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Community-acquired pneumonia (CAP) is one of the leading causes of morbidity and mortality in children worldwide. In this study, we aimed to describe the aetiology of viral infection of pediatric CAP in Chinese mainland. During November 2014 to June 2016, the prospective study was conducted in 13 hospitals. The hospitalized children under 18 years old who met the criteria for CAP were enrolled. The throat swabs or nasopharyngeal aspirates (NPAs) were collected which were then screened 18 respiratory viruses using multiplex PCR assay. Viral pathogens were present in 56.6% (1539/2721) of the enrolled cases, with the detection rate of single virus in 39.8% of the cases and multiple viruses in 16.8% of the cases. The most frequently detected virus was respiratory syncytial virus (RSV) (15.2%, 414/2721). The highest detection rate of virus was in

Free-range chickens might mediate the spread of Cryptosporidium oocysts to humans and other animals. Few studies have evaluated the prevalence of Cryptosporidium species in domestic free-range poultry in China. Here, we characterized the prevalence and distribution of species and genotypes of Cryptosporidium in domestic free-range chickens, ducks, and geese in Anhui Province, China. A total of 1910 fresh fecal samples from three poultry species were examined from 18 free-range poultry farms by nested PCR and analysis of the Cryptosporidium SSU rRNA gene. The overall prevalence of Cryptosporidium species was 2.9% (55/1910), with infection rates of 1.3% (11/829) in chickens, 7.3% (36/487) in ducks, and 1.4% (8/594) in geese. C. baileyi (0.6%), C. meleagridis (0.2%), C. galli (0.2%), and C. xiaoi-like genotype (0.2%) were identified in chickens, and only C. baileyi was identified in ducks and geese, with infection rates of 7.4% and 1.3%, respectively. C. baileyi was the most prevalent species. Sequencing of the GP60 gene revealed that the C. meleagridis isolates belonged to the IIIbA26G1R1b subtype. This is the first study to document C. galli and C. xiaoi-like genotype in domestic free-range chickens in China. These findings expand the range of avian hosts known for Cryptosporidium and highlight the need for additional studies to characterize the diversity of Cryptosporidium in avian species.

The etiology of primary Sjogren's syndrome (pSS) remains unknown, and there is no complete curative drug. In this study, we treated a mouse model of the submandibular gland (SG) protein-immunized experimental Sjogren's syndrome (ESS) with paeoniflorin-6'-O-benzene sulfonate (termed CP-25) to evaluate the potential therapeutic effects of CP-25. Through in vivo experiments, we found that CP-25 increased saliva flow, alleviated the salivary gland indexes, and improved tissue integrity in the ESS model. The viability of splenocytes and B-lymphocyte migration from spleen were reduced in ESS mice. Furthermore, CP-25 decreased the expression of IgG antibodies, anti-SSA and anti-SSB antibodies and modulated the levels of cytokines in the serum of SS mice. The numbers of total B lymphocytes, plasma cells (PCs), and memory B cells diminished in the salivary gland. Increased expression of the JAK1-STAT1-CXCL13 axis and IFNα was found in human tissue isolated from pSS patients. In vitro, after stimulation with IFNα, the levels of CXCL13 mRNA and CXCL13 in human salivary gland epithelial cells (HSGEC) increased, while CP-25 counteracted the secretion of CXCL13 and downregulated the expression of CXCL13. IFN-α activated the JAK1-STAT1/2-CXCL13 signaling pathway in HSGEC, which was negatively regulated by additional CP-25. As a consequence, B-cell migration was downregulated in coculture with IFN-α-stimulated HSGEC. Taken together, this study demonstrated that the therapeutic effects of CP-25 were associated with the inhibition of the JAK1-STAT1/2-CXCL13 signaling pathway in HSGEC, which impedes the migration of B cells into the salivary gland. We identified the underlying mechanisms of the therapeutic effect of CP-25 and provided an experimental foundation for CP-25 as a potential drug in the treatment of the human autoimmune disorder pSS.

Background Emerging evidence has revealed that circular RNAs (circRNAs) participated in hepatocellular carcinoma (HCC) development. However, the roles of most circRNAs have not been studied. Methods CircZNF609, miR-342-3p and RAP2C expressions were assessed by qPCR or Western blot. Loss-of-function experiments were performed using si-circZNF609 transfection, followed by CCK-8 assay, flow cytometry, wound healing assay and transwell assay. Informatic tools and rescue experiments were carried out to investigate the underlying mechanisms. Results We showed that circZNF609 was overexpressed in HCC tissues and cells, as well as associated with poor clinical characteristics. Depletion of circZNF609 restrained HCC cell viability, migration and invasion while enhanced cell apoptosis. As to mechanism, miR-342-3p was sponged by circZNF609, and RAP2C was targeted by miR-342-3p. The effects on HCC cells induced by si-circZNF609 could be reversed by miR-342-3p inhibitor or RAP2C. In vivo, circZNF609 knockdown inhibited tumorigenesis of HCC mice, confirming the findings in vitro. Conclusion CircZNF609 was highly expressed in HCC tissues and driven HCC progression by sponging miR-342-3p and upregulating RAP2C. This study may provide new potential therapeutic targets for HCC treatment.

To explore the potential clinical value of platelet parameters in early pregnancy in predicting gestational diabetes mellitus (GDM).A total of 1188 singleton pregnant women were included in the regular antenatal examination and delivered in the First Affiliated Hospital of Sun Yat-Sen University from January 2016 to December 2018, who had no pre-pregnancy diabetes, no factors leading to elevated blood glucose level, no medical complications and no other obstetrical complications. Blood routine examination was performed at the 11-13There were significant differences in platelet count (PLT), platelet distribution width (PDW), mean platelet volume (MPV), and plateletcrit (PCT) between the GDM group and the normal group (MPV and PCT in early pregnancy are potential indicators in predicting gestational diabetes mellitus.

Bioactive peptides are an emerging area of biomedical research in the study of numerous human diseases, including acute alcoholic liver injury (AALI). To study the role and mechanism of the milk‑derived hexapeptide Pro‑Gly‑Pro‑Ile‑Pro‑Asn (PGPIPN) in preventing and reducing AALI, the present study established a mouse model of AALI. PGPIPN was used as a therapeutic drug, and glutathione (GSH) was used as a positive control. The body and liver weights of mice were measured, and the liver indexes were calculated to observe mice health. The pathological morphology of liver tissues stained with hematoxylin and eosin were examined to analyze hepatic injury, and hepatocyte apoptosis was measured with a TUNEL assay. The concentrations or activities of alanine aminotransferase (ALT), aspartate aminotransferase, tumor necrosis factor‑α, interleukin (IL)‑1β, IL‑6, triglyceride, total cholesterol, malondialdehyde, superoxide dismutase and GSH peroxidase (GSH‑PX) were detected in serum and/or liver homogenates. The 78 kDa glucose‑regulated protein (GRP78), protein kinase R‑like (PKR) endoplasmic reticulum kinase (PERK), phosphorylated (p)‑PERK, eukaryotic initiation factor 2α (eIF‑2α), p-eIF-2α, inositol‑requiring enzyme 1α (IRE‑1α), spliced X‑box binding protein 1 (XBP‑1s), C/EBP homologous protein (CHOP), caspase‑3 and cleaved caspase‑3 proteins associated with endoplasmic reticulum stress in hepatocytes were assessed by western blotting, and RNA levels of XBP‑1s, CHOP and caspase‑3 genes were assessed by reverse transcription‑quantitative PCR. The results suggested that PGPIPN attenuated alcoholic hepatocyte damage in animal models and reduced hepatocyte oxidative stress in a dose‑dependent manner. Moreover, PGPIPN reduced endoplasmic reticulum stress by regulating the expression levels of p‑PERK, p‑eIF‑2α, XBP‑1s, CHOP, caspase‑3 and cleaved caspase‑3. Collectively, the present results indicated that PGPIPN, as a potential therapeutic drug for AALI, exerted a protective effect on the liver and could reduce liver damage.

