Comparative diagnostic accuracy of the IOTA SRR and LR2 scoring systems for discriminating between malignant and Benign Adnexal masses by junior physicians in Chinese patients: a retrospective observational study.

BACKGROUND: The accuracy of ultrasound in distinguishing benign from malignant adnexal masses is highly correlated with the experience of ultrasound physicians. In China, most of ultrasound differentiation is done by junior physicians. PURPOSE: To compare the diagnostic performance of the International Ovarian Tumour Analysis (IOTA) Simple Rules Risk (SRR) and IOTA Logistic Regression Model 2 (LR2) scoring systems in Chinese patients with adnexal masses. METHODS: Retrospective analysis of ovarian cancer tumor patients who underwent surgery at a hospital in China from January 2016 to December 2021. Screening patients with at least one adnexal mass on inclusion and exclusion criteria. Two trained junior physicians evaluated each mass using the two scoring systems. A receiver operating characteristic curve was used to test the diagnostic performance of each system. RESULTS: A total of 144 adnexal masses were retrospectively collected. Forty masses were histologically diagnosed as malignant. Compared with premenopausal women, postmenopausal women had a much higher rate of malignant masses. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of the SRR was 97.5% (95% CI: 86.8 -99.9%), 82.7% (95% CI: 74.0 -89.4%), 68.4% (95% CI: 58.7 -76.8%) and 98.9% (95% CI: 92.5 -99.8%). The sensitivity, specificity, PPV, NPV of the LR2 were 90.0% (95% CI: 76.5 -97.2%), 89.4% (95% CI: 81.9 -94.6%), 76.6% (95% CI: 65.0 -85.2%), and 95.9% (95% CI: 90.2 -98.3%). There was good agreement between two scoring systems, with 84.03% total agreement and a kappa value of 0.783 (95% CI: 0.70-0.864). The areas under the curve for predicting malignant tumours using SRR and LR2 were similar for all patients (P > 0.05 ). CONCLUSION: The two scoring systems can effectively distinguish benign from malignant adnexal masses. Both scoring systems have high diagnostic efficacy, and diagnostic efficacy is stable, which can provide an important reference for clinical decision making.

Transcriptome analysis revealed detoxification gene expression changes in Tetranychus cinnabarinus challenged with ethyl oleate.

Natural acaricides are potential biorational mite control alternatives to conventional chemical acaricides. However, little is known about the molecular mechanism of defense response to natural acaricides in mites. We previously reported significant acaricidal properties of ethyl oleate (EO) against Tetranychus cinnabarinus (here referred to as a sibling species of two-spotted spider mite, Tetranychus urticae), a highly polyphagous pest devastating crops in fields and greenhouses worldwide. In this study, we explored the molecular responses of T. cinnabarinus exposed to EO using RNA-Seq and differentially expressed gene (DEG) analysis. A total of 131, 185, and 154 DEGs were identified in T. cinnabarinus after 1, 6, and 24 h of EO treatment. In addition, 36 putative detoxification-related DEGs, including 10 cytochrome P450s (P450s), three glutathione S-transferases (GSTs), nine UDP-glycosyltransferases (UGTs), eight esterases (ESTs), and six ATP-binding cassette transporters (ABC transporters), were identified. Interestingly, the upregulation of these detoxification-related genes might be the main defense response of T. cinnabarinus exposed to EO. A quantitative real-time PCR analysis indicated that the expression profiles of 19 random DEGs were consistent with the RNA-Seq results. These findings serve as valuable information for a better understanding of the acaricide-mite interaction and molecular mechanisms involved in the defense response of T. cinnabarinus against EO.

Effect of MT2A on apoptosis and proliferation in HL60 cells.

