Ultrasensitive amplification-free detection of circulating miRNA via droplet-based processing of SERS tag-miRNA-magnetic nanoparticle sandwich nanocomplexes on a paper-based electrowetting-on-dielectric platform.

MicroRNAs (miRNAs) have emerged as a promising class of biomarkers for early detection of various cancers, including ovarian cancer. However, quantifying miRNAs in human blood samples is challenging owing to the issues of sensitivity and specificity. In this study, hsa-miR-200a-3p of the miR-200a sub-family, which is a biomarker of ovarian cancer, was used as the analyte to demonstrate the analytical capability of an integrated biosensing platform using an extremely sensitive surface-enhanced Raman scattering (SERS) nanotag-nanoaggregate-embedded beads (NAEBs), magnetic nanoparticles (MNPs), a pair of highly specific locked nucleic acid (LNA) probes, and a semi-automated paper-based electrowetting-on-dielectric (pEWOD) device to provide labor-less and thorough sample cleanup and recovery. A sandwich approach where NAEBs are modified by one LNA-1 probe and MNPs are modified by another LNA-2 probe was applied. Then, the target analyte miRNA-200a-3p was introduced to form a sandwich nanocomplex through hybridization with the pair of LNA probes. The pEWOD device was used to achieve short cleanup time and good recovery of the nanocomplex, bringing the total analysis time to less than 30 min. The detection limit of this approach can reach 0.26 fM through SERS detection. The versatility of this method without the need for RNA extraction from clinical samples is expected to have good potential in detecting other miRNAs.

Unraveling the Fluoride-Induced Interface Reconstruction Across Lead-Based Hierarchical MnO2 Anode in Zinc Electrowinning.

Although the hierarchical manganese dioxide film electrode shows promise as a durable and catalytically active anode for zinc electrowinning, it often fails and deactivates when it is exposed to fluoride-rich environments. The lack of understanding regarding the mechanism behind fluoride-induced irreversible interface reconstruction hinders their practical application in large-scale energy-saving and pollution-reduction efforts. Here, we conducted multidimensional operando investigations to gain insights into the dynamic evolution across the film electrode interface with temporal and spatial resolution. Our findings reveal that electroosmosis of F- initially triggers structural collapse and subsequent reconstruction of [MnO6] units, followed by interaction with the spontaneous oxide film at the surface of lead substrate. Experimental studies and theoretical calculations indicate that F- facilitates the irreversible transformation of γ-MnO2 into more stable yet protective catalytic dual-defective α-MnO2. Additionally, lower levels of F- at the interface promote a change in microenvironmental pH within porous PbSO4, triggering the development of microporous corrosion-resistant ß-PbO2 as the dominant phase. The combined effects of MnO2 and interphase evolution effectively explain the abnormally elevated oxygen evolution overpotential. Then, the proposed appropriate application scenarios based on the corrosion behavior will serve as a practical guide for the implementation of the hierarchical manganese dioxide film electrode.

Influence of occupational exposure on hyperuricemia in steelworkers: a nested case-control study.

BACKGROUND: Occupational exposure may be associated with an increased risk of developing hyperuricemia. This study sheds lights on the association between occupational exposure and hyperuricemia in steelworkers. METHOD: A nested case-control study was conducted within a cohort of workers in steel companies to explore the association between occupational exposure and hyperuricemia. The case group consisted of a total of 641 cases of hyperuricemia identified during the study period, while 641 non-hyperuricemia subjects with the same age and gender distribution were randomly selected from the cohort as the control group. RESULTS: The incidence rate of hyperuricemia among workers in the steel company was 17.30%, with an incidence density of 81.32/1,000 person-years. In comparison to the reference group, the risks of developing hyperuricemia for steelworkers undergoing ever shifts, current shifts, heat exposure, and dust exposure were 2.18 times, 1.81 times, 1.58 times and 1.34 times higher respectively. The odds ratios (ORs) and 95% confidence intervals (CIs) were 1.87(1.12-3.13) and 2.02(1.21-3.37) for the cumulative number of days of night work at 0-1,972.80 and ≥ 1,972.80 (days), respectively. Compared to the group with the cumulative heat exposure of 0 (°C/year), the ORs (95% CI) for the risk of developing hyperuricemia in the groups with the cumulative heat exposure of 0-567.83 and ≥ 567.83 (°C/year) were 1.50(1.02-2.22) and 1.64(1.11-2.43), respectively. The OR (95% CI) for the risk of developing hyperuricemia was 1.56(1.05-2.32) at the cumulative dust exposure of ≥ 30.02 (mg/m3/year) compared to that at the cumulative dust exposure of 0 (mg/m3/year). Furthermore, there was a multiplicative interaction between heat exposure and dust exposure in the development of hyperuricemia. CONCLUSION: Shift work, heat, and dust are independent risk factors for the development of hyperuricemia in steelworkers. Additionally, there is a multiplicative interaction between heat exposure and dust exposure in the development of hyperuricemia. Interventions for shift work, heat and dust may help to reduce the incidence rate of hyperuricemia and improve the health of steelworkers.

