Phosphorylated α-synuclein deposited in Schwann cells interacting with TLR2 mediates cell damage and induces Parkinson's disease autonomic dysfunction.

Despite the significant frequency of autonomic dysfunction (AutD) in Parkinson's disease (PD) patients, its pathogenesis and diagnosis are challenging. Here, we aimed to further explore the mechanism of phosphorylated α-synuclein (p-α-syn) deposited in vagus nerve Schwann cells (SCs) causing SCs damage and PD AutD. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 20 mg/kg) was administrated to C57BL/6 mice twice a week for 35 days. Following the final injection, locomotor functions, gastrointestinal symptoms, urine functions, and cardiovascular system functions were evaluated. Meanwhile, we examined p-α-syn deposited in vagus nerve SCs, Toll-like receptor 2 (TLR2) activation, and SCs loss using immunofluorescence, western blot, and Luxol fast blue staining. In vitro, the rat SCs line RSC96 cells were exposed to α-synuclein preformed fibril (α-syn PFF), and cell viability was detected by CCK8. Co-IP was used to identify the interaction between p-α-syn and TLR2. Furthermore, the role of TLR2 in p-α-syn-mediated SCs damage was confirmed by the administration of CU-CPT22, a specific blocker of TLR2. In vivo, apart from dyskinesia, MPTP mice exhibited constipation, urinary dysfunction, and cardiovascular failure, which were associated with the deposition of p-α-syn in vagus nerve SCs, TLR2 activation, and vagus nerve demyelination. In vitro, stimulation of α-syn PFF induced a time-dependent loss of viability, and p-α-syn deposited in RSC96 cells induced a cellular inflammatory response by interacting with TLR2, resulting in cell dysfunction and apoptosis. However, both SCs inflammatory response and cell viability were alleviated after inhibition of TLR2. Furthermore, 1 h fecal pellets and water content, the frequency of 1 h urine, blood pressure, heart rate, and heart rate variability of mice in the MPTP + CU-CPT22 group were also improved. Our results support the perspective that p-α-syn interacts with TLR2 induced SCs damage and is involved in PD AutD, which sheds fresh light on the mechanism of PD AutD and indicates a promising treatment for PD AutD targeting SCs p-α-syn/ TLR2 signaling pathway.

Impacts of Climate Changes on Geographic Distribution of Primula filchnerae, an Endangered Herb in China.

Primula filchnerae, an endangered plant endemic to China, has drawn people's attention in recent years due to its ornamental value in flower. It was rarely recorded since being described in 1902, but it was rediscovered in 2009 and is now known from a limited number of sites located in Hubei and Shaanxi Provinces. Since the species is still poorly known, a number of unanswered questions arise related to it: How has P. filchnerae responded to past climate change and how might it respond in the future? Why was P. filchmerae so rarely collected during the past century? We assembled geographic coordinates for P. filchnerae through the field surveys and website searches, and then used a maximum entropy model (MaxEnt) to simulate its potential suitable distribution in six periods with varied carbon emission levels by combining bioclimatic and environmental factors. MaxEnt showed that Min Temperature of the Coldest Month (bio6) and Precipitation of the Coldest Quarter (bio19) affected P. filchnerae's distribution most, with an aggregate contribution >60% and suitable ranges above -5 °C and below 40 mm, respectively. We also analyzed potential habitat distribution in various periods with differing impacts of climate change compared to today's suitable habitats, and in most cases, Shaanxi and Sichuan remained the most stable areas and with possible expansion to the north under various carbon emission scenarios, but the 2050s SSP5-8.5 scenario may be an exception. Moreover, we used MaxEnt to evaluate population shifts, with various scenarios indicating that geometric center would be concentrated in Sichuan Province in China. Finally, conservation strategies are suggested, including the creation of protected areas, long-term monitoring, raising public awareness of plant conservation, situ conservation measures, assisted migration, and species introduction. This study demonstrates how P. filchnerae may have adapted to changes in different periods and provides a scientific basis for germplasm conservation and management.

