Understanding the inter-city causality and regional transport of atmospheric PM2.5 pollution in winter in the Harbin-Changchun megalopolis in China: A perspective from local and regional.

Harbin-Changchun megalopolis (HCM) is the typical cold urban agglomeration in China, where PM2.5 pollution is still serious in winter against the backdrop of continuous improvement in annual air quality in China. To further understand interactions of atmospheric pollution among HCM cities, inter-city causality and regional transport of PM2.5 in the winter in the HCM were comprehensively investigated by using convergent cross mapping (CCM) and CMAQ-BFM methods. CCM analysis results suggest strong bidirectional causal relationships between cities in the HCM, and the causality during polluted episodes were significantly larger than that during clean period. In addition, the influence on local PM2.5 from the HCM western cities were larger than that from cities in the southeast. Inter-city and regional transport contributions results demonstrated that although local emission were the largest contributors among 14 sub-regions for most HCM cities, interactions among cities were strong. Regional transport (42.8%-77.4%) largely contributes to HCM cities' PM2.5 concentrations. Among three regions outside the HCM, NMG (including part of inner Mongolia and Baicheng city in Jilin, 9.1%) was the largest contributor to the PM2.5 concentration in the whole HCM, followed by JLS (including Liaoning Province, Tonghua and Baishan cities in Jilin province, 5.1%) and HLJ (including cities of Heihe, Yichun, Jiamusi, Hegang, Shuangyashan, Jixi, Qitaihe in the Heilongjiang province, 3.8%). Regional transport contribution to the most HCM cities increased significantly from excellent to heavily polluted days. Furthermore, close relationships between transport paths/intensity and wind direction/speed in studied region suggests that we can quantitatively guide the regional joint emergency prevention and control before and during heavily polluted events based on regional weather forecasts in the future.

A duplex nested RT-PCR method for monitoring porcine epidemic diarrhea virus and porcine delta-coronavirus.

BACKGROUND: Porcine epidemic diarrhea virus (PEDV) and porcine delta-coronavirus (PDCoV) are economically important pathogens that cause diarrhea in sows and acute death of newborn piglets. Moreover, the emerging PDCoV was reported to infect children. The current situation is that vaccine prevention has not met expectations, and emergency containment strategies following outbreaks cannot prevent the damages and losses already incurred. Therefore, a more sensitive detection method, that is both convenient and enables accurate and effective sequencing, that will provide early warning of PEDV and PDCoV is necessary. This will enable active, effective, and comprehensive prevention and control, which will possibly reduce disease occurrences. RESULTS: Duplex nested RT-PCR (dnRT-PCR) is an ideal method to achieve early warning and monitoring of PEDV and PDCoV diseases, and to additionally investigate any molecular epidemiological characteristics. In this study, two pairs of primers were designed for each virus based upon the highly conserved N protein sequences of both PEDV and PDCoV strains retrieved from the NCBI Genbank. After optimization of the reaction conditions, the dnRT-PCR assay amplified a 749-bp fragment specific to PEDV and a 344-bp fragment specific to PDCoV. Meanwhile, the specificity and sensitivity of the primers and clinical samples were tested to verify and establish this dnRT-PCR method. The limit of detection (LoD)for both PEDV and PDCoV was 10 copies/µL. The results showed that among 251 samples, 1 sample contained PEDV infection, 19 samples contained a PDCoV infection, and 8 samples were infected with both viruses, following the use of dnRT-PCR. Subsequently, the positive samples were sent for sequencing, and the sequencing results confirmed that they were all positive for the viruses detected using dnRT-PCR, and conventional RT-PCR detection was conducted again after the onset of disease. As these results were consistent with previous results, a detection method for PEDV and PDCoV using dnRT-PCR was successfully established. In conclusion, the dnRT-PCR method established in this study was able to detect both PEDV and PDCoV, concomitantly. CONCLUSIONS: The duplex nested RT-PCR method represents a convenient, reliable, specific, sensitive and anti-interference technique for detecting PEDV and PDCoV, and can additionally be used to simultaneously determine the molecular epidemiological background.

Occurrence, distribution and human exposure of phthalic esters in road dust samples across China.

