Identification of a murder caused by brodifacoum poisoning based on clinical examinations and LC-MS/MS results.

Brodifacoum exerts its antagonistic effect against the metabolism of vitamin K, an essential component in the synthesis of blood coagulation factors. This effect ultimately hinders the blood's capacity to clot effectively, rendering it a commonly employed rodenticide. Instances of lethal poisonings are exceedingly rare owing to expeditious medical intervention and treatment. Within this report, we present a case of brodifacoum-induced homicide, wherein the patient exhibited distinct clinical examinations and symptoms. Moreover, the patient's blood sample exhibited a noteworthy brodifacoum concentration of 0.681 µg/mL even after a period of 43 days following the incident of poisoning. Although an autopsy was not conducted due to religious restrictions, we endeavor to reasonably deduce the cause of death and furnish corroborative evidence for clinical diagnosis, treatment, and forensic examination in instances involving brodifacoum poisoning.

Mutagenesis-Derived Resistance to Black Point in Wheat.

Black point, a severe global wheat disease, necessitates deploying resistant cultivars for effective control. However, susceptibility remains prevalent among most wheat cultivars. Identifying new sources of resistance and understanding their mechanisms are crucial for breeding resistant cultivars. This study pinpointed black point resistance in an ethyl methane sulfonate (EMS)-mutagenized wheat population of Wanyuanbai 1 (WYB) and analyzed resistant mutants using RNA-Seq. The findings revealed the following: (i) wyb-18, among 10,008 EMS-mutagenized lines, exhibited robust resistance with significantly lower black point incidence under artificial Bipolaris sorokiniana inoculation in 2020 and 2021 (average incidence of 5.2% over 2 years), markedly reduced compared with WYB (50.9%). (ii) wyb-18 kernels displayed black point symptoms at 12 days after inoculation (dai), 3 days later than WYB. At 15 dai, wyb-18 kernels had isolated black spots, unlike WYB kernels, where the entire embryo turned black. (iii) wyb-18 showed heightened antioxidant enzyme activity, including peroxidase, catalase, and superoxide dismutase. (iv) Analysis of 543 differentially expressed genes between wyb-18 and WYB at 9 dai identified enrichment in the MAPK signaling pathway through KEGG analysis. Ten genes in this pathway exhibited upregulated expression, while one was downregulated in wyb-18. Among these genes, PR1, WRKY11, SAPK5, and TraesCS1A02G326800 (chitin recognition protein) consistently showed upregulation in wyb-18, making them potential candidates for black point resistance. These results offer valuable germplasm resources for breeding and novel insights into the mechanisms of black point resistance.

Screening and Resistance Locus Identification of the Mutant fcrZ22 Resistant to Crown Rot Caused by Fusarium pseudograminearum.

Crown rot caused by Fusarium pseudograminearum is a devastating wheat disease worldwide. In addition to yield losses, the fungi causing Fusarium crown rot (FCR) also deteriorate the quality and safety of food because of the production of mycotoxins. Planting resistant cultivars is an effective way to control FCR. However, most wheat cultivars are susceptible to FCR. Therefore, development of new sources and detection of loci for FCR resistance are necessary. In the present study, a resistant mutant, fcrZ22, was identified from an ethyl methane sulfonate (EMS)-mutagenized population of the cultivar Zhoumai 22, and then fcrZ22 was crossed with the wild type to produce an F2 population. Genetic analysis of the F2 population was carried out by the mixed inheritance model of major genes plus polygenes, and 20 resistant and 20 susceptible plants were selected to assemble mixed pools. Combining 660K SNP arrays, the resistance loci were detected by bulked segregant analysis. The resistance to FCR caused by F. pseudograminearum in the F2 population was in accordance with the "mixed model with two major genes of additive-epistasis effect + additive-dominant polygenes," and the heritability of the major gene was 0.92. Twenty-one loci were detected, which were located on 10 chromosomes, namely, 1B (1), 1D (1), 2A (3), 1B (1), 3A (3), 3B (3), 4A (2), 5A (2), 7A (3), and 7B (2). Among the 21 loci, eight were new loci for FCR resistance. This is the first report of detecting loci for FCR resistance from a mutant. The results of the present study provided excellent germplasm resources for breeding wheat cultivars with FCR resistance and laid the foundation for fine mapping of FCR resistance loci.

A rare case of respiratory failure associated with spinal cord injury caused by stabbing with scissors and a knife.