Elevated branched-chain amino acid promotes atherosclerosis progression by enhancing mitochondrial-to-nuclear H2O2-disulfide HMGB1 in macrophages.

As the essential amino acids, branched-chain amino acid (BCAA) from diets is indispensable for health. BCAA supplementation is often recommended for patients with consumptive diseases or healthy people who exercise regularly. Latest studies and ours reported that elevated BCAA level was positively correlated with metabolic syndrome, diabetes, thrombosis and heart failure. However, the adverse effect of BCAA in atherosclerosis (AS) and its underlying mechanism remain unknown. Here, we found elevated plasma BCAA level was an independent risk factor for CHD patients by a human cohort study. By employing the HCD-fed ApoE-/- mice of AS model, ingestion of BCAA significantly increased plaque volume, instability and inflammation in AS. Elevated BCAA due to high dietary BCAA intake or BCAA catabolic defects promoted AS progression. Furthermore, BCAA catabolic defects were found in the monocytes of patients with CHD and abdominal macrophages in AS mice. Improvement of BCAA catabolism in macrophages alleviated AS burden in mice. The protein screening assay revealed HMGB1 as a potential molecular target of BCAA in activating proinflammatory macrophages. Excessive BCAA induced the formation and secretion of disulfide HMGB1 as well as subsequent inflammatory cascade of macrophages in a mitochondrial-nuclear H2O2 dependent manner. Scavenging nuclear H2O2 by overexpression of nucleus-targeting catalase (nCAT) effectively inhibited BCAA-induced inflammation in macrophages. All of the results above illustrate that elevated BCAA promotes AS progression by inducing redox-regulated HMGB1 translocation and further proinflammatory macrophage activation. Our findings provide novel insights into the role of animo acids as the daily dietary nutrients in AS development, and also suggest that restricting excessive dietary BCAA consuming and promoting BCAA catabolism may serve as promising strategies to alleviate and prevent AS and its subsequent CHD.

Inhibition of TLR4 Alleviates Heat Stroke-Induced Cardiomyocyte Injury by Down-Regulating Inflammation and Ferroptosis.

Inflammatory response and cell death play key roles in the mechanism of myocardial cell injury induced by heat stroke (HS) in rats. Ferroptosis is a newly discovered regulatory type of cell death, which is involved in the occurrence and development of various cardiovascular diseases. However, the role of ferroptosis in the mechanism of cardiomyocyte injury caused by HS remains to be clarified. The purpose of this study was to investigate the role and potential mechanism of Toll-like receptor 4 (TLR4) in cardiomyocyte inflammation and ferroptosis under HS conditions at the cellular level. The HS cell model was established by exposing H9C2 cells at 43 °C for 2 h and then recovering at 37 °C for 3 h. The association between HS and ferroptosis was investigated by adding the ferroptosis inhibitor, liproxstatin-1, and the ferroptosis inducer, erastin. The results show that the expressions of ferroptosis-related proteins recombinant solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were decreased, the contents of glutathione (GSH) were decreased, and the contents of malondialdehyde (MDA), reactive oxygen species (ROS), and Fe2+ were increased in H9C2 cells in the HS group. Moreover, the mitochondria of the HS group became smaller and the membrane density increased. These changes were consistent with the effects of erastin on H9C2 cells and were reversed with liproxstatin-1. The addition of TLR4 inhibitor TAK-242 or NF-κB inhibitor PDTC reduced the expressions of NF-κB and p53, increased the expressions of SLC7A11 and GPX4, reduced the contents of TNF-α, IL-6 and IL-1ß, increased the content of GSH and reduced MDA, ROS, and Fe2+ levels in H9C2 cells under the HS condition. TAK-242 may improve the mitochondrial shrinkage and membrane density of H9C2 cells induced by HS. In conclusion, this study illustrated that inhibition of the TLR4/NF-κB signaling pathway can regulate the inflammatory response and ferroptosis induced by HS, which provides new information and a theoretical basis for the basic research and clinical treatment of cardiovascular injuries caused by HS.

Mitochondria-derived H2O2 triggers liver regeneration via FoxO3a signaling pathway after partial hepatectomy in mice.