203 road dust samples were conducted across China covering 28 provinces from January to February，2016 to comprehensively investigates the occurrence, distribution and human exposure of 21 phthalic esters (PAEs). The concentration of Σ21PAEs in road dust ranged from 2.3 to 531 mg/kg, with a mean concentration of 64.1 ± 57.2 mg/kg. DEHP, DnBP and DiBP were the dominant components accounting for 63.3-97.9% (mean: 92.1%) of the Σ21PAEs. Significant Pearson correlation (r = 0.51, p < 0.0001) between Σ21PAEs concentrations and longitude demonstrated a distinguished geographical trend. Higher concentration of PAEs in sidewalk (SW) and trunk road (TR) may reflect influence of human activities such as shoe wear and traffic load. Significant differences were found among different human activities area (urban commercial, urban residential, and suburbs/rural). For total daily intake of Æ©21PAEs via street dust, children had the highest exposure risk followed by teenagers and adults with the median values of 160.8, 43.6, and 37.7 ng/kg-bw/day, respectively. The maximum exposure risk of PAEs calculated based on measurement and simulation were all far below reference values. The sensitivity analysis results demonstrated that concentrations, ingestion rate (IR) and fraction of PAEs absorbed in the skin (AF) were most important parameters on the assessment of exposure risk of PAEs via street dust. Specific parameters based on China and Chinses population is needed to obtain more reliable exposure risk via street dust.

Body weight management in overweight and obese breast cancer survivors.

BACKGROUND: Studies suggest that overweight and obese breast cancer survivors are at increased risk of cancer recurrence and have higher all-cause mortality. Obesity has an impact on breast cancer survivor's quality of life (QOL) and increases the risk of longer-term morbidities such as type 2 diabetes mellitus and cardiovascular disease. Many cancer guidelines recommend survivors maintain a healthy weight but there is a lack of evidence regarding which weight loss method to recommend. OBJECTIVES: To assess the effects of different body weight loss approaches in breast cancer survivors who are overweight or obese (body mass index (BMI) ≥ 25 kg/m2). SEARCH METHODS: We carried out a search in the Cochrane Breast Cancer Group's (CBCG's) Specialised Register, the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 6), MEDLINE (2012 to June 2019), Embase (2015 to June 2019), the World Health Organisation International Clinical Trials Registry Platform (WHO ICTRP) and Clinicaltrials.gov on 17 June 2019. We also searched Mainland Chinese academic literature databases (CNKI), VIP, Wan Fang Data and SinoMed on 25 June 2019. We screened references in relevant manuscripts. SELECTION CRITERIA: We included randomised controlled trials (RCTs), quasi-RCTs and randomised cross-over trials evaluating body weight management for overweight and obese breast cancer survivors (BMI ≥ 25 kg/m2). The aim of the intervention had to be weight loss. DATA COLLECTION AND ANALYSIS: Two review authors independently performed data extraction and assessed risk of bias for the included studies, and applied the quality of the evidence using the GRADE approach. Dichotomous outcomes were analysed as proportions using the risk ratio (RR) as the measure of effect. Continuous data were analysed as means with the measure of effect being expressed as the mean differences (MDs) between treatment groups in change from baseline values with 95% confidence intervals (CIs), when all studies reported exactly the same outcomes on the same scale. If similar outcomes were reported on different scales the standardised mean difference (SMD) was used as the measure of effect. Quality of life data and relevant biomarkers were extracted where available. MAIN RESULTS: We included a total of 20 studies (containing 23 intervention-comparisons) and analysed 2028 randomised women. Participants in the experimental groups received weight loss interventions using the core element of dietary changes, either in isolation or in combination with other core elements such as 'diet and exercise', 'diet and psychosocial support' or 'diet, exercise and psychosocial support'. Participants in the controls groups either received usual care, written materials or placebo, or wait-list controls. The duration of interventions ranged from 0.5 months to 24 months. The duration of follow-up ranged from three months to 36 months. There were no time-to-event data available for overall survival, breast cancer recurrence and disease-free survival. There was a relatively small amount of data available for breast cancer recurrence (281 participants from 4 intervention-comparisons with 14 recurrence events; RR 1.95, 95% CI 0.68 to 5.60; low-quality evidence) and the analysis was likely underpowered. Overall, we found low-quality evidence that weight loss interventions for overweight and obese breast cancer survivors resulted in a reduction in body weight (MD: -2.25 kg, 95% CI: -3.19 to -1.3 kg; 21 intervention-comparisons; 1751 women), body mass index (BMI) (MD: -1.08 kg/m2, 95% CI: -1.61 to -0.56 kg/m2; 17 intervention-comparisons; 1353 women), and waist circumference (MD:-1.73 cm, 95% CI: -3.17 to -0.29 cm; 13 intervention-comparisons; 1193 women), and improved overall quality of life (SMD: 0.74; 95% CI: 0.20 to 1.29; 10 intervention-comparisons; 867 women). No increase was seen in adverse events for women in the intervention groups compared to controls (RR 0.94, 95% CI: 0.76 to 1.17; 4 intervention-comparisons; 394 women; high-quality evidence). Subgroup analyses revealed that decreases in body weight, BMI and waist circumference were present in women regardless of their ethnicity and menopausal status. Multimodal weight loss interventions (which referred to 'diet, exercise and psychosocial support') appeared to result in greater reductions in body weight (MD: -2.88 kg, 95% CI: -3.98 to -1.77 kg; 13 intervention-comparisons; 1526 participants), BMI (MD: -1.44 kg/m2, 95% CI: -2.16 to -0.72 kg/m2; 11 studies; 1187 participants) and waist circumference (MD:-1.66 cm, 95% CI: -3.49 to -0.16 cm; 8 intervention-comparisons; 1021 participants) compared to dietary change alone, however the evidence was low quality. AUTHORS' CONCLUSIONS: Weight loss interventions, particularly multimodal interventions (incorporating diet, exercise and psychosocial support), in overweight or obese breast cancer survivors appear to result in decreases in body weight, BMI and waist circumference and improvement in overall quality of life. There was no increase in adverse events. There is a lack of data to determine the impact of weight loss interventions on survival or breast cancer recurrence. This review is based on studies with marked heterogeneity regarding weight loss interventions. Due to the methods used in included studies, there was a high risk of bias regarding blinding of participants and assessors. Further research is required to determine the optimal weight loss intervention and assess the impact of weight loss on survival outcomes. Long-term follow-up in weight loss intervention studies is required to determine if weight changes are sustained beyond the intervention periods.

