Cadmium in Market Pork Kidneys: A Study on Cadmium Bioavailability and the Health Effects Based on Mouse Models.

Edible offal of farmed animals can accumulate cadmium (Cd). However, no studies have investigated Cd bioavailability and its health effects. Here, based on mouse models, market pork kidney samples exhibited high Cd relative bioavailability of 74.5 ± 11.2% (n = 26), close to 83.8 ± 7.80% in Cd-rice (n = 5). This was mainly due to high vitamin D3 content in pork kidney, causing 1.7-2.3-fold up-regulated expression of duodenal Ca transporter genes in mice fed pork kidney compared to mice fed Cd-rice, favoring Cd intestinal absorption via Ca transporters. However, although pork kidney was high in Cd bioavailability, subchronic low-dose (5% in diet) consumption of two pork kidney samples having 0.48 and 0.97 µg Cd g-1 dw over 35 d did not lead to significant Cd accumulation in the tissue of mice fed Cd-free rice but instead remarkably decreased Cd accumulation in the tissue of mice fed Cd-rice (0.48 µg Cd g-1) by ∼50% and increased abundance of gut probiotics (Faecalibaculum and Lactobacillus). Overall, this study contributed to our understanding of the bioavailability and health effects associated with Cd in edible offal, providing mechanistic insights into pork kidney consumption safety based on Cd bioavailability.

Machine learning analysis of serum cholesterol's impact on knee osteoarthritis progression.

The controversy surrounding whether serum total cholesterol is a risk factor for the graded progression of knee osteoarthritis (KOA) has prompted this study to develop an authentic prediction model using a machine learning (ML) algorithm. The objective was to investigate whether serum total cholesterol plays a significant role in the progression of KOA. This cross-sectional study utilized data from the public database DRYAD. LASSO regression was employed to identify risk factors associated with the graded progression of KOA. Additionally, six ML algorithms were utilized in conjunction with clinical features and relevant variables to construct a prediction model. The significance and ranking of variables were carefully analyzed. The variables incorporated in the model include JBS3, Diabetes, Hypertension, HDL, TC, BMI, SES, and AGE. Serum total cholesterol emerged as a significant risk factor for the graded progression of KOA in all six ML algorithms used for importance ranking. XGBoost algorithm was based on the combined best performance of the training and validation sets. The ML algorithm enables predictive modeling of risk factors for the progression of the KOA K-L classification and confirms that serum total cholesterol is an important risk factor for the progression of KOA.

Convenient two-step method constructed silicon-based microfluidic chip for fast CYP2C19 SNPs detection.

The rapid detection of single nucleotide polymorphisms (SNPs) in the CYP2C19 gene is crucial for precise clopidogrel usage. Quantitative real-time polymerase chain reaction (qPCR), as a powerful amplification tool, has been widely employed for CYP2C19 SNPs detection. However, traditional qPCR suffers from long amplification times and high reagent consumption. To address these challenges, this work presents a microfluidic SNPs detection device based on on-chip qPCR. The device includes a rapid thermal cycling system, an optical detection system, a control system, and a complementary silicon-glass chip for CYP2C19 SNPs detection. Compared to commercial qPCR instruments that take 1 hour for testing, this device completes the test in just 15 minutes (40 PCR cycles). The resulting linearity is similar to that found using commercial qPCR instruments but with higher amplification efficiency. Additionally, compared with other silicon-based qPCR chips, this chip is constructed by using a convenient two-step method and offers low manufacturing costs, which potentially reduces single-test costs to an acceptable level. This makes our chip promising for point-of-care testing (POCT).

Fast and efficient remediation of antimony-contaminated surface water and field soil using alumina supported Fe-Mn binary oxide.