Reactive oxygen species (ROS) can induce oxidative injury and are generally regarded as toxic byproducts, although they are increasingly recognized for their signaling functions. Increased ROS often accompanies liver regeneration (LR) after liver injuries, however, their role in LR and the underlying mechanism remains unclear. Here, by employing a mouse LR model of partial hepatectomy (PHx), we found that PHx induced rapid increases of mitochondrial hydrogen peroxide (H2O2) and intracellular H2O2 at an early stage, using a mitochondria-specific probe. Scavenging mitochondrial H2O2 in mice with liver-specific overexpression of mitochondria-targeted catalase (mCAT) decreased intracellular H2O2 and compromised LR, while NADPH oxidases (NOXs) inhibition did not affect intracellular H2O2 or LR, indicating that mitochondria-derived H2O2 played an essential role in LR after PHx. Furthermore, pharmacological activation of FoxO3a impaired the H2O2-triggered LR, while liver-specific knockdown of FoxO3a by CRISPR-Cas9 technology almost abolished the inhibition of LR by overexpression of mCAT, demonstrating that FoxO3a signaling pathway mediated mitochondria-derived H2O2 triggered LR after PHx. Our findings uncover the beneficial roles of mitochondrial H2O2 and the redox-regulated underlying mechanisms during LR, which shed light on potential therapeutic interventions for LR-related liver injury. Importantly, these findings also indicate that improper antioxidative intervention might impair LR and delay the recovery of LR-related diseases in clinics.

Anodal high-definition transcranial direct current stimulation reduces heart rate and modulates heart-rate variability in healthy young people: A randomized cross-controlled trial.

Objective: To investigate whether anodal high-definition transcranial current stimulation (HD-tDCS) over the left dorsolateral pre-frontal cortex (DLPFC) could modulate the heart rate (HR) and heart-rate variability (HRV) in healthy young people. Methods: Forty healthy young people were enrolled in this randomized crossover trial. The participants were randomized to receive anodal HD-tDCS (n = 20) or sham HD-tDCS (n = 20) over the left DLPFC with a washout period of 1 week. Electrocardiogram (ECG) data were continuously recorded 20 min before the stimulation, during the session (20 min), and 20 min after the session. HR and the time- and frequency-domain indices of the HRV were measured to investigate the activity of the sympathetic and parasympathetic nervous systems. Results: Anodal HD-tDCS over the left DLPFC induced a significant decrease in HR and a significant increase in the average of normal-to-normal intervals (AVG NN), low-frequency (LF) power, total power (TP), and LF/high-frequency (HF) ratio in comparison with the sham stimulation and the baseline. However, sham HD-tDCS over the left DLPFC had no significant effect on HR or HRV. Conclusions: Anodal HD-tDCS over the left DLPFC could reduce HR and modulate the HRV in healthy young people. HD-tDCS may show some potential for acutely modulating cardiovascular function.

Phase stability and the interface structure of a nanoscale Si crystallite in Al-based alloys.

An atomic-scale understanding of the role of strain on the microstructural properties of nanoscale precipitates will be helpful to explore the precipitation behavior as well as the structure-property relationships in crystalline multi-phase systems. Nanoscale Si precipitates are formed in Al-based alloys prepared by selective laser melting. The phase structure and the nature of heterointerface have been characterized using advanced electron microscopy. The nanocrystalline Si mainly contains two polymorphs, diamond-cubic Si (DC-Si) and 4H hexagonal Si (4H-Si). Heteroepitaxy occurs at the DC-Si(111)/Al(100) and 4H-Si(0001)/Al(100) interfaces in terms of a coincidence-site lattice model. The nanocrystalline Si undertakes tensile strain superposed by the matrix through heterointerfaces, facilitating the formation of 4H-Si in the nanoscale crystallite, which provides a strategy for designing Si polymorphic materials by strain engineering.

Preconditioning with Cathodal High-Definition Transcranial Direct Current Stimulation Sensitizes the Primary Motor Cortex to Subsequent Intermittent Theta Burst Stimulation.