Although accumulating evidence has revealed that metallothioneins (MTs) and its family member MT2A are strongly linked to the risk of various solid tumors, researches on the occurrence and development of acute myeloid leukemia (AML) have rarely been investigated. Here, we constructed a lentiviral vector with MT2A over-expression and the interfering plasmids with MT2A expression inhibition to study the influence of MT2A on the bioactivities of HL60 cells. After cells were infected with a lentiviral vector containing the MT2A gene, both transcription and translation levels of MT2A were significantly increased in the over-expressed group in comparison with control groups. In vitro experiments, all results demonstrated that cell reproductive capacity was inhibited, but cell apoptosis rate was significantly increased. Together, the expression of apoptosis-related protein Bcl2 was remarkably reduced, while a high expression level of Bax protein was detected. Further experiments revealed that up-regulation of MT2A induced cell apoptosis and promoted G2/M phase arrest. The mechanism may be associated with down-regulated p-IκB-α and cyclinD1 expression and up-regulated IκB-α expression in the nuclear factor-kappaB (NF-κB) pathway. On the contrary, MT2A expression was down-regulated by interfering plasmids. We found that cell proliferative potential was notably increased in the interfering group compared with the negative and untreated group. What's more, MT2A may be closely related to AML cell proliferation and function via the NF-κB signal pathway.

Soluble cpg15 from Astrocytes Ameliorates Neurite Outgrowth Recovery of Hippocampal Neurons after Mouse Cerebral Ischemia.

The present study focuses on the function of cpg15, a neurotrophic factor, in ischemic neuronal recovery using transient global cerebral ischemic (TGI) mouse model and oxygen-glucose deprivation (OGD)-treated primary cultured cells. The results showed that expression of cpg15 proteins in astrocytes, predominantly the soluble form, was significantly increased in mouse hippocampus after TGI and in the cultured astrocytes after OGD. Addition of the medium from the cpg15-overexpressed astrocytic culture into the OGD-treated hippocampal neuronal cultures reduces the neuronal injury, whereas the recovery of neurite outgrowths of OGD-injured neurons was prevented when cpg15 in the OGD-treated astrocytes was knocked down, or the OGD-treated-astrocytic medium was immunoadsorbed by cpg15 antibody. Furthermore, lentivirus-delivered knockdown of cpg15 expression in mouse hippocampal astrocytes diminishes the dendritic branches and exacerbates injury of neurons in CA1 region after TGI. In addition, treatment with inhibitors of MEK1/2, PI3K, and TrkA decreases, whereas overexpression of p-CREB, but not dp-CREB, increases the expression of cpg15 in U118 or primary cultured astrocytes. Also, it is observed that the Flag-tagged soluble cpg15 from the astrocytes transfected with Flag-tagged cpg15-expressing plasmids adheres to the surface of neuronal bodies and the neurites. In conclusion, our results suggest that the soluble cpg15 from astrocytes induced by ischemia could ameliorate the recovery of the ischemic-injured hippocampal neurons via adhering to the surface of neurons. The upregulated expression of cpg15 in astrocytes may be activated via MAPK and PI3K signal pathways, and regulation of CREB phosphorylation.SIGNIFICANCE STATEMENT Neuronal plasticity plays a crucial role in the amelioration of neurological recovery of ischemic injured brain, which remains a challenge for clinic treatment of cerebral ischemia. cpg15 as a synaptic plasticity-related factor may participate in promoting the recovery process; however, the underlying mechanisms are still largely unknown. The objective of this study is to reveal the function and mechanism of neuronal-specific cpg15 expressed in astrocytes after ischemia induction, in promoting the recovery of injured neurons. Our findings provided new mechanistic insight into the neurological recovery, which might help develop novel therapeutic options for cerebral ischemia via astrocytic-targeting interference of gene expression.

Isolation of Bactrian Camel Single Domain Antibody for Parathion and Development of One-Step dc-FEIA Method Using VHH-Alkaline Phosphatase Fusion Protein.

A heavy chain variable fragment of heavy chain only antibodies derived from camelids termed VHH shows beneficial characteristics for immunoassay in terms of high sensitivity, outstanding stability and ease in expression. In the present study, we isolated six VHHs from phage display library against parathion, which is a widely used organophosphorus pesticide with high toxicity and persistence. One of six selected VHHs named VHH9, showed highest specificity and superior thermo-stability. A VHH9-alkaline phosphatase (AP) fusion was constructed and used to establish a one-step direct competitive fluorescence enzyme immunoassay (dc-FEIA) with a half maximal inhibitory concentration (IC50) of 1.6 ng/mL and a limit of detection of 0.2 ng/mL which was 4-fold or 3-fold higher sensitivity than direct competitive enzyme-linked immunoassay (dc-ELISA) and indirect competitive enzyme-linked immunoassay (ic-ELISA) for parathion. Furthermore, our assay indicated a 50% reduction on operation time compared with the ic-ELISA method. The presented immunoassay was validated with spiked Chinese cabbage, cucumber, and lettuce samples, and confirmed by UPLC-MS/MS. The results indicated that the VHH-AP-based dc-FEIA is a reproducible detection assay for parathion residues in vegetable samples.