The mechanisms of arsenic-induced ovotoxicity, ultrastructural alterations, and autophagic related paths: An enduring developmental study in folliculogenesis of mice.

Arsenic (As) exerts a wide range of adverse effects on biological systems, including the reproductive organs in males and females. However, the mechanisms of As-induced reproductive toxicity are mostly obscure. Recently, we showed that autophagy is an essential route for As2O3-induced reprotoxicity through the hypothalamic-pituitary-gonadal-sperm (HPG-S) axis in pubertal and matured F1-male mice. However, the role of autophagy in As2O3- induced ovarian toxicity is mostly unknown. Hence, this study aimed to elucidate the role of oxidative stress, mitochondrial impairment, and autophagic processes in the ovary of As-exposed female mice. For this purpose, mature female mice were challenged with 0, low (0.2), medium (2), and high (20 ppm) As2O3 from 35-days before mating till weaning their pups, and the F1- females from weaning until maturity. Then, all the mice were sacrificed, and oxidative stress parameters, mitochondrial indices, electron microscopic evaluation of the ovaries, expression of autophagic-related genes and proteins, and autophagosome formation were assessed. It was shown that medium and high As2O3 doses were a potent inducer of oxidative stress, mitochondrial dysfunction, and autophagy in the ovary of F1-generation. A dose-dependent increment in the gene expression of PDK1, PI3K, TSC2, AMPK, ULK1, ATG13, Beclin1, ATG12, ATG5, LC3, P62, ATG3, ATG7, and p62, as well as protein expression of Beclin1, and LC3- I, II, was evident in the ovaries of the As-treated animals. Moreover, a dose-dependent decrease in the expression of mTOR and Bcl-2 genes, and mTOR protein was detected with increasing doses of As, suggesting that As treatment-induced autophagy. Along with a dose-dependent increase in the number of MDC-labeled autophagic vacuoles, transmission electron microscopy also confirmed more autophagosomes and injured mitochondria in medium and high As2O3 doses groups. As2O3 also negatively affected the mean body weight, litter size, organ coefficient, and stereological indices in female mice. Finally, in physiological conditions, arsenic trioxide (As2O3) leads to an increased level of autophagy in the oocyte when many oocytes were being lost. These findings indicated that an imbalance in the oxidant-antioxidant system, mitochondrial impairment, and the autophagic process, through inhibition of mTOR, dependent and independent pathways, and Bcl-2, as well as activation of AMPK/PI3K/Beclin1/LC3 routes, could play a pivotal role in As-induced reproductive toxicity through ovarian dysfunction in females.

Integration of a Thermoelectric Heating Unit with Ionic Wind-Induced Droplet Centrifugation Chip to Develop Miniaturized Concentration Device for Rapid Determination of Salmonella on Food Samples Using Antibody-Functionalized SERS Tags.

When a centrifugation-enriched sample of 100 µL containing the surface-enhanced Raman scattering (SERS) tag-bound bacteria (Salmonella in this study) is siphoned onto a glass slide next to an embedded thermoelectric heating chip, such a sessile droplet is quickly evaporated. As the size of the sample droplet is significantly reduced during the heating process, ionic wind streams from a corona discharge needle, stationed above the sample, sweep across the liquid surface to produce centrifugal vortex flow. Tag-bound Salmonella in the sample are then dragged and trapped at the center of droplet bottom. Finally, when the sample is dried, unlike the "coffee ring" effect, the SERS tag-bound Salmonella is concentrated in one small spot to allow sensitive detection of a Raman signal. Compared with our previous electrohydrodynamic concentration device containing only a corona discharge needle, this thermoelectric evaporation-assisted device is more time-effective, with the time of concentrating and drying about 100 µL sample reduced from 2 h to 30 min. Hence, sample throughput can be accelerated with this device for practical use. It is also more sensitive, with SERS detection of a few cells of Salmonella in neat samples achievable. We also evaluated the feasibility of using this device to detect Salmonella in food samples without performing the culturing procedures. Having spiked a few Salmonella cells into ice cubes and lettuce leaves, we use filtration and ultracentrifugation steps to obtain enriched tag-bound Salmonella samples of 200 µL. After loading an aliquot of 100 µL of sample onto this concentration device, the SERS tag signals from samples of 100 g ice cubes containing two Salmonella cells and 20 g lettuce leaf containing 5 Salmonella cells can be successfully detected.