Noninvasive brain stimulation techniques such as transcranial magnetic stimulation (TMS) and transcranial direct current stimulation (tDCS) can induce long-term potentiation-like facilitation, but whether the combination of TMS and tDCS has additive effects is unclear. To address this issue, in this randomized crossover study, we investigated the effect of preconditioning with cathodal high-definition (HD) tDCS on intermittent theta burst stimulation- (iTBS-) induced plasticity in the left motor cortex. A total of 24 healthy volunteers received preconditioning with cathodal HD-tDCS or sham intervention prior to iTBS in a random order with a washout period of 1 week. The amplitude of motor evoked potentials (MEPs) was measured at baseline and at several time points (5, 10, 15, and 30 min) after iTBS to determine the effects of the intervention on cortical plasticity. Preconditioning with cathodal HD-tDCS followed by iTBS showed a greater increase in MEP amplitude than sham cathodal HD-tDCS preconditioning and iTBS at each time postintervention point, with longer-lasting after-effects on cortical excitability. These results demonstrate that preintervention with cathodal HD-tDCS primes the motor cortex for long-term potentiation induced by iTBS and is a potential strategy for improving the clinical outcome to guide therapeutic decisions.

Does renewable energy electricity and economic complexity index help to achieve carbon neutrality target of top exporting countries?

Despite extensive studies into international trade and CO2 pollution, a panel of only top exporting economies was not considered. As the current research is an endeavor to address the gap by exploring the impact of economic complexity index (ECI) and renewable energy electricity (RELC) on consumption-based carbon emissions (CCO2) in the presences of exports, imports, and gross domestic product (GDP) in the top exporting countries from 1990 to 2019. The results confirm a relationship between consumption-based carbon emissions, economic complexity index, renewable energy electricity, exports, imports, and economic growth in both the short-run and long run. Empirical evidence for cross-sectional auto-regressive distributed lags (CS-ARDL) estimates, confirms that economic complexity index imports and GDP are positively associated and have significant adverse short-run and long-run impacts on consumption-based carbon emission. Whereas, export and renewable energy electricity significantly reduce carbon emissions. To achieve carbon neutrality target, policymakers should make economic complexity reforms, import green products, and promote investments in sustainable sources.

Effects of varying intensities of heat stress on neuropeptide Y and proopiomelanocortin mRNA expression in rats.

The aim of the present study was to investigate the effects of varying intensities of heat stress on the mRNA expression levels of neuropeptide Y (NPY), proopiomelanocortin (POMC) and stress hormones in rats. To establish a rat model of heat stress, the temperature and time were adjusted in a specialized heating chamber. Sprague-Dawley (SD) rats were randomly divided into four groups; control (CN; temperature, 24±1˚C); moderate strength 6 h (MS6; temperature, 32±1˚C time, 6 h), moderate strength 24 h (MS24; temperature, 32±1˚C; time, 24 h) and high strength 6 h (HS6; temperature, 38±1˚C; time, 6 h) groups. SD rats were exposed to heat for 14 consecutive days. The levels of heat stress-related factors, including corticotropin-releasing hormone (CRH), cortisol (COR), epinephrine (EPI) and heat shock protein 70 (HSP70), were measured in the rat blood using ELISA. In addition, the weight of the spleen, thymus, hypophysis and hypothalamus were determined. The mRNA expressions levels of NPY and POMC were detected using quantitative PCR. The results showed that the CRH, COR and HSP70 levels were increased in the three heat stress groups compared with the CN group. Notably, the levels of CRH, EPI and HSP70 were increased in the HS6 group compared with the CN and MS6 groups (P<0.05). Furthermore, the weights of the hypophysis and hypothalamus in the HS6 group were significantly lower compared with the CN group (P<0.05). In addition, NPY and POMC expression levels were downregulated in the MS24 group compared with the CN group. The mRNA expression levels of NPY and POMC were altered in response to different intensities of heat stress. Therefore, their levels were downregulated and upregulated following long-time and moderate-time heat exposure, respectively. The results of the present study suggested that the reduced mRNA expression levels of NPY may be partially responsible for the heat-induced injuries in rats following long-time heat exposure.

