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Lepista nuda is a wild edible fungus that is valued for its odor and taste. Recent studies identified intraspecific morphological and genetic differences in L. nuda. Although single-nucleotide polymorphisms (SNPs) are useful for revealing intraspecific differences, the traditional methods used for investigating SNPs are time consuming and expensive, and they only locate a limited number of SNPs. This study used a 'restriction-site associated DNA' (RAD) method combined with high throughput sequencing to efficiently identify a large number of SNPs in two samples of L. nuda. A total of 7 and 9 billion bp of raw data were obtained from the two collections. A total of 712 SNPs were found. These SNPs will be useful for the further analysis of the genetic variation within L. nuda. The study also confirms that the RAD method can be used to identify SNPs in a nonmodel macrofungus for which a reference genome is unavailable. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Baicalin and scutellarin are the principal bioactive components of Scutellaria baicalensis Georgi which has extensively been incorporated into heat-clearing and detoxification formulas for the treatment of Helicobacter pylori -related gastrointestinal disorders in traditional Chinese medicine. However, the mechanism of action remained to be defined. Aim of the study To explore the inhibitory effect, kinetics and mechanism of Helicobacter pylori urease (the vital pathogenetic factor for Helicobacter pylori infection) inhibition by baicalin and scutellarin, for their therapeutic potential. Materials and methods The ammonia formations, indicator of urease activity, were examined using modified spectrophotometric Berthelot (phenol–hypochlorite) method. The inhibitory effect of baicalin and scutellarin was characterized with IC 50 values, compared to acetohydroxamic acid (AHA), a well known Helicobacter pylori urease inhibitor. Lineweaver–Burk and Dixon plots for the Helicobacter pylori urease inhibition of baicalin and scutellarin was constructed from the kinetic data. SH-blocking reagents and competitive active site Ni 2+ binding inhibitors were employed for mechanism study. Molecular docking technique was used to provide some information on binding conformations as well as confirm the inhibition mode. Moreover, cytotoxicity experiment using Gastric Epithelial Cells (GES-1) was evaluated. Results Baicalin and scutellarin effectively suppressed Helicobacter pylori urease in dose-dependent and time-independent manner with IC 50 of 0.82±0.07 mM and 0.47±0.04 mM, respectively, compared to AHA (IC 50 =0.14±0.05 mM). Structure-activity relationship disclosed 4′-hydroxyl gave flavones an advantage to binding with Helicobacter pylori urease. Kinetic analysis revealed that the types of inhibition were non-competitive and reversible with inhibition constant K i of 0.14±0.01 mM and 0.18±0.02 mM for baicalin and scutellarin, respectively. The mechanism of urease inhibition was considered to be blockage of the SH groups of Helicobacter pylori urease, since thiol reagents ( l , d -dithiothreitol, l -cysteine and glutathione) abolished the inhibitory action and competitive active site Ni 2+ binding inhibitors (boric acid and sodium fluoride) carried invalid effect. Molecular docking study further supported the structure-activity analysis and indicated that baicalin and scutellarin interacted with the key residues Cys321 located on the mobile flap through S H· π interaction, but did not interact with active site Ni 2+ . Moreover, Baicalin (at 0.59–1.05 mM concentrations) and scutellarin (at 0.23–0.71 mM concentrations) did not exhibit significant cytotoxicity to GES-1. Conclusions Baicalin and scutellarin were non-competitive inhibitors targeting sulfhydryl groups especially Cys321 around the active site of Helicobacter pylori urease, representing potential to be good candidate for future research as urease inhibitor for treatment of Helicobacter pylori infection. Furthermore, our work gave additional scientific support to the use of Scutellaria baicalensis in traditional Chinese medicine (TCM) to treat gastrointestinal disorders. [ABSTRACT FROM AUTHOR]

Two new species, Melanoleuca leucopoda and M. porphyropoda , are described based on collections made from Shenyang City, Liaoning Province, China. Melanoleuca leucopoda is mainly characterized by its whitish stipe with fibrils and oblong spores with elongated warts. Melanoleuca porphyropoda differs from all other Melanoleuca species in lacking cystidia and in having decurrent gills and a purplish stipe. The sequences of internal transcribed spacer regions (ITS1-5.8S-ITS2) of Melanoleuca species were analyzed and the results indicated that two new species clustered into two clades and differed from the other species of the genus. The combination of morphological and molecular data confirmed that the two fungi are new species. The morphological similarity of the new species to other species of Melanoleuca and the systematic position of the two species based on molecular data are also discussed. [ABSTRACT FROM AUTHOR]