Development of Coplanar Electro-Wetting Based Microfluidic Sorter to Select Micro-Particles in High Volume Throughput at Milliliter Amount within Twenty Minutes.

This paper reports the work of developing one coplanar microfluidic sorter while using the electro-wetting on dielectrics (EWOD) technique. When connected with delivery capillary to receive sample solution containing micro-particles, this device can select about 10 micro-particles in high volume throughput of milliliter amount within 20 min, to potentially match the requirement of efficiently determining the low amounts of bacteria in concentrated food and environmental samples, of which the typical bacteria density is 10 colony forming unit or less, much smaller than that of clinical pathogen samples. This coplanar T-shape EWOD device contains two fluidic channels, one inlet channel and the other collection channel stemmed from the middle of inlet channel. When the solution droplet falls from the delivery capillary to the entrance end of inlet channel, the droplet is driven to the intersection of two channels. The droplet containing fluorescent particle will be diverted to the lower channel to collect. Otherwise, the non-fluorescent droplet keeps moving toward the other end of inlet channel to waste zone. The particle fluorescence is collected through microscope lens to detect with one photomultiplier tube. The detected signals trigger the personal computer control board to active each EWOD electrode to direct the droplet moving directions. When the solution of 1 mL containing about 10 fluorescent micro-particles is delivered into this sorting device, nearly all the particles were correctly directed into collection zone in 20 min.

A case control study on the relationship between occupational stress and genetic polymorphism and dyslipidemia in coal miners.

Dyslipidemia is one of the known risk factors for cardiovascular disease, and its prevalence is increasing worldwide. At present, the study of dyslipidemia has gradually shifted from simple environmental or genetic factors to environment-gene interactions. In order to further explore the etiology and mechanism of dyslipidemia, we used occupational stress(OS) and LYPLAL1, APOC3 and SOD2 gene as research variables to explore their association with dyslipidemia.Here we used a case-control study to include Han workers from a coal mining enterprise in China to determine the association between study variables and dyslipidemia. Monofactor analysis showed that smoking, drinking, physical activity level, DASH diet score, sleep quality, BMI, hypertension, hyperuricemia, shift work, OS were significantly different between the two groups (P < 0.05). In the APOC3 rs2854116 dominant model, patients with CT/CC genotype had a higher risk of dyslipidemia than those with TT genotype. In SOD2 rs4880 recessive model, patients with GG genotype had a lower risk of dyslipidemia than those with AA/AG genotype, and the difference was statistically significant. We found that rs12137855 and OS, rs2854116 and OS, rs4880 and OS had joint effects, but no interaction based on the multiplication and addition model was found (Pinteraction > 0.05). GMDR model showed that the rs12137855-rs2854116-rs4880-OS four-factor model had the highest cross-validation consistency and training-validation accuracy (P < 0.05), suggesting that there was a high-order interaction between them associated with dyslipidemia. We found that dyslipidemia in coal miners was related to OS and genetic factors. Through this study, we revealed the dual regulation of environmental factors and genetic factors on dyslipidemia. At the same time, this study provides clues for understanding the etiology and mechanism of dyslipidemia.

Association of Chinese Visceral Adiposity Index and Carotid Atherosclerosis in Steelworkers: A Cross-Sectional Study.

The Chinese Visceral Adiposity Index (CVAI) is an indicator of visceral adiposity dysfunction used to evaluate the metabolic health of the Chinese population. Steelworkers are more likely to be obese due to their exposure to special occupational factors, and have a higher prevalence of carotid atherosclerosis (CAS). This study aimed to analyze the special relationship between CVAI and CAS among steelworkers. A total of 4075 subjects from a northern steel company were involved in the cross-sectional study. Four logistic regression models were developed to analyze the correlation between CVAI and CAS. In addition, the restricted cubic spline was applied to fit the dose-response association between CVAI and CAS risk. In the study, the prevalence of CAS was approximately 25.94%. After adjustment for potential confounders, we observed a positive correlation between CVAI and CAS risk. Compared to the first CVAI quartile, the effect value odds ratio (OR) and 95% CI in the second, third, and fourth CVAI quartile were 1.523 (1.159-2.000), 2.708 (2.076-3.533), and 4.101 (3.131-5.372), respectively. Additionally, this positive correlation was stable in all subgroups except for female. Furthermore, we also found a non-linear relationship between CVAI and CAS risk (p nonlinear < 0.05). Notably, CVAI could increase the risk of CAS when higher than 106. In conclusion, our study showed that CVAI might be a reliable indicator to identify high-risk populations of CAS among steelworkers.