Functional analysis of nuclear receptor genes in molting and metamorphosis of the cigarette beetle, Lasioderma serricorne.

Nuclear receptors (NRs) are ligand-regulated transcription factors that are important for the normal growth and development of insects. However, systematic function analysis of NRs in the molting process of Lasioderma serricorne has not been reported. In this study, we identified and characterized 16 NR genes from L. serricorne. Spatiotemporal expression analysis revealed that six NRs were mainly expressed in 3-d-old 4th-instar larvae; five NRs were primarily expressed in 5-d-old adults and four NRs were predominately expressed in prepupae. All the NRs were highly expressed in epidermis, fat body and foregut. RNA interference (RNAi) experiments revealed that knockdown of 15 NRs disrupted the larva-pupa-adult transitions and caused 64.44-100 % mortality. Hematoxylin-eosin staining showed that depletion of 12 NRs prevented the formation of new cuticle and disrupted apolysis of old cuticle. Silencing of LsHR96, LsSVP and LsE78 led to newly formed cuticle that was thinner than the controls. The 20E titer and chitin content significantly decreased by 17.67-95.12 % after 15 NR dsRNA injection and the gene expression levels of 20E synthesis genes and chitin metabolism genes were significantly reduced. These results demonstrated that 15 NR genes are essential for normal molting and metamorphosis of L. serricorne by regulating 20E synthesis and chitin metabolism.

Mechanisms and protective measures for radiation-induced brachial plexus nerve injury.