Background Effects of prenatal Manganese (Mn) exposure at an environmental relevant level on neonatal neurodevelopment remains unclear. Objectives In the multi-center study, we assessed the impact of low level prenatal Mn exposure on neonatal behavioral neurological assessments (NBNA), and explore a threshold umbilical cord blood Mn on neonatal neurological development. Methods We investigated 933 mother-newborn pairs in Shanghai, China, from 2008 through 2009. Umbilical cord serum concentrations of Mn were measured and NBNA tests were conducted. The NBNA contains five clusters: behavior, active tone, passive tone, primary reflexes and general assessment with a maximal total score of 40. The score<37 is defined as low. Results The median serum Mn concentration was 4.0 μg/L. Of the 933 infants, 44 (4.7%) had low NBNA. After adjusting for potential confounders, a high level of Mn (≥75th percentile ) was associated with a lower NBNA score (adjusted ß =−1.1, 95% CI: −1.4–0.7, p <0.01) and a higher risk of low NBNA (adjusted OR=9.4, 95% CI: 3.4–25.7, p <0.01). A nonlinear relationship was observed between cord serum Mn and NBNA after adjusting for potential confounders. NBNA score decreased with increasing Mn levels after 5.0 μg/L(LgMn≥0.7). The cord serum Mn≥5.0 μg/L had adverse effects on behavior, active tone and general reactions of clusters ( p <0.001). Conclusions High prenatal Mn exposure even at an environmental relevant level, is associated with poor fetal neurobehavioral development in a nonlinear pattern. A threshold cord serum Mn of 5.0 μg/L existed for lower neonatal behavioral neurological assessments. [ABSTRACT FROM AUTHOR]

Abstract: In the present study, the inactivation effect of scutellarin (SL) on jack bean urease was investigated to elucidate the inhibitory potency, kinetics and mechanism of inhibition. It was revealed that SL acted as a concentration- and time-dependent inactivator of urease characteristic of slow-binding inhibition with an IC50 of 1.35±0.15mM. The rapid formation of the initial SL–urease complex with an inhibition constant of Ki =5.37×10−2 mM was followed by a slow isomerization into the final complex with the overall inhibition constant of Ki* =3.49×10−3 mM. High effectiveness of thiol protectors, such as L-cysteine (L-cys), 2-mercaptoethanol (2-ME) and dithiothreitol (DTT) significantly slowed down the rate of inactivation, indicating the strategic role of the active site sulfhydryl group in the blocking process. While the insignificant protection by boric acid and fluoride from the inactivation further confirmed that the active site cysteine should be obligatory for urease inhibition, which was also rationalized by the molecular docking study. The inhibition of SL on urease proved to be reversible since SL-blocked urease could be reactivated by DTT application and multidilution. The results obtained indicated that urease inactivation resulted from the reaction between SL and the sulfhydryl group. [Copyright &y& Elsevier]

Abstract: Background: Epidemiological studies examining the effects of prenatal toxic heavy metals exposure on neonatal development at the population-based level are limited. Objective: To explore prenatal exposure levels to toxic heavy metals and examine whether exposure is associated with neonatal development. Methods: A stratified multi-stage cluster sampling methodology was used to investigate 1652 mother–infant pairs from 2008 through 2009 in Shanghai. Cord blood concentrations of Pb, Hg, Cd, As, Tl and NBNA (Neonatal Behavioral Neurological Assessments) were tested. We defined three different exposure levels (low, medium and high) according to the considered safe values. Results: The median blood Pb, Hg, Cd, As and Tl concentrations were 41μg/L, 1.88μg/L, 0.03μg/L, 0.86μg/L and 0.02μg/L, respectively, which all were in the level considered safe. Increasing exposure to Cd, Hg, As and Tl during pregnancy was associated with decreasing NBNA scores. High level-exposure (exceeding the level considered safe) of Hg, Cd and Tl had lower NBNA scores compared to medium and low levels (both in the level considered safe), which implied that the level considered safe of these heavy metals was safe for the newborns'' development. The mean decreasing scores of NBNA was 0.61, 1.50 and 0.84 (total score=40) with high-level exposure of Hg, Cd and Tl, respectively. The medium-level exposure (in the level considered safe) to As had lower NBNA scores compared to low-level exposure, which implied that level of considered safe for As was not safe to the newborns'' development. However, prenatal Pb high-exposure did not affect NBNA scores either by single or multiple factor analysis. In addition, important contribution factors for heavy metals pollutants were diet, lifestyle and housing renovation. Conclusions: Prenatal heavy metals except Pb exposures were associated with NBNA. The adverse effect of medium-level As warrants the need to further investigate the safe range of As. [Copyright &y& Elsevier]

Abstract: As a transcription factor family dependent of oxygen, hypoxia-inducible factors (HIFs) play important roles in organic and cellular homeostasis, which are master regulators of tumorigenesis, angiogenesis and embryogenesis. Up to date, some known posttranslational modifications, such as hydroxylation, acetylation, phosphorylation and S-nitrosylation, have been proved to influence protein stability and transcriptional activity of HIFs. On the other hand, HIFs can be physiologically degraded through ubiquitin-dependent or -independent proteasome pathway. Two biochemical processes involve in many modulators, which are correlated and compose an intricate mechanism to control the biological function of HIFs systemically. Herein, we review and discuss the diverse roles of involved modulators in both HIFs stability and degradation, describe the potential link between the two molecular scenarios. [Copyright &y& Elsevier]