Objective: To compare curative effect and safety of omeprazole under different treatment courses in treatment of children with peptic ulcer (PU, diameter ≤1.0cm) and helicobacter pylori (HP) infection and its influence on inflammatory cytokines. Methods: The study was a randomized controlled study and conducted at Baoding children’s hospital from June 2015 to June 2018. In this study 100 PU children with positive HP were chosen and classified into two groups at random. The 58 cases in the observation group were given omeprazole + amoxicillin + clarithromycin, and the antibiotics were not used two weeks later. Then, omeprazole was used to treat for two weeks. 42 cases in the control group were given omeprazole + amoxicillin + clarithromycin for two weeks. Curative effect, HP eradication rate, clinical symptoms, incidence of adverse reactions, level of serum inflammatory cytokine interleukin-6 (IL-6) and level of tumor necrosis factor-a (TNF-a) in two groups were compared. Results: Total effective rate, HP eradication rate and clinical symptom relief of observation group were better than those of control group, and the differences showed statistical significance (P

The trichomonad species Tetratrichomonas buttreyi and Pentatrichomonas hominis have been reported in the bovine digestive tract in only a few studies, and the prevalence and pathogenicity of these two protists in cattle herds remain unknown. In this study, the prevalence of T. buttreyi and P. hominis in yellow cattle, dairy cattle, and water buffalo in Anhui Province, China, was determined with a PCR analysis of the small subunit ribosomal RNA genes. The overall infection rates for T. buttreyi and P. hominis were 8.1% and 5.4%, respectively. Double infections were found in 15 (1.6%) samples from four farms. The prevalence of P. hominis in cattle with abnormal feces was significantly higher than that in cattle with normal feces (χ2 = 13.0, p

Annular syphilis may range from mildly raised lesions with scaly borders to verrucous plaques. Localized annular syphilis on the genitalia has been rarely reported in HIV-negative cases. This paper reports a case of annular secondary syphilis on the penis. Dermoscopy showed peripheral dotted and short linear vessels and white scaling with a relatively clear central area in an erythematous annular plaque. Histopathology revealed mild hyperkeratosis, parakeratosis, psoriasiform acanthosis, and focal basal vacuolar degeneration with lichenoid, perivascular, and periadnexal infiltrate of lymphohistiocytes and plasma cells in the superficial dermis. Silver stain showed several spirochetes in the lower epidermis and superficial dermis. Electron microscopy revealed a few intercellular and intracytoplasmic spirochetes in the basal epidermis and free spirochetes in the papillary dermis. Rapid plasma reagin and Treponema pallidum particle agglutination assays were positive. The lesions disappeared after intramuscular benzathine penicillin, with no relapse at six-month follow-up.

The cat is a reported reservoir for several zoonotic pathogens, including Cryptosporidium spp., Giardia duodenalis, and Blastocystis sp. These parasites represent a significant, but often neglected, threat to humans and animals. Furthermore, Tritrichomonas foetus has been described inhabiting the digestive tract of cats, and may be causative agents of gastrointestinal symptoms. However, scant data are available concerning the molecular epidemiology of these parasites in domestic cats in China. This study examined fecal samples from domestic cats in Eastern China to unravel the molecular epidemiology of four protozoans. Of the 346 samples examined, 47 (13.6%) were positive for the detected pathogens, including 8 (2.3%), 5 (1.4%), 2 (0.6%), and 35 (10.1%) samples positive for Cryptosporidium spp., G. duodenalis, Blastocystis sp., and T. foetus, respectively. Co-infection with Cryptosporidium spp. and T. foetus was detected in three cats, no other mixed infections were observed. No age, sex or fecal condition predisposition was observed with any of the four pathogens. The species/assemblages/subtypes/genotypes were C. felis, Assemblage A and F, ST1, and cat genotype for Cryptosporidium spp., G. duodenalis, Blastocystis sp., and T. foetus detected in this study, respectively. The presence of zoonotic species/assemblages/subtypes/genotypes poses a threat to public health. These findings provide useful information for the design of prevention and control strategies to reduce the burden of protozoal infections in cats.

Identifying molecular mediators of neural circuit development and/or function that contribute to circuit dysfunction when aberrantly reengaged in neurological disorders is of high importance. The role of the TWEAK/Fn14 pathway, which was recently reported to be a microglial/neuronal axis mediating synaptic refinement in experience-dependent visual development, has not been explored in synaptic function within the mature central nervous system. By combining electrophysiological and phosphoproteomic approaches, we show that TWEAK acutely dampens basal synaptic transmission and plasticity through neuronal Fn14 and impacts the phosphorylation state of pre- and postsynaptic proteins in adult mouse hippocampal slices. Importantly, this is relevant in two models featuring synaptic deficits. Blocking TWEAK/Fn14 signaling augments synaptic function in hippocampal slices from amyloid-beta–overexpressing mice. After stroke, genetic or pharmacological inhibition of TWEAK/Fn14 signaling augments basal synaptic transmission and normalizes plasticity. Our data support a glial/neuronal axis that critically modifies synaptic physiology and pathophysiology in different contexts in the mature brain and may be a therapeutic target for improving neurophysiological outcomes.

Lactobacillus casei (L. casei), a kind of probiotics, is known as a "healthy triple benefit bacterium" along with Lactobacillus acidophilus and Bifidobacterium. L. casei is associated with the alteration of the intestinal flora population, and the gut microbiota-brain axis has been demonstrated to play an important role in many central nervous system diseases. The aim of this study was to evaluate the effectiveness of L. casei intervention in ameliorating mental disorders and potential mechanisms using a rat model with depression-like behaviour induced by chronic unpredictable mild stress (CUMS). L. casei intervention improved the CUMS-induced depression-like behaviors of rats, including a reduced body growth rate, decreased sucrose preference, increased immobility time, and lowered moving distance and velocity. In addition, L. casei intervention amended the gut microbiota structure changes induced by CUMS in rats. Furthermore, L. casei intervention reversed the CUMS-induced protein expression changes of monoamines dopamine, noradrenaline and 5-hydroxytryptamine, brain-derived neurotrophic factor (BDNF) and its receptor tyrosine kinase receptor B (TrkB), and N-methyl-d-aspartic acid receptor 1, as well as CUMS-induced activations of ERK1/2 and p38 MAPK signal pathways. These findings suggested that L. casei could significantly protect against depression in rats, which was possibly associated with the alterations in the gut microbiota composition and mediation of BDNF-TrkB signaling.

Several Cryptosporidium species that infect reptiles, especially squamates, are well described, but there is limited data about Cryptosporidium species infecting crocodilians. In this study, we assess the occurrence of intestinal parasites using traditional microscopic examination and describe the prevalence and Cryptosporidium species in the captive-bred Chinese alligators (Alligator sinensis) in eastern China using molecular methods. The results of microscopic examination showed that no intestinal parasites were detected among the 491 fecal samples examined from the Chinese alligators. The overall prevalence for Cryptosporidium was 0.41% (2/491) by PCR detection using the SSU rRNA locus. Sequence and phylogenetic analysis of the SSU rRNA, COWP, and actin genes revealed the presence of Cryptosporidium testudinis, which has been isolated primarily from chelonians. This is the first detection of the specific DNA of C. testudinis in the feces of the Chinese alligator. This study expands our knowledge of the Cryptosporidium species involved in crocodiles, and more extensive studies are necessary to confirm the validity of C. testudinis in crocodiles.