VHH antibodies: emerging reagents for the analysis of environmental chemicals.

A VHH antibody (or nanobody) is the antigen binding fragment of heavy chain only antibodies. Discovered nearly 25 years ago, they have been investigated for their use in clinical therapeutics and immunodiagnostics, and more recently for environmental monitoring applications. A new and valuable immunoreagent for the analysis of small molecular weight environmental chemicals, VHH will overcome many pitfalls encountered with conventional reagents. In the work so far, VHH antibodies often perform comparably to conventional antibodies for small molecule analysis, are amenable to numerous genetic engineering techniques, and show ease of adaption to other immunodiagnostic platforms for use in environmental monitoring. Recent reviews cover the structure and production of VHH antibodies as well as their use in clinical settings. However, no report focuses on the use of these VHH antibodies to detect small environmental chemicals (MW < 1500 Da). This review article summarizes the efforts made to produce VHHs to various environmental targets, compares the VHH-based assays with conventional antibody assays, and discusses the advantages and limitations in developing these new antibody reagents particularly to small molecule targets. Graphical Abstract Overview of the production of VHHs to small environmental chemicals and highlights of the utility of these new emerging reagents.

Heavy chain single-domain antibodies to detect native human soluble epoxide hydrolase.

The soluble epoxide hydrolase (sEH) is a potential pharmacological target for treating hypertension, vascular inflammation, pain, cancer, and other diseases. However, there is not a simple, inexpensive, and reliable method to estimate levels of active sEH in tissues. Toward developing such an assay, a polyclonal variable domain of heavy chain antibody (VHH) sandwich immunoassay was developed. Ten VHHs, which are highly selective for native human sEH, were isolated from a phage-displayed library. The ten VHHs have no significant cross-reactivity with human microsomal epoxide hydrolase, rat and mouse sEH, and denatured human sEH. There is a high correlation between protein levels of the sEH determined by the enzyme-linked immunosorbent assay (ELISA) and the catalytic activity of the enzyme in S9 fractions of human tissues (liver, kidney, and lung). The VHH-based ELISA appears to be a new reliable method for monitoring the sEH and may be useful as a diagnostic tool for diseases influenced by sEH. This study also demonstrates the broad utility of VHH in biochemical and pharmacological research.

Nucleopolyhedrovirus infection and/or parasitism by Microplitis pallidipes Szepligeti affect hemocyte apoptosis of Spodoptera exigua (Hübner) larvae.

We determined the effects of parasitism by the endoparasitoid Microplitis pallidipes Szepligeti and/or nucleopolyhedrovirus (NPV) infection on hemocyte apoptosis of Spodoptera exigua (Hübner) larvae. Compared to healthy (control) larvae, larvae that were parasitized, virus-infected, or both all showed a significant increase in hemocyte apoptosis during 48-h observation period. The peaks of hemocyte apoptosis in parasitized, virus-infected and parasitized+infected larvae were at 12, 24 and 48 h after treatment, and were 86.7±1.9, 87.4±3.6 and 76.5±1.6%, respectively. Meanwhile, compared to parasitized larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 12.9%, 18.7% and 2.8% at 8, 36 and 48 h respectively, and decreased by 39.0% and 9.1% at 12 and 24h. Compared to virus-infected larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 13.4%, 2.4% and 15.3% at 8, 36 and 48 h, respectively, and decreased by 4.0% and 29.9% at 12 and 24h. Our study found that joint and separate parasitism and SeNPV infection induced hemocyte apoptosis of S. exigua larvae. It also revealed that NPV infection promoted host hemocyte apoptosis induced by parasitism at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae, and that parasitism promoted host hemocyte apoptosis induced by NPV infection at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae.

Papillary breast lesions on contrast-enhanced ultrasound: morphological enhancement patterns and diagnostic strategy.