The ability to differentiate stimuli that predict fear is critical for survival; however, the underlying molecular and circuit mechanisms remain poorly understood. We combined transgenic mice, in vivo transsynaptic circuit-dissecting anatomical approaches, optogenetics, pharmacological methods, and electrophysiological recording to investigate the involvement of specific extended amygdala circuits in different fear memory. We identified the projections from central lateral amygdala (CeL) protein kinase C δ (PKCδ)–positive neurons and somatostatin (SST)-positive neurons to GABAergic (gamma-aminobutyric acidergic) and glutamatergic neurons in the ventral part of the bed nucleus of stria terminalis (vBNST). Prolonged optogenetic activation or inhibition of the PKCδCeL-vBNST pathway specifically reduced context fear memory, whereas the SSTCeL-vBNST pathway mainly reduced tone fear memory. Intriguingly, optogenetic manipulation of vBNST neurons that received the projection from PKCδCeL neurons exerted bidirectional regulation of context fear, whereas manipulation of vBNST neurons that received the projection from SSTCeL neurons could bidirectionally regulate both context and tone fear memory. We subsequently demonstrated the presence of δ and κ opioid receptor protein expression within the CeL-vBNST circuits, potentially accounting for the discrepancy between prolonged activation of GABAergic circuits and inhibition of downstream vBNST neurons. Finally, administration of an opioid receptor antagonist cocktail on the PKCδCeL-vBNST or SSTCeL-vBNST pathway successfully restored context or tone fear memory reduction induced by prolonged activation of the circuits. Together, these findings establish a functional role for distinct CeL-vBNST circuits in the differential regulation and appropriate maintenance of fear. [ABSTRACT FROM AUTHOR]

Three new species,Calocybe aurantiaca, C. convexa, andC. decolorata, are described based on collections made in Shenyang City, Liaoning Province, China. The main characters ofC. aurantiacaare its orange-yellow sporocarps and small and smooth basidiospores.Calocybe convexais characterized by its orange-buff pileus, very small basidiospores, and tortuous stipe, whereasC. decoloratais mainly characterized by its gills that turn blue when bruised. The sequences of nuc rDNA ITS1-5.8S-ITS2 (ITS) and the 28S D1–D5 region of theCalocybespecies were analyzed, and the results indicated that the three new species belonged to the genusCalocybeand differed from other species ofCalocybe. The morphological similarities of the new species to otherCalocybespecies and the classification system within the genusCalocybebased on molecular data are also discussed. A key is provided for theCalocybespecies as reported from China in order to facilitate future studies of the genus. [ABSTRACT FROM PUBLISHER]

The wild, edible mushroom Rugiboletus extremiorientalis has an excellent taste and also medicinal effects, and is very popular in China. In this study, hot water extraction is used to isolate polysaccharides from R. extremiorientalis fruiting bodies, and response surface methodology is used to optimize the extraction conditions in terms of temperature, liquid–solid ratio, and time. The crude extract (REP) is purified in turn by DEAE‐52 chromatography and Sephadex G‐100 chromatography, resulting in three fractions (REP‐1, REP‐2, and REP‐3) with recovery rates of 16.7%, 57.65%, and 6.63%, respectively. In vitro antioxidant activity is highest for REP‐2, and the subsequent research is therefore focused on that fraction. High‐performance liquid chromatography reveals that REP‐2 is comprised of fucose, arabinose, galactose, glucose, xylose, and mannose. Fourier‐transform infrared spectroscopy of REP‐2 indicates the presence of functional groups typical for polysaccharides. High‐performance gel filtration chromatography indicates the REP‐2 has a molecular weight of 342.4 kDa. The results of this study provide a foundation for future research concerning the antioxidant activity of polysaccharides in R. extremiorientalis. [ABSTRACT FROM AUTHOR]

Coprinus comatus, widely known as "Jituigu", is an important commodity and food in China. The yield of C. comatus, however, is substantially reduced by the autolysis of the fruiting bodies after harvest. To gain insight into the molecular mechanism underlying this autolysis, we divided the growth of C. comatus fruiting bodies into four stages: infant stage (I), mature stage (M), discolored stage (D), and autolysis stage (A). We then subjected these stages to de novo transcriptomic analysis using high-throughput Illumina sequencing. A total of 12,946 unigenes were annotated and analyzed with the Gene Ontology (GO), Clusters of Orthologous Groups of proteins (COG), and Kyoto Encyclopedia of Genes and Genomes (KEGG). We analyzed the differentially expressed genes (DEGs) between stages I and M, M and D, and D and A. Because the changes from M to D are thought to be related to autolysis, we focused on the DEGs between these two stages. We found that the pathways related to metabolic activity began to vary in the transition from M to D, including pathways named as autophagy—yeast, peroxisome, and starch and sucrose metabolism. This study also speculates the possible process of the autolysis of Coprinus comatus. In addition, 20 genes of interest were analyzed by quantitative real-time PCR to verify their expression profiles at the four developmental stages. This study, which is the first to describe the transcriptome of C. comatus, provides a foundation for future studies concerning the molecular basis of the autolysis of its fruiting bodies. [ABSTRACT FROM AUTHOR]