Antimony (Sb) pollution in surface water and soil has earned extensive attention. Our previous study synthesized a new class of alumina supported Fe-Mn binary oxide (Fe-Mn@Al2O3) and found that MnO2 in the composite oxidized Sb(III) to Sb(V) and FeOOH and Al2O3 played an indispensable role in adsorption of Sb(III) and Sb(V). This study further explored the removal of Sb in surface water and in situ sequestration of Sb in Sb-contaminated field soil via Fe-Mn@Al2O3. Sb removal from water was pH independent and the removal efficiencies of Sb(III) and total Sb kept constant at 95.4% and 60.5%, respectively, over a pH range of 5.0-10.0. Increasing dissolved organic matter (DOM) from 0 to 22.8 mg/L had negligible effect on Sb(III) removal whereas inhibited the total Sb removal from 60.5% to 51.2%. Dissolved oxygen cannot oxidize aqueous Sb(III), yet, enhanced the Sb(III) removal whereas decreased the total Sb removal. The composite performed well in natural surface water with high DOM and inorganic ligands. In addition, the composite effectively immobilized Sb in field soil. 5% of the composite significantly inhibited the H2SO4 and HNO3 leachable Sb by 93.6% after 30 d. The amendment transformed the Sb speciation from more easily available fractions (i.e., exchangeable, carbonate-bound, and Fe-Mn oxides-bound species) into more stable fractions (i.e., organic material bound and residual species), leading to declined Sb bioaccessibility and reduced environmental risk. The composite facilitated a long-term stability of Sb in soil. The study demonstrated an easy, fast, and effective strategy for efficient immobilization of Sb in water and soil.

Influence of soil pH and organic carbon content on the bioaccessibility of lead and copper in four spiked soils.

Exploration of the association between heavy metal bioaccessibility (BAc) and soil properties is essential for rationalization of risk assessment and remediation of contaminated soil; however, the high complexity of soil systems often yield conflicting outcomes. To avoid erroneous conclusions, individual comparisons of soil properties is essential. Herein, we determined the changes in the BAc of Pb and Cu with the variation in soil pH and SOC content using Unified Bioaccessibility Research Group of Europe method, and validated these findings with in vivo mouse bioassays. Results indicated that the BAc of Pb and Cu in gastric and intestinal phases decreased by 1.76%-3.92% and 0.90%-3.27%, and by 0.41%-6.01% and 0.67%-1.59%, respectively, with every unit increase in soil pH. Furthermore, with every 1% increase in the absolute content of SOC, the BAc of Pb and Cu decreased by 4.04%-13.94% and 4.01%-34.7%, and by 8.98%-30.15% and 9.58%-20.03%, respectively. The in vivo bioassays results confirmed decrease in Pb concentrations in the liver, kidney, and blood of mice with the increase in Ferralosol pH and SOC content. These findings revealed that the health risks associated with accidental exposures to Pb- and Cu-contaminated soils with high pH and SOC level were relatively low, and the consistent in vivo and in vitro results for the BAc of Pb and Cu suggest the requirement for a swift and simple approach for assessing the risks of heavy metal contaminated soils. Thus, this study enhanced our understanding of the variations in risk assessments with soil properties of Pb- and Cu-contaminated soils, highlighting the role of soil characteristics in health risk assessment and remediation of contaminated soils.

High Tibial Osteotomy with Posterior Medial Meniscal Root Reconstruction Yields Improved Radiographic and Functional Outcomes and Healing Rates Compared to Osteotomy Alone.