In forensic practice, spinal cord injury (SCI) resulting in death has rarely been reported. Here, we present the case of a 65-year-old woman who on admission was conscious without dyspnea or dysphagia. Physical examination revealed two sharp objects penetrating the neck: a pair of scissors lodged in the neck on the right side of the thyroid cartilage and a knife embedded in the nuchal region accompanied by minor seepage of bloody exudate. Radiography showed that the scissors and knife were retained in the cervical spine. Despite a series of medical interventions, the patient died 26 days later. METHOD: A systematic forensic autopsy was performed. RESULTS: The cause of death was confirmed to be respiratory failure associated with SCI, which was caused by the combination of scissors and a knife. CONCLUSION: Based on this case, we believe that when there are multiple causes of death, forensic pathologists should determine the primary, immediate, contributory, and other causes of death to ascertain criminal responsibility.

Genome-wide identification and expression analysis of the plant-specific PLATZ gene family in Tartary buckwheat (Fagopyrum tataricum).

BACKGROUND: Plant AT-rich sequence and zinc-binding (PLATZ) proteins belong to a novel class of plant-specific zinc-finger-dependent DNA-binding proteins that play essential roles in plant growth and development. Although the PLATZ gene family has been identified in several species, systematic identification and characterization of this gene family has not yet been carried out for Tartary buckwheat, which is an important medicinal and edible crop with high nutritional value. The recent completion of Tartary buckwheat genome sequencing has laid the foundation for this study. RESULTS: A total of 14 FtPLATZ proteins were identified in Tartary buckwheat and were classified into four phylogenetic groups. The gene structure and motif composition were similar within the same group, and evident distinctions among different groups were detected. Gene duplication, particularly segmental duplication, was the main driving force in the evolution of FtPLATZs. Synteny analysis revealed that Tartary buckwheat shares more orthologous PLATZ genes with dicotyledons, particularly soybean. In addition, the expression of FtPLATZs in different tissues and developmental stages of grains showed evident specificity and preference. FtPLATZ3 may be involved in the regulation of grain size, and FtPLATZ4 and FtPLATZ11 may participate in root development. Abundant and variable hormone-responsive cis-acting elements were distributed in the promoter regions of FtPLATZs, and almost all FtPLATZs were significantly regulated after exogenous hormone treatments, particularly methyl jasmonate treatment. Moreover, FtPLATZ6 was significantly upregulated under all exogenous hormone treatments, which may indicate that this gene plays a critical role in the hormone response of Tartary buckwheat. CONCLUSIONS: This study lays a foundation for further exploration of the function of FtPLATZ proteins and their roles in the growth and development of Tartary buckwheat and contributes to the genetic improvement of Tartary buckwheat.

Exogenous 24-epibrassinolide boosts plant growth under alkaline stress from physiological and transcriptomic perspectives: The case of broomcorn millet (Panicum miliaceum L.).

Land alkalization is an abiotic stress that affects global sustainable agricultural development and the balance of natural ecosystems. In this study, two broomcorn millet cultivars, T289 (alkaline-tolerant) and S223 (alkaline-sensitive), were selected to investigate the response of broomcorn millet to alkaline stress and the role of brassinolide (BR) in alkaline tolerance. Phenotypes, physiologies, and transcriptomes of T289 and S223 plants under only alkaline stress (AS) and alkaline stress with BR (AB) were compared. The results showed that alkaline stress inhibited growth, promoted the accumulation of soluble sugars and malondialdehyde, enhanced electrolyte leakage, and destroyed the integrity of broomcorn millet stomata. In contrast, BR lessened the negative effects of alkaline stress on plants. Transcriptome sequencing analysis showed that relative to control groups (CK, nutrient solution), in AS groups, 21,113 and 12,151 differentially expressed genes (DEGs) were identified in S223 and T289, respectively. Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) revealed various terms and pathways related to metabolism. Compared to S223, alkaline stress strongly activated the brassinosteroid biosynthesis pathway in T289. Conversely, ARF, TF, and TCH4, associated with cell growth and elongation, were inhibited by alkaline stress in S223. Moreover, alkaline stress induced the activation of the mitogen-activated protein kinase (MAPK) pathway, the abscisic acid signaling pathway that initiates stomatal closure, as well as the starch and sucrose metabolism. The EG and BGL genes, which are associated with cellulose degradation, were notably activated. BR enhanced alkaline tolerance, thereby alleviating the transcriptional responses of the two cultivars. Cultivar T289 is better in alkalized regions. Taken together, these results reveal how broomcorn millet responds to alkaline stress and BR mitigates alkaline stress, thus promoting agriculture in alkalized regions.