Exercise and dietary intervention ameliorate high-fat diet-induced NAFLD and liver aging by inducing lipophagy.

Exercise and dietary intervention are currently available strategies to treat nonalcoholic fatty liver disease (NAFLD), while the underlying mechanism remains controversial. Emerging evidence shows that lipophagy is involved in the inhibition of the lipid droplets accumulation. However, it is still unclear if exercise and dietary intervention improve NAFLD through regulating lipophagy, and how exercise of skeletal muscle can modulate lipid metabolism in liver. Moreover, NAFLD is associated with aging, and little is known about the effect of lipid accumulation on aging process. Here in vivo and in vitro models, we found that exercise and dietary intervention reduced lipid droplets formation, decreased hepatic triglyceride in the liver induced by high-fat diet. Exercise and dietary intervention enhanced the lipophagy by activating AMPK/ULK1 and inhibiting Akt/mTOR/ULK1 pathways respectively. Furthermore, exercise stimulated FGF21 production in the muscle, followed by secretion to the circulation to promote the lipophagy in the liver via an AMPK-dependent pathway. Importantly, for the first time, we demonstrated that lipid accumulation exacerbated liver aging, which was ameliorated by exercise and dietary intervention through inducing lipophagy. Our findings suggested a new mechanism of exercise and dietary intervention to improve NAFLD through promoting lipophagy. The study also provided evidence to support that muscle exercise is beneficial to other metabolic organs such as liver. The FGF21-mediated AMPK dependent lipophagy might be a potential drug target for NAFLD and aging caused by lipid metabolic dysfunction.

Recombinase polymerase amplification with polymer flocculation sedimentation for rapid detection of Staphylococcus aureus in food samples.

Currently, rapid, sensitive, and convenient visual detection methods for Staphylococcus aureus (S. aureus) are scarce. In this study, a novel detection method based on recombinase polymerase amplification (RPA) and polymer flocculation sedimentation (PFS) was developed. Twelve effective primer combinations derived from four forward primers F1, F2, F3, F4, and three reverse primers R1, R2, R3 targeting the nuc gene of S. aureus were designed and screened by a polymerase chain reaction and RPA methods. RPA reaction conditions, including temperature, time, and volume as well as PEG8000 and NaCl concentrations range, were optimized. Moreover, the specificity and sensitivity of the RPA-PFS assay were further analyzed. Finally, the potential use of the RPA-PFS assay was evaluated using artificially S. aureus contaminated food samples, including pork, beef, shrimp, fish, cheese, cabbage, leftover rice, egg, milk, and orange juice. Results showed that the SA5 (F2/R2) combination was the optimal primer candidate. The optimal temperature range, the shortest time and the minimal volume of RPA reaction were 40-42 °C, 10 min and 10 µL, respectively and the optimal PEG8000/NaCl concentrations were 0.2 g/mL and 2.5 M, respectively, for the adsorption between magnetic beads and RPA products. The RPA-PFS method could detect as little as 13 fg genomic DNA of S. aureus and was also specific for five target S. aureus as well as twenty-seven non-target foodborne bacteria. The limit of detection of RPA-PFS for S. aureus in artificially contaminated food samples was 38 CFU/mL (g). Besides, RPA-PFS has directly been judged by the naked eye and has totally taken less than 20 min. In short, the assay RPA-PFS developed in this study is a rapid, sensitive, and specific visual detection method for S. aureus.

Identification of a Sixteen-gene Prognostic Biomarker for Lung Adenocarcinoma Using a Machine Learning Method.