PURPOSE: To compare pre- and post-operative clinical and radiological outcomes between patients undergoing high tibial osteotomy (HTO) with medial meniscus posterior root tear (MMPRT) reconstruction using gracilis tendon graft versus those without reconstruction MMPRT. METHODS: Patients with MMPRT who underwent HTO between January 2018 and December 2021 with minimum 2-year follow-up were included. All the patients were divided into 2 groups based on whether underwent meniscus root reconstruction with tendon graft, HTO alone (33 cases) and HTO with reconstruction MMPRT (21 cases). Clinical evaluation included Lysholm score, international knee documentation committee (IKDC) score, and visual analogue scale (VAS) score. The functional recovery and radiologically outcome of the knee were evaluated preoperatively and at the latest follow-up. Meniscus root healing rates and medial meniscal extrusion (MME) according to the second MRI were compared between the two groups at the latest follow-up. RESULTS: The results showed a statistically significant improvement in the postoperative Lysholm score, IKDC score, and VAS score in both groups at the latest follow-up (P<0.001). The analysis of Minimal Clinically Important Difference (MCID) for postoperative outcomes revealed that the percentage of patients who reached MCID thresholds was 100% for Lysholm, 100% for IKDC, and 100% for VAS in the HTO with reconstruction MMPRT group. In comparison, the percentages were 87.9% for Lysholm, 90.9% for IKDC, and 100% for VAS in the HTO alone group. Additionally, compared with the HTO alone group, the HTO with medial meniscus posterior root reconstruction using gracilis tendon graft significantly improved the meniscus root healing rates (Complete healing 85.7% vs. 45.4%, 95%CI: 0.003-0.007, P=0.001) and functional recovery (P<0.005 ) at the final follow-up. Additionally, the HTO with reconstruction MMPRT had a significantly better change in K-L grade (improved knees K-L grade: 10/21 vs. 6/33, P=0.033) and MME (2.1±1.0mm vs. 3.1±1.6mm, 95%CI: 0.3-1.7, P=0.007) compared to the HTO alone group. CONCLUSIONS: HTO with reconstruction of the meniscal root using a tendon graft resulted in improved radiographic and patient-reported outcomes as well as improved healing rates compared to the HTO alone.

Recent advances in the pathogenesis and prevention strategies of dental calculus.

Dental calculus severely affects the oral health of humans and animal pets. Calculus deposition affects the gingival appearance and causes inflammation. Failure to remove dental calculus from the dentition results in oral diseases such as periodontitis. Apart from adversely affecting oral health, some systemic diseases are closely related to dental calculus deposition. Hence, identifying the mechanisms of dental calculus formation helps protect oral and systemic health. A plethora of biological and physicochemical factors contribute to the physiological equilibrium in the oral cavity. Bacteria are an important part of the equation. Calculus formation commences when the bacterial equilibrium is broken. Bacteria accumulate locally and form biofilms on the tooth surface. The bacteria promote increases in local calcium and phosphorus concentrations, which triggers biomineralization and the development of dental calculus. Current treatments only help to relieve the symptoms caused by calculus deposition. These symptoms are prone to relapse if calculus removal is not under control. There is a need for a treatment regime that combines short-term and long-term goals in addressing calculus formation. The present review introduces the mechanisms of dental calculus formation, influencing factors, and the relationship between dental calculus and several systemic diseases. This is followed by the presentation of a conceptual solution for improving existing treatment strategies and minimizing recurrence.

Microplastic co-exposure elevates cadmium accumulation in mouse tissue after rice consumption: Mechanisms and health implications.

Rice cadmium (Cd) and microplastics are prevalent contaminants, posing a co-exposure threat to humans by means of dietary intake. To assess whether co-exposure of microplastics affects the bioavailability of rice Cd, mice were exposed to Cd-contaminated rice with microplastic co-exposure. We found that polyethylene (PE), polystyrene (PS), polypropylene (PP), and polyamide (PA) microplastic co-exposure via diet consumption (2 µg g-1) caused 1.17-1.38-fold higher Cd accumulation in tissue of mice fed by Cd-rice. For mice with co-exposure of PE microplastics, the higher rice-Cd bioavailability corresponded to colonization of Lactobacillus reuteri (38.9 % vs 17.5 %) in the gut compared to control mice, which caused higher production of gut metabolites particularly peptides, likely causing a 'side effect' of elevating Cd solubility in the intestinal lumen. In addition, abundance of sphingosine 1-phosphate in the gut of mice was reduced under PE microplastic exposure, which may reduce intracellular calcium ions (Ca2+) in enterocytes and form a weaker competition in pumping of intracellular Ca2+ and Cd2+ across the basolateral membrane of enterocytes, leading to higher Cd2+ transport efficiency. The results suggest elevated Cd exposure risk from rice consumption with microplastic co-exposure at environmentally relevant low concentrations.

Common metabolism and transcription responses of low-cadmium-accumulative wheat (Triticum aestivum L.) cultivars sprayed with nano-selenium.