Radiation therapy is a common treatment modality for patients with malignant tumors of the head and neck, chest and axilla. However, radiotherapy inevitably causes damage to normal tissues at the irradiated site, among which damage to the brachial plexus nerve(BP) is a serious adverse effect in patients receiving radiation therapy in the scapular or axillary regions, with clinical manifestations including abnormal sensation, neuropathic pain, and dyskinesia, etc. These adverse effects seriously reduce the living quality of patients and pose obstacles to their prognosis. Therefore, it is important to elucidate the mechanism of radiation induced brachial plexus injury (RIBP) which remains unclear. Current studies have shown that the pathways of radiation-induced BP injury can be divided into two categories: direct injury and indirect injury, and the indirect injury is closely related to the inflammatory response, microvascular damage, cytokine production and other factors causing radiation-induced fibrosis. In this review, we summarize the underlying mechanisms of RIBP occurrence and possible effective methods to prevent and treat RIBP.

[An investigation of the statistical power of the effect size in randomized controlled trials for the treatment of patients with type 2 diabetes mellitus using Chinese medicine].

OBJECTIVE: To investigate whether the power of the effect size was based on adequate sample size in randomized controlled trials (RCTs) for the treatment of patients with type 2 diabetes mellitus (T2DM) using Chinese medicine. METHODS: China Knowledge Resource Integrated Database (CNKI), VIP Database for Chinese Technical Periodicals (VIP), Chinese Biomedical Database (CBM), and Wangfang Data were systematically recruited using terms like "Xiaoke" or diabetes, Chinese herbal medicine, patent medicine, traditional Chinese medicine, randomized, controlled, blinded, and placebo-controlled. Limitation was set on the intervention course > or = 3 months in order to identify the information of outcome assessement and the sample size. Data collection forms were made according to the checking lists found in the CONSORT statement. Independent double data extractions were performed on all included trials. The statistical power of the effects size for each RCT study was assessed using sample size calculation equations. RESULTS: (1) A total of 207 RCTs were included, including 111 superiority trials and 96 non-inferiority trials. (2) Among the 111 superiority trials, fasting plasma glucose (FPG) and glycosylated hemoglobin HbA1c (HbA1c) outcome measure were reported in 9% and 12% of the RCTs respectively with the sample size > 150 in each trial. For the outcome of HbA1c, only 10% of the RCTs had more than 80% power. For FPG, 23% of the RCTs had more than 80% power. (3) In the 96 non-inferiority trials, the outcomes FPG and HbA1c were reported as 31% and 36% respectively. These RCTs had a samples size > 150. For HbA1c only 36% of the RCTs had more than 80% power. For FPG, only 27% of the studies had more than 80% power. CONCLUSIONS: The sample size for statistical analysis was distressingly low and most RCTs did not achieve 80% power. In order to obtain a sufficient statistic power, it is recommended that clinical trials should establish clear research objective and hypothesis first, and choose scientific and evidence-based study design and outcome measurements. At the same time, calculate required sample size to ensure a precise research conclusion.

[Systematic review on methodology of randomized controlled trials of post-marketing Chinese patent drugs for treatment of type 2 diabetes].