Three Entamoeba spp. including E. suis, zoonotic E. polecki, and E. histolytica, have been described in pigs to date. However, little is known about the molecular epidemiology of these neglected parasites in pigs globally. In this study we surveyed the occurrence and molecular epidemiology of porcine Entamoeba spp. in pigs in eastern China and evaluated their zoonotic potential. Five hundred fresh fecal samples, collected from seven pig farms in Anhui province, eastern China,were examined for the presence of E. histolytica, E. suis, and E. polecki ST1 and ST3 infections by a combination of nested PCR targeting the small subunit ribosomal DNA gene and subsequent sequencing.The overall occurrence of Entamoeba spp. was 45.8% (229/500). Infection with E. polecki ST1 (38.2%; 191/500) was the most common, followed by E. polecki ST3 (10.0%; 50/500), and E. suis (0.8%; 4/500). No E. histolytica infection was detected. Double infections with E. polecki ST1 and E. suis, and with E. polecki ST1 and ST3 were found in two (0.4%) and 14 (2.8%) samples, respectively. No age predisposition to infection with Entamoeba spp. was observed. PCR and subsequent sequencing confirmed the validity and feasibility of the nested PCR method used in this study in identifying species/subtypes of porcine Entamoeba spp.This is the first report to describe the occurrence and molecular epidemiology of Entamoeba species in pigs in China. The presence of two zoonotic E. polecki subtypes implies that pigs can be reservoirs for human E. polecki infections. More studiess are needed to better understand the transmission and public health significance of porcine Entamoeba spp.

The Brazil-Falklands/Malvinas Confluence (BFMC), a highly energetic convergence of surface currents in the western South Atlantic, has shifted southward in recent years and this shift is projected to progress in the future. Palaeoecological insights documenting past changes of these currents may help in anticipating future impacts on the environment. We used dinoflagellate cyst analyses from a marine sediment core to reconstruct environmental changes in the Argentine continental margin, western South Atlantic, during the last ca. 12,600 years. The dynamics of the BFMC were reconstructed using the relative frequency of warm-water indicators for the Brazil Current (BC) versus cold-water taxa thriving in the Falklands/Malvinas Current (FMC). We found that the latitudinal position of the BFMC was relatively stable with only minor amplitude migrations between 12.6 and 8.7 cal kyr BP, followed by periods with stronger shifts to the south and the north until 0.66 cal kyr BP. After that, the BFMC shifted continuously to the south. The increase in freshwater algae abundance after ca. 5.7 cal kyr BP suggests an increase in precipitation over the adjacent Rio de la Plata drainage basin in south-eastern South America. As previously documented, such an increase in precipitation was probably related to a higher El Nino-Southern Oscillation frequency and longer, stronger El Nino events since the mid-Holocene. The dinoflagellate cyst record indicates a phase of the enhanced presence of nutrient-rich waters over the core site between ca. 6.3 and 5.7 cal kyr BP, as well as from 0.66 cal kyr BP to the recent. The highest eutrophication in the ocean surface occurred during the last ca. 100 years, most probably due to stronger human impact in the area of the Rio de la Plata drainage basin.

Preeclampsia (PE), a common obstetrical disorder, is one of the leading causes of pregnancy associated death. PE is closely linked with impaired migration and invasion ability of trophoblastic cells. miR-362-3p recently received our particular attention due not only to its aberrant expression in the placentas of patients with PE, but also to its important roles in regulating migration and invasion of various cells. This study was thus conducted to investigate the roles of miR-362-3p in PE and the related mechanism. The expression of miR-362-3p and Pax3 was examined in placentas of patients with PE and in normal placentas. HTR8/SVneo cells were cultured under hypoxia and transfected with miR-362-3p mimics, miR-362-3p inhibitors or Pax3 over-expression vectors. Results showed up-regulation of miR-362-3p but down-regulation of Pax3 in placentas of preeclamptic pregnancies. Luciferase report assay confirmed that Pax3 is a direct target of miR-362-3p. Although Pax3 was predicted to be targeted by miR-30a-3p and miR-181a-5p as well, their expression either had no difference between placentas of PE patients and normal placentas or showed less increment in placentas of PE patients than miR-362-3p. Exposure to hypoxia inhibited cell viability, migration and invasion of HTR8/SVneo cells. Increasing miR-362-3p by the mimics conferred improved effects on the inhibition. However, deletion of miR-362-3p or overexpression of Pax3 abolished the inhibiton. These results suggest that miR-362-3p/Pax3 axis regulates cell viability, migration and invasion of HTR8/SVneo cells under hypoxia. The present study adds to the further understanding of the pathogenesis of PE.

Oxidative stress is a major cause of adverse outcomes in preeclampsia (PE). Ferroptosis, i.e. programmed cell death from iron-dependent lipid peroxidation, likely mediates PE pathogenesis. We evaluated specific markers for ferroptosis in normal and PE placental tissues, using in vitro (trophoblasts) and in vivo (rat) models. Increase in malondialdehyde content and total Fe2+ along with reduced the glutathione content and glutathione peroxidase activity was observed in PE placenta. While the trophoblasts experienced death under hypoxia, inhibitors of ferroptosis, apoptosis, autophagy, and necrosis increased the cell viability. Microarrays, bioinformatic analysis, and luciferase reporter assay revealed that upregulation of miR-30b-5p in PE models plays a pivotal role in ferroptosis, by downregulating Cys2/glutamate antiporter and PAX3 and decreasing ferroportin 1 (an iron exporter) expression, resulting in decreased GSH and increased labile Fe2+. Inhibition of miR-30b-5p expression and supplementation with ferroptosis inhibitors attenuated the PE symptoms in rat models, making miR-30b-5p a potential therapeutic target for PE. Keywords: miR-30-5p, Ferroptosis, Preeclampsia, SLC7A11, Ferroportin 1

Enterocytozoon bieneusi, the dominant zoonotic species of microsporidia, has been reported in various animals and humans. However, relatively little information is available concerning the worldwide prevalence and genotypes of E. bieneusi in ruminants, such as sheep and goats. The present study was conducted to evaluate the prevalence and genetic characteristics of E. bieneusi in sheep and goats in east-central China. Fresh fecal samples from 832 sheep and 781 goats were evaluated for the presence of E. bieneusi using PCR and sequencing of the ribosomal internal transcribed spacer. 28 sheep specimens (3.4%) and 32 goat specimens (4.1%) were positive for E. bieneusi. A total of 12 ITS genotypes were identified, including 7 known genotypes (COS-I, OEB1, BEB6, CHG1, CHG3, COS-II, and CHC8) and 5 novel genotypes (named AHS1, AHS2, JSS1, AHG1, and AHG2). The dominant genotypes in sheep and goats were BEB6 and CHG3, respectively. Phylogenetic analysis revealed that all genotypes obtained in this study belonged to the so-called “cattle-specific” Group 2. These findings show a relatively low occurrence and genetic diversity of E. bieneusi in sheep and goats in east-central China. Further studies are required to elucidate the role of sheep and goats in the epidemiology of microsporidia.