Cadmium (Cd) contamination in soils threatens food security, while cultivating low-Cd-accumulative varieties, coupled with agro-nanotechnology, offers a potential solution to reduce Cd accumulation in crops. Herein, foliar application of selenium nanoparticles (SeNPs) was performed on seedlings of two low-Cd-accumulative wheat (Triticum aestivum L.) varieties grown in soil spiked with Cd at 3 mg/kg. Results showed that foliar application of SeNPs at 0.16 mg/plant (SeNPs-M) significantly decreased the Cd content in leaves of XN-979 and JM-22 by 46.4 and 40.8 %, and alleviated oxidative damage. The wheat leaves treated with SeNPs-M underwent significant metabolic and transcriptional reprogramming. On one hand, four specialized antioxidant metabolites such as L-Tyrosine, beta-N-acetylglucosamine, D-arabitol, and monolaurin in response to SeNPs in JM-22 and XN-979 is the one reason for the decrease of Cd in wheat leaves. Moreover, alleviation of stress-related kinases, hormones, and transcription factors through oxidative post-translational modification, subsequently regulates the expression of defense genes via Se-enhanced glutathione peroxidase. These findings indicate that combining low-Cd-accumulative cultivars with SeNPs spraying is an effective strategy to reduce Cd content in wheat and promote sustainable agricultural development.

Lappanolides A-N, fourteen undescribed sesquiterpenoids from Saussurea costus (Syn. Saussurea lappa) and their anti HBV activity.

Lappanolides A-N (1-14), 14 undescribed sesquiterpenoids, along with 23 known ones (15-37), were isolated from the roots of Saussurea costus, which were primarily categorized into eudesmane, guaiane, and germacrane types. Lappanolide A (1) possessed an unprecedented pseudo-disesquiterpenoids. Their structures and absolute configurations were established using physical data analyses (HRESIMS, IR, 1D and 2D NMR) and ECD calculations. All isolated compounds were tested for anti-hepatitis B virus (anti-HBV) activity. Ten compounds (1, 9, 11, 12, 19, 22, 28, 29, 31, and 36) exhibited activities against HBsAg secretions as determined by ELISA assay, with IC50 values ranging from 5.2 to 45.7 µM. In particular, compounds 28 and 29 showed inhibition of HBsAg secretion with IC50 values of 5.28 and 5.30 µM, and CC50 values of 9.85 and 6.37 µM, respectively, though they all exhibited low selectivity. Several compounds displayed cytotoxicity in the MTT assay. Among them, compound 28 was the most notable and was chosen for further study using flow cytometry. The result showed that it significantly induced HepG2 cell arrest in the S phase and induced apoptosis.

Rhamnolipids-based bio-supramolecular solvents as green and sustainable media for extraction of pyrethroid insecticides in water and food matrices.

A novel bio-supramolecular solvent (bio-SUPRAS) based on rhamnolipids (RLs) was designed for efficient extraction of pyrethroid insecticides in water and food matrices. Benefiting from RLs as amphiphiles equipped with the attractive properties of bio-degradable, low toxicity and high stability, bio-SUPRAS was spontaneously generated through salt induced coagulation. The bio-SUPRAS was characterized by cryo-scanning electron microscope and main factors influencing the extraction performance were investigated in detail. Under the optimized conditions, the method was found to have desirable limits of detection (5∼10 µg l-1), good precision (RSDs<16.9 %) and satisfactory recovery (75.2 %∼94.3 %). More importantly, the extraction mechanism was studied by density functional theory systematically. Following greenness assessment, the technique was successfully used for enrichment of pyrethroid pesticides in real samples before HPLC-UV analysis. Thus, the method showed the outstanding merits of eco-efficient, green, time-saving, and had favorable application prospect to remove trace analytes from intricate sample matrices.

Multi-Step Two-Dimensional Ultrasonic-Assisted Grinding of Silicon Carbide: An Experimental Study on Surface Topography and Roughness.