OBJECTIVES: To identify features which determine the papillary nature of breast lesions by contrast-enhanced ultrasound (CEUS) based on morphological and pathological correlations, and predict which papillomas are malignant. METHODS: A retrospective review of 47 papillary lesions and 51 hypervascular benign abnormalities was performed. Enhancement patterns and diagnostic performance of CEUS were assessed. RESULTS: The most frequent finding in papillary lesions was perilesional linear ductal enhancement (87.2%) followed by heterogeneous enhancement (80.9%), and the presence of perfusion defects (66%). Among all CEUS morphological descriptors, a perilesional linear ductal enhancement pattern was the most effective feature to discriminate between papillary and hypervascular benign lesions. Using this criterion, the positive and negative predictive value for papilloma were 100% and 89.5%, respectively. A perilesional linear ductal pattern correlated with a ductal origin. The distinctive features associated with atypical or malignant papillomas were non-confluent enhancement, late overall wash-out, the presence of regional perfusion defect, clumped vascular pattern, and the presence of peripheral radial or penetrating vessels. The sensitivity, specificity, and positive and negative predictive values for predicting malignancy were 91.7%, 82.6%, 84.6% and 90.5%, respectively. CONCLUSIONS: CEUS provided characteristic enhancement patterns which could be helpful for identifying papillary lesions and for predicting potentially malignant papillomas. KEY POINTS: Papillary breast lesions display a characteristic appearance on CEUS. Adding CEUS to conventional US helps identify the papillary nature of papillomas. CEUS may be helpful for predicting potential malignancies among papillomas.

Codon modification for the DNA sequence of a single-chain Fv antibody against clenbuterol and expression in Pichia pastoris.

The expression efficiency was improved for the recombinant single-chain variable fragment (scFv) against clenbuterol (CBL) obtained from mouse and expressed in the methylotrophic yeast Pichia pastoris GS115, by redesigning and synthesizing the DNA sequence encoding for CBL-scFv based on the codon bias of P. pastoris. The codons encoding 124 amino acids were optimized, in which a total of 156 nucleotides were changed, and the G+C ratio was simultaneously decreased from 53 to 47.2 %. Under the optimized expression conditions, the yield of the recombinant CBL-scFv (41 kDa) antibodies was 0.223 g L⁻¹ in shake culture. Compared to the non-optimized control, the expression level of the optimized recombinant CBL-scFv based on preferred codons in P. pastoris demonstrated a 2.35-fold higher yield. Furthermore, the recombinant CBL-scFv was purified by Ni-NTA column chromatography, and the purity was 95 %. The purified CBL-scFv showed good CBL recognition by a competitive indirect enzyme-linked immunoassay. The average concentration required for 50 % inhibition of binding and the limit of detection for the assay were 5.82 and 0.77 ng mL⁻¹, respectively.

Contrast-enhanced ultrasonography for evaluation of cystic renal mass: in comparison to contrast-enhanced CT and conventional ultrasound.

PURPOSE: To assess the value of contrast-enhanced ultrasonography (CEUS) in evaluating cystic renal lesions compared with conventional ultrasound (US) and contrast-enhanced computed tomography (CECT). METHODS: One hundred and three patients with complex cystic renal masses underwent preoperative US and CEUS, among which 70 conducted CECT at our institution. The images were analyzed with the number of septa, septa and wall thickness and the presence of solid component, and final diagnosis was made. RESULTS: In malignancies, CEUS demonstrated more septa, thicker wall or septa, and more solid components than US and CECT. CEUS permitted categorization of 51.7% (30/58) and 28.6% (10/35) of malignant tumors in higher grade than by US and CECT, respectively. In benign lesions, CEUS detected more septa than CECT and correctly diagnosed benign cysts which appeared as solid lesions in US. CEUS permitted downgrading of 71.1% (32/45) and 17.1% (6/35) of benign lesions compared to US and CECT. The diagnostic performance of CEUS was better than US for benign cystic lesions. The phenomenon that solid-like component by US did not enhance by CEUS was a strong predictor of benign disease, with a positive predictive value (PPV) of 100%. Enhancement of solid, soft tissue by CEUS was highly predictive of malignancy, with a PPV of 100%. CONCLUSIONS: CEUS was superior to US and CECT in visualizing the number of septa septa and wall thickness, and the presence of solid component of cystic renal lesions. CEUS may play a similar role to CECT in the diagnosis of renal cystic lesions, and better than US.

