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Contrast-enhanced ultrasound (CEUS) for the differentiation of benign and malignant peripheral pulmonary lesions has been considered experimental for many years. This study was aimed at evaluating the feasibility of CEUS as a diagnostic modality in this area of discussion. CEUS diagnostic accuracy was explored by comparison with contrast-enhanced computed tomography (CECT). The collective included 449 patients with 449 definitive diagnoses (benign, 178; malignant, 271). Logistic regression analysis of CEUS data revealed that delayed time to enhancement, chaotic pattern of distribution of vessels and mild extent of enhancement were independent risk factors for predicting malignancy. Time to wash-out and homogeneity of enhancement did not differ between the two groups (p > 0.05). Based on histopathology or clinical follow-up as a reference standard, CEUS and CECT had similar diagnostic accuracies of 80.16% and 81.75%, respectively. CEUS is a potentially useful imaging tool for diagnosing peripheral pulmonary lesions.

Objective to evaluate the role of Contrast-Enhanced Ultrasound (CEUS) of the Simple Rules (SR) from the International Ovarian Tumor Analysis group for the differential diagnosis of benign and malignant adnexal tumors. Methods A total of 180 patients with suspected malignant adnexal masses admitted to Lanzhou University Second Hospital were included, all of whom received conventional ultrasound (US) and CEUS examination before surgery. All masses were assessed using SR in US and SR in CEUS respectively. To compare the diagnostic performance , the sensitivity, specificity, negative and positive predictive values , Youden index and area under the curves values of SR in US and SR in CEUS were obtained. Results SR in CEUS was better than SR in US with its sensitivity, negative predictive values , Youden index and area under the curves . The specificity and negative predictive value of SR in CEUS were lower than that of SR in US. Compared with SR in US, SR in CEUS significantly reduced the number of uncertain masses(21, 36.7% vs 66, 11.7%). Conclusion We concluded that SR in CEUS performed well in the diagnosis of malignant adnexal masses. SR in CEUS is a supplement to SR in US.

© The Author(s), under exclusive licence to Springer Nature Limited 2021, corrected publication 2022, The COVID-19 pandemic has increased negative emotions and decreased positive emotions globally. Left unchecked, these emotional changes might have a wide array of adverse impacts. To reduce negative emotions and increase positive emotions, we tested the effectiveness of reappraisal, an emotion-regulation strategy that modifies how one thinks about a situation. Participants from 87 countries and regions (n = 21,644) were randomly assigned to one of two brief reappraisal interventions (reconstrual or repurposing) or one of two control conditions (active or passive). Results revealed that both reappraisal interventions (vesus both control conditions) consistently reduced negative emotions and increased positive emotions across different measures. Reconstrual and repurposing interventions had similar effects. Importantly, planned exploratory analyses indicated that reappraisal interventions did not reduce intentions to practice preventive health behaviours. The findings demonstrate the viability of creating scalable, low-cost interventions for use around the world., This project was supported by funds from: the Amazon Web Services (AWS) Imagine Grant (to E.M.B.); the Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (JSPS KAKENHI; 16h03079, 17h00875, 18k12015, and 20h04581 to Y.Y.); the research programme Dipartimenti di Eccellenza from the Ministry of Education University and Research (MIUR to N. Cellini and G.M. and the Department of General Psychology of the University of Padua); statutory funds of the University of Wroclaw (to A. Sorokowska); the Charles University Research Programme PROGRES (Q18 to M. Vranka); the Knut and Alice Wallenberg Foundation (2016:0229 to J.K.O.); the Rubicon Grant (019.183sg.007 to K.v.S.) from the Netherlands Organisation for Scientific Research; the Australian Research Council (dp180102384 to R.M.R.); the US National Institutes of Health (NIMH111640 to M.N.-D.), the Huo Family Foundation to N.J.; the NSF Directorate for Social, Behavioral and Economic Sciences, Division of Social and Economic Sciences (1559511 to J.S.L.); the US National Institutes of Health (RO1-CA-224545 to J.S.L.); Eesti Teadusagentuur–Estonian Research Council (PSG525 to A. Uusberg); the J. William Fulbright Program (to F. Azevedo); the HSE Basic Research Program (to D. Dubrov); Dominican University (a Faculty Development Grant to A. Krafnick); and the French National Research Agency Investissements d’avenir supporting PSF (ANR-15-IDEX-02 to H.I.); the Slovak Research and Development Agency (project no. APVV-20-0319 to M. Adamkovič); the programme FUTURE LEADER of Lorraine Université d’Excellence within the French National Research Agency Investissements d’avenir (ANR-15-IDEX-04-LUE to S.M.). Computation for this research was assisted by: the Harvard Business School compute cluster (HBSGrid); and the Open Science Grid. The Open Science Grid is supported by the National Science Foundation award 1148698 and the US Department of Energy’s Office of Science, as well as by the compute resources and assistance of the UW-Madison Center For High Throughput Computing (CHTC) in the Department of Computer Sciences. The CHTC is supported by UW-Madison, the Advanced Computing Initiative, the Wisconsin Alumni Research Foundation, the Wisconsin Institutes for Discovery, and the National Science Foundation, and is an active member of the Open Science Grid, which is supported by the National Science Foundation and the U.S. Department of Energy’s Office of Science.