Two-dimensional ultrasonic-assisted grinding (2D-UAG) has exhibited advantages in improving the machining quality of hard and brittle materials. However, the grinding mechanism in this process has not been thoroughly revealed due to the complicated material removal behaviors. In this study, multi-step 2D-UAG experiments of silicon carbide are conducted to investigate the effects of machining parameters on surface quality. The experimental results demonstrate that the tool amplitude and the workpiece amplitude have similar effects on surface roughness. In the rough grinding stage, the surface roughness decreases continuously with increasing ultrasonic amplitudes and the material is mainly removed by brittle fracture with different surface defects. Under semi-finishing and finishing grinding steps, the surface roughness first declines and then increases as the tool amplitude or workpiece amplitude grows from 0 µm to 8 µm and the inflection point appears around 4 µm. The surface damage contains small-sized pits with band-like distribution and localized grooves. Furthermore, the influences of cutting parameters on surface quality are similar to those in conventional grinding. Discussions of the underlying mechanisms for the experimental phenomena are also provided based on kinematic analysis. The conclusions gained in this study can provide references for the optimization of machining parameters in 2D-UAG of hard and brittle materials.

Nomogram for predicting outcomes in elderly women with mucinous breast cancer: A retrospective study combined with external validation in southwest China.

OBJECTIVE: Mucinous breast cancer (MBC) is a kind of breast cancer (BC), which is rare in clinic, mainly for women, because of the low incidence rate, so there is no unified standard treatment protocol. Elderly patients have a poor prognosis due to their combined comorbidities. This study aims to investigate the effect of surgery and chemoradiotherapy on the prognosis of elderly female MBC patients and construct nomograms for predicting the OS and CSS in elderly female MBC patients. METHODS: Data for female MBC patients over 65 years are obtained from the Surveillance, Epidemiology and End Results (SEER) database, patients were divided into two groups: the training set and the validation set. External validation data of the prediction model were provided by Kunming Hospital of Traditional Chinese Medicine. We used Cox regression modeling, which was used to identify independent risk factors affecting patient prognosis. After avoiding confounding bias according to the multifactorial Cox regression model, we used these screened statistically significant results to construct column-line plots. The performance of the model was tested using the consistency index (c-index), the calibration curve, and the area under the operating characteristic curve of the receiver (AUC). Subsequently, we used decision curve analysis (DCA) to examine the potential clinical value of our nomograms. RESULTS: A total of 8103 elderly MBC female patients were extracted from the database SEER and were assigned to the training and validation set, randomly. A total of 83 patients from Kunming Hospital of Traditional Chinese Medicine were used in the external verification set. After multifactorial Cox regression analysis, we found that age, race, T-stage, M-stage, surgical approach, radiotherapy, and tumor size were independent risk factors for OS in elderly MBC patients. Similarly, independent risk factors of CSS included age, marital status, N stage, M stage, surgical approach, chemotherapy, and tumor size. The C-index for the OS training, validation, and external verification set were 0.731 (95%CI 0.715-0.747), 0.738 (95%CI 0.724-0.752), and 0.809 (95%CI 0.731-0.8874). The C-index of the training set, the validation set, and external verification set for CSS were 0.786 (95%CI 0.747-0.825), 0.776 (95%CI 0.737-0.815), and 0.84 (95%CI0.754-0.926), respectively. The AUC, calibration curves and DCA also showed good accuracy. CONCLUSIONS: In this study, we construct a new nomogram to predict the prognosis of elderly patients with MBC. The nomograms have undergone internal and external validation and have been confirmed to have good clinical applicability. At the same time, we found that for elderly female MBC patients, surgery and radiotherapy significantly benefit their survival, but chemotherapy is not conducive to patient survival.

A real-world study of foreign body aspiration in children with 4227 cases in Western China.