A method for determination of concentrations of polychlorinated biphenyl congeners (PCB-28, 52, 101, 118, 138, 153, 156, and 187) and organochlorine pesticides (hexachlorobenzene, alpha-hexachlorocyclohexane, beta-hexachlorocyclohexane, gamma-hexachlorocyclohexane, delta-hexachlorocyclohexane, p,p'-dichlorodiphenyl dichloroethylene, o,p'-dichlorodiphenyl trichloroethane, p,p'-dichlorodiphenyl dichloroethane, p,p'-dichlorodiphenyl trichloroethane, alpha-chlordane, gamma-chlordane, heptachlor, heptachlor epoxide, and aldrin) in human serum is developed. Recovery is assessed with artificial serum, in which PCBs and OCPs could not be detected. The method is then confirmed with pooled human serum. Experiments are performed by adding two concentrations of analytes (0.5 µg/L and 1.0 µg/L) to both matrices. The sample pretreatment process involves denaturing with a mixture of water–1-propanol (v:v, 85:15), extraction with a C–18 cartridge, and cleanup with an Alumina B cartridge. This process required about 2 mL of serum. The limit of detection ranged from 0.05–0.35 µg/L for all the analytes. Recovery of analytes at low and high spiking concentrations varied from 63–122% and 61–124% for artificial serum and pooled human serum, respectively. Relative standard deviation was lower than 16% and 18% for artificial serum and pooled human serum, respectively. Stability of the method, expressed as relative standard deviation, was lower than 14%. The method has been applied in epidemiological research. [ABSTRACT FROM PUBLISHER]

Abstract: The use of most polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) has been restricted in China; however, their use remains a concern because of their adverse effects on human health, especially on fetuses and infants. To date, there is no data regarding the exposure levels of pregnant women to PCBs and OCPs in Shanghai. In order to evaluate PCB and OCP exposure levels and the contribution of dietary habits to these levels, we determined the concentrations of 8 PCBs and 14 OCPs in the umbilical cord blood serum of healthy pregnant women in Shanghai. Dietary habits of the pregnant women were obtained from a self-administered questionnaire. Results showed that p, p′-DDE, HCB and β-HCH were the major pollutants present in the serum samples; PCBs were detected in a few samples at low concentrations. Age, weight and body mass index before delivery were positively associated with serum levels of p, p′-DDE and β-HCH. Women and their husbands who had higher education levels, higher income levels, tended to have higher levels of p, p′-DDE and β-HCH. Spearman correlation analysis results suggested that consumption of foods such as milk, eggs, meat, fish, and shrimp may contribute to higher serum levels of p, p′-DDE and β-HCH. Furthermore, multiple linear regression analyses indicated that the age and educational levels of the pregnant women and their intake of fried/flamed food and shellfish were positively associated with β-HCH levels, and that the age and educational levels of the pregnant women and their intake of parity, beef, pork, mutton, and shrimp were positively associated with p, p′-DDE levels. This is the first study to investigate the exposure levels of pregnant women to PCBs and OCPs in Shanghai, and it should provide useful information for future related research. [Copyright &y& Elsevier]

Recognizing the affective states of social counterparts and responding appropriately fosters successful social interactions. However, little is known about how the affective states are expressed and perceived and how they influence social decisions. Here, we show that male and female mice emit distinct olfactory cues after experiencing distress. These cues activate distinct neural circuits in the piriform cortex (PiC) and evoke sexually dimorphic empathic behaviors in observers. Specifically, the PiC → PrL pathway is activated in female observers, inducing a social preference for the distressed counterpart. Conversely, the PiC → MeA pathway is activated in male observers, evoking excessive self-grooming behaviors. These pathways originate from non-overlapping PiC neuron populations with distinct gene expression signatures regulated by transcription factors and sex hormones. Our study unveils how internal states of social counterparts are processed through sexually dimorphic mechanisms at the molecular, cellular, and circuit levels and offers insights into the neural mechanisms underpinning sex differences in higher brain functions. [Display omitted] • Male and female mice display different empathic behaviors in response to others' pain • Female mice show social preference, whereas male mice display excessive self-grooming • Distinct behaviors arise from neural circuits involving two populations of PiC neurons • The two PiC neuron populations exhibit distinct gene expression signatures Fang et al. discovered that male and female mice display different empathic behaviors in response to others' pain. Specifically, males engage in self-grooming, whereas females demonstrate social approaching. Processing information about others' pain involves distinct neural circuits for each sex. Importantly, these circuits inherently differ in gene expression patterns. [ABSTRACT FROM AUTHOR]