Enzymatic browning is a major issue affecting the quality of processed potato (Solanum tuberosum L.). To understand the molecular mechanism of browning, transcriptional analyses were performed by employing potatoes that differed in browning. Coexpression analysis indicated that 9 out of 15 upregulated genes in browning-less groups encoded for potato protease inhibitors (StPIs). In addition, gene otology analysis showed that the enriched terms were mainly involved in protease inhibitors. Overexpression of cysteine StPI 143 and StPI 146 individually reduced browning and lowered protease activities and tyrosine and total free amino acid (FAA) contents, but they could not decrease polyphenol oxidase activity. Moreover, supplementing exogenous tyrosine or total FAAs into transgenic potato mash to wild-type amounts promoted mash browning, browning with total FAAs, more than with tyrosine, resembling wild-type levels. These results implied that cysteine StPIs reduced browning via lowering the accumulation of FAAs in addition to tyrosine. Our findings have enriched the knowledge about the roles and mechanisms of protease inhibitors in regulating enzymatic browning of potato, which provide new ways for controlling potato browning.

Objective: Overactive bladder (OAB) is a disease characterized by the presence of urinary urgency. We carried out a meta-analysis to assess the effectiveness and safety of trigonal-involved injection of onabotulinumtoxinA (BoNT-A) in comparison with the trigonal-sparing technique in cases with OAB [neurogenic detrusor overactivity (NDO) and idiopathic detrusor overactivity (IDO)].Methods: Randomized controlled trials (RCTs) of BoNT-A injection for OAB were searched systematically by using EMBASE, MEDLINE, and the Cochrane Controlled Trials Register. The datum was calculated by RevMan version 5.3.0. The original references of relating articles were also reviewed.Results: In total, six RCTs involving 437 patients were included in our analysis. For OAB, the trigone-including group showed a different patient symptom score (p = 0.03), complete dryness rate (p = 0.002), frequency of incontinence episodes (p = 0.01), detrusor pressure at maximum flow rate (p = 0.01), and volume at the first desire to void (p = 0.0004) compared with the trigone-sparing group. Also, a trigone-including intradetrusor injection demonstrated a significant improvement in the patient symptom score (p = 0.0004), complete dryness rate (p = 0.0002), frequency of incontinence episodes (p = 0.0003), detrusor pressure at maximum flow rate (p = 0.01), and volume at the first desire to void (p = 0.00006) compared with the trigone-sparing group for treatment of NDO. The adverse events rates were similar in both groups.Conclusions: The meta-analysis has demonstrated that trigone-including BoNT-A injection was more effective compared with the trigone-sparing injection for the treatment of OAB, especially for NDO.

In this paper, for the first time the binding behavior of cadmium (Cd2+) to rice proteins (RPs) was studied. The results showed that the equilibrium of binding was reached within 30 min at 303 K with a maximum binding amount (q) of 15.26 mg/g, and the pH was an important factor positively influencing q. At both 308 K and 313 K, the binding of Cd2+ to RPs belonged to spontaneous, endothermic interactions with high-affinity, assigned to a multidentate coordination. Except for acetate, all the investigated competing coordination agents, such as edetate, pyrophosphate and citrate, showed inhibitory effects on RPs-Cd2+ binding, and edetate seemed to be the most effective one. At pH 6.5, calcium, copper and zinc began to restrict RPs-Cd2+ binding when the metal ion concentration reached 500 mg/kg, and the decreasing of pH would strengthen the inhibitory effects of the investigated metal ions including ferric ions.