A Design of Electromagnetic Velocity Sensor with High Sensitivity Based on Dual-Magnet Structure.

The most permanent magnets in current electromagnetic velocity sensors are magnet cylinders that have been axially magnetized, with magnetic boots changing the propagation direction of the magnetic induction lines of the magnet cylinders. However, the magnetic field generated by the magnet cylinders is not fully utilized, which leads to uneven magnetic field intensity of the working air-gap and high magnetic field intensity of the nonworking air-gap. We propose a novel dual-magnet structure (DM) mainly consisting of two magnet loops that are magnetized radially and a magnetic conductive shaft, adopting a concentric nested configuration. The dual-magnet structure can make the magnetic induction lines enter the working air-gap directly from the magnet and increase the effective magnetic field, which is perpendicular to the coils in the working air-gap. This design can further improve the sensitivity of a velocity sensor and enhance its ability to detect weak signals in microtremor exploration. The validity of the dual-magnet structure has been established by numerical simulations and verified by experiments. The results reveal that the magnetic field intensity is increased by 29.18% and the sensitivity is improved by 23.9%, when the total volume and material of the magnet are unchanged. The full utilization of the material is achieved without increasing the complexity of the structure.

Dynamic and Comparative Transcriptome Analyses Reveal Key Factors Contributing to Cadmium Tolerance in Broomcorn Millet.

Broomcorn millet (Panicum miliaceum L.) has great potential in Cd phytoextraction, but its mechanisms are largely unknown. Two contrasting broomcorn millet varieties, 'Ningmi6' (Cd-sensitive variety) and '4452' (Cd-tolerant variety), were investigated through morphological, physiological, and transcriptomic analyses to determine the factors responsible for their differential Cd tolerance and translocation. The Cd-tolerant variety can accumulate more Cd, and its cell wall and vacuole component Cd proportions were higher compared with the Cd-sensitive variety. Under Cd stress, the glutathione content and peroxidase activity of the Cd-tolerant variety were significantly higher than those of the Cd-sensitive variety. Additionally, weighted gene co-expression network analysis (WGCNA) revealed hub modules that were associated with Cd stress and/or variety. Notably, genes involved in these hub modules were significantly enriched for roles in glutathione metabolism, phenylpropanoid biosynthesis, ABC transport, and metal ion transport process. These results suggested that regulation of genes associated with cell wall precipitation and vacuole compartmentalization may increase Cd tolerance and reduce Cd translocation in the Cd-tolerant variety, although it can absorb more Cd. This study provides a foundation for exploring molecular mechanisms of Cd tolerance and transport in broomcorn millet and new insights into improving Cd phytoremediation with this crop through genetic engineering.

Conuping BSA-Seq and RNA-Seq Reveal the Molecular Pathway and Genes Associated with the Plant Height of Foxtail Millet (Setaria italica).

Foxtail millet (Setaria italica) plays an important role in C4 crop research and agricultural development in arid areas due to its short growth period, drought tolerance, and barren tolerance. Exploration of the dwarfing mechanism and the dwarf genes of foxtail millet can provide a reference for dwarf breeding and dwarf research of other C4 crops. In this study, genetic analysis was performed using phenotypic data, candidate genes were screened by bulk segregant analysis sequencing (BSA-Seq); differentially expressed genes and metabolic pathways in different strains of high samples were analyzed by RNA sequencing (RNA-Seq). The association analysis of BSA-Seq and RNA-Seq further narrowed the candidate range. As a result, a total of three quantitative trait loci (QTLs) and nine candidate genes related to plant height were obtained on chromosomes I and IX. Based on the functional prediction of the candidate genes, we propose a hypothetical mechanism for the formation of millet dwarfing, in which, metabolism and MAPK signaling play important roles in the formation of foxtail millet plant height.

Endogenous bioactive gibberellin/abscisic acids and enzyme activity synergistically promote the phytoremediation of alkaline soil by broomcorn millet (Panicum miliaceum L.).