The effect and safety assessment of monitoring ethanol concentration in exhaled breath combined with intelligent control of renal pelvic pressure on the absorption of perfusion fluid during flexible ureteroscopic lithotripsy.

PURPOSE: Previously, we designed a ureteral access sheath with the capability of renal pelvic pressure (RPP) measurement and a medical perfusion and aspiration platform, allowing for the intelligent control of RPP. However, the effect of different RPP levels on perfusion fluid absorption remains unclear. This randomized controlled trial aimed to investigate the effects of exhaled ethanol concentration monitoring and intelligent pressure control on perfusion fluid absorption during flexible ureteroscopic lithotripsy. METHODS: Eighty patients scheduled for flexible ureteroscopic lithotripsy were randomly divided into four groups. In groups A, B, and C, the RPPs were set at 0, - 5, and - 10 mmHg, respectively. Group D was regarded as the controls with unfixed RPP. Isotonic saline containing 1% ethanol was used as the irrigation fluid, with an average irrigation flow rate of 100 mL/min. The primary outcome of this study was the absorption of perfusion fluid that was calculated based on the exhaled ethanol concentration. The secondary outcomes included duration of operation and amounts of perfusion fluid used. Postoperative complications, pre- and postoperative renal function, infection markers, and blood gas analysis were also recorded for safety assessment. RESULTS: In all, 76 patients were involved in this study, whose demographic characteristics and preoperative conditions were comparable among groups. Under the same perfusion flow rate, the groups with fixed RPP exhibited reduced absorption of perfusion fluid, duration of operation, and perfusion volume. In particular, the lowest values were observed in group C (RPP = - 10 mmHg). In contrast to the unfixed RPP group, no considerable difference were observed in levels of BUN, Scr, WBC, CRP, and blood gas values among the fixed RPP groups. Moreover, postoperative complications showed no significant difference among groups. CONCLUSION: In flexible ureteroscopic lithotripsy, the groups with fixed RPP had less absorption of perfusion fluid and perfusion volume, shorter duration of surgery, and higher safety than the unfixed group.

Identification and expression profile of odorant-binding proteins in the parasitic wasp Microplitis pallidipes using PacBio long-read sequencing.

Microplitis pallidipes Szépligeti (Hymenoptera: Braconidae) is an important parasitic wasp of second and third-instar noctuid larvae such as the insect pests Spodoptera exigua, Spodoptera litura, and Spodoptera frugiperda. As in other insects, M. pallidipes has a chemosensory recognition system that is critical to foraging, mating, oviposition, and other behaviors. Odorant-binding proteins (OBPs) are important to the system, but those of M. pallidipes have not been determined. This study used PacBio long-read sequencing to identify 170,980 M. pallidipes unigenes and predicted 129,381 proteins. Following retrieval of possible OBP sequences, we removed those that were redundant or non-full-length and eventually cloned five OBP sequences: MpOBP2, MpOBP3, MpOBP8, MpOBP10, and MpPBP 429, 429, 459, 420, and 429 bp in size, respectively. Each M. pallidipes OBP had six conserved cysteine residues. Phylogenetic analysis revealed that the five OBPs were located at different branches of the phylogenetic tree. Additionally, tissue expression profiles indicated that MpOBP2 and MpPBP were mainly expressed in the antennae of male wasps, while MpOBP3, MpOBP8, and MpOBP10 were mainly expressed in the antennae of female wasps. MpOBP3 was also highly expressed in the legs of female wasps. Temporal profiles revealed that the expression of each M. pallidipes OBP peaked at different days after emergence to adulthood. In conclusion, we identified five novel odorant-binding proteins of M. pallidipes and demonstrated biologically relevant differences in expression patterns.