Yanfang Wang,1 Fang Nie,1 Guojuan Wang,1 Ting Liu,1 Tiantian Dong,1 Yamin Sun2 1Medical Center of Ultrasound, Lanzhou University Second Hospital, Lanzhou, Gansu, People’s Republic of China; 2Department of Pathology, Lanzhou University Second Hospital, Lanzhou, Gansu, People’s Republic of ChinaCorrespondence: Fang NieMedical Center of Ultrasound, Lanzhou University Second Hospital, Chengguan District, Lanzhou, Gansu Province, 730030, People’s Republic of ChinaTel +86 0931 8942545Email ery_nief@lzu.edu.cnPurpose: Early and accurate preoperative diagnosis of central lymph node metastasis (CLNM) is crucial to improve surgical management of patients with clinical lymph node-negative papillary thyroid carcinoma (PTC). Towards improving diagnosis of CLNM, we assessed the value of combining preoperative clinical characteristics, conventional ultrasound, and contrast-enhanced ultrasound (CEUS) in preoperative prediction of CLNM of different sized PTCs.Patients and Methods: Patients were divided according to tumor size: a PTC group (> 10 mm) and a papillary thyroid microcarcinoma (PTMC) group (≤ 10 mm). We retrospectively analyzed the clinical and ultrasonographic features of 120 PTC patients and 165 PTMC patients. Multivariate logistic regression analysis was used to screen independent risk factors and establish prediction models. Receiver operating characteristic curves were used to determine the best cut-off values for continuous variables and assess the performance of prediction models.Results: Independent risk predictors of CLNM for the PTC group were extrathyroidal extension in CEUS (OR=7.923), tumor size > 14 mm (OR=5.491), and multifocality (OR=3.235). For the PTMC group, the independent risk factors were the distance from the thyroid capsule =0 mm (OR=4.629), male (OR=3.315), tumor size > 5 mm (OR=3.304), and microcalcification (OR=2.560). The predictive model of combined method had better performance in predicting CLNM of PTC compared with models based on CEUS and conventional ultrasound alone (area under the curve: 0.832 vs 0.739, P=0.0011; 0.832 vs 0.678, P=0.0012). For PTMC, comparing with CEUS, the combined method and conventional ultrasound performed better than CEUS alone in predicting CLNM (area under the curve: 0.783 vs 0.636, P=0.0016; 0.738 vs 0.636, P=0.0196).Conclusion: The predictive models of combined method obtained from significant preoperative clinical and ultrasonographic features can potentially improve the preoperative diagnosis and individual treatment of CLNM in patients with PTC and PTMC. CEUS may be helpful in predicting CLNM of PTC, but CEUS would be ineffective in predicting CLNM of PTMC.Keywords: papillary thyroid carcinoma, lymph node metastasis, prophylactic central neck dissection, ultrasonography, contrast-enhanced ultrasound

ObjectiveThis study aims to investigate the value of contrast-enhanced ultrasound (CEUS) in the differential diagnosis and risk stratification of ACR TI-RADS category 4 and 5 thyroid nodules with non-hypovascular.MethodsFrom January 2016 to December 2019 in our hospital, 217 ACR TI-RADS category 4 and 5 nodules with non-hypovascular in 210 consecutive patients were included for a derivation cohort. With surgery and/or fine-needle aspiration (FNA) as a reference, conventional ultrasound (US) features and CEUS features were analyzed. Multivariate logistic regression analysis was used to screen the independent risk factors and establish a risk predictive model. Between January 2020 and March 2021, a second cohort of 100 consecutive patients with 101 nodules were included for an external validation cohort. The model was converted into a simplified risk score and was validated in the validation cohort. The area under the receiver operating characteristic curves (AUC) were used to assess the models’ diagnostic performance.ResultsMicro-calcification, irregular margin, earlier wash-out, centripetal enhancement, and absence of ring enhancement were independent risk factors and strongly discriminated malignancy in the derivation cohort (AUC = 0.921, 95% CI 0.876–0.953) and the validation cohort (0.900, 0.824–0.951). There was no significant difference (P = 0.3282) between the conventional US and CEUS in differentiating malignant non-hypovascular thyroid nodules, but a combination of them (the predictive model) had better performance than the single method (all P ConclusionsCEUS is helpful in combination with conventional US in differentiating ACR TI-RADS category 4 and 5 nodules with non-hypovascular. The risk score in this study has the potential to improve the diagnosis and risk stratification of non-hypovascular thyroid nodules.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has become worldwide pandemic and posed a serious public health risk. It was proved that lung ultrasound (LUS) can be extremely valuable in the diagnosis and treatment of the disease, which could also minimize number of exposed healthcare workers and equipment. Since in the department of ultrasound, the healthcare workers are close contacted with patients who might be infected or virus carrying, it is extremely important to offer sufficient protection of them. Too aggressive protection should also be avoided because all the hospital might encounter lack of protection equipment. Guidance about the proper protection management should be provided in detailed, for example, how to choose personal protective equipment, or how to disinfect the environment. To address those problems, on behalf of Chinese Ultrasound Doctors Association, Chinese PLA Professional Committee of Ultrasound in Medicine, Beijing Institute of Ultrasound in Medicine, and Chinese Research Hospital Association Ultrasound Professional Committee, the authors summarized the recommendations for effective protection according to the exist hygienic standards, their experience, and available literature. After the recommendations were finished, two online conferences were held on January 31st 2020 and February 7th 2020 respectively, and the recommendations were discussed in detail part by part. A modified version of the work was circulated and finally approved by all the authors, which was the present Chinese Experts Consensus on Protection for Ultrasound Healthcare Workers against COVID-19.