Broomcorn millet (Panicum miliaceum L.), an important food crop, grows in arid and semi-arid areas that face soil saline-alkalization. To date, no studies have investigated the mechanisms by which broomcorn millet seeds respond to and tolerate alkali stress. In this study, six broomcorn millet genotypes (B102, B220, B269, B279, B289, and B297) were selected to explore the physiological and molecular mechanisms of alkali stress at the germination stage. The results showed that alkali stress delayed the germination of broomcorn millet, and α-amylase activity was positively correlated with the germination rate. After alkali stress, the genotypes with lower alkali damage rates exhibited stronger antioxidant defenses. Real-time polymerase chain reaction analysis showed that alkali stress downregulated gibberellic acid (GA) synthesis genes but upregulated GA inactivation and abscisic acid (ABA) synthesis genes. Similarly, seeds displayed lower GA concentrations and higher ABA concentrations after alkali stress. Therefore, the ratios of various GAs/ABA decreased within the range of 35.77% to approximately 96.45%. Additionally, genotypes associated with lower alkali damage rates had higher GA/ABA ratios. These findings indicate that the alkali tolerance of broomcorn millet at the germination stage may be attributed to higher GA/ABA ratios, higher α-amylase activity, and stronger antioxidant defense, which synergistically resist alkali stress. This study will contribute to molecular breeding aiming to enhance alkali-tolerance and restoration of alkaline soils.

Optimization of DOTAP/chol Cationic Lipid Nanoparticles for mRNA, pDNA, and Oligonucleotide Delivery.

Lipid nanoparticles (LNPs) can be used as delivery vehicles for nucleic acid biotherapeutics. In fact, LNPs are currently being used in the Pfizer/BioNTech and Moderna COVID-19 vaccines. Cationic LNPs composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP)/cholesterol (chol) LNPs have been classified as one of the most efficient gene delivery systems and are being tested in numerous clinical trials. The objective of this study was to examine the effect of the molar ratio of DOTAP/chol, PEGylation, and lipid to mRNA ratio on mRNA transfection, and explore the applications of DOTAP/chol LNPs in pDNA and oligonucleotide transfection. Here we showed that PEGylation significantly decreased mRNA transfection efficiency of DOTAP/chol LNPs. Among non-PEGylated LNP formulations, 1:3 molar ratio of DOTAP/chol in DOTAP/chol LNPs showed the highest mRNA transfection efficiency. Furthermore, the optimal ratio of DOTAP/chol LNPs to mRNA was tested to be 62.5 µM lipid to 1 µg mRNA. More importantly, these mRNA-loaded nanoparticles were stable for 60 days at 4 °C storage without showing reduction in transfection efficacy. We further found that DOTAP/chol LNPs were able to transfect pDNA and oligonucleotides, demonstrating the ability of these LNPs to transport the cargo into the cell nucleus. The influence of various factors in the formulation of DOTAP/chol cationic LNPs is thus described and will help improve drug delivery of nucleic acid-based vaccines and therapies.

A promising crop for cadmium-contamination remediation: Broomcorn millet.

Cadmium (Cd) pollution highly threatens food security and human health, and phytoremediation with Cd-tolerant plants is a cost-effective in situ method for remediation of Cd contamination. Broomcorn millet is known for its strong abiotic stress resistance and can be used as a pioneer crop in both marginal regions and newly reclaimed land. To evaluate their potential in remediation of Cd contamination, a total of 288 broomcorn millet core collections were investigated under hydroponic conditions to compare their capabilities in Cd tolerance, translocation, and accumulation. The core collections varied considerably in their growth parameters, Cd concentration, Cd translocation factor, Cd bioaccumulation factor, and Cd accumulation under Cd stress. According to the Cd tolerance index (TI) values, 160 varieties were Cd tolerant. The Cd TI was significantly positively correlated with Cd accumulation, and the shoot Cd concentrations of five Cd-tolerant varieties were more than 100 mgkg-1, the threshold for being Cd hyperaccumulators. Moreover, the concentrations of essential metal elements were significantly decreased in shoots, and Cd concentration had a significantly positive relationship with magnesium (Mg) and zinc (Zn) concentrations in roots under Cd stress. These results demonstrate that broomcorn millet shows considerable tolerance to Cd stress and great differences in Cd accumulation abilities among varieties. Accordingly, broomcorn millet is a promising plant species for Cd bioremediation, with valuable varieties that have been identified for further study on Cd tolerance mechanisms and the remediation of Cd contamination.