Scientific projections point to the increase of greenhouse gas levels in the atmosphere will turn the oceans warmer and, consequently, the upper layer of the thermohaline circulation will get thicker, restricting the productivity areas. Oligotrophic conditions will tend to prevail, and expanding the range of proxy data in oligotrophic oceans, across different climate scenarios, has the potential to improve predictions of climate models about how primary productivity will respond. Still, the variability of past primary production in the equatorial Atlantic remains elusive. We used organic-walled dinoflagellate cysts (dinocysts) assemblages to reconstruct changes in the sea surface hydrography and productivity in the western equatorial Atlantic over the last ~130 kyr. Four dinocysts assemblages were defined: open ocean, river outflow, neritic and nutricline assemblages. Multiple proxies were used to support the environmental changes evidenced by the observed variations in the dinocyst assemblages, such as the X-ray fluorescence (XRF) Ti/Ca ratio, and planktonic foraminifera abundances. We observed glacial-interglacial as well as the millennial-scale changes in the dinocyst assemblages. A clear glacial-interglacial pattern was marked by more autotrophs dinocysts over the interglacials and the prevalence of heterotrophs dinocysts during the glacial period. We pointed out the interchange of autotrophs assemblages (nutricline and open ocean) over the interglacials intervals, denoting divergences in nutrient availability. During the last glacial period, precipitation events occurred in northeastern Brazil due to southward displacements of the Intertropical Convergence Zone (ITCZ). The increase of precipitation boosted the freshwater input to the adjacent ocean, and the dinocyst assemblages oscillated in response to such freshwater/salinity changes. Over the glacial period, dinocysts with different nutritional requirements (autotrophs and heterotrophs) varied following the ITCZ displacements. Thus, in this study, we showed the influence of ocean-atmospheric processes on the ocean and investigated the productivity in an oligotrophic region over distinct past climatic backgrounds.

Fatty acid biosynthesis is essential to bacterial growth in Gram-negative pathogens. Several small molecules identified through a combination of high-throughput and fragment screening were cocrystallized with FabH (β-ketoacyl-acyl carrier protein synthase III) from Escherichia coli and Streptococcus pneumoniae. Structure-based drug design was used to merge several scaffolds to provide a new class of inhibitors. After optimization for Gram-negative enzyme inhibitory potency, several compounds demonstrated antimicrobial activity against an efflux-negative strain of Haemophilus influenzae. Mutants resistant to these compounds had mutations in the FabH gene near the catalytic triad, validating FabH as a target for antimicrobial drug discovery.

A new type of photocurable resin composite containing nano-MgO was synthesized in order to reduce the occurrence of secondary caries. Different mass ratios (0 %, 1 %, 2 %, 4 %, 8 %) of nano-MgO were added into resin composites. The antibacterial properties of nano-MgO powder and modified resin composites against Streptococcus mutans (S. mutans) were detected by antibacterial ring test and film contact test, respectively. Compressive strength (CS) and wear resistance were determined by a universal testing machine and an abrasion test machine. The results indicated that antibacterial activity and wear resistance of resin composites containing nano-MgO were superior to the control group (p

Fasudil, a Rho kinase (ROCK) inhibitor, effectively inhibits disease severity in a mouse model of Alzheimer's disease (AD). However, given its significant limitations, including a relatively narrow safety window and poor oral bioavailability, Fasudil is not suitable for long-term use. Thus, screening for ROCK inhibitor(s) that are more efficient, safer, can be used orally and suitable for long-term use in the treatment of neurodegenerative disorders is required. The main purpose of the present study is to explore whether FSD-C10, a novel ROCK inhibitor, has therapeutic potential in amyloid precursor protein/presenilin-1 transgenic (APP/PS1 Tg) mice, and to determine possible mechanisms of its action. The results showed that FSD-C10 effectively improved learning and memory impairment, accompanied by reduced expression of amyloid-β 1-42 (Aβ1-42), Tau protein phosphorylation (P-tau) and β-site APP-cleaving enzyme in the hippocampus and cortex area of brain. In addition, FSD-C10 administration boosted the expression of synapse-associated proteins, such as postynaptic density protein 95, synaptophsin, α-amino 3-hydroxy-5-methyl-4-isoxa-zolep-propionate receptor and neurotrophic factors, e,g., brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor. Taken together, our results demonstrate that FSD-C10 has therapeutic potential in the AD mouse model, possibly through inhibiting the formation of Aβ1-42 and P-tau, and promoting the generation of synapse-associated proteins and neurotrophic factors.

Vegetation and climate changes in southern Brazil, as well as the dynamics of the adjacent South Atlantic were investigated through the analyses of pollen, spores and dinocysts from marine sediment core GeoB6211-2 that covers the last 19.3 kyr. The pollen record indicates the dominance of grassland (campos) in southeastern South America (SESA), reflecting cold and/or dry conditions during the Last Glacial Maximum. Forests, mainly gallery forests, expanded slightly during Heinrich Stadial 1, suggesting slightly wetter conditions. A stronger expansion of the Atlantic lowland rainforest is noticed in the record after ca. 5.5 cal kyr BP, likely due to wetter conditions. The relatively high amount of exotic Nothofagus pollen, transported by wind, rivers and then by oceanic currents northwards to the study site, as well as the dinocyst Brigantedinium spp., indicate a noticeable influence of the Brazilian Coastal Current from the south between 19.3 and 14.8 cal kyr BP. After that, the decrease in Nothofagus and Brigantedinium spp. together with the increase in dinocyst Operculodinium centrocarpum indicate that the Brazil Current from the north dominated the coring site. The abundance of freshwater algae between ca. 19.3 and 17.0 cal kyr BP suggests that the Rio de la Plata mouth was located close to the coring site during this period, and its discharge of nutrient-rich freshwaters strongly affected the upper water column. Sea level rise decreased this impact during the late glacial phase by moving the coastline further away from the core site. The presence of the Brazil Current at the core site became stronger after ca. 15 cal kyr BP and strongest after 9 cal kyr BP. In summary, the pollen, spores and dinocyst records from core GeoB6211-2 provide important climatic records to reconstruct the environmental changes in SESA.

Sulbactam is a class A β-lactamase inhibitor with intrinsic whole-cell activity against certain bacterial species, including Acinetobacter baumannii . The clinical use of sulbactam for A. baumannii infections is of interest due to increasing multidrug resistance in this pathogen. However, the molecular drivers of its antibacterial activity and resistance determinants have yet to be precisely defined. Here we show that the antibacterial activities of sulbactam vary widely across contemporary A. baumannii clinical isolates and are mediated through inhibition of the penicillin-binding proteins (PBPs) PBP1 and PBP3, with very low frequency of resistance; the rare pbp3 mutants with high levels of resistance to sulbactam are attenuated in fitness. These results support further investigation of the potential clinical utility of sulbactam.

Inhibition of the rennin-angiotensin system (RAS) could reduce insulin resistance in patients with hypertension and diabetic kidney disease (DKD), but whether the effect of losartan on insulin resistance is associated with reduction of oxidative stress and enhancement of insulin signaling transduction has not been fully elucidated.130 patients with type 2 DKD were randomly assigned into 2 groups, the losartan group (n=65, 100 mg orally daily for 12 months) and the amlodipine group (n=65, 10 mg orally daily for 12 months). Oxidative stress markers in plasma, urine concentrations of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and nitrotyrosine (NT) as well as SOD activity were measured by ELISA. After in vitro treatment with different doses of losartan (10, 100 μmol/L) or amlodipine for 48 h, the size of H2O2-induced adipocytes and glucose consumption were measured. Western blot was performed to investigate IRS-1 serine phosphorylation level as well as the protein expressions of phosphorylated insulin receptor (pIR), phosphatidylinositol 3- kinase (PI3K) and insulin receptor substrate 1 (IRS-1) in 3T3-L1 adipocytes.After 12-month treatment, there were no significant differences in systolic and diastolic blood pressures decreases, plasma fasting blood glucose and HbA1c between the 2 groups. Compared with amlodipine group, fasting blood insulin levels and insulin resistance index (HOMA-IR) were significantly decreased in losartan group, and in addition, the circulating levels of 8-OHdG and NT were significantly decreased in losartan group, while the serum SOD activity was enhanced. There were significant positively correlations of HOMA-IR with inflammatory oxidative stress markers. In vitro study showed that losartan could increase glucose uptake in 3T3-L1 adipocytes (P0.01) and decrease adipocyte size (P0.01), while amlodipine can't. Losartan can also enhance adiponectin (P0.05) and decrease TNF-α (P0.05) and IL-6 (P0.01) secretion, while amlodipine can't. The protein expressions of pIR, IRS-1 and PI3K were significantly increased after treatment with losartan (P0.01), while the level of IRS-1 serine phosphorylation was decreased (P0.01), which could be blocked by specific PI3K inhibitor wortmannin.These results suggest that the effect of losartan on insulin resistance is associated with the reduction of oxidative stress and inflammation in patients with type 2 DKD as well as the activation of insulin signal pathway in insulin-resistance 3T3-L1 adipocytes through modulation of PI3K pathway. (Clinical Trials. gov number, NCT 00774904).