Purpose: To explore the diagnostic value of Contrast-enhanced Ultrasound Liver Imaging Reporting and Data System version 2017 (CEUS LI-RADS v2017) in differentiating alpha-fetoprotein (AFP)-negative hepatocellular carcinoma (HCC) from other primary malignancies (OM) of the liver. Methods: The data of 99 patients with primary liver malignant tumors confirmed by surgical pathology and AFP-negative from January 2018 to January 2021 were retrospectively analyzed, and the lesions were divided into 61 cases in the AFP-negative HCC group and 38 cases in the OM group according to the pathological findings, the CEUS features of the lesions were analyzed and the lesions were classified according to the CEUS LI-RADS v2017. Comparison of CEUS features between the two groups was performed using the χ2 test. The sensitivity, specificity, positive predictive value, negative predictive value, and coincidence rate of CEUS LI-RADS v2017 for the diagnosis of AFP-negative HCC and OM were calculated using pathological findings as the gold standard. Results: The differences in features of arterial phase enhancement and wash-out between the HCC and OM groups were statistically significant (p < 0.05). The sensitivity of diagnosing HCC by LR-5 was 62.3% and the specificity was 92.1%. The sensitivity of diagnosing OM by LR-M was 92.1% and the specificity was 83.6%. Conclusions: When AFP is negative in patients with intrahepatic focal lesions, LR-5 has high specificity but low sensitivity in the diagnosis of HCC, and LR-M has high sensitivity and specificity in the diagnosis of OM. CEUS LI-RADS is a tool to differentiate AFP-negative HCC and OM effectively.

Enzymatic browning is an important issue affecting the quality and shelf life of fresh-cut potato. Previous research showed post-cut sodium chloride (NaCl) dipping treatment doesn’t have anti-browning effect on fresh-cut potato. In this research, the influence of pre-cut NaCl solution treatment on browning, polyphenol oxidase (PPO) activity and amino acids contents in fresh-cut potato was investigated. The results showed that pre-cut NaCl solution treatment at 5 % for 3 h improved the overall visual quality, reduced browning and surface dehydration and extended shelf-life by 3−4 d at 5 °C. The PPO activity in pre-cut NaCl treated potato tubers and slices was lower than that in control and water treated ones. Compared with control, the contents of glutamic acid and proline in NaCl treated potato were increased, whereas tyrosine and arginine decreased. Exogenous differential compensation of glutamic acid reduced the browning of control potato mash, while the compensation of tyrosine and arginine as well as their mixture aggravated the browning of pre-cut NaCl treated potato mash. The compensation of proline showed no influence on the browning of control potato mash, whereas pre-cut proline solution treatment inhibited the browning of fresh-cut potato. This research showed that pre-cut NaCl solution treatment can effectively inhibit the browning of fresh-cut potato by influencing polyphenol oxidase activity and several free amino acids contents, which provides a new way to prevent the browning of fresh-cut potato and elucidates the anti-browning mechanism of pre-cut NaCl treatment.