BACKGROUND: Randomized controlled trial (RCT) is considered as the gold standard for the efficacy assessment of medicines. With the increasing number of Chinese patent drugs for treatment of type 2 diabetes, the methodology of post-marketing RCTs evaluating the efficacy and specific effect has become more important. OBJECTIVE: To investigate post-marketing Chinese patent drugs for treatment of type 2 diabetes, as well as the methodological quality of post-marketing RCTs. SEARCH STRATEGY: Literature was searched from the books of Newly Compiled Traditional Chinese Patent Medicine and Chinese Pharmacopeia, the websites of the State Food and Drug Administration and the Ministry of Human Resources and Social Security of the People's Republic of China, China National Knowledge Infrastructure Database, Chongqing VIP Chinese Science and Technology Periodical Database, Chinese Biomedical Database (SinoMed) and Wanfang Data. The time period for searching ran from the commencement of each database to August 2011. INCLUSION CRITERIA: RCTs of post-marketing Chinese patent drugs for treatment of type 2 diabetes with intervention course no less than 3 months. DATA EXTRACTION AND ANALYSIS: Two authors independently evaluated the research quality of the RCTs by the checklist of risk bias assessment and the data collection forms based on the CONSORT Statement. Independent double data-extraction was performed. RESULTS: The authors identified a total of 149 Chinese patent drugs for treatment of type 2 diabetes. According to different indicative syndromes, the Chinese patent drugs can be divided into the following types, namely, yin deficiency and interior heat (n=48, 32%), dual deficiency of qi and yin (n=58, 39%) and dual deficiency of qi and yin combined with blood stasis (n=22, 15%). A total of 41 RCTs meeting the inclusion criteria were included. Neither multicenter RCTs nor endpoint outcome reports were found. Risk bias analysis showed that 81% of the included studies reported randomization for grouping without sequence generation, 98% of these studies did not report concealment of random numbers, 5% used placebo, 10% reported outcome attrition bias and no study employed the analysis of intention-to-treat and 98% reported the diagnostic criteria for type 2 diabetes. The participants mainly consisted of outpatients without complications (76%). The minimum and maximum sample size was 40 and 300 (106 ± 60), respectively. CONCLUSION: The inclusion and exclusion criteria and outcome measures did not match the purposes and contents of post-marketing research in the included studies. They also failed to reflect the basic principles of traditional Chinese medicine in the process of diagnosis and treatment. The demographic characteristics of the patients, the indications for medicine and the syndrome differentiation process were not reported sufficiently and transparently. In order to improve the post-marketing research and promote the rational use of Chinese patent drugs, it is recommended that phase IV clinical trials should establish clear research purpose as well as hypothesis first, and choose scientific and evidence-based study design and outcome measures. In addition, guidelines for implementation of post-marketing research should be developed.

Phosphorylation of covalent organic framework nanospheres for inhibition of amyloid-ß peptide fibrillation.

The development and exploration of new nanostructural inhibitors against Alzheimer's disease (AD)-associated amyloid-ß (Aß) fibrillation have attracted extensive attention and become a new frontier in nanomedicine. However, focusing on finding an effective nanostructure is one of the most challenging parts of the therapeutics task. Herein, nanoscale spherical covalent organic frameworks (COFs) via post-synthetic functionalization with sodium phosphate (SP) groups on the channel networks were found to efficiently inhibit Aß fibrillation. The as-prepared uniform SP-COF nanospheres with high surface area, good crystallinity, and chemical stability were characterized by multifarious microscopic and spectroscopic techniques. Moreover, molecular dynamics simulation together with fibrillation kinetics and cytotoxicity assay experiments shows that there were restricted-access adsorption channels in the SP-COFs which were formed by the cavities with size and functional groups accommodated to the Aß peptide sequence and significantly affected the fibrillation and cytotoxicity of Aß. Transmission electron microscopy (TEM), dynamic light scattering (DLS) monitoring, isothermal titration calorimetry (ITC), Fourier transform infrared (FT-IR) and circular dichroism (CD) spectra measurements, and confocal imaging observation were performed to understand the inhibition mechanism and influencing factors of the SP-COFs. To our knowledge, our strategy is the first exploration of COF-based anti-amyloidogenic nanomaterials with high affinity and specific targeting, which are crucial for the inhibition of Aß fibrillation for AD prevention and treatment.

Aspartic Acid-Assisted Size-Controllable Synthesis of Nanoscale Spherical Covalent Organic Frameworks with Chiral Interfaces for Inhibiting Amyloid-ß Fibrillation.

Covalent organic framework nanospheres (COF NSs) have garnered special attention due to their uniform sphere morphology, adjustable particle size, and mesoporous microenvironment. However, methods to control an optimal particle size scale while achieving solution dispersibility and specific surface properties remain underdeveloped, which precludes many of the biomedical applications. Here, we propose and develop a general strategy to access simultaneous size control and surface functionalization of uniform spherical COF NSs in a single step using aspartic acid (d-/l-Asp) that plays center roles in an acid catalyst, hydrophilicity, size-controllable synthesis, and chiral enantiomer. In this study, for the first time, we have employed a surface chemistry engineering study to create a variety of nanoscale spherical COFs and subsequently measure parameters to evaluate the effectiveness of Asp in the regulation of the particle size. Moreover, the potential utilization of the d/l-enantiomeric Asp-COF NSs in preventing ß-amyloid (Aß) aggregation is investigated by analyzing their interactions with Aß amyloids using a multitechnique experimental approach. To our knowledge, our strategy is the first synthesis of hydrophilic COF NSs with an optimal length scale and a chiral-selective targeting surface, which are crucial for the inhibition of Aß fibrillation for Alzheimer's disease prevention.