Risk Prediction for the Development of Hyperuricemia: Model Development Using an Occupational Health Examination Dataset.

OBJECTIVE: Hyperuricemia has become the second most common metabolic disease in China after diabetes, and the disease burden is not optimistic. METHODS: We used the method of retrospective cohort studies, a baseline survey completed from January to September 2017, and a follow-up survey completed from March to September 2019. A group of 2992 steelworkers was used as the study population. Three models of Logistic regression, CNN, and XG Boost were established to predict HUA incidence in steelworkers, respectively. The predictive effects of the three models were evaluated in terms of discrimination, calibration, and clinical applicability. RESULTS: The training set results show that the accuracy of the Logistic regression, CNN, and XG Boost models was 84.4, 86.8, and 86.6, sensitivity was 68.4, 72.3, and 81.5, specificity was 82.0, 85.7, and 86.8, the area under the ROC curve was 0.734, 0.724, and 0.806, and Brier score was 0.121, 0.194, and 0.095, respectively. The XG Boost model effect evaluation index was better than the other two models, and similar results were obtained in the validation set. In terms of clinical applicability, the XG Boost model had higher clinical applicability than the Logistic regression and CNN models. CONCLUSION: The prediction effect of the XG Boost model was better than the CNN and Logistic regression models and was suitable for the prediction of HUA onset risk in steelworkers.

Development and Internal Validation of Risk Assessment Models for Chronic Obstructive Pulmonary Disease in Coal Workers.

Coal workers are more likely to develop chronic obstructive pulmonary disease due to exposure to occupational hazards such as dust. In this study, a risk scoring system is constructed according to the optimal model to provide feasible suggestions for the prevention of chronic obstructive pulmonary disease in coal workers. Using 3955 coal workers who participated in occupational health check-ups at Gequan mine and Dongpang mine of Hebei Jizhong Energy from July 2018 to August 2018 as the study subjects, random forest, logistic regression, and convolutional neural network models are established, and model performance is evaluated to select the optimal model, and finally a risk scoring system is constructed according to the optimal model to achieve model visualization. The training set results show that the logistic, random forest, and CNN models have sensitivities of 78.55%, 86.89%, and 77.18%; specificities of 85.23%, 92.32%, and 87.61%; accuracies of 81.21%, 85.40%, and 83.02%; Brier scores of 0.14, 0.10, and 0.14; and AUCs of 0.76, 0.88, and 0.78, respectively, and similar results are obtained for the test set and validation set, with the random forest model outperforming the other two models. The risk scoring system constructed according to the importance ranking of random forest predictor variables has an AUC of 0.842; the evaluation results of the risk scoring system shows that its accuracy rate is 83.7% and the AUC is 0.827, and the established risk scoring system has good discriminatory ability. The random forest model outperforms the CNN and logistic regression models. The chronic obstructive pulmonary disease risk scoring system constructed based on the random forest model has good discriminatory power.

A Predictive Model for Abnormal Bone Density in Male Underground Coal Mine Workers.

The dark and humid environment of underground coal mines had a detrimental effect on workers' skeletal health. Optimal risk prediction models can protect the skeletal health of coal miners by identifying those at risk of abnormal bone density as early as possible. A total of 3695 male underground workers who attended occupational health physical examination in a coal mine in Hebei, China, from July to August 2018 were included in this study. The predictor variables were identified through single-factor analysis and literature review. Three prediction models, Logistic Regression, CNN and XG Boost, were developed to evaluate the prediction performance. The training set results showed that the sensitivity of Logistic Regression, XG Boost and CNN models was 74.687, 82.058, 70.620, the specificity was 80.986, 89.448, 91.866, the F1 scores was 0.618, 0.919, 0.740, the Brier scores was 0.153, 0.040, 0.156, and the Calibration-in-the-large was 0.104, 0.020, 0.076, respectively, XG Boost outperformed the other two models. Similar results were obtained for the test set and validation set. A two-by-two comparison of the area under the ROC curve (AUC) of the three models showed that the XG Boost model had the best prediction performance. The XG Boost model had a high application value and outperformed the CNN and Logistic regression models in prediction.