Nitrogen metabolism in citrus has received increased attention due to its effects on plant growth and productivity. However, little is known about the effects of nitrogen fertilization on nitrogen metabolism in young trees of citrus cultivar 'Huangguogan' (Citrus reticulata × Citrus sinensis). Here, genes encoding nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate dehydrogenase (GDH), and asparagine synthetase (AS), represented as HgNR, HgNiR, HgGS, HgGDH, and HgAS, respectively, were cloned from Huangguogan. Deduced protein sequences were analyzed and proteins were confirmed to be localized in their respective cellular organelles. Moreover, pot-cultured 'Huangguogan' seedlings were fertilized with 0 (N1), 1.36 (N2), 1.81 (N3), 2.26 (N4), or 2.72 (N5) kg N/year, for 12 months. Enzyme activity and enzyme-gene expression were studied in roots, leaves, and fruits at different stages. Finally, the effects of N application rate on root activity, leaf N, soluble protein, yield, and fruit quality at the ripening stage were measured. The results showed that: 1) HgNR, HgNiR, HgGDH, and HgAS gene products were found mainly in the cytoplasm and plasma membrane, while HgGS gene product was found mainly in cytoplasm and mitochondria. 2) Gene expression and enzyme activity differed among plant organs. As the root is in permanent direct contact with the soil we suggest that root gene expression and enzyme activity can be used as reference to determine N application rate. 3) Yield, fruit quality, enzyme activity, and enzyme-related gene expression were considerably lower at low than at high-N supply. However, they were all inhibited by excess nitrogen (i.e., 2.72 kg/year). Therefore, we recommend 1.81 kg N/year as the optimal N application rate for young 'Huangguogan' trees.

Bitterness, which is mainly attributed to limonoids, seriously affects the taste of pomelo fruits and hinders their competitiveness in the market. Therefore, a high-performance liquid chromatography method was developed for simultaneous isolation and determination of limonin, nomilin, and limonin glucoside contents in pomelo fruit, and the method showed good precision (RSD ≤ 2.85 %), repeatability (RSD ≤ 3.37 %) and high sample recovery rate (95.8 %–99.7 %). Four pomelo varieties were used to investigate the accumulation and distribution of limonoids in different stages of fruit development and in different plant tissues and varieties, as well as to reveal the synthesis mechanism and provide a basis for the utilisation of pomelo resources in different industries. Results showed that flavedo was the tissue richest in limonoids, and the maximum limonoid content could reach 5959 μg/g fresh weight. During fruit growth and development, the limonin and nomilin contents in juice sacs and albedo first increased and then decreased. Although there were some fluctuations, the limonin and nomilin contents in the flavedo generally showed a decreasing trend with fruit development. Our investigation of the LGT gene expression pattern provided the basis for exploring natural debittering mechanisms in pomelo. Expression of the LGT gene was positively correlated with limonin glucoside content.

Enzymatic browning greatly affects the quality of fresh-cut potatoes (Solanum tuberosum L.). The transcript level of a novel aspartic protease inhibitor gene, StASPI, was found significantly higher in browning-less potatoes than in browning-susceptible potatoes, and exogenous aspartic protease inhibitor Pepstatin A inhibited the browning of potato pulp. Therefore, we speculated that StASPI plays important roles in browning-resistance of potatoes. The effects of overexpressed-StASPI on browning, total free amino acids (FAAs), antioxidant enzyme activity and ROS accumulation in potatoes were investigated in this study. Results showed that overexpression of StASPI effectively reduced enzymatic browning after cutting, significantly decreased protease activity, and reduced the accumulation of total FAAs in potatoes. Browning degrees of transgenic potato mash were increased by supplementing exogenous FAAs, but the degrees were still significantly lower than that of wild-type (WT) mash. Furthermore, overexpression of StASPI decreased PPO activity, enhanced the activities of antioxidant enzymes SOD and CAT, and reduced H2O2 and O2˙― contents. These results indicated that overexpression of StASPI inhibited the enzymatic browning of potatoes, decreasing FAAs accumulation, reducing PPO activity, and enhancing activity of antioxidant enzymes. This study provides a new perspective on the strategies for inhibiting enzymatic browning of potatoes.

The authors describe a dual signal amplification electrochemiluminescent (ECL) immunoassay for ultrasensitive determination of the β-adrenergic agonist salbutamol (SAL). It is based on the use of gold nanoparticles (AuNPs) and silica-coated quantum dots of the type CdSe@SiO2. The CdSe@SiO2 nanoparticles (NPs) were synthesized by water-in-oil reversed-phase microemulsion. Amino-modified NPs were then prepared and used as an ECL signal probe for detection of SAL in the presence of K2S2O8. The gold NPs with their large specific surface carry the antigen, facilitate electron transfer and improve the electrochemical reaction efficiency of QDs and K2S2O8. The ECL immunosensor was fabricated by coating the AuNPs with antigen, and then SAL competed with SAL antigen for the binding sites of the antibody on the CdSe@SiO2 NPs. Under optimized conditions, the ECL intensity is linearly related to the logarithm of the SAL concentration in the range from 0.001 to 1000 ng mL−1, with a lower detection limit of 0.17 pg mL−1 (at an S/N ratio of 3). The assay has good operational stability, is highly sensitive and selective. It is judged to possess a wider scope in that it may be applied to many other small molecule analytes for which adequate antibodies are available.