Free amino acid, 5'-Nucleotide, and lipid distribution in different tissues of blue mussel (Mytilis edulis L.) determined by mass spectrometry based metabolomics.

Blue mussel (Mytilus edulis L.) is a popular, nutritional, and tasty mollusk. To better understand the composition of nutrients and improve further processing of the mussels, metabolomic approaches were used to analyze the free amino acids, 5'-nucleotides, and lipid compositions of different tissues. Our results showed that the viscera and gonad were rich in glutamine and glycine. Adenosine 5'-monophosphate, uridine 5'-monophosphate, guanosine 5'-monophosphate, and inosine 5'-monophosphate were abundant in the mantle, foot, and adductor muscle. Three main types of lipids, phospholipids (PLs), glycerides, and fatty acids (FAs), were semi-quantified. PLs were mainly distributed in the gonad of male mussels and viscera, gonad, and mantles of female mussels. FAs were relatively high in the viscera of males and in the gonad and viscera of females. The viscera of females were rich in phosphatidylcholine, such as 16:0/22:6 and 16:0/20:5. Triglycerides were the key lipids for distinguishing different tissues, especially 16:0/18:1/18:3 and 16:0/18:4/20:5.

A visual method for direct selection of high-producing Pichia pastoris clones.

BACKGROUND: The methylotrophic yeast, Pichia pastoris, offers the possibility to generate a high amount of recombinant proteins in a fast and easy way to use expression system. Being a single-celled microorganism, P. pastoris is easy to manipulate and grows rapidly on inexpensive media at high cell densities. A simple and direct method for the selection of high-producing clones can dramatically enhance the whole production process along with significant decrease in production costs. RESULTS: A visual method for rapid selection of high-producing clones based on mannanase reporter system was developed. The study explained that it was possible to use mannanase activity as a measure of the expression level of the protein of interest. High-producing target protein clones were directly selected based on the size of hydrolysis holes in the selected plate. As an example, the target gene (9elp-hal18) was expressed and purified in Pichia pastoris using this technology. CONCLUSIONS: A novel methodology is proposed for obtaining the high-producing clones of proteins of interest, based on the mannanase reporter system. This system may be adapted to other microorganisms, such as Saccharomyces cerevisiae for the selection of clones.

SPOP negatively regulates Toll-like receptor-induced inflammation by disrupting MyD88 self-association.

Toll-like receptor (TLR) signaling pathways need to be tightly controlled to avoid excessive inflammation and unwanted damage to the host. Myeloid differentiation primary response gene 88 (MyD88) is a critical adaptor of TLR signaling. Here, we identified the speckle-type POZ protein (SPOP) as a MyD88-associated protein. SPOP was recruited to MyD88 following TLR4 activation. TLR4 activation also caused the translocation of SPOP from the nucleus to the cytoplasm. SPOP depletion promoted the aggregation of MyD88 and recruitment of the downstream signaling kinases IRAK4, IRAK1 and IRAK2. Consistently, overexpression of SPOP inhibited the TLR4-mediated activation of NF-κB and production of inflammatory cytokines, whereas SPOP depletion had the opposite effects. Furthermore, knockdown of SPOP increased MyD88 aggregation and inflammatory cytokine production upon TLR2, TLR7 and TLR9 activation. Our findings reveal a mechanism by which MyD88 is regulated and highlight a role for SPOP in limiting inflammatory responses.

Identification of a new orthonairovirus associated with human febrile illness in China.