Title: Identification et profil d'expression des protéines de liaison aux odeurs chez la guêpe parasite Microplitis pallidipes à l'aide du séquençage à lecture longue PacBio. Abstract: Microplitis pallidipes Szépligeti (Hymenoptera : Braconidae) est une importante guêpe parasite des larves de noctuelles de deuxième et troisième stades telles que les insectes ravageurs Spodoptera exigua, Spodoptera litura et Spodoptera frugiperda. Comme d'autres insectes, M. pallidipes possède un système de reconnaissance chimiosensoriel, essentiel à la recherche de nourriture, à l'accouplement, à la ponte et à d'autres comportements. Les protéines de liaison aux odeurs (PLO) sont importantes pour le système, mais celles de M. pallidipes n'ont pas été déterminées. Cette étude a utilisé le séquençage à lecture longue PacBio pour identifier 170 980 unigènes de M. pallidipes et prédit 129 381 protéines. Après la récupération des séquences de PLO possibles, nous avons supprimé celles qui étaient redondantes ou pas de pleine longueur et avons finalement cloné cinq séquences de PLO, MpOBP2, MpOBP3, MpOBP8, MpOBP10 et MpPBP, respectivement de taille 429, 429, 459, 420 et 429 pb. Chaque PLO de M. pallidipes avait six résidus de cystéine conservés. L'analyse phylogénétique a révélé que les cinq PLO étaient situés à différentes branches de l'arbre phylogénétique. De plus, les profils d'expression tissulaire ont indiqué que MpOBP2 et MpPBP étaient principalement exprimés dans les antennes des guêpes mâles, tandis que MpOBP3, MpOBP8 et MpOBP10 étaient principalement exprimés dans les antennes des guêpes femelles. MpOBP3 était également fortement exprimé dans les pattes des guêpes femelles. Les profils temporels ont révélé que l'expression de chaque PLO de M. pallidipes culminait à différents jours après l'émergence à l'âge adulte. En conclusion, nous avons identifié cinq nouvelles protéines de liaison aux odeurs de M. pallidipes et démontré des différences biologiquement pertinentes dans les profils d'expression.

Nuclear magnetic resonance-based metabolomics enable detection of the effects of a whole grain rye and rye bran diet on the metabolic profile of plasma in prostate cancer patients.

Prostate cancer (PC) is the most common cancer in the Western world and the second most important cancer causing male deaths, after lung cancer, in the United States and Britain. Lifestyle and dietary changes are recommended for men diagnosed with early-stage PC. It has been shown that a diet rich in whole grain (WG) rye reduces the progression of early-stage PC, but the underlying mechanism is not clear. This study sought to identify changes in the metabolic signature of plasma in patients with early-stage PC following intervention with a diet rich in WG rye and rye bran product (RP) compared with refined white wheat product (WP) as a tool for mechanistic investigation of the beneficial health effects of RP on PC progression. Seventeen PC patients received 485 g RP or WP in a randomized, controlled, crossover design during a period of 6 wk with a 2-wk washout period. At the end of each intervention period, plasma was collected after fasting and used for (1)H NMR-based metabolomics. Multilevel partial least squares discriminant analysis was used for paired comparisons of multivariate data. A metabolomics analysis of plasma showed an increase in 5 metabolites, including 3-hydroxybutyric acid, acetone, betaine, N,N-dimethylglycine, and dimethyl sulfone, after RP. To understand these metabolic changes, fasting plasma homocysteine, leptin, adiponectin, and glucagon were measured separately. The plasma homocysteine concentration was lower (P = 0.017) and that of leptin tended to be lower (P = 0.07) after RP intake compared to WP intake. The increase in plasma 3-hydroxybutyric acid and acetone after RP suggests a shift in energy metabolism from anabolic to catabolic status, which could explain some of the beneficial health effects of WG rye, i.e., reduction in prostate-specific antigen and reduced 24-h insulin secretion. In addition, the increase in betaine and N,N-dimethylglycine and the decrease in homocysteine show a favorable shift in homocysteine metabolism after RP intake.

[Relationship between the mutation of leptin receptor gene and tumorigenesis of breast cancer].