Lepista nuda is a popular wild edible mushroom that grows in China. In this study, we used ISSR and SRAP molecular markers to analyze the genetic diversity of 72 samples of L. nuda from eight populations in Northeast China. In total, six ISSR primers and five pairs of SRAP primers that produced clear and polymorphic banding profiles were selected for assessing L. nuda genetic diversity. The results revealed a high level of genetic variation among the 72 samples (94.4% polymorphism) but a low degree of gene flow among the populations. Among L. nuda populations, genetic distance was not correlated significantly with geographic distance. The antioxidant activity of the samples from each population was also tested and the result showed that all the selected samples had more than 60% DPPH scavenging activities. Nonetheless, the antioxidant activity diversity is not coincident with both the genetic diversity and the geographic distribution. The results indicate that ISSR and SRAP molecular markers are useful for studying the genetic diversity of L. nuda. The results also suggest that L. nuda populations in Northeast China require protection. [ABSTRACT FROM AUTHOR]

Pogostemonis Herba is an important Chinese medicine widely used in the treatment of gastrointestinal dysfunction. Patchouli alcohol (PA), a tricyclic sesquiterpene, is the major active constituent of Pogostemonis Herba. This study aimed to investigate the possible anti-ulcerogenic potential of PA and the underlying mechanism against ethanol, indomethacin and water immersion restraint-induced gastric ulcers in rats. Gross and histological gastric lesions, biochemical and immunological parameters were taken into consideration. The gastric mucus content and the antisecretory activity were analyzed through pylorus ligature model in rats. Results indicated that oral administration with PA significantly reduced the ulcer areas induced by ethanol, indomethacin and water immersion restraint. PA pretreatment significantly promoted gastric prostaglandin E 2 (PGE 2 ) and non-protein sulfhydryl group (NP-SH) levels, upregulated the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) mRNA expression, and considerably boosted the gastric blood flow (GBF) and gastric mucus production in comparison with vehicle. In addition, PA modulated the levels of interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α). The levels of glutathione (GSH), catalase (CAT) and malonaldehyde (MDA) were also restored by PA. However, the gastric secretion parameters (pH, volume of gastric juice and pepsin) did not show any significant alteration. These findings suggest that PA exhibited significant gastroprotective effects against gastric ulceration. The underlying mechanisms might involve the stimulation of COX-mediated PGE 2 , improvement of antioxidant and anti-inflammatory status, preservation of GBF and NP-SH, as well as boost of gastric mucus production. [ABSTRACT FROM AUTHOR]

Mycorrhizal association is known to be important to orchid species, and a complete understanding of the fungi that form mycorrhizas is required for orchid ecology and conservation. Liparis japonica (Orchidaceae) is a widespread terrestrial photosynthetic orchid in Northeast China. Previously, we found the genetic diversity of this species has been reduced recent years due to habitat destruction and fragmentation, but little was known about the relationship between this orchid species and the mycorrhizal fungi. The Rhizoctonia-like fungi are the commonly accepted mycorrhizal fungi associated with orchids. In this study, the distribution, diversity and specificity of culturable Rhizoctonia-like fungi associated with L. japonica species were investigated from seven populations in Northeast China. Among the 201 endophytic fungal isolates obtained, 86 Rhizoctonia-like fungi were identified based on morphological characters and molecular methods, and the ITS sequences and phylogenetic analysis revealed that all these Rhizoctonia-like fungi fell in the same main clade and were closely related to those of Tulasnella calospora species group. These findings indicated the high mycorrhizal specificity existed in L. japonica species regardless of habitats at least in Northeast China. Our results also supported the wide distribution of this fungal partner, and implied that the decline of L. japonica in Northeast China did not result from high mycorrhizal specificity. Using culture-dependent technology, these mycorrhizal fungal isolates might be important sources for the further utilizing in orchids conservation. [ABSTRACT FROM AUTHOR]

This study aims to investigate the four vitamin D receptor (VDR) gene single nucleotide polymorphisms and their possible relationship with bone mineral density (BMD) in Chinese 0-6-year-old Han children. Two hundred four 0-6-year-old Han children without metabolic bone disease were randomly recruited in Shanghai, China. The BMD of the middle tibia was measured by an ultrasonic bone density instrument. VDR genotypes were determined by polymerase chain reaction restriction fragment length polymorphism using endonuclease ApaI, BsmI, TaqI and FokI. The alleles of a, T, b and F and the genotypes of aa, TT, bb and Ff were predominant. The frequency alleles of a, T, b and F were, respectively, 70.6, 95.8, 95.3 and 57.6%. When the influences of confounders such as serum 25(OH)D, serum zinc and outdoor activities on BMD were removed, the genotypes of BsmI and FokI were found apparently to be related to BMD. The BMD of the Bb carrier was much lower than that of the bb carrier (22.00 ± 27.84 and 43.14 ± 31.98, P < 0.05). The BMD of the ff carrier was lower than that of the Ff or FF carrier (26.97 ± 34.22 and 37.95 ± 29.70 and 53.52 ± 30.35, P < 0.001), while the genotypes of ApaI and TaqI have no relation with BMD in 0-6-year-old Han children. These findings show that the Bb and ff genotypes of the VDR BsmI and FokI variants are significantly associated with a decreased BMD in Chinese Han children aged 0-6 years, while the VDR ApaI and TaqI polymorphisms are not significantly associated with it. [ABSTRACT FROM AUTHOR]