The early diagnosis and treatment of foreign body aspiration (FBA) can significantly improve the overall prognosis of children. There are significant differences in the epidemiology and clinical characteristics of FBA in different regions. Therefore, we conducted a real-world study in the western region of China with over 4000 patients. The aim of this study was to improve the understanding of FBA in terms of its types, the specific months of its occurrence, and the distribution of primary caregiver characteristics in western China. We collected the clinical and epidemiological data of children who were diagnosed with FBA in our hospital over the past 20 years through a big data centre. We matched the data of healthy children who underwent routine physical examinations at the paediatric health clinic during the same period to analyse the differences in the data of actual guardians. A total of 4227 patients from five provinces were included in this study. Foreign bodies were removed by rigid bronchoscopy in 99.4% (4202/4227) of patients, with a median age of 19 months and a median surgical duration 16 min. January was the most common month of onset for 1725 patients, followed by February, with 1027 patients. The most common types of foreign objects were melon peanuts, seeds and walnuts, accounting for 47.2%, 15.3%, and 10.2%, respectively. In the FBA group, the proportion of grandparents who were primary caregivers was 70.33% (2973/4227), which was significantly greater than the 63.05% in the healthy group (2665/4227) (P < 0.01). FBA most commonly occurs in January and February. More than 60% of FBAs occur between the ages of 1 and 2 years, and the incidence of FBA may be greater in children who are cared for by grandparents. A rigid bronchoscope can be used to remove most aspirated foreign bodies in a median of 16 min.

Detecting Internal Defects in FRP-Reinforced Concrete Structures through the Integration of Infrared Thermography and Deep Learning.

This study represents a significant advancement in structural health monitoring by integrating infrared thermography (IRT) with cutting-edge deep learning techniques, specifically through the use of the Mask R-CNN neural network. This approach targets the precise detection and segmentation of hidden defects within the interfacial layers of Fiber-Reinforced Polymer (FRP)-reinforced concrete structures. Employing a dual RGB and thermal camera setup, we captured and meticulously aligned image data, which were then annotated for semantic segmentation to train the deep learning model. The fusion of the RGB and thermal imaging significantly enhanced the model's capabilities, achieving an average accuracy of 96.28% across a 5-fold cross-validation. The model demonstrated robust performance, consistently identifying true negatives with an average specificity of 96.78% and maintaining high precision at 96.42% in accurately delineating damaged areas. It also showed a high recall rate of 96.91%, effectively recognizing almost all actual cases of damage, which is crucial for the maintenance of structural integrity. The balanced precision and recall culminated in an average F1-score of 96.78%, highlighting the model's effectiveness in comprehensive damage assessment. Overall, this synergistic approach of combining IRT and deep learning provides a powerful tool for the automated inspection and preservation of critical infrastructure components.

Radionuclide therapy of bevacizumab-based PNA-mediated pretargeting.

BACKGROUND: The radionuclide-labeled bevacizumab (BV) is a potential therapeutic approach for vascular endothelial growth factor overexpressed tumors. Because of its large molecular weight, BV is cleared slowly in vivo, which caused damage to healthy tissues and organs. On account of this situation, using the pretargeting strategy with DNA/RNA analogs, such as peptide nucleic acid (PNA), is an effective way of treating solid tumors. METHODS: The BV-PNA conjugate (BV-PNA-1) was injected intravenously as the pretargeted probe, which was specifically accumulated in a solid tumor and gradually metabolically cleared. Then the [177Lu]Lu-labeled complementary PNA strand ([177Lu]Lu-PNA-2) as the second probe was injected, and bound with BV-PNA-1 by the base complementary pairing. In this study, the BV-based PNA-mediated pretargeting strategy was systematically studied, including stability of probes, specific binding ability, biodistribution in animal model, evaluation of single photon emission computed tomography/computed tomography imaging, and therapeutic effect. RESULTS: Compared with group A ([177Lu]Lu-BV), the group B (BV-PNA-1 + [177Lu]Lu-PNA-2) showed lower blood radiotoxicity (22.55 ±1.62 vs. 5.18 ±â€…0.40%, %ID/g, P < 0.05), and similar accumulation of radioactivity in tumor (5.32 ±â€…0.66 vs. 6.68 ± 0.79%, %ID/g, P > 0.05). Correspondingly, there was no significant difference in therapeutic effect between groups A and B. CONCLUSION: The PNA-mediated pretargeting strategy could increase the tumor-to-blood ratio, thereby reducing the damage to normal tissues, while having a similar therapeutic effect to solid tumor. All the experiments in this study showed the potential and effectiveness of pretargeting radioimmunotherapy.