Litopenaeus vannamei has a salinity tolerance range of 0.5–78 ppt. In China, cultivation of L. vannamei in high salinity waters in the Circum-Bohai Region resulted in low survival rates and yields because high salinity has toxic effects on shrimp. Thus, identifying differential gene expression in different tissues of L. vannamei under high salinity stress would help revealing its salt tolerance mechanisms. In the present study, L. vannamei was cultivated under two salinity levels (34 and 58 ppt) for 90 days. Gill, muscle, and hepatopancreatic tissues were then extracted from the shrimp for transcriptome sequencing and quantitative real-time PCR, to analyze the differential expression of genes at the transcriptome level. The 1659 significantly differentially expressed genes comprised 436, 318, and 364 upregulated genes and 428, 317, and 156 downregulated genes in gill, hepatopancreatic, and muscle tissues, respectively. Higher differential expression was found in genes related to osmoregulation, ion transport, and antioxidation in gill tissue; immune defense, bacterial lysis, and apoptosis in hepatopancreatic tissue; and development and nutrient metabolism in muscle tissue. The results of this study can enrich the basic knowledge of shrimp stress biology, and provide reference for understanding the molecular mechanism of shrimp salt tolerance.

'Huangguogan' (Citrus reticulata × C. sinensis) is a new cultivar of mandarin citrus in China, and the research on fertilization of 'Huangguogan' is very limited. In this study, the effect of N fertilization on 'Huangguogan' fruit quality was determined at ripening. Sugars (sucrose, fructose, and glucose), organic acids (pyruvic, oxalic, citric acid, etc.), and vitamin components were measured at six stages of fruit development, and eight enzymes related to the glycolytic and Krebs cycle were assessed. The 1.81 kg N y-1 treatment group showed the highest total soluble solids concentration and total soluble solids/titratable acidity ratio but the lowest titratable acidity (acid content) at ripening, while the N1 treatment (0 kg N y-1) showed the opposite trend. Sucrose and citric acid accumulated to the largest extent during fruit development. Sucrose and ascorbic acid content increased (8.46 to 50.97 mg g-1 and 8.16 to 27.39 mg g-1, respectively), while citric acid content decreased (90.81 to 0.02 mg g-1). Aconitase was the key enzyme responsible for the observed changes in citric acid. The N concentrations in ripening fruit ranged from 2.25% to 4.15%. Curve estimation and principal component analysis revealed that fruit N was positively correlated with the sugars and vitamin components and negatively correlated with the organic acids. The accumulation of these metabolites seemed closely related to the dynamic changes in fruit N concentration at the five N levels tested. In conclusion, we suggest that the 1.81 kg N y-1 treatment represents the most suitable N fertilizer treatment for 'Huangguogan' citrus fruit.

Nitrogen metabolism in citrus has received increased attention due to its effects on plant growth and productivity. However, little is known about the effects of nitrogen fertilization on nitrogen metabolism in young trees of citrus cultivar 'Huangguogan' (Citrus reticulata × Citrus sinensis). Here, genes encoding nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), glutamate dehydrogenase (GDH), and asparagine synthetase (AS), represented as HgNR, HgNiR, HgGS, HgGDH, and HgAS, respectively, were cloned from Huangguogan. Deduced protein sequences were analyzed and proteins were confirmed to be localized in their respective cellular organelles. Moreover, pot-cultured 'Huangguogan' seedlings were fertilized with 0 (N1), 1.36 (N2), 1.81 (N3), 2.26 (N4), or 2.72 (N5) kg N/year, for 12 months. Enzyme activity and enzyme-gene expression were studied in roots, leaves, and fruits at different stages. Finally, the effects of N application rate on root activity, leaf N, soluble protein, yield, and fruit quality at the ripening stage were measured. The results showed that: 1) HgNR, HgNiR, HgGDH, and HgAS gene products were found mainly in the cytoplasm and plasma membrane, while HgGS gene product was found mainly in cytoplasm and mitochondria. 2) Gene expression and enzyme activity differed among plant organs. As the root is in permanent direct contact with the soil we suggest that root gene expression and enzyme activity can be used as reference to determine N application rate. 3) Yield, fruit quality, enzyme activity, and enzyme-related gene expression were considerably lower at low than at high-N supply. However, they were all inhibited by excess nitrogen (i.e., 2.72 kg/year). Therefore, we recommend 1.81 kg N/year as the optimal N application rate for young 'Huangguogan' trees.