Proso millet (Panicum miliaceum L.): A potential crop to meet demand scenario for sustainable saline agriculture.

Proso millet (Panicum miliaceum L.) is resilient to abiotic stress, especially to land degradation caused by soil salinization. However, the mechanisms by which its roots adapt and tolerate salt stress are obscure. In this study, plants of a salt-sensitive cultivar (SS 212) and a salt-tolerant cultivar (ST 47) of proso millet were exposed to severe salt stress and subsequent re-watering. ST 47 exhibited greater salt tolerance than SS 212, as evidenced by higher increases in total root length (TRL), root surface area (RSA), root tip number (RTN). Moreover, microstructural analysis showed that relative to SS 212, the roots of ST 47 could maintain more intact internal structures and thicker cell walls under salt stress. Digital RNA sequence analysis revealed that ST 47 maintained better Na+/K+ balance to resist Na+ toxicity via a higher capability to restrict Na+ uptake, vacuolar Na+ sequestration, and Na+ exclusion. The mechanism for Na+ toxicity resistance in ST 47 involved promoting cell wall composition changes via efficient regulation of galactose metabolism and biosynthesis of cellulose and phenylpropanoids. Overall, this study provides valuable salt-tolerant cultivar resources and mechanisms for regulating salt tolerance, which could be applied for the rehabilitation of saline lands.

Exogenous Melatonin Modulates the Physiological and Biochemical Mechanisms of Drought Tolerance in Tartary Buckwheat (Fagopyrum tataricum (L.) Gaertn).

Tartary buckwheat is one of the nutritious minor cereals and is grown in high-cold mountainous areas of arid and semi-arid zones where drought is a common phenomenon, potentially reducing the growth and yield. Melatonin, which is an amphiphilic low molecular weight compound, has been proven to exert significant effects in plants, under abiotic stresses, but its role in the Tartary buckwheat under drought stress remains unexplored. We evaluated the influence of melatonin supplementation on plant morphology and different physiological activities, to enhance tolerance to posed drought stress by scavenging reactive oxygen species (ROS) and alleviating lipid peroxidation. Drought stress decreased the plant growth and biomass production compared to the control. Drought also decreased Chl a, b, and the Fv/Fm ratio by 54%, 70%, and 8%, respectively, which was associated with the disorganized stomatal properties. Under drought stress, H2O2, O2•-, and malondialdehyde (MDA) contents increased by 2.30, 2.43, and 2.22-folds, respectively, which caused oxidative stress. In contrast, proline and soluble sugar content were increased by 84% and 39%, respectively. However, exogenous melatonin (100 µM) could improve plant growth by preventing ROS-induced oxidative damage by increasing photosynthesis, enzymatic antioxidants (superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase), secondary metabolites like phenylalanine ammonialyase, phenolics, and flavonoids, total antioxidant scavenging (free radical DPPH scavenging), and maintaining relative water content and osmoregulation substances under water stress. Therefore, our study suggested that exogenous melatonin could accelerate drought resistance by enhancing photosynthesis and antioxidant defense in Tartary buckwheat plants.

Effect of ligand-receptor interaction of osteopontin-CD44 on the expression of hyaluronic acid in human knee osteoarthritic chondrocytes in vitro. / 骨桥蛋白-CD44é ä½“å—ä½“ååº”å¯¹äººè†éª¨å ³èŠ‚ç‚Žè½¯éª¨ç»†èƒžé€æ˜Žè´¨é ¸è¡¨è¾¾çš„å½±å“.