Abstract: GGA (geranylgeranylacetone) may induce Hsp70 synthesis, thus contributing to the protective effects of GGA in several disease states. This study evaluated the protective effects of GGA against heat injury to rat and striatum neurons in terms of mechanisms. Rats were exposed to 41.5°C for 35min to induce heatstroke; the protective effects of GGA were then evaluated by change in rectal temperature (Tre) during heat exposure and survival time after heatstroke. Primary cultured striatum neurons were incubated with GGA for 24h, and then heat-treated at 43°C for a further 1h. The viability, membrane surface ultrastructure and Hsp70 expression of striatum neurons were all observed. Furthermore, the effects of quercetin an inhibitor of Hsp70 synthesis were also investigated. Compared to the heatstroke group, GGA delayed Tre in reaching 42.1°C (P <0.05) and prolonged the survival time after heatstroke (P <0.01). The LDH releasing percentage decreased in GGA groups (P <0.05, P <0.01) compared to the heat-treatment group and increased in quercetin groups (P <0.05) compared to GGA group. Results from AFM showed that GGA protected membrane surface ultrastructure against heat injury. In addition, results from Western blot showed that GGA-induced Hsp70 expression of neurons both in normal and heat-treatment conditions (P <0.01, P <0.05) and quercetin inhibited GGA-induced Hsp70 expression (P <0.05). Therefore, GGA had protective effects against heat injury in striatum neurons and rat heatstroke. Quercetin inhibited GGA-induced Hsp70 expression and prevented GGA-protective effects, which indicated that this protection was dependent on the Hsp70 synthesis. [Copyright &y& Elsevier]

Abstract: Estrogen receptor α (ERα) plays an important role in the development and progression of breast cancer, and recent studies showed that ERα expression is associated with resistance to hormonal therapy. Therefore, a number of studies have explored ways to deplete ERα from breast cancer cells as a new therapy especially for hormone-refractory breast cancer. We reported here that suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, effectively depletes ERα in breast cancer MCF-7 cells. However, the intrinsic mechanisms by which SAHA decreases ERα levels are not clear. Our present data demonstrated that both inhibition of ERα mRNA level and promotion of ERα degradation by the proteasome contribute to SAHA-induced ERα depletion, indicating that SAHA may exert its effects through transcriptional and posttranslational mechanisms. Furthermore, the decrease of ERα protein level in MCF-7 cells after SAHA treatment is mainly the result of its rapid degradation by the ubiquitin-proteasome pathway rather than transcriptional inhibition. In addition, we showed that inactivation of the heat shock protein-90 (Hsp90) is involved in SAHA-induced ERα degradation, and ubiquitin ligase CHIP (C-terminal Hsc70 interacting protein) enhances SAHA-induced ERα degradation. SAHA-induced ERα depletion is paralleled with reduction of transcriptional activity of ERα and SAHA is able to effectively inhibit cell proliferation and induce apoptosis of MCF-7 cells. Taken together, our results revealed a mechanism for SAHA-induced ERα degradation and indicated that SAHA is a suitable pharmacological agent for depletion of ERα and a potential choice for breast cancer expressing high ERα. [Copyright &y& Elsevier]

Abstract: Exposure to hazardous Hg can adversely affect children's neurodevelopment. However, few data are available on either Hg levels in neonates and their mothers or the impact of prenatal exposure to Hg on neonates’ neurobehavioral development in the Chinese population. Therefore, this study examined Hg levels in neonates and their mothers and the relationship between prenatal exposure to Hg and neonates’ neurobehavioral development in Zhoushan City, Zhejiang Province, China. Between August and October 2004, 417 women who delivered their babies at Zhoushan Women''s and Children''s Health Hospital, an island city in east China were invited to take part in this study. A total of 408 complete questionnaires, 405 maternal hair samples, and 406 umbilical cord samples were collected. Neonatal behavioral neurological assessments (NBNA) were conducted for 384 neonates. The geometric mean (GM) of Hg level in cord blood was 5.58μg/L (interquartile range: 3.96–7.82μg/L), and the GM of maternal hair Hg level was 1246.56μg/kg (interquartile range: 927.34–1684.67μg/kg), a level much lower than other reported fish-eating populations, indicating Hg exposure in Zhoushan city is generally below those considered hazardous. However, according to the reference dose of Hg levels (RfD 5.8μg/L) derived by EPA, 69.9% of newborns had levels at or above the RfD, an estimated level assumed to be without appreciable harm. There was a strong correlation between maternal hair and cord blood Hg levels (r=0.82, P<0.01). Frequency of fish consumption was associated with hair Hg (r=0.48, P<0.01) and cord blood Hg levels (r=0.54, P<0.01). Increased prenatal Hg exposure was associated with decreased behavioral ability for males (OR=1.235, 95%CI of OR=1.078–1.414, P<0.001), but not for females. Our results provide some support for the hypothesis that there is neurodevelopmental risk for males from prenatal MeHg exposure resulting from fish consumption. But the findings of this study may be due to chance, and long-term follow-up research is needed to evaluate cumulative effects of exposure to mercury. [Copyright &y& Elsevier]