Long-term changes in vegetation and climate of southern Brazil, as well as ocean dynamics of the adjacent South Atlantic, were studied by analyses of pollen, spores and organic-walled dinoflagellate cysts (dinocysts) in marine sediment core GeoB2107-3 collected offshore southern Brazil covering the last 73.5 cal kyr BP. The pollen record indicates that grasslands were much more frequent in the landscapes of southern Brazil during the last glacial period if compared to the late Holocene, reflecting relatively colder and/or less humid climatic conditions. Patches of forest occurred in the lowlands and probably also on the exposed continental shelf that was mainly covered by salt marshes. Interestingly, drought-susceptible Araucaria trees were frequent in the highlands (with a similar abundance as during the late Holocene) until 65 cal kyr BP, but were rare during the following glacial period. Atlantic rainforest was present in the northern lowlands of southern Brazil during the recorded last glacial period, but was strongly reduced from 38.5 until 13.0 cal kyr BP. The reduction was probably controlled by colder and/or less humid climatic conditions. Atlantic rainforest expanded to the south since the Lateglacial period, while Araucaria forests advanced in the highlands only during the late Holocene. Dinocysts data indicate that the Brazil Current (BC) with its warm, salty and nutrient-poor waters influenced the study area throughout the investigated period. However, variations in the proportion of dinocyst taxa indicating an eutrophic environment reflect the input of nutrients transported mainly by the Brazilian Coastal Current (BCC) and partly discharged by the Rio Itajai (the major river closest to the core site). This was strongly related to changes in sea level. A stronger influence of the BCC with nutrient rich waters occurred during Marine Isotope Stage (MIS) 4 and in particular during the late MIS 3 and MIS 2 under low sea level. Evidence of Nothofagus pollen grains from the southern Andes during late MIS 3 and MIS 2 suggests an efficient transport by the southern westerlies and Argentinean rivers, then by the Malvinas Current and finally by the BCC to the study site. Major changes in the pollen/spore and dinocyst assemblages occur with similar pacing, indicating strongly interlinked continental and marine environmental changes. Proxy comparisons suggest that the changes were driven by similar overarching factors, of which the most important was orbital obliquity.

LptA is a soluble periplasmic component of the lipopolysaccharide (LPS) transport system of Gram-negative bacteria that transports newly synthesized LPS from the inner membrane to the outer leaflet of the outer membrane. LptA links the inner membrane components (LptBFGC) to the outer membrane components (LptDE), but it is uncertain whether LptA is a freely moving LPS shuttle or part of a stable trans-periplasm structure. Escherichiacoli LptA forms highly polymerized head-to-tail oligomers in solution, but dimers in vivo. We studied the oligomerization of purified Pseudomonasaeruginosa LptA. Size-exclusion chromatography showed that P. aeruginosa LptA, unlike E. coli LptA, is a dimer over a wide range of concentrations. Chemical crosslinking with bis(sulfosuccinimidyl) suberate confirmed that dimers were the predominant species even at sub-micromolar LptA concentrations, which was unaffected by LPS binding. Mass spectrometry of crosslinked dimers showed that crosslinks occurred between the N-terminal α-amino group and either Lys-172 or Lys-173 near the C-terminus. These results support a hypothetical structure for the dimer of isolated P. aeruginosa LptA in which the N-terminus of one monomer is in close proximity to the C-terminus of the other, and the same surface of each monomer forms the interface between them, preventing further oligomerization.

A new enzyme-free and ultrasensitive electrochemical Pb 2+ biosensor was developed. By coupling the DNA-assisted cascade of hybridization reaction with the quantum dots (QDs) for signal amplification, a detection limit as low as 6.1 pM can be obtained for Pb 2+ . In this study, the “8-17” DNAzyme was used for specific recognition of Pb 2+ . In the presence of Pb 2+ , the DNAzyme was activated and cleaved the substrate strand. And then, the hybridization between the linker probe and signal probe was initiated, which resulted in formation of a long cascade DNA structure as well as assemble of numerous QDs at last. By the use of magnetic beads, the free signal probe can be easily removed by external magnetic field. After acid lysis, a great amount of redox cations can be released from the QDs and eventually result in significantly amplified electrochemical signals. This method is highly sensitive, selective and simple without the participation of any protein based enzyme (nuclease), thereby holds great potential for real sample analysis.

Aim To assess the prognostic value of mediastinal lymph node metastases (N2 disease), carcinoembryonic antigen (CEA) levels and C-reactive protein (CRP) in non-small cell lung cancer (NSCLC), according to the 7th edition of the TNM classification. Methods Newly diagnosed stage III-IV NSCLC were enrolled, including 75 patients with malignant pleural effusion. The relationship between serum CRP levels and other relevant variables such as sex, Eastern Cooperative Oncology Group status, smoking status, initial staging, N2 disease, serum albumin, white blood cell count, platelet count, CEA, comorbidity and pathology were analyzed. Univariate and multivariate analyses were performed to find prognostic markers using Cox's proportional hazards model. Results Of the 127 patients enrolled, 55 (43%) had elevated CRP levels. There was a significant correlation between serum CRP level and platelet count (P = 0.011). Median overall survival (OS) in the normal CRP group was significantly longer than in the high CRP group (15.7 months vs 9.1 months, P = 0.013). Hypoalbuminemia (P = 0.047), higher CEA (P = 0.043) and N2 disease (P = 0.040) were additional prognostic factors on univariate analysis. On multivariate analysis an elevated CRP serum level (HR = 1.796; P = 0.005), higher CEA (HR = 1.563; P = 0.031) and N2 disease (HR = 1.723; P = 0.012) were independent prognostic factors for poor survival. Conclusion High levels of serum CRP and CEA, and N2 disease are independent prognostic indicators for the survival of patients with stage III-IV NSCLC.

A simple and sensitive method, based on surface-enhanced Raman scattering (SERS), for immunoassay and label-free protein detection is reported. A series of bowl-shaped silver cavity arrays were fabricated by electrodeposition using a self-assembled polystyrene spheres template. The reflection spectra of these cavity arrays were recorded as a function of film thickness, and then correlated with SERS enhancement using sodium thiophenolate as the probe molecule. The results reveal that SERS enhancement can be maximized when the frequency of both the incident laser and the Raman scattering approach the frequency of the localized surface plasmon resonance. The optimized array was then used as the bottom layer of a silver nanoparticle-protein-bowl-shaped silver cavity array sandwich. The second layer of silver was introduced by the interactions between the proteins in the middle layer of the sandwich architecture and silver nanoparticles. Human IgG bound to the surface of this microcavity array can retain its recognition function. With the Raman reporter molecules labeled on the antibody, a detection limit down to 0.1 ng mL(-1) for human IgG is easily achieved. Furthermore, the SERS spectra of label-free proteins (catalase, cytochrome C, avidin and lysozyme) from the assembled sandwich have excellent reproducibility and high quality. The results reveal that the proposed approach has potential for use in qualitative and quantitative detection of biomolecules.

