Somatosensory neurons express specific sets of lincRNAs, and lincRNA CLAP promotes itch sensation in mice.

Somatosensory neurons are highly heterogeneous with distinct types of neural cells responding to specific stimuli. However, the distribution and roles of cell-type-specific long intergenic noncoding RNAs (lincRNAs) in somatosensory neurons remain largely unexplored. Here, by utilizing droplet-based single-cell RNA-seq (scRNA-seq) and full-length Smart-seq2, we show that lincRNAs, but not coding mRNAs, are enriched in specific types of mouse somatosensory neurons. Profiling of lincRNAs from single neurons located in dorsal root ganglia (DRG) identifies 200 lincRNAs localized in specific types or subtypes of somatosensory neurons. Among them, the conserved cell-type-specific lincRNA CLAP associates with pruritus and is abundantly expressed in somatostatin (SST)-positive neurons. CLAP knockdown reduces histamine-induced Ca2+ influx in cultured SST-positive neurons and in vivo reduces histamine-induced scratching in mice. In vivo knockdown of CLAP also decreases the expression of neuron-type-specific and itch-related genes in somatosensory neurons, and this partially depends on the RNA binding protein MSI2. Our data reveal a cell-type-specific landscape of lincRNAs and a function for CLAP in somatosensory neurons in sensory transmission.

Structure Simulation and Equilibrium Evaluation Analysis of Regional Water Resources, Society, Economy and Ecological Environment Complex System.

Currently, the implementation of water resource spatial equilibrium strategy is a fundamental policy of water resource integrated management in China; it is also a considerable challenge to explore the relationship structure features of water resources, society, economy and ecological environment (WSEE) complex system. For this purpose, firstly, we applied information entropy, ordered degree and connection number coupling method to reveal the membership characteristics between different evaluation indicators and grade criterion. Secondly, the system dynamics approach was introduced to describe the relationship features among different equilibrium subsystems. Finally, the ordered degree, connection number, information entropy and system dynamics integrated model was established to conduct relationship structure simulation and evolution trend evaluation of the WSEE system. The application results in Hefei city, Anhui Province, China, demonstrated that: (1) the variation of overall equilibrium conditions of WSEE system in Hefei city, 2020-2029 was higher compared to that of 2010-2019, though the increasing rate of ordered degree and connection number entropy (ODCNE) became slower after 2019; and (2) the annual ODCNE value from 2020 to 2029 of WSEE system under dry year scenarios increased about 0.0812, which indicated that the construction of Yangtze-Huaihe Diversion (YHD) project could play significant positive role in mitigating the equilibrium situation of WSEE system in Hefei city in the future. On the whole, this study is capable of providing the guidance basis for constructing a theoretical framework of structure simulation and equilibrium evaluation analysis of WSEE complex system.

Drought identification based on Palmer drought severity index and return period analysis of drought characteristics in Huaibei Plain China.

The threshold level method for drought identification is challenging due to the problems of selection of drought index reflecting the drought process associated with water supply and demand as well as the underlying physical meaning of drought thresholds. The frequently used hydrological drought indices (e.g., runoff) are susceptible to being affected by human activities, and drought characteristics are incapable of revealing spatial and temporal comparability. Furthermore, the drought process with the same severity but a longer duration is more likely to be evaluated as a more severe event, which contradicts the actual drought situation. In this study, the Palmer drought severity index (PDSI) method, in which the meteorological factors less influenced by human activities were taken as the input, was adopted to determine the dry/wet states and the PDSI value at each period firstly. The dry/wet states were defined with dry period, wet period, transition period, transition period in dry spell, and transition period in a wet spell. Following that, drought identification criteria were established through the dry/wet states and PDSI value according to the consistency of the identified results and the actual drought situations. Particularly, drought severity and peak intensity were taken as drought characteristics in this paper, and the joint return periods of the characteristics were estimated based on the Gumbel-Hougaard copula function. And eventually, a case study was conducted in Huaibei Plain, China. The results showed that the most severe droughts identified by PDSI had a good consistency with the actual drought situations, drought severity and peak intensity were applicable to reflect the drought impacts. It is worth noting that the implications of