In the present work, a simple and sensitive flow injection chemiluminescent competitive immunoassay was developed for the determination of mercury(II) ion (Hg(2+)) based on carboxylic resin beads, a novel specific monoclonal antibody (McAb) and HRP enzyme-amplification. Resin beads with carboxyl groups were creatively employed as supports for immobilizing more coating antigen through acylamide bonds. With a competitive-type assay mode, the Hg(2+) in solution competed with the immobilized coating antigen for the limited McAb. Then, the second antibody labeled with HRP was introduced, and an effectively increased CL was obtained, which was ascribed to the catalytic activity of HRP for the luminol-PIP-H2O2 reaction. With increasing concentration of Hg(2+), the CL of this system decreases because less HRP is present in the CL reaction. At optimal conditions, the CL signal displayed a good linear relation toward Hg(2+) in the range of 0.05-200 ng mL(-1) with a detection limit (3σ) of 0.015 ng mL(-1). The immunosensor possessed high specificity, acceptable accuracy and reproducibility, and was examined in real samples with favorable results. This immunoassay will have intriguing application prospects for the determination of other heavy metal ions and environmental contaminants.

Objectives: Evasion of apoptosis and avoidance of immune surveillance represent two key hallmarks of cancer. Members of the inhibitor of apoptosis (IAP) gene play important interconnecting roles in both of these characteristic pathways of tumorigenesis, providing a critical nexus in targeting of cancer. Small-molecule antagonists of the IAPs, known as Smac mimetic compounds (SMCs), are in quick clinical development for cancer therapy. We disclose here a novel IAP antagonist TQB3728 and its preclinical evaluation on tumor therapy. Methods: The biochemical activities of TQB3728 for cIAP1 BIR3, cIAP2 BIR3 and XIAP BIR3 were measured by the corresponding biochemical assays. The in vitro anti-proliferative activity was evaluated on MDA-MB-231 and EMT6 cell lines. The in vivo antitumor activity of TQB3728 was evaluated in two cell-derived xenograft (CDX) models of TNBC (MDA-MB-231) and lymphoma (Karpas 422), and one syngeneic model of TNBC (EMT6). PK/PD was assessed by western blot analysis of cIAP1 expression and cleaved caspase 3 level in the treated tumor tissues. Results: TQB3728 displayed potent biochemical activities for cIAP1 BIR3, cIAP2 BIR3 and XIAP BIR3 with IC50 of 0.7, 11.8 and 22.5 nM, respectively. The expected anti-proliferative activities were also observed in both tumor cell lines, with a IC50 of 14.3 Nm for MDA-MB-231, and IC50 of 149.5 nM for EMT6 in the presence of exogenous TNFα (1 ng/ml). TQB3728 showed in vivo antitumor activity in the MDA-MB-231 CDX model with 89.8% TGI @30 mpk QD, which is better than LCL-161 (71.9% TGI @30 mpk, QD). TQB3728 also showed good antitumor activity in vivo in the Karpas 422 lymphoma CDX model with 112.2% TGI @100 mpk Q3D. In EMT6 tumor-bearing mice, the combination of TQB3728 and anti-PD1 antibody significantly regressed the tumor growth (TGI = 106.3%), whereas monotherapies only displayed moderate activities ( 60.6% and 0.4% TGI for anti-PD1 and TQB3728, respectively). Western blot analysis of the tumor samples showed that TQB3728 promoted caspase 3 cleavage, indicating that the tumor growth inhibition could be mediated by cell apoptosis signaling. Conclusions: We have discovered a potent orally active IAP antagonist, TQB3728, which shows activity of cIAP1 inhibition and strong antitumor activity in both in vitro and in vivo preclinical tumor models. These results are highly promising and warrant moving the compound forward to clinical investigation. Citation Format: Yingchun Liu, Xiquan Zhang, Ling Yang, Xin Tian, Tiantian Dong, Charles Z. Ding, Lihong Hu, Zhaobing Xu, Yuanfeng Xia, Haijuan Tong, Wenxi Li, Lijuan Hou, Chichung Chan, Shuhui Chen. Preclinical evaluation of TQB3728, a potent orally active IAP antagonist [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 724.