Background— Liraglutide, a glucagon-like peptide-1 analog, was reported to reduce reperfusion injury in mice. We planned to evaluate the effects of liraglutide on reperfusion injury in patients with acute ST-segment–elevation myocardial infarction treated with primary percutaneous coronary intervention. Methods and Results— A total of 96 patients with ST-segment–elevation myocardial infarction undergoing emergency primary percutaneous coronary intervention were randomized to receive either subcutaneous liraglutide or placebo. Study treatment was commenced 30 minutes before intervention (1.8 mg) and maintained for 7 days after the procedure (0.6 mg for 2 days, 1.2 mg for 2 days, followed by 1.8 mg for 3 days). The salvage index was calculated from myocardial area at risk, measured during the index admission (35±12 hours), and final infarct size measured at 91±5 days after primary percutaneous coronary intervention by cardiac magnetic resonance. At 3 months, the primary end point, a higher salvage index was found in the liraglutide group than in the placebo group in 77 patients evaluated with cardiac magnetic resonance (0.66±0.14 versus 0.55±0.15; P =0.001). The final infarct size was lower in the liraglutide group than that in the placebo group (15±12 versus 21±15 g; P =0.05). Serum high-sensitivity C-reactive protein level was lower in the liraglutide group ( P Conclusions— Our study provides evidence that liraglutide improves myocardial salvage and infarct size after ST-segment–elevation myocardial infarction, possibly by reducing reperfusion injury, making it a promising treatment for evaluation in larger trials. Clinical Trial Registration— URL: https://www.clinicaltrials.gov . Unique identifier: NCT02001363.

Vascular endothelial growth factor (VEGF-A) stimulating angiogenesis is required for tumor growth and progression. The conventional VEGF-A isoforms have been considered as pro-angiogenic factors. Another family of VEGF-A isoforms generated by alternative splicing, termed VEGFxxxb isoforms, has anti-angiogenic property, exemplified by VEGF165b. Here, we identify a new number of VEGFxxx family-VEGF111b induced by mitomycin C, although not detected in mitomycin C-unexposed ovarian cancer cells. SKOV3 cells were transfected with pcDNA3.1 empty vector, pcDNA3.1-VEGF111b or pcDNA3.1-VEGF165b to collect conditioned mediums respectively. VEGF111b overexpression inhibits proliferation, migration and tube formation of endothelial cell by inhibiting VEGF-R2 phosphorylation and its downstream signaling, similar to VEGF165b but slightly lower than VEGF165b. The anti-angiogenic property depends on the six amino acids of exon 8b of the VEGFxxxb isoforms. Our results show that VEGF111b is a novel potent anti-angiogenic agent that can target the VEGF-R2 and its signaling pathway to inhibit ovarian tumor growth.

Ghrelin is an acylated peptide originally identified in the rat stomach as the endogenous ligand for growth hormone secretagogue receptor (GHSR) that promotes gastric motility. Our aims were to explore the effects of ghrelin on gastric-distension-sensitive neurons in the hippocampus and the potential for ghrelin to regulate gastric motility through the arcuate nucleus (Arc). Single-unit discharges in the hippocampus were recorded extracellularly, and gastric motility in conscious rats was monitored. The expression of GHSR-1a in the hippocampus was determined by PCR, Western blot and fluo-immunohistochemistry staining. Retrograde tracing and fluo-immunohistochemistry staining were used to determine ghrelin neuron projection. Ghrelin-Fluoro-Gold double-labelled neurons and GHSR-1a expression were observed in the Arc and hippocampus, respectively. There were gastric-distension-sensitive neurons in the hippocampus that could be excited by ghrelin or by electrical stimulation of the Arc. The excitatory effects could be blocked completely or partly by pretreatment with the ghrelin receptor antagonist [d-Lys-3]-GHRP-6. Gastric motility was significantly promoted by the administration of ghrelin into the hippocampus in a dose-dependent manner that could be completely abolished by [d-Lys-3]-GHRP-6. Electrical stimulation of the Arc could promote gastric motility as well. Nevertheless, these effects could be mitigated by pretreatment with [d-Lys-3]-GHRP-6. Electrical lesioning of the hippocampus diminished the excitatory effects on gastric motility that were induced by electrical stimulation the Arc. Our findings suggest that ghrelin plays an important role in promoting gastric motility via the hippocampus. The Arc may be involved in regulation of the influence of the hippocampus on gastric motility.

Recombinant human erythropoietin (rHuEPO) reduces serum insulin levels, increases insulin sensitivity, and reduces insulin resistance (IR). However, the mechanisms behind these effects are unclear. This study aimed to investigate the mechanism by which rHuEPO effects IR in 3T3L1 adipocytes. After treatment with different concentrations of rHuEPO, glucose consumption, and tumor necrosis factor (TNF-α), adiponectin, and leptin levels were assayed with a commercial enzyme-linked immunosorbent assays. Endogenous erythropoietin receptor (EPOR) expression was inhibited using small interfering RNA (siRNA). EPOR protein and mRNA expression was detected via immunofluorescence and real-time PCR analyses, respectively. The expression of pAKT/AKT and p-STAT5/STAT5 was determined via Western blot analysis. rHuEPO treatment improved glucose uptake, increased adiponectin levels, and reduced TNF-α and leptin levels in 3T3L1 adipocytes with dexamethasone-induced IR. Whereas EPOR protein and gene expression was absent in preadipocytes, it was observed in mature 3T3L1 adipocytes. However, the expression of EPOR in insulin resistant 3T3L1 adipocytes was significantly decreased (p

MLAA-34 is a novel acute monocytic leukemia (M5)-associated antigen (MLAA) that plays a role in the apoptosis of U937 cells. However, the expression and molecular mechanism of MLAA-34 in U937 cells remain largely unclear. Here, we utilized three strategies to gain insight into the expression and molecular functions of MLAA-34 and to identify its interacting proteins and pathways involved in the fine-tuning of the MLAA-34 response. Western blot analysis was performed to assess the expression of MLAA-34 in 41 cell lines and five mixed cell types, which revealed that MLAA-34 is most strongly expressed in U937 cells. Immunostaining indicated that MLAA-34 is localized in the cytoplasm and cell membrane. Furthermore, lentivirus-mediated overexpression of MLAA-34 in the U937 cell line led to significant suppression of apoptosis and increased the potential of tumorigenicity. Co-immunoprecipitation (Co-IP), shotgun and bioinformatic analysis identified 256 proteins and 225 of them were annotated by gene ontology categories. This analysis revealed 71 proteins involved in cell apoptosis or proliferation of biological processes and signaling pathways. Moreover, the effect of MLAA-34 apoptosis may be through interaction with the Ras, Wnt, calcium and chemokine signaling pathways and thirteen of the annotated proteins may interact with MLAA-34 and participate in carcinogenesis directly. This study provides a basis for a better understanding of the molecular mechanism and proteomics in the inhibition of apoptosis by MLAA-34 in U937 cells and indicates that MLAA-34 may be a potential candidate for the early diagnosis and therapeutic application of M5.