OBJECTIVE: The aim of this study was to investigate the relationship of the mutations of leptin receptor gene exon 4, exon 6, exon9, and exon20 with the tumorigenesis of breast cancer. METHODS: Genomic DNA was extracted from breast cancer tissues of 155 patients, benign lesions of 56 patients and normal tissues and blood samples from 100 health control subjects. The leptin receptor genes were assayed with polymerase chain reaction (PCR) amplification and direct sequence analysis. RESULTS: Nucleotide substitutions no mutations were found at exon 4, and nucleotide substitutions occurred at codon 1029 in exon 9, no significant difference among the three groups (P = 0.574). The nucleotide substitutions at codon 668 in exon 6 resulted in Gln223Arg polymorphisms. The occurring frequencies of GG, GA, AA in breast cancer, breast benign lesions tissues and health tissues control group were 70.9% and 17.4%, 12.3%; 80.4%, 14.3% and 5.4%; and 81.0%, 16.0%, and 3.0%, respectively. Alleles of G and A in the three groups were 79.1% and 20.8%, 87.5% and 12.5%, and 89.0% and 11.0%, respectively. Compared the Gln223Arg genotype with the three allele groups, there were significant differences (χ(2) = 16.11, P < 0.005 and χ(2) = 11.41, P < 0.01), respectively. The nucleotide substitutions at codon 3057 in exon 20 resulted in Pro1019Pro polymorphisms. The occurrence frequencies of GG, GA, AA in the breast cancer, benign disease and health control groups were 11.6%, 30.3% and 56.1%; 32.1%, 44.0% and 28.5%; and 32.0%, 45.0% And 23.0%, respectively. Alleles of G and A in the three groups were 26.8% and 73.2%, 51.8% and 48.2%, and 54.5% and 45.5%, respectively. There are significant differences among the three groups (χ(2) = 6.56, P < 0.03 and χ(2) = 5.45, P < 0.05), respectively. Nucleotide substitutions occurred at relatively high frequencies at exon 6 and exon 20 in obese and overweight breast cancer patients compared with those in normal weight breast cancer patients, there were significant differences (P < 0.05 and P < 0.01). CONCLUSIONS: Our findings show that there is no relationship between the variations of leptin receptor gene exon 9 and tumorigenesis of breast cancer. The variation rate of leptin receptor gene exon 6 and exon 20 are significantly increased in the obese and overweight breast cancer patients.

Plants impact cellular immunity of caterpillars to an entomovirus.

BACKGROUND: Tri-trophic interactions among plants, insect herbivores and entomopathogens are one of the hot topics in ecology. Although plants have been shown to impact the interactions between herbivores and entomopathogens, it is still unclear how plants affect the cellular immunity of herbivores to entomopathogens. RESULTS: The number of hemocytes and the proportion of two main cell types (granular hemocytes and plasmatocytes), plasmatocyte-spreading rate, apoptosis rate, two Spodoptera exigua caspase (SeCasp-1, SeCasp-5) activities and gene expressions were all higher and the activities and gene expression of S. exigua inhibitor of apoptosis protein (SeIAP) were lower in nucleopolyhedrovirus (NPV)-infected caterpillars fed Ipomoea aquatica than those fed other plants or artificial diet. Scanning electron microscopy images were consistent with molecular patterns of immune responses. CONCLUSION: This study suggests that host plants affect the immune responses of herbivores to entomopathogens by manipulating the composition, morphology and apoptosis of herbivore hemocytes, which sheds light on the mechanisms that allow host plants to influence multi-trophic interactions. © 2021 Society of Chemical Industry.

Highly specific nanobody against herbicide 2,4-dichlorophenoxyacetic acid for monitoring of its contamination in environmental water.

2,4-dichlorophenoxyacetic acid (2,4-D), a widely used herbicide, is a small organic chemical pollutant in the environment. To develop a nanobody-based immunoassay for monitoring trace levels of 2,4-D, a step-wise strategy for the generation of nanobodies highly specific against this small chemical was employed. Firstly, we synthesized three novel haptens mimicking 2,4-D and assessed their influence on the sensitivity and specificity of the existing antibody-based assay. Polyclonal antibodies (pAb) from rabbits showed good sensitivity and moderate specificity for 2,4-D, pAb from llama based on selected haptens showed similar performance when compared to those from rabbits. Secondly, nanobodies derived from llama were generated for 2,4-D by an effective procedure, including serum monitoring and one-step library construction. One nanobody, NB3-9, exhibited good sensitivity against 2,4-D (IC50 = 29.2 ng/mL) had better specificity than the rabbit pAb#1518, with no cross-reactivities against the 2,4-D analogs tested. Thirdly, one-step fluorescent enzyme immunoassay (FLEIA) for 2,4-D based on a nanobody-alkaline phosphatase (AP) fusion was developed with IC50 of 1.9 ng/mL and a linear range of 0.4-8.6 ng/mL. Environmental water samples were analyzed by FLEIA and LC-MS/MS for comparison, and the results were consistent between both methods. Therefore, the proposed step-wise strategy from hapten design to nanobody-AP fusion production was successfully conducted, and the resulting nanobody based FLEIA was demonstrated as a convenient tool to monitor 2,4-D residuals in the environment.