A universal fluorescence biosensor based on rolling circle amplification and locking probe for DNA detection.

A stable DNA signal amplification sensor was developed on account of rolling circle amplification (RCA). This sensor includes target DNA-controlled rolling circle amplification technology and locking probe DNA replacement technology, which can be used to detect DNA fragments with genetic information, thus constructing a biosensor for universal detection of DNA. This study takes the homologous DNA of human immunodeficiency virus (HIV) and let-7a as examples to describe this biosensor. The padlock probe is first cyclized by T4 DNA ligase in response to the target's reaction with it. Then, rolling cycle amplification is initiated by Phi29 DNA polymerase, resulting in the formation of a lengthy chain with several triggers. These triggers can open the locked probe LP1 with the fluorescence signal turned off, so that it can continue to react with H2 to form a stable H1-H2 double strand. This regulates the distance between B-DNA modified by the quenching group and H1 modified by fluorescent group, and the fluorescence signal is recovered.

The U-Box E3 Ubiquitin Ligase PUB35 Negatively Regulates ABA Signaling through AFP1-mediated Degradation of ABI5.

Abscisic acid (ABA) signaling is crucial for plant responses to various abiotic stresses. The Arabidopsis (Arabidopsis thaliana) transcription factor ABA INSENSITIVE 5 (ABI5) is a central regulator of ABA signaling. ABI5 BINDING PROTEIN 1 (AFP1) interacts with ABI5 and facilitates its 26S-proteasome-mediated degradation, although the detailed mechanism has remained unclear. Here, we report that an ABA-responsive U-box E3 ubiquitin ligase, PLANT U-BOX 35 (PUB35), physically interacts with AFP1 and ABI5. PUB35 directly ubiquitinated ABI5 in a bacterially reconstituted ubiquitination system and promoted ABI5 protein degradation in vivo. ABI5 degradation was enhanced by AFP1 in response to ABA treatment. Phosphorylation of the T201 and T206 residues in ABI5 disrupted the ABI5-AFP1 interaction and affected the ABI5-PUB35 interaction and PUB35-mediated degradation of ABI5 in vivo. Genetic analysis of seed germination and seedling growth showed that pub35 mutants were hypersensitive to ABA as well as to salinity and osmotic stresses, whereas PUB35 overexpression lines were hyposensitive. Moreover, abi5 was epistatic to pub35, whereas the pub35-2 afp1-1 double mutant showed a similar ABA response to the two single mutants. Together, our results reveal a PUB35-AFP1 module involved in fine-tuning ABA signaling through ubiquitination and 26S-proteasome-mediated degradation of ABI5 during seed germination and seedling growth.

Detection of MicroRNA-155 based on lambda exonuclease selective digestion and CRISPR/cas12a-assisted amplification.

In numerous malignancies, miRNA-155 is overexpressed and has oncogenic activity because it is one of the most efficient microRNAs for inhibiting apoptosis in human cancer cells. As a result, the highest sensitive detection of the miRNA-155 gene is a technological instrument that can enable early cancer screening. In this study, a miRNA-155 biosensor was created to create a hairpin probe that can bind to the miRNA-155 gene using lambda nucleic acid exonuclease, which can cut the 5' phosphorylated double strand, and by the DNA probe is recognized by the Cas12a enzyme, which then activates Cas12a to catalyze trans-cutting produces strong fluorescence. Research finding, the target concentration's logarithm and corresponding fluorescence intensity have a strong linear connection, and the limit of detection (LOD) of the sensing system was determined to be 8.3 pM. In addition, the biosensor displayed exceptional specificity, low false-positive signal, and high sensitivity in detecting the miRNA-155 gene in serum samples. This study's creation of a biosensor that has high sensitivity, good selectivity, and is simple to operate provides promising opportunities for research into biosensor design and early cancer detection.