Electrochemiluminescent (ECL) immunosensor with multiple signal amplification was designed based on gold nanoparticles (AuNPs), polyamidoamine dendrimers (PAMAM) and silver-cysteine hybrid nanoribbon (SNR). Low toxic l -cysteine capped CdSe QDs was chosen as the ECL signal probe. To verify the proposed ultrasensitive ECL immunosensor for β-adrenergic agonists (β-AA), we detected Brombuterol (Brom) as a proof-of-principle analyte. Therein, AuNPs as the substrate can simplify the experiment process, accelerate the electron transfer rate, and carry more coating antigen (Ag-OVA) to enlarge ECL signal. On one hand, SNR on the surface of electrode can avoid the aggregation of AuNPs, and SNR-PAMAM-AuNPs also can be acted as a good accelerator for electron transfer. On the other hand, PAMAM (16 -NH 2 ) functionalized SNR (SNR-PAMAM) with numerous amino groups could be employed to bond abundant actived QDs to further amplify ECL signal. The new immunosensor can offer a simple, reliable, rapid, and selective detection for Brom, which have a dynamic range of 0.005–700 ng mL −1 with a low detection limit at 1.5 pg mL −1 . The proposed biosensor will extend the application of nanomaterials in ECL immunoassays and open a new road for the detection of Brom and other β-AA in the future.

Objective: Heat Shock Protein 90 (Hsp90) is a molecular chaperone that mediates the post-translational stability of its protein substrates, many of which are onco-proteins. Hsp90 is emerging as an important target for cancer therapy because its inactivation may abrogate multiple signaling pathways simultaneously, irrespective of the mutational status of proteins involved. We disclose here our clinical candidate WXFL1024H251, a novel second-generation small-molecule Hsp90 inhibitor with a chemical structure unrelated to the first generation, the ansamycin family of Hsp90 inhibitors. Method: The anti-proliferative activity of WXFL1024H251 was evaluated in a variety of cancer cell lines in a 96-hour viability assay. QW, Q2W or TIW intravenous administration of at doses of 5~50 mg/kg was used to evaluate the in vivo anti-tumor activity of WXFL1024H251 in BT474 cell line-derived xenograft (CDX) breast cancer model. Effects of WXFL1024H251 on HSP90 function were assessed by western blot analysis of PD markers HSP70, Her2 and p-Her2 in the BT474 tumors. Result: A panel of 20 cancer cell lines were screened for sensitivity to WXFL1024H251. Proliferation of 2 cancer cell lines A549 and BT474 were potently inhibited by WXFL1024H251, with IC50 of 0.007 μM and 0.009 μM, respectively. A549 cell line has KRAS mutation and medium to high EGFR expression. BT474 is an ER+ and ERBB2+ breast cancer cell line. In the BT474 tumor model, 3 week treatment with WXFL1024H251 showed good antitumor efficacy with T/C values of 36% at 7.5 mg/kg Q2W and 45% at 10 mg/kg QW; toxicity were acceptable with both dosing regiments. Conclusion: These preclinical studies showed good antitumor efficacy of WXFL1024H251, which was associated with client protein maturation and HSP70 up-regulation in the tumor tissue. WXFL1024H251 may represent a novel promising clinical antitumor agent for treating breast cancer and other cancer types with high expression of HSP90. Citation Format: Xiaobing yan, Charles Z. Ding, Lihong Hu, Wei Huang, Shuhui Chen, Dongdong Wu, Jiahu Wu, Guoqiang Ma, Qiyao Zhang, Rui Gao, Yusong Zhu, Tiantian Dong, Yizhong Zhu, Caixia Yan, Yan Yan. Preclinical evaluation of WXFL1024H251, a potent second-generation HSP90 inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1672.

This study describes a novel electrochemical immunosensor to amplify the electrochemiluminescence (ECL) signal for the ultrasensitive detection of salbutamol (SAL) using quantum dots (QDs) and gold nanoparticle (AuNP) conjugated horseradish peroxidase (HRP). The electrochemical detection was based on the HRP catalyzed consumption of self-produced H2O2, which has been extensively used as a co-reactant of QDs, by o-phenylenediamine (OPD). The enzymatic reaction rate is proportional to the amount of HRP bound to the electrode. In the presence of a SAL standard solution, the immobilized SAL coating antigens competed with the SAL solution for the Ab-AuNPs-HRP complexes. With an increase in the SAL concentration, the amount of immobilized HRP decreases, which leads to an increase in the ECL intensity. Under optimized conditions, the ECL intensity changes linearly with the logarithm of the SAL concentration in the range of 0.05-500 ng mL(-1) with a detection limit of 0.017 ng mL(-1) (S/N = 3). The ECL immunosensor possesses high sensitivity, satisfactory reproducibility and selectivity, and may provide a feasible route for practical application.