Background ACFP is an anti-cancer fusion peptide derived from bovine milk protein. This study was to investigate the anti-cancer function and underlying mechanisms of ACFP in ovarian cancer. Methods Fresh ovarian tumor tissues were collected from 53 patients who underwent initial debulking surgery, and primary cancer cells were cultured. Normal ovarian surface epithelium cells (NOSECs), isolated from 7 patients who underwent surgery for uterine fibromas, were used as normal control tissue. Anti-viabilities of ACFP were assessed by WST-1 (water-soluble tetrazolium 1), and apoptosis was measured using a flow cytometry-based assay. Gene expression profiles of ovarian cancer cells treated with ACFP were generated by cDNA microarray, and the expression of apoptotic-specific genes, such as bcl-xl, bax, akt, caspase-3, CDC25C and cyclinB1, was assessed by real time PCR and western blot analysis. Results Treatment with ACFP inhibited the viability and promoted apoptosis of primary ovarian cancer cells but exhibited little or no cytotoxicity toward normal primary ovarian cells. Mechanistically, the anti-cancer effects of ACFP in ovarian cells were shown to occur partially via changes in gene expression and related signal pathways. Gene expression profiling highlighted that ACFP treatment in ovarian cancer cells repressed the expression of bcl-xl, akt, CDC25C and cyclinB1 and promoted the expression of bax and caspase-3 in a time- and dose-dependent manner. Conclusions Our results suggest that ACFP may represent a potential therapeutic agent for ovarian cancer that functions by altering the expression and signaling of cancer-related pathways in ovarian cancer cells. Electronic supplementary material The online version of this article (doi:10.1186/s12885-016-2281-6) contains supplementary material, which is available to authorized users.

Background: Some bioactive peptides derived from natural resources or synthesized by rational design have been shown to have very good anticancer effects. We designed an anticancer fusion peptide (ACFP) based on the structure of bovine lactoferricin (LfcinB) and hexapeptide (PGPIPN) derived from bovine milk protein. Objective: To prepare ACFP through genetic engineering and study its antiovarian cancer activity. Method: ACFP gene was produced by a flexible link arm connecting LfcinB and PGPIPN. ACFP was inductively expressed in Escherichia coli by the recombinant plasmid pGEX-KG-ACFP. ACFP was prepared and purified by affinity chromatography, and identified by polyacrylamide gel electrophoresis (PAGE), high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Recombinant lentivirus vectors were produced by cotransfecting 293T cells with constructed plasmid pLJM1-ACFP, envelope plasmid Δ8.91 and pVSVG using Lipofectamine. ACFP gene was transfected into ovarian cancer cells by pLJM1-ACFP lentivirus. Cell Viability was assayed by the methyl thiazolyl tetrazolium (MTT). The apoptosis of ovarian cancer SKOV3 cells was measured by flow cytometry and observed by Hoechst33258 staining. Results: ACFP was successfully prepared and purified by genetic engineering. ACFP more effectively inhibited the viability of human ovarian cancer SKOV3 cells than the single parent peptides in vitro. ACFP was found to have no cytotoxicity towards untransformed cells. The ACFP gene in cancer cells infected with pLJM1-ACFP lentivirus could significantly inhibit the viability of SKOV3 cells and induce their apoptosis. Conclusion: ACFP is a potential therapeutic agent for the treatment of ovarian cancer.

Liensinine and neferine, a kind of isoquinoline alkaloid, can antagonize the ventricular arrhythmias. The human ether-a-go-go-related gene (hERG) is involved in repolarization of cardiac action potential. We investigated the effects of liensinine and neferine on the biophysical properties of hERG channel and the underlying structure-activity relationships. The effects of liensinine and neferine were examined on the hERG channels in the stable transfected HEK293 cells using a whole-cell patch clamp technique, western blot analysis and immunofluorescence experiment. The pharmacokinetics and tissue distribution determination of liensinine and neferine in rats were determined by a validated RP-HPLC method. Liensinine and neferine induced decrease of current amplitude in dose-dependent. Liensinine reduced hERG tail current from 70.3±6.3 pA/pF in control group to 56.7±2.8 pA/pF in the 1 μM group, 53.0±2.3 pA/pF (3 μM) and 17.8±0.7 pA/pF (30 μM); the corresponding current densities of neferine-treated cells were 41.9±3.1 pA/pF, 32.3±3.1 pA/pF and 16.2±0.6 pA/pF, respectively. Neferine had binding affinity for the open and inactivated state of hERG channel, liensinine only bound to the open state. The inhibitory effects of liensinine and neferine on hERG current were attenuated in the F656V or Y652A mutant channels. Neferine distributed more quickly than liensinine in rats, which was found to be in higher concentration than liensinine. Both liensinine and neferine had no effect on the generation and expression of hERG channels. In conclusion, neferine is a more potent blocker of hERG channels than liensinine at low concentration (

Some bioactive peptides derived from natural resources or synthesized by rational design have been proved to have very good anticancer effect. We studied the inhibition of PGPIPN, a hexapeptide derived from bovine β-casein, on the invasion and metastasis of human ovarian cancer cells in vitro and its molecular mechanism. The human ovarian cancer cells studied include the cell line SKOV3 as well as the primary ovarian cancer cells from ovarian tumor tissues of 37 patients at initial debulking surgery, diagnosed as serous ovarian adenocarcinoma. We showed that PGPIPN inhibited the invasion of ovarian cancer cells with Transwell chamber assay, the migration of ovarian cancer cells with cell scratch assay and colony formation of ovarian cancer cells. The expression (mRNAs and proteins) of genes relevant to invasion and metastasis, MTA1, and NM23H1 were analyzed by real-time PCR and western blotting. PGPIPN repressed the expression of MTA1, and promoted NM23H1. The effects of PGPIPN were dose-dependent. Thus, our study suggests that PGPIPN is a potential therapeutic agent for adjuvant therapy of human malignant ovarian tumors.

To identify differentially expressed microRNAs (miRNAs) and expression patterns of specific miRNAs during meiosis in human oocytes.To identify differentially expressed miRNAs, GV oocytes and MII oocytes matured at conventional FSH levels (5.5 ng/ml) were analyzed by miRNA microarray. Real-time RT-PCR was used to confirm the changed miRNAs. To validate the dynamic changes of miRNAs from GV to MII stages, oocytes were divided into four groups (#1-4), corresponding to GV oocytes, MI oocytes, MII oocytes matured in conventional FSH level and MII oocytes matured in high FSH level (2,000 ng/ml) respectively.Compared with GV oocytes, MII oocytes exhibited up-regulation of 4 miRNAs (hsa-miR-193a-5p, hsa-miR-297, hsa-miR-625 and hsa-miR-602), and down-regulation of 11 miRNAs (hsa-miR-888*, hsa-miR-212, hsa-miR-662, hsa-miR-299-5p, hsa-miR-339-5p, hsa-miR-20a, hsa-miR-486-5p, hsa-miR-141*, hsa-miR-768-5p, hsa-miR-376a and hsa-miR-15a). RT-PCR analysis of hsa-miR-15a and hsa-miR-20a expression revealed concordant dynamic changes in oocytes from group 1 to group 4.Specific miRNAs in human oocytes had dynamic changes during meiosis. High-concentration FSH in IVM medium led to reverse effect on the expression of hsa-miR-15a and hsa-miR-20a.

Background/Aims: Human ether-a-go-go-related gene (hERG) has an important role in the repolarization of the cardiac action potential. Our studies were to investigate the effects of oxymatrine (one of the natural constituents extracted from Chinese herb Sophora flavescens Ait) on hERG-encoded K+ channels at different temperatures and its underlying mechanism. Methods: The effects of oxymatrine were examined on hERG channels stably expressed in HEK293 cells using a whole-cell patch clamp technique. Results: At the temperature 30°C, oxymatrine inhibited hERG current in a concentration-dependent manner and the IC50 was ∼665 µM. However at the temperature of 20°C, low concentration oxymatrine C≤100 µM increased hERG current density. However, high concentration oxymatrine C>100 µM inhibited the hERG current density significantly. Oxymatrine only affected the activation kinetic of hERG channels at all temperatures and had a high binding affinity for open state of hERG channels except the 300 µM-20°C group which had a high binding affinity for inactive state of hERG channels. Conclusion: Oxymatrine is a low potency blocker of hERG K+ channels at 30°C, low concentration oxymatrine affect the hERG activation gating with accelerating hERG tail current at 20°C, oxymatrine is a potential hERG activator at low temperatures.