OBJECTIVES: To investigate the effect of ligand-receptor interaction of osteopontin (OPN)-CD44 on the expression of hyaluronic acid (HA) in the cultured human knee osteoarthritis (OA) chondrocytes via interfering the reaction between OPN and CD44 ligand-receptor. METHODS: The OA chondrocytes and normal chondrocytes were obtained from knee joint cartilage tissues in the patients with knee OA and malignant tumor respectively. The normal chondrocytes and OA chondrocytes were detected and analyzed, and then the intervention analysis of OA chondrocytes was carried out. The OA chondrocytes were divided into 4 groups: a negative control group, which was cultured with complete medium without any molecular intervention reagent; an OPN intervention group, which was cultured with recombinant human OPN (rhOPN) for 24 hours; a CD44 blocking group, which were pretreated with CD44 receptor specific antagonist for 1 hour to block the binding of OPN-CD44, and then treated with rhOPN for 23 hours; a CD44 homotype group, which was pretreated with CD44 for 1 hour and then treated with rhOPN for 23 hours. In addition, the study for OPN-CD44 axis was also divided into 4 groups: an OA-negative control group (OA-NC group), a si-OPN intervention group, a rhOPN intervention group, and a rhOPN + CD44 antibody (Ab) group. Western blotting, real-time PCR, and enzyme linked immunosorbent assay (ELISA) were used to detect the protein and mRNA expression levels of OPN, CD44, hyaluronate synthase (HAS), and HA, respectively. RESULTS: The protein expression levels of OPN, CD44, and HAS1 and the secretion levels of HA in the OA chondrocytes were higher than those in the normal chondrocytes. Compared with the OPN intervention group, the expression levels of HAS1, HAS2, HAS3 and HA in the CD44 blocking group were lower than those in OPN intervention group (all P<0.05); but there was no significant difference in the expression levels of HAS1, HAS2, HAS3 and HA between the CD44 homotype group and the OPN intervention group (all P>0.05). The results of OPN-CD44 axis study showed that: compared with the OA-NC group, the expression of CD44 in the rhOPN intervention group was slightly lower, but the protein and mRNA levels of HAS1 were significantly increased (all P<0.05); compared with the OA-NC group, the expression of CD44 was up-regulated, but the protein and mRNA level of HAS1 were significantly inhibited in the si-OPN intervention group (all P<0.05); compared with the OA-NC group, the protein and mRNA levels of HAS1 in the rhOPN+CD44 Ab group were also significantly inhibited (all P<0.05). CONCLUSIONS: The OPN in OA chondrocytes can promote the expression of HAS1, and the OPN can stimulate the secretion of HAS and induce HA expression by reacting with CD44 ligand receptor. They constitute the axis of OPN/CD44/HAS1, which plays an important role in regulating the expression of HA in chondrocytes.

A retrospective study of 29 fatal cases of insulin overdose.

PURPOSE: To summarize recent cases of fatal insulin poisoning both domestically and internationally, thereby offering valuable insights for the forensic identification of insulin overdose cases. METHODS: Literature published since 2000 on fatal insulin overdose were systematically searched and screened. Data encompassing variables such as year, age, sex, cause of death, scene conditions, occupations, medical histories of victims and perpetrators, autopsy timing, dosage and administration methods, forensic pathology, and toxicological analysis, were compiled for rigorous statistical analysis. RESULTS: Among the 29 fatal cases of insulin poisoning, suicides and homicides accounted for 55.2 % and 41.4 %, respectively. Precisely 34.5 % of victims or perpetrators were associated with the medical industry, 27.6 % had diabetes, and 24.1 % had mental illnesses such as depression. Intravenous injection resulted in quicker death than did subcutaneous injection. In some cases, immunohistochemical staining of insulin and protamine at injection sites yielded positive results. The average molar ratio of insulin to C-peptide in post-mortem blood was 13.76 ± 5.167, indicating a significant diagnostic value for insulin poisoning. CONCLUSION: Assessment of cases of fatal insulin overdose should be thorough, incorporating case investigation, scene examination, medical records review, autopsy findings, pathological examinations, and laboratory tests, alongside considering the condition of the body and timing of death autopsy. Using mass spectrometry to detect insulin proves valuable, particularly in cases of poor body preservation.

Review of the lethal mechanism of insulin poisoning and the characteristic of forensic identification.

Insulin, as the only hypoglycemic hormone in the body, plays a key role in blood sugar control. However, excessive insulin intake can lead to insulin poisoning and even death, which often occurs in clinical and forensic work. At present, some researches on insulin poisoning have been carried out at home and abroad, however, it seems that the mechanism and forensic characteristics of insulin poisoning are not clear and complete. Therefore, in this paper, we reviewed the potential mechanism of insulin poisoning, the methods of insulin detection and the forensic identification of poisoning cases, aiming at providing services for the forensic identification of insulin poisoning.

Engineering an injectable gellan gum-based hydrogel with osteogenesis and angiogenesis for bone regeneration.