Objectives: Lung adenocarcinoma (LUAD) accounts for a majority of cancer-related deaths worldwide annually. The identification of prognostic biomarkers and prediction of prognosis for LUAD patients is necessary. Materials and Methods: In this study, LUAD RNA-Seq data and clinical data from the Cancer Genome Atlas (TCGA) were divided into TCGA cohort I (n = 338) and II (n = 168). The cohort I was used for model construction, and the cohort II and data from Gene Expression Omnibus (GSE72094 cohort, n = 393; GSE11969 cohort, n = 149) were utilized for validation. First, the survival-related seed genes were selected from the cohort I using the machine learning model (random survival forest, RSF), and then in order to improve prediction accuracy, the forward selection model was utilized to identify the prognosis-related key genes among the seed genes using the clinically-integrated RNA-Seq data. Second, the survival risk score system was constructed by using these key genes in the cohort II, the GSE72094 cohort and the GSE11969 cohort, and the evaluation metrics such as HR, p value and C-index were calculated to validate the proposed method. Third, the developed approach was compared with the previous five prediction models. Finally, bioinformatics analyses (pathway, heatmap, protein-gene interaction network) have been applied to the identified seed genes and key genes. Results and Conclusion: Based on the RSF model and clinically-integrated RNA-Seq data, we identified sixteen key genes that formed the prognostic gene expression signature. These sixteen key genes could achieve a strong power for prognostic prediction of LUAD patients in cohort II (HR = 3.80, p = 1.63e-06, C-index = 0.656), and were further validated in the GSE72094 cohort (HR = 4.12, p = 1.34e-10, C-index = 0.672) and GSE11969 cohort (HR = 3.87, p = 6.81e-07, C-index = 0.670). The experimental results of three independent validation cohorts showed that compared with the traditional Cox model and the use of standalone RNA-Seq data, the machine-learning-based method effectively improved the prediction accuracy of LUAD prognosis, and the derived model was also superior to the other five existing prediction models. KEGG pathway analysis found eleven of the sixteen genes were associated with Nicotine addiction. Thirteen of the sixteen genes were reported for the first time as the LUAD prognosis-related key genes. In conclusion, we developed a sixteen-gene prognostic marker for LUAD, which may provide a powerful prognostic tool for precision oncology.

Rapid analysis of Escherichia coli O157:H7 using isothermal recombinase polymerase amplification combined with triple-labeled nucleotide probes.

Rapid analytical methods are urgently needed to evaluate Escherichia coli (E. coli) O157:H7 in food. In this work, a novel recombinase polymerase amplification (RPA)-based lateral flow dipstick (LFD) method was developed to detect E. coli. Briefly, suitable primers and probes were designed and screened. Then, RPA reaction parameters, including volume, time, and temperature, were optimized. The specificity and sensitivity of RPA-LFD were analyzed, and a contaminated milk sample was used to test the detection performance of the proposed method. The optimal RPA reaction conditions included a minimum volume of 10 µL, incubation time of 10 min, temperature range of 39-42 °C, the primer pair EOF4/EOR3, and the probe EOProb. RPA-LFD was highly sensitive, it could detect as little as 1 fg of the genomic DNA of E. coli O157:H7, and 19 nontarget DNA of foodborne bacteria did not yield amplification products. Finally, the limit of detection of RPA-LFD for E. coli O157:H7 in artificially contaminated raw milk was 4.4 CFU/mL. In summary, the RPA-LFD assay developed in this study is an effective tool for the rapid investigation of E. coli O157:H7 contamination in raw milk samples.

Preparation and Evaluation of Exhaust-Purifying Cement Concrete Employing Titanium Dioxide.

To address the increasing air pollution caused by vehicle exhaust, environment-friendly pavement materials that possesses exhaust-purifying properties were prepared using common cement concrete and porous cement concrete as the base of photocatalyst nano-titanium dioxide (TiO2), respectively. Firstly, Fe3+-doped TiO2 powder was prepared by applying planetary high-energy ball milling in order to improve the efficiency of the semiconductor photocatalyst for degrading vehicle exhausts. Two nano-TiO2, namely the original and modified nanomaterials, were adopted to produce the photocatalytic cement concretes subsequently. The physicochemical properties of the modified powder, as well as the mechanical and photocatalytic properties of TiO2-modified concrete, were characterized using a suite of complementary techniques, including X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), compressive strength and degradation efficiency tests. The results show that the ball milling method not only successfully doped Fe3+ into catalysts but also caused significant changes in: (1) decreased particle sizes, (2) more amorphous morphology, (3) decreased percentage of the most thermodynamically stable crystal facet, and (4) increased percentage of other high gas sensing crystal facets. Both the original and modified nano-TiO2 can improve the concrete strength while the strengthening effect of modified nanomaterials is superior. It is pronounced that the photocatalytic property of the modified nano-TiO2 is much better than that of the original nano particles, and the degradation rate of porous concrete is also better than common concrete when exposed to the same photocatalyst content. In a comprehensive consideration of both mechanical performance and degradation efficiency, the recommended optimum dosage of TiO2 is 3% to 4% for exhaust-purifying concrete.