The genus Orthonairovirus, which is part of the family Nairoviridae, includes the important tick-transmitted pathogens Crimean-Congo hemorrhagic fever virus and Nairobi sheep disease virus, as well as many other poorly characterized viruses found in ticks, birds and mammals1,2. In this study, we identified a new orthonairovirus, Songling virus (SGLV), from patients who reported being bitten by ticks in Heilongjiang Province in northeastern China. SGLV shared similar genomic and morphological features with orthonairoviruses and phylogenetically formed a unique clade in Tamdy orthonairovirus of the Nairoviridae family. The isolated SGLV induced cytopathic effects in human hepatoma cells in vitro. SGLV infection was confirmed in 42 hospitalized patients analyzed between 2017 and 2018, with the main clinical manifestations being headache, fever, depression, fatigue and dizziness. More than two-thirds (69%) of patients generated virus-specific antibody responses in the acute phase. Taken together, these results suggest that this newly discovered orthonairovirus is associated with human febrile illness in China.

Postmortem nucleotide degradation in turbot mince during chill and partial freezing storage.

Nucleotide degradation in fish is an important biochemical change after death, which is closely related to freshness and sensory quality. However, except ATP-relative nucleotides, it remains unclear about changes in other nucleotide metabolites during postmortem stage. In this study, a strategy for the simultaneous quantification of 28 nucleobases, nucleosides, and nucleotides using hydrophilic interaction chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) with positive/negative ion switching was developed. This method showed good linearity, precision, repeatability, and recovery. Furthermore, it was successfully applied to monitor the postmortem nucleotide degradation of turbot mince during chill (4 °C) and partial freezing (-3 °C) storage for 168 h. It was noted that the patterns of the changes in nucleotide metabolites differed considerably depending on the storage temperature. Meanwhile, the different pathway and speed of nucleotide catabolism in turbot mince was summarized based on the quantification data.

Polycyclic aromatic hydrocarbons and their nitro-derivatives in urban road dust across China: Spatial variation, source apportionment, and health risk.

As an essential carrier of hazardous substances, fugitive road dust has become a severe issue in China. In this study, 212 road dust samples from 53 cities in China were collected to comprehensively investigate the spatial variations, potential sources, and cancer risk of 16 polycyclic aromatic hydrocarbons (PAHs) and 16 nitro-PAHs. The total PAHs concentrations ranged from 0.07 to 345 µg/g dry weight, which is at a moderate level compared to other regions in the world. The mean concentration of Σ16nitro-PAHs was 111 ± 115 ng/g, which is 2-3 orders of magnitude lower than that of Σ16PAHs. A clear geographical trend of dust PAHs and nitro-PAHs was observed in the northeast, north, and east coastal regions of China at a higher level. Moreover, a significant correlation between latitude and PAHs/nitro-PAHs revealed the influences of outdoor temperature and coal combustion for heating in the different regions on the emission and reaction of PAHs and nitro-PAHs. The secondary formation of most nitro-PAHs increases with a decrease in latitude indicated that solar radiation and temperature are important factors on secondary formation of nitro-PAHs. The average concentration of total PAHs and their derivatives in trunk road samples were statistically higher than those in other road samples (p < 0.05), indicating the influence of traffic load on target compound concentration. Generally, the primary sources of PAHs in the road dust samples were coal combustion (23.9%), vehicles (57.1%), and wood/biomass combustion (19.0%). For nitro-PAHs, the main sources were secondary formation (30.9%), biomass/coal combustion (28.4%), and vehicles (44.9%). Furthermore, a moderate potential carcinogenic risk due to PAHs and nitro-PAHs in the dust samples was found in China.

[Stoichiometry of soil extracellular enzymes and its seasonal variation in natural forests with different altitudes in northern Greater Khingan Mountains, China]. / å¤§å ´å®‰å²­åŒ—éƒ¨ä¸åŒæµ·æ‹”å¤©ç„¶æž—åœŸå£¤èƒžå¤–é ¶åŒ–å­¦è®¡é‡ç‰¹å¾åŠå ¶å­£èŠ‚åŠ¨æ€.