We studied the effects of isoquinoline alkaloid neferine (Nef) extracted from the seed embryo of Nelumbo nucifera Gaertn on Human ether-a-go-go-related gene (HERG) channels stably expressed in human embryonic kidney (HEK293) cells using whole-cell patch clamp technique, western blot analysis and immunofluorescence experiment. Nef induced a concentration-dependent decrease in current amplitude according to the voltage steps and tail currents of HERG with an IC50 of 7.419 μM (n H −0.5563). Nef shifted the activation curve in a significantly negative direction and accelerated recovery from inactivation and onset of inactivation, however, slowed deactivation. In addition, it had no significant influence on steady-state inactivation curve. Western blot and immunofluorescence results suggested Nef had no significant effect on the expression of HERG protein. In summary, Nef can block HERG K+ channels that functions by changing the channel activation and inactivation kinetics. Nef has no effect on the generation and trafficking of HERG protein. A blocked-off HERG channel was one mechanism of the anti-arrhythmic effects by Nef.

The novel rice (Oryza sativa L.) line YSBR1 is an intersubspecies hybrid with high level of partial resistance to sheath blight (SB). For the purpose of assessing the resistant level of YSBR1 using multiple approaches, the disease score of YSBR1 was investigated after artificial inoculation in the field and greenhouse in 2006 and 2007. The inhibition rate SB-toxin to the embryo root was also used as a criterion of SB resistance. Compared with other rice genotypes, YSBR1 had significantly higher resistance to SB and the resistance was stable and reliable. The average SB disease score of YSBR1 was 2.39±0.23 and felt in “resistant” grade in the “0–9” rating system when the susceptible control Lemont showed a mean disease score of 7.55±0.26. Correlation analysis showed that the SB-toxin inhibition rate was significantly correlated with disease score from the field test. The SB-toxin inhibition rate of YSBR1 was 57.55%, which was similar to that of Jasmine 85 but significantly lower than that of other tested varieties and Lemont (97.39%). In addition to the high resistance to SB, YSBR1 is also in good performance of plant type and height, growth duration, and grain yield, indicating its prosperous application in variety improvement of SB resistance in rice breeding programs.

Human ether-à-go-go-related gene (hERG) has an important role in the repolarization of the cardiac action potential. Sophocarpine and sophoridine are quinolizidine alkaloids and their structures are similar. Our aim was to investigate the effects of sophocarpine or sophoridine on hERG-encoded K(+) channels and the underlying structure-activity relationships. The effects of sophocarpine and sophoridine were examined on stably expressed hERG channels in HEK293 cells using a whole-cell patch clamp technique and Western blot analysis. The oil-water partition coefficients of sophocarpine and sophoridine were determined by a validated RP-HPLC method. At 300 microM, fractional block was 60.9+/-1.4% for sophocarpine versus 41.9+/-2.0% for sophoridine. Compared with sophocarpine, voltage-dependence of hERG channels inhibition by sophoridine was more notable. Sophoridine altered the activation properties, but not sophocarpine. Sophocarpine shifted the inactivation curve in a negative direction, but not sophoridine. Both drugs had no significant effect on the expression of hERG protein. The partition coefficients for the n-octanol/water system of sophocarpine and sophoridine at 37 degrees C were 16.03+/-0.42 and 1.94+/-0.03, respectively. In summary, sophocarpine and sophoridine are low potency blockers of hERG channels that functions by changing the channel kinetics, and sophocarpine is a more potent blocker of hERG K(+) channels than sophoridine, which may be due to higher hydrophobic nature of sophocarpine compared with sophoridine. Sophocarpine may have a higher binding affinity for the inactivate state. In contrast, sophoridine has a higher binding affinity for the open state. Both drugs have no effect on the generation and trafficking of hERG protein.

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism (AFLP) markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects (E-1-0 model). The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I. In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.

The study was devised to investigate the effect of retinoic acid trifloromethyl chalcone (RAFC) on mammary carcinogenesis in female rats. The data revealed a significant decrease in number of rats with mammary tumor, number of tumors per rat and tumor volume by 54, 72 and 75% respectively in RAFC group compared to control group. The ibuprofen treated rats also showed a significant decrease in number of rats with tumor, number of tumors per rat and tumor volumes by 43%, 55%, and 59%, respectively. Treatment of rats with RAFC also increased the latency period of tumor induction significantly. Median detection period (50% of tumors) was 92, 83 and 56 days respectively in the rats from RAFC, ibuprofen and control groups respectively after DMBA induction. These results demonstrate that RAFC possesses strong chemopreventive activity against mammary carcinogenesis.

Objective: To explore the effects of matrine on HCCR1 and HCCR2 expression in cultural human hepatocellular carcinomas (HCC) cells at the level of gene and protein. Methods: Three methods, representational difference analysis (RDA) of cDNA, microarrays and fluorescence in situ hybridization (FISH) were used to detect levels of mRNA and protein expression of HCCR1 and HCCR-2 before and after treatment of matrine. Results: Matrine had inhibitory effect on the mRNA and protein expression of HCCR1 and HCCR2 in cultural HCC cells. Conclusion: Matrine has inhibitory effect on gene transcription, protein expression of HCCR1 and HCCR2 in cultural HCC cells.

Objective: To identify aberrant gene expression in primary human T-cell acute lymphoblastic leukemia (T-ALL) and to evaluate the differently expressed level of Lyl1 and LMO2 genes in human LMO2 positive/TAL1 negative T-ALL tumors. Methods: Three methods, representational difference analysis (RDA) of cDNA, cDNA microarrays and RT-PCR were used to detect if Lyl1 and LMO2 genes were differently expressed in human LMO2 positive/tal1 negative T-ALL tumors. Results: The results of cDNA RDA and cDNA array shown that Lyl1 and LMO2 genes are differently expressed in human T-cell tumors. The result of RT-PCR also shown Lyl1 and LMO2 are high expressed in human T-cell tumors and very low level or no expressed in normal group. Conclusion: We have found that cDNA RDA and cDNA microarray can be successfully used to identify aberrant gene expression in T-ALL cells. In the study described in this manuscript we found that Lyl1 and LMO2 are aberrantly expressed in human T-ALL LMO2 positive/TAL1 negative T-ALL tumors.

Noninvasive in situ detection of suspected cariogenic bacterial species within dental biofilms could facilitate monitoring of the dynamic change of oral microbial flora and assist in the assessment of the treatment efficacy of therapeutic agents. In this study, we explore the possibility to use three well-characterized monoclonal antibodies (MAbs) against Streptococcus mutans, Actinomyces naeslundii, and Lactobacillus casei to identify these three important members of the oral microbial community in the complex environment of oral biofilms. These MAbs, which were conjugated to different fluorescent labels and visualized with confocal laser scanning microscopy (CLSM), proved to be an useful tool to identify the three species of interest (S. mutans, A. naeslundii, and L. casei) under various experimental conditions including in vitro and in vivo derived oral biofilms. Manifold addition of the MAbs on consecutive days did not alter the biofilm structure thus allowing monitoring of the same biofilm over extended time periods. Using this MAb-based method the effect of sucrose challenge on the biofilm composition and the distribution of S. mutans, A. naeslundii, and L. casei were examined. S. mutans was found to be the predominant species under the various biofilm conditions tested. These studies indicate that MAbs based bacterial detection with CLSM is a versatile tool which permits new insights into the ecology of oral biofilm development.