Injectable hydrogels are currently a topic of great interest in bone tissue engineering, which could fill irregular bone defects in a short time and avoid traditional major surgery. Herein, we developed an injectable gellan gum (GG)-based hydrogel for bone defect repair by blending nano-hydroxyapatite (nHA) and magnesium sulfate (MgSO4). In order to acquire an injectable GG-based hydrogel with superior osteogenesis, nHA were blended into GG solution with an optimized proportion. For the aim of endowing this hydrogel capable of angiogenesis, MgSO4 was also incorporated. Physicochemical evaluation revealed that GG-based hydrogel containing 5% nHA (w/v) and 2.5 mM MgSO4 (GG/5%nHA/MgSO4) had appropriate sol-gel transition time, showed a porosity-like structure, and could release magnesium ions for at least 14 days. Rheological studies showed that the GG/5%nHA/MgSO4 hydrogel had a stable structure and repeatable self-healing properties. In-vitro results determined that GG/5%nHA/MgSO4 hydrogel presented superior ability on stimulating bone marrow mesenchymal stem cells (BMSCs) to differentiate into osteogenic linage and human umbilical vein endothelial cells (HUVECs) to generate vascularization. In-vivo, GG/5%nHA/MgSO4 hydrogel was evaluated via a rat cranial defect model, as shown by better new bone formation and more neovascularization invasion. Therefore, the study demonstrated that the new injectable hydrogel, is a favorable bioactive GG-based hydrogel, and provides potential strategies for robust therapeutic interventions to improve the repair of bone defect.

Microenvironment-responsive metal-phenolic network release platform with ROS scavenging, anti-pyroptosis, and ECM regeneration for intervertebral disc degeneration.

Intervertebral disc degeneration (IVDD) can be caused by aging, injury, and genetic factors. The pathological changes associated with IVDD include the excessive accumulation of reactive oxygen species (ROS), cellular pyroptosis, and extracellular matrix (ECM) degradation. There are currently no approved specific molecular therapies for IVDD. In this study, we developed a multifunctional and microenvironment-responsive metal-phenolic network release platform, termed TMP@Alg-PBA/PVA, which could treat (IL-1ß)-induced IVDD. The metal-phenolic network (TA-Mn-PVP, TMP) released from this platform targeted mitochondria to efficiently scavenge ROS and reduce ECM degradation. Pyroptosis was suppressed through the inhibition of the IL-17/ERK signaling pathway. These findings demonstrate the versatility of the platform. And in a rat model of IVDD, TMP@Alg-PBA/PVA exhibited excellent therapeutic effects by reducing the progression of the disease. TMP@Alg-PBA/PVA, therefore, presents clinical potential for the treatment of IVDD.

High-content stimulated Raman histology of human breast cancer.

Histological examination is crucial for cancer diagnosis, however, the labor-intensive sample preparation involved in the histology impedes the speed of diagnosis. Recently developed two-color stimulated Raman histology could bypass the complex tissue processing to generates result close to hematoxylin and eosin staining, which is one of the golden standards in cancer histology. Yet, the underlying chemical features are not revealed in two-color stimulated Raman histology, compromising the effectiveness of prognostic stratification. Here, we present a high-content stimulated Raman histology (HC-SRH) platform that provides both morphological and chemical information for cancer diagnosis based on un-stained breast tissues. Methods: By utilizing both hyperspectral SRS imaging in the C-H vibration window and sparsity-penalized unmixing of overlapped spectral profiles, HC-SRH enabled high-content chemical mapping of saturated lipids, unsaturated lipids, cellular protein, extracellular matrix (ECM), and water. Spectral selective sampling was further implemented to boost the speed of HC-SRH. To show the potential for clinical use, HC-SRH using a compact fiber laser-based stimulated Raman microscope was demonstrated. Harnessing the wide and rapid tuning capability of the fiber laser, both C-H and fingerprint vibration windows were accessed. Results: HC-SRH successfully mapped unsaturated lipids, cellular protein, extracellular matrix, saturated lipid, and water in breast tissue. With these five chemical maps, HC-SRH provided distinct contrast for tissue components including duct, stroma, fat cell, necrosis, and vessel. With selective spectral sampling, the speed of HC-SRH was improved by one order of magnitude. The fiber-laser-based HC-SRH produced the same image quality in the C-H window as the state-of-the-art solid laser. In the fingerprint window, nucleic acid and solid-state ester contrast was demonstrated. Conclusions: HC-SRH provides both morphological and chemical information of tissue in a label-free manner. The chemical information detected is beyond the reach of traditional hematoxylin and eosin staining and heralds the potential of HC-SRH for biomarker discovery.