In this study, we examined the characteristics and influence mechanism of soil extracellular enzyme activity (EEA) and enzymatic stoichiometry in different soils in forests at different altitudes (750-1420 m) in Aokelidui Mountains in the north of the Greater Khingan Mountains. The results showed that altitude, season and their interactions significantly affected the activities of ß-glucosidase (BG), ß-1,4-N-acetylglucosaminidase (NAG), L-leucine aminopeptidase (LAP), and acid phosphatase (AP). In May, BG and NAG activities gradually increased with increasing altitude, while AP activities increased first and then decreased with increasing altitude. In July, NAG activity increased with altitude, while AP activity increased first and then decreased. In September, NAG activity changed significantly in different altitudes, with the highest activity at 1420 m (124.22 nmol·h-1·g-1). With the increases of altitude, ln(BG): ln(NAG+LAP) showed a decreasing trend. Except for the altitude of 830 m, stoichiometric ratio in all altitudes was the highest in July. The ratio of logarithmic conversion of soil C, N, and P invertase activity was 1:1.25:0.82. Altitude and soil temperature were the main factors affecting soil extracellular enzyme activities. There was a significant positive correlation between soil temperature and BG, NAG, and AP. Enzymatic stoichiometry ln(BG):ln(NAG+LAP) and ln(NAG+LAP):ln(AP) showed significant positive and negative correlations with soil pH, and had a negative and positive relationship with DOC. The ratio of ln(BG):ln(AP) was greatly affected by soil bulk density.

Construction of siRNA/miRNA expression vectors based on a one-step PCR process.

BACKGROUND: RNA interference (RNAi) has become a powerful means for silencing target gene expression in mammalian cells and is envisioned to be useful in therapeutic approaches to human disease. In recent years, high-throughput, genome-wide screening of siRNA/miRNA libraries has emerged as a desirable approach. Current methods for constructing siRNA/miRNA expression vectors require the synthesis of long oligonucleotides, which is costly and suffers from mutation problems. RESULTS: Here we report an ingenious method to solve traditional problems associated with construction of siRNA/miRNA expression vectors. We synthesized shorter primers (< 50 nucleotides) to generate a linear expression structure by PCR. The PCR products were directly transformed into chemically competent E. coli and converted to functional vectors in vivo via homologous recombination. The positive clones could be easily screened under UV light. Using this method we successfully constructed over 500 functional siRNA/miRNA expression vectors. Sequencing of the vectors confirmed a high accuracy rate. CONCLUSION: This novel, convenient, low-cost and highly efficient approach may be useful for high-throughput assays of RNAi libraries.

Cell-surface display of the active mannanase in Yarrowia lipolytica with a novel surface-display system.

A novel surface-display system was constructed using the cell-wall anchor protein Flo1p from Saccharomyces cerevisiae, the mannanase (man1) from Bacillus subtilis fused with the C-terminus of Flo1p and the 6xHis tag was inserted between Flo1p and man1. The fusion protein was displayed on the cell surface of Yarrowia lipolytica successfully, and it was confirmed by immunofluorescence. In succession, the surface-displayed mannanase was characterized. The optimum catalytic conditions for the recombinant mannanase were 55 degrees C at pH 6.0, and it exhibited high stability against pH variation. The highest activity of the recombinant mannanase reached 62.3 IU/g (dry cell weight) after the recombinant was cultivated for 96 h in YPD medium [1% (w/v) yeast extract/2% (w/v) peptone/2% (w/v) glucose]. To our knowledge, the present paper is the first to report that high-activity mannanase is displayed on the cell surface of Y. lipolytica with Flo1p.

Flavin-Based Fluorescent Protein EcFbFP Auto-Guided Surface Display of Methyl Parathion Hydrolase in Escherichia coli.

Methyl parathion hydrolase (MPH) plays an important role in degrading a range of organophosphorus compounds. In order to display MPH on the cell surface of Escherichia coli strain RosettaBlue™, the Flavin-based fluorescent protein EcFbFP was severed as an auto-anchoring matrix. With net negative charges of EcFbFP supplying the driving forces, fusion protein MPH-EcFbFP through a two-step auto-surface display process was finally verified by (a) inner membrane translocation and (b) anchoring at outer membrane. Cells with surface-displayed MPH obtained activity of 0.12 U/OD600 against substrate methyl parathion. MPH when fused with engineered EcFbFP containing 20 net negative charges exhibited fivefold higher anchoring efficiency and tenfold higher enzymatic catalytic activity of 1.10 U/OD600. The above result showed that MPH was successfully displayed on cell surface and can be used for biodegradation of methyl parathion.