Abstract: Some pan-histone-deacetylase (HDAC) inhibitors have recently been reported to exert their anti-leukemia effect by inhibiting the activity of class IIB HDAC6, which is the deacetylase of Hsp90 and α-tubulin, thereby leading to hyperacetylation of Hsp90, disruption of its chaperone function and apoptosis. In this study, we compared the effect of a class I HDAC inhibitor FK228 with the pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) on the Hsp90 chaperone function of K562 cells. We demonstrated that, although having a weaker inhibitory effect on HDAC6, FK228 mediated a similar disruption of Hsp90 chaperone function compared to SAHA. Unlike SAHA, FK228 did not mediate hyperacetylation of Hsp90, instead the acetylation of Hsp70 was increased and Bcr-Abl was increasingly associated with Hsp70 rather than Hsp90, forming an unstable complex that promotes Bcr-Abl degradation. These results indicated that FK228 may disrupt the function of Hsp90 indirectly through acetylation of Hsp70 and inhibition of its function. [Copyright &y& Elsevier]

Abstract: Exposure to lead in utero and in infancy is associated with a risk of impaired cognitive development. Increasing evidence suggests that the family of metabotropic glutamate receptors (mGluRs) plays an important role in synaptic plasticity and memory formation. We determined whether mGluRs subtypes 1, 3, and 7 (mGluR1, mGluR3, and mGluR7) were involved in developmental neurotoxicity due to lead. Embryonic rat hippocampal neurons were cultured for 21 days and exposed to lead chloride beginning on the fourth day of incubation. We investigated levels of mGluR1, mGluR3, and mGluR7 mRNA expression by using quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR) with lead exposure at 10nM, 1μM, and 100μM. Lead exposure in vitro downregulated the expression of mGluR1 mRNA and upregulated the expression of mGluR3 and mGluR7 mRNA in a dose-dependent manner. We speculate that mGluRs may be involved in lead neurotoxicity. Pathways that likely contribute to lead neurotoxicity by means of mGluRs are impairment of long-term potentiation, effects on N-methyl-d-aspartate (NMDA) receptor functions, and depotentiation. [Copyright &y& Elsevier]

• This is the first anti-tumor study on Nano-realgar and metabolic reprogramming. • Nano-realgar inhibits LCSCs viability and tumor growth. • Nano-realgar induces glucose metabolism and inhibits related protein expression. • Nano-realgar downregulated the expression of HIF-1α and PI3K/Akt/mTOR pathways. • Nano-realgar inhibits the constitutive activation of HIF-1α signaling. Recent studies in cancer biology suggest that metabolic glucose reprogramming is a potential target for cancer treatment. However, little is known about drug intervention in the glucose metabolism of cancer stem cells (CSCs) and its related underlying mechanisms. The crude realgar powder was Nano-grinded to meets the requirements of Nano-pharmaceutical preparations, and Nano-realgar solution (NRS) was prepared for subsequent experiments. Isolation and characterization of lung cancer stem cells (LCSCs) was performed by magnetic cell sorting (MACS) and immunocytochemistry, respectively. Cell viability and intracellular glucose concentration were detected by MTT assay and glucose oxidase (GOD) kit. Protein expressions related to metabolic reprogramming was detected by ELISA assay. Determination of the expression of HIF-1α and PI3K/Akt/mTOR pathways was carried out by RT-PCR and western blotting analysis. A subcutaneous tumor model in BALB/c-nu mice was successfully established to evaluate the effects of Nano-realgar on tumor growth and histological structure, and the expression of HIF-1α in tumor tissues was measured by immunofluorescence. Nano-realgar inhibits cell viability and induces glucose metabolism in LCSCs, and inhibits protein expression related to metabolic reprogramming in a time- and dose-dependent manner. Nano-realgar downregulated the expression of HIF-1α and PI3K/Akt/mTOR pathways in vitro and in vivo. Nano-realgar inhibits tumor growth and changes the histological structure of tumors through in vivo experiments and consequently inhibits the constitutive activation of HIF-1α signaling. These results reveal that Nano-realgar inhibits tumor growth in vitro and in vivo by repressing metabolic reprogramming. This inhibitory effect potentially related to the downregulation HIF-1α expression via PI3K/Akt/mTOR pathway. [ABSTRACT FROM AUTHOR]