Decline of three farmland pest species in rapidly urbanizing landscapes.

Urbanization is a pressing challenge for earth's humans because it is changing not only natural environments but also agricultural lands. Yet, the consequences of cropland loss on pest insect populations that largely depend on these habitats remain largely unclear. We used a 17-year data set to investigate the dynamics of three moth pest species (i.e., striped stem borer, yellow stem borer, and pink stem borer) and their driving forces across the largest mega-urban region of China. Total abundance of three pest species is declined by about 80%, which was strongly associated with cropland loss during rapid urbanization. Our findings indicate that not only the increasing conversion of natural areas to human-dominated landscapes but also that of agricultural lands to urban landscapes can be critical to insect populations. It is therefore essential to monitor and understand the insect dynamics in rapidly urbanizing regions, which are currently found in many developing countries worldwide.

Broad-specificity immunoassay for O,O-diethyl organophosphorus pesticides: application of molecular modeling to improve assay sensitivity and study antibody recognition.

A monoclonal antibody (mAb) against 4-(diethoxyphosphorothioyloxy)benzoic acid (hapten 1) was raised and used to develop a broad-specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) for 14 O,O-diethyl organophosphorus pesticides (OPs). Computer-assisted molecular modeling was used to model two-dimensional (2D) and three-dimensional (3D) quantitative structure-activity relationships (QSARs) to study antibody recognition. On the basis of insights obtained from the QSAR models, two heterologous coating haptens, 4-(diethoxyphosphorothioylamino)butanoic acid (hapten 2) and 4-(diethoxyphosphorothioyloxy)-2-methylbenzoic acid (hapten 3) were designed, synthesized, and used to develop heterologous ciELISAs with significantly improved sensitivity. The heterologous ciELISA using hapten 2 as the coating hapten showed good sensitivity in a broad-specific manner for eight O,O-diethyl OPs and may be used as a screening method for the determination of these OPs. Our studies demonstrated that molecular modeling can provide insights into the spatial and electronic effects of molecular structures that are important for antibody activity, which can then be used to improve immunoassay sensitivity.

Rye whole grain and bran intake compared with refined wheat decreases urinary C-peptide, plasma insulin, and prostate specific antigen in men with prostate cancer.

Rye whole grain and bran intake has shown beneficial effects on prostate cancer progression in animal models, including lower tumor take rates, smaller tumor volumes, and reduced prostate specific antigen (PSA) concentrations. A human pilot study showed increased apoptosis after consumption of rye bran bread. In this study, we investigated the effect of high intake of rye whole grain and bran on prostate cancer progression as assessed by PSA concentration in men diagnosed with prostate cancer. Seventeen participants were provided with 485 g rye whole grain and bran products (RP) or refined wheat products with added cellulose (WP), corresponding to ~50% of daily energy intake, in a randomized controlled, crossover design. Blood samples were taken from fasting men before and after 2, 4, and 6 wk of treatment and 24-h urine samples were collected before the first intervention period and after treatment. Plasma total PSA concentrations were lower after treatment with RP compared with WP, with a mean treatment effect of -14% (P = 0.04). Additionally, fasting plasma insulin and 24-h urinary C-peptide excretion were lower after treatment with RP compared with WP (P < 0.01 and P = 0.01, respectively). Daily excretion of 5 lignans was higher after the RP treatment than after the WP treatment (P < 0.001). We conclude that whole grain and bran from rye resulted in significantly lower plasma PSA compared with a cellulose-supplemented refined wheat diet in patients with prostate cancer. The effect may be related to inhibition of prostate cancer progression caused by decreased exposure to insulin, as indicated by plasma insulin and urinary C-peptide excretion.