with 3-MA or rapamycin increased the ratio of apoptotic cells to normal hepatic stellate cells.

KRAS (G12D) cooperates with AML1/ETO to initiate a mouse model mimicking human acute myeloid leukemia.

BACKGROUND/AIMS: It has been demonstrated that KRAS mutations represent about 90% of cancer-associated mutations, and that KRAS mutations play an essential role in neoplastic transformation. Cancer-associated RAS mutations occur frequently in acute myeloid leukemia (AML), suggesting a functional role for Ras in leukemogenesis. METHODS: We successfully established a mouse model of human leukemia by transplanting bone marrow cells co-transfected with the K-ras (G12D) mutation and AML1/ETO fusion protein. RESULTS: Mice transplanted with AML/ETO+KRAS co-transduced cells had the highest mortality rate than mice transplanted with AML/ETO- or KRAS-transduced cells (115d vs. 150d). Upon reaching a terminal disease stage, EGFP-positive cells dominated their spleen, lymph nodes, peripheral blood and central nervous system tissue. Immunophenotyping, cytologic analyses revealed that AML/ETO+KRAS leukemias predominantly contained immature myeloid precursors (EGFP(+)/c-Kit(+)/Mac-1(-)/Gr-1(-)). Histologic analyses revealed that massive leukemic infiltrations were closely packed in dense sheets that effaced the normal architecture of spleen and thymus in mice transplanted with AML1/ETO + KRAS co-transduced cells. K-ras mRNA and protein expression were upregulated in bone marrow cells of the K-ras group and AML1/ETO + Kras group. The phosphorylation of MEK/ERK was significantly enhanced in the AML1/ETO + Kras group. The similar results of the AML1/ETO + Nras group were consistent with those reported previously. CONCLUSION: Co-transduction of Kras(G12D) and AML1/ETO induces acute monoblastic leukemia. Since expression of mutant K-ras alone was insufficient to induce leukemia, this model may be useful for investigating the multi-step leukemogenesis model of human leukemia.

The protective role of autophagy in Heterocephalus glaber hepatic stellate cells exposed to H2O2 or nutritional stress.

BACKGROUND/AIMS: Naked mole rats (NMRs) survive and thrive in dark, dank environments with low levels of oxygen and poor quality nutrition. Their long lifespan is attributed to sustained good health and pronounced resistance to cancer. Physiological and biochemical processes, such as autophagy, may contribute to the successful aging of this exceptionally long-lived species. We demonstrated that NMRs have higher levels of autophagy than short-lived C57BL/6 mice, and this may play an important role in the maintenance of cellular protein quality and the defense of cells against intracellular and extracellular aggressors in NMRs. The present study assesses autophagy as a means for cells to flexibly respond to environmental changes (H2O2 treatment and a shortage of nutrients). METHODS: Primary NMR HSCs were isolated from liver and treated with serum-free medium. Cells in the experimental group were incubated with different concentrations of hydrogen peroxide (H2O2) in the presence and / or absence of 3-MA (5 mM).The LC3-II/LC3-I ratio was determined by western blot analysis. Western blotting was performed to analyze the expression level of Beclin 1 protein. Apoptosis and cell-cycle progression were analyzed by flow cytometry. RESULTS: Our data reveal that both poor quality nutrition and H2O2 treatment induces apoptosis and autophagy in NMR hepatic stellate cells(HSCs). CONCLUSION: NMR cells have the capacity to induce cell death through apoptosis and downregulate the energy consuming processes through inhibition of proliferation when they become superfluous or irreversibly damaged.

The significant diagnostic value of human telomerase RNA component (hTERC) gene detection in high-grade cervical lesions and invasive cancer.

Gains of 3q26 chromosome region, where the human telomerase RNA gene (hTERC) is located, have been previously documented in cervical carcinomas. However, published data on this subject are inconclusive. Therefore, we performed a meta-analysis to evaluate the diagnostic value of hTERC in high-grade cervical lesions and invasive cancer. We searched all the eligible studies through PubMed, EMBASE, and the Cochrane Library database without language limitation. Studies were assessed for quality using quality assessment of diagnostic accuracy studies (QUADAS). Positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were pooled separately and compared with overall accuracy measures of diagnostic odds ratio (DOR) and symmetric summary receiver operating characteristic (SROC). The PLR and NLR and their 95 % confidence interval (CI) were calculated using a fixed effects model according to the Mantel-Haensed method and random effects model based on the work of Der Simonian and laird, respectively. A total of 12 studies were included for the analysis. The pooled sensitivity was 0.81 (95 % CI, 0.80-0.82). The pooled specificity was 0.83 (95 % CI, 0.82-0.84). The DOR estimate was performed, and the result was 17.37. Our meta-analysis showed that the detection of genomic amplification of hTERC is a noninvasive and effective approach for high-grade cervical lesions and invasive cancer.

Basic mechanism leading to stimulation of glycogenolysis by isoproterenol, EGF, elevated extracellular K+ concentrations, or GABA.

Glycogenolysis, in brain parenchyma an astrocyte-specific process, has changed from being envisaged as an emergency procedure to playing central roles during brain response to whisker stimulation, memory formation, astrocytic K(+) uptake and stimulated release of ATP. It is activated by several transmitters and by even very small increases in extracellular K(+) concentration, and to be critically dependent upon an increase in free cytosolic Ca(2+) concentration ([Ca(2+)]i), whereas cAMP plays only a facilitatory role together with increased [Ca(2+)]i. Detailed knowledge about the signaling pathways eliciting glycogenolysis is therefore of interest and was investigated in the present study in well differentiated cultures of mouse astrocytes. The ß-adrenergic agonist isoproterenol stimulated glycogenolysis by a ß1-adrenergic effect, which initiated a pathway in which cAMP/protein kinase A activated a Gi/Gs shift, leading to Ca(2+)-activated glycogenolysis. Inhibition of this pathway downstream of cAMP but upstream of the Gi/Gs shift abolished the glycogenolysis. However, inhibitors operating downstream of the Ca(2+)-sensitive step, but preventing transactivation-mediated epidermal growth factor (EGF) receptor stimulation, a later step in the activated pathway, also caused inhibition of glycogenolysis. For this reason the effect of EGF was investigated and it was found to be glycogenolytic. Large increases in extracellular K(+) activated glycogenolysis by a nifedipine-inhibited L-channel opening allowing influx of Ca(2+), known to be glycogenolysis-dependent. Small increases (addition of 5 mM KCl) caused a smaller effect by a similarly glycogenolysis-reliant opening of an IP3 receptor-dependent ouabain signaling pathway. The same pathway could be activated by GABA (also in brain slices) due to its depolarizing effect in astrocytes.