• L. paracasei FZU103 has the potential to protect against NAFLD associated with hyperlipidemia. • The probiotic L. paracasei FZU103 can regulate weight and glycolipid levels in mice. • L. paracasei FZU103 alters the intestinal microecological environment of mice fed with HFD. • Energy, amino acid, fatty acid and lipid metabolisms play a major role in the preventive effect. • The mRNA expression of lipid metabolism were influenced by L. paracasei FZU103 intervention. Lactobacillus paracasei FZU103, a probiotic previously isolated from the traditional brewing process of Hongqu rice wine, may have the beneficial effect of improving the disorder of lipid metabolism. This study aimed to determine the beneficial effects of L. paracasei FZU103 on improving hepatic lipid accumulation associated with hyperlipidemia. Results indicated that L. paracasei FZU103 intervention significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, ameliorated the biochemical parameters of serum and liver related to lipid metabolism in HFD-fed mice. Histological analysis also showed that the excessive accumulation of lipid droplets in the livers induced by HFD-feeding was greatly alleviated by L. paracasei FZU103 intervention. In addition, L. paracasei FZU103 also promoted the excretion of bile acids (BAs) through feces. Metagenomic analysis revealed that oral supplementation with L. paracasei FZU103 significantly increased the relative abundance of Ruminococcus , Alistipes , Pseudoflavonifractor and Helicobacter , but decreased the levels of Blautia , Staphylococcos and Tannerella in HFD-fed mice. The relationships between lipid metabolic parameters and intestinal microbial phylotypes were also revealed by correlation heatmap and network. Furthermore, ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS)-based liver metabolomics demonstrated that L. paracasei FZU103 had a significant regulatory effect on the metabolic pathways of glycerophospholipid metabolism, fatty acid degradation, fatty acid elongation, unsaturated fatty acids biosynthesis, riboflavin metabolism, glycerolipid metabolism, primary bile acid biosynthesis, arachidonic acid metabolism, etc. Additionally, L. paracasei FZU103 intervention regulated expression of hepatic genes involved in lipid metabolism and bile acid homeostasis, and promoted fecal excretion of intestinal BAs. These findings present new evidence supporting that L. paracasei FZU103 has the potential to improve lipid metabolism, and could be used as a potential functional food for the prevention of hyperlipidemia. [ABSTRACT FROM AUTHOR]

Background: Cladribine or 2-chlorodeoxyadenosine (2-CDA) is a well-known purine nucleoside analog with particular activity against lymphoproliferative disorders, such as hairy cell leukemia (HCL). Its benefits in multiple myeloma (MM) remain unclear. Here we report the inhibitory effects of cladribine on MM cell lines (U266, RPMI8226, MM1.S), and its therapeutic potential in combination with a specific inhibitor of the signal transducer and activator of transcription 3 (STAT3).Methods: MTS-based proliferation assays were used to determine cell viability in response to cladribine. Cell cycle progression was examined by flow cytometry analysis. Cells undergoing apoptosis were evaluated with Annexin V staining and a specific ELISA to quantitatively measure cytoplasmic histone-associated DNA fragments. Western blot analyses were performed to determine the protein expression levels and activation.Results: Cladribine inhibited cell proliferation of MM cells in a dose-dependent manner, although the three MM cell lines exhibited a remarkably different responsiveness to cladribine. The IC50 of cladribine for U266, RPMI8226, or MM1.S cells was approximately 2.43, 0.75, or 0.18 μmol/L, respectively. Treatment with cladribine resulted in a significant G1 arrest in U266 and RPMI8226 cells, but only a minor increase in the G1 phase for MM1.S cells. Apoptosis assays with Annexin V-FITC/PI double staining indicated that cladribine induced apoptosis of U266 cells in a dose-dependent manner. Similar results were obtained with an apoptotic-ELISA showing that cladribine dramatically promoted MM1.S and RPMA8226 cells undergoing apoptosis. On the molecular level, cladribine induced PARP cleavage and activation of caspase-8 and caspase-3. Meanwhile, treatment with cladribine led to a remarkable reduction of the phosphorylated STAT3 (P-STAT3), but had little effect on STAT3 protein levels. The combinations of cladribine and a specific STAT3 inhibitor as compared to either agent alone significantly induced apoptosis in all three MM cell lines.Conclusions: Cladribine exhibited inhibitory effects on MM cells in vitro. MM1.S is the only cell line showing significant response to the clinically achievable concentrations of cladribine-induced apoptosis and inactivation of STAT3. Our data suggest that MM patients with the features of MM1.S cells may particularly benefit from cladribine monotherapy, whereas cladribine in combination with STAT3 inhibitor exerts a broader therapeutic potential against MM. [ABSTRACT FROM AUTHOR]