Sensitive and rapid visual detection of Salmonella Typhimurium in milk based on recombinase polymerase amplification with lateral flow dipsticks.

Salmonella Typhimurium (S. Typhimurium) can cause serious foodborne diseases. In this study, an assay combining recombinase polymerase amplification (RPA) with lateral flow dipsticks (LFD) was developed to detect S. Typhimurium in milk. The RPA forward primers STF1, STF2, STF3, the reverse primer STR labeled with digoxin, and the probe STProb labeled with FAM were designed and screened to produce RPA products for LFD detection. The RPA reaction volume, temperature, and time were then optimized, and the sensitivity and specificity of the developed method were analyzed. Finally, the RPA-LFD method was evaluated using milk artificially contaminated with S. Typhimurium. Results indicated that the primer pair STF1/STR is the optimal combination for detecting the bacterium. The minimum volume, shortest time, and optimal temperature of the RPA reaction were 10 µL, 10 min, and 40-42 °C, respectively. The limit of detection of RPA-LFD for detecting the genomic DNA of S. Typhimurium was 1 fg, which is 5 and 10 times lower than the corresponding limits of RPA-agarose gel electrophoresis (AGE) and PCR-AGE, respectively. Testing with 29 other foodborne bacteria as controls revealed that RPA-LFD was highly specific for S. Typhimurium. RPA-LFD can detect S. Typhimurium at concentrations as low as 1.95 CFU/mL in artificially inoculated milk samples and is thus 10 times more sensitive than PCR. Hence, the RPA-LFD assay established in this study could be a potential point-of-care/need test for S. Typhimurium, especially in areas with limited resources.

Development of duplex PCR-ELISA for simultaneous detection of Salmonella spp. and Escherichia coli O157: H7 in food.

In the current study, a duplex PCR-ELISA method was developed targeting the specific genes, invA of Salmonella spp. and rfbE of Escherichia coli O157: H7, to detect one or both bacteria in food. In brief, PCR product amplified by PCR primer labeled with digoxin at the 5'-end and a probe labeled with biotin at the 3'-end can form dimer by nucleic acid hybridization which can be captured by binding of biotin to streptomycin coated in ELISA plate before using enzyme-labeled anti-digoxin antibody and substrate to develop color. Also, evaluation of the duplex PCR-ELISA method was conducted in different food samples including milk, juice, cabbage, shrimp, chicken, pork and beef. Results indicated that the duplex PCR-ELISA developed here was specific when using 25 non-target bacteria strains as controls and was sensitive with a limit of detection (LOD) of 1 CFU/mL, 1, 000 times higher than that of the duplex PCR method and was repeatable regardless of inter- and intra-batch variations. The duplex PCR-ELISA method established in the present study has proven to be highly specific, sensitive and repeatable. It has the potential to be applied in such fields as clinical diagnosis of food-borne diseases, food hygiene monitoring and pathogen detection in food.

Effect of dehydration heat exposure on thoracic aorta reactivity in rats.