Trace Determination of Grouper Nervous Necrosis Virus in Contaminated Larvae and Pond Water Samples Using Label-Free Fiber Optic Nanoplasmonic Biosensor.

We developed a fast (<20 min), label-free fiber optic particle plasmon resonance (FOPPR) immunosensing method to detect nervous necrosis virus (NNV), which often infects high-value economic aquatic species, such as grouper. Using spiked NNV particles in a phosphate buffer as samples, the standard calibration curve obtained was linear (R2 = 0.99) and the limit of detection (LOD) achieved was 2.75 × 104 TCID50/mL, which is superior to that obtained using enzyme-linked immunosorbent assay (ELISA). By using an enhancement method called fiber optic nanogold-linked immunosorbent assay (FONLISA), the LOD can be further improved to <1 TCID50/mL, which is comparable to that found by the conventional qPCR method. Employing the larvae homogenate samples of NNV-infected grouper, the results obtained by the FOPPR biosensor agree with those obtained by the quantitative polymerase chain reaction (qPCR) method. We also examined pond water samples from an infected container in an indoor aquaculture facility. The lowest detectable level of NNV coat protein was found to be 0.17 µg/mL, which is one order lower than the LOD reported by ELISA. Therefore, we demonstrated the potential of the FOPPR biosensor as an outbreak surveillance tool, which is able to give warning indication even when the trend of larvae death toll increment is still not clear.

Apoptosis-inducing factor plays a role in the pathogenesis of hepatic and renal injury during cholestasis.

Cholestasis is a clinical complication with different etiologies. The liver is the primary organ influenced in cholestasis. Renal injury is also a severe clinical complication in cholestatic/cirrhotic patients. Several studies mentioned the importance of oxidative stress and mitochondrial impairment as two mechanistically interrelated events in cholestasis-induced organ injury. Apoptosis-inducing factor (AIF) is a flavoprotein located in the inner mitochondrial membrane. This molecule is involved in a distinct pathway of cell death. The current study aimed to evaluate the role of AIF in the pathophysiology of cholestasis-associated hepatic and renal injury. Bile duct ligation (BDL) was used as an animal model of cholestasis. Serum, urine, and tissue samples were collected at scheduled time intervals (3, 7, 14, and 28 days after BDL surgery). Tissues' AIF mRNA levels, as well as serum, urine, and tissue activity of AIF, were measured. Moreover, markers of DNA fragmentation and apoptosis were assessed in the liver and kidney of cholestatic animals. A significant increase in liver and kidney AIF mRNA levels, in addition to increased AIF activity in the liver, kidney, serum, and urine, was detected in BDL rats. DNA fragmentation and apoptosis were raised in the liver and kidney of cholestatic animals, especially at the early stage of the disease. The apoptotic mode of cell death in the liver and kidney was connected to a higher AIF level. These data mention the importance of AIF in the pathogenesis of cholestasis-induced organ injury, especially at the early stage of this disease. Mitochondrial release of apoptosis-inducing factor (AIF) seems to play a pathogenic role in cholestasis-associated hepatic and renal injury. AIF release is directly connected to oxidative stress and mitochondrial impairment in cholestatic animals.

The activation of nuclear factor-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling blunts cholestasis-induced liver and kidney injury.

Cholestasis is a severe clinical complication that severely damages the liver. Kidneys are also the most affected extrahepatic organs in cholestasis. The pivotal role of oxidative stress has been mentioned in the pathogenesis of cholestasis-induced organ injury. The activation of the nuclear factor-E2-related factor 2 (Nrf2) pathway is involved in response to oxidative stress. The current study was designed to evaluate the potential role of Nrf2 signaling activation in preventing bile acids-induced toxicity in the liver and kidney. Dimethyl fumarate was used as a robust activator of Nrf2 signaling. Rats underwent bile duct ligation surgery and were treated with dimethyl fumarate (10 and 40 mg/kg). Severe oxidative stress was evident in the liver and kidney of cholestatic animals (P < 0.05). On the other hand, the expression and activity of Nrf2 and downstream genes were time-dependently decreased (P < 0.05). Moreover, significant mitochondrial depolarization, decreased ATP levels, and mitochondrial permeabilization were detected in bile duct-ligated rats (P < 0.05). Histopathological alterations included liver necrosis, fibrosis, inflammation and kidney interstitial inflammation, and cast formation. It was found that dimethyl fumarate significantly decreased hepatic and renal injury in cholestatic animals (P < 0.05). Based on these data, the activation of the cellular antioxidant response could serve as an efficient therapeutic option for managing cholestasis-induced organ injury.