The aim of the present study was to investigate the effect of one week dehydration heat exposure on thoracic aorta reactivity in rats. Eighteen Male Sprague-Dawley rats were randomly divided into 3 groups (n=6 each group): Control group (CN), heat exposure group (HE), dehydration heat exposure group (DHE). The CN group was exposed to a room temperature of 24°C, while the HE and DHE groups were exposed to a heat temperature of 32°C. After 7 days of heat exposure, the heart rate and blood pressure of the rats were measured, and the noradrenaline (NA)-induced contraction on the aorta rings was measured by tension recording. The average contents of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were detected using ELISA. The expression of apoptotic genes in the thoracic aorta was measured using RT-PCR. Compared with CN, the heart rate in the HE and DHE groups had a tendency to become retarded, but there was no significant difference (P>0.05). In the HE group, the systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) of the rats were significantly higher than that of the CN (P<0.05). In the DHE group, the SBP of rats was significantly higher than that of the CN (P<0.05), while the SBP, DBP, and MAP of the rats were decreased compared to the rats in the HE group, although there was no statistical significance (P>0.05). In the HE and DHE groups, the NA-induced contraction on the rats thoracic aorta ring was larger than that of the CN (P<0.05), albeit there was no significant difference between the HE and DHE groups (P>0.05). The serum SOD content decreased in the HE and DHE groups, however, the reduction was significant only in the DHE group (P<0.05). The content of MDA in serum was significantly increased in the DHE group (P<0.05). The expression of BAX was significantly upregulated whereas Bcl2 expression was decreased in the DHE group (P<0.05). The results showed that a high temperature was harmful to the body, especially in the case of lack of food and water. Additionally, the heat exposure elevated blood pressure, and increased arterial reactivity, which were related to the elevated production of MDA, led to the impaired production of SOD, and an increase of cell apoptosis. These findings are useful to understand the influence of dehydrated heat exposure on the vascular function, and they provide certain theoretical and experimental guidance for protection under high temperature.

Ginsenoside Rd attenuates Aß25-35-induced oxidative stress and apoptosis in primary cultured hippocampal neurons.

One of the most common pathological changes in Alzheimer's disease (AD) brain is the large number of amyloid ß (Aß) peptides accumulating in lesion areas. Ginsenosides are the most active components extracted from ginseng. Ginsenoside Rd (GRd) is a newly discovered saponin that has a stronger pharmacological activity than other ginsenosides, especially in neuroprotection. Here we examined the neuroprotective effects of GRd against neuronal insults induced by Aß25-35 in primary cultured hippocampal neurons. A 10µM GRd treatment significantly prevented the loss of hippocampal neurons induced by Aß25-35. In addition, GRd significantly ameliorated Aß25-35-induced oxidative stress by decreasing the reactive oxygen species (ROS) production and malondialdehyde (MDA) level, and increasing the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px); which is similar in treatments with 10µM of probucol (PB) and 100µM of edaravone (EDA). Moreover, our present study demonstrated that GRd significantly enhanced the expression of Bcl-2 mRNA, and decreased the expressions of Bax mRNA and Cyt c mRNA. GRd also downregulated the protein level of cleaved Caspase-3 compared to controls. These results highlighted the neuroprotective effects of GRd against Aß25-35-induced oxidative stress and neuronal apoptosis, suggesting that this may be a promising therapeutics against AD.

Parent Ratings of ADHD Symptoms in Chinese Urban Schoolchildren: Assessment With the Chinese ADHD Rating Scale-IV: Home Version.

OBJECTIVE: The objective of this study was to assess the psychometric properties of the Chinese ADHD Rating Scale-IV (ADHD RS-IV): Home Version and to explore parent ratings of ADHD symptoms in a large sample of urban schoolchildren in China. METHOD: Parents of a representative sample of 1,616 schoolchildren (aged 6-17) in 12 Chinese cities completed the ADHD RS-IV: Home Version. RESULTS: The Chinese ADHD RS-IV: Home Version demonstrated satisfactory internal consistency, test-retest reliability, parent-teacher correlation, discriminant validity, and convergent validity. Factor analysis revealed the DSM-IV two-factor model with "inattention" and "hyperactivity-impulsivity" dimensions, accounting for equal variances. Parent ratings revealed lower/similar scores for Chinese schoolchildren compared with the U.S. CONCLUSION: The ADHD RS-IV: Home Version is a reliable and valid ADHD rating scale in China. The factor structure is similar but not identical to the U.S. STUDY: Normative data reveal cultural differences in some aspects of the parent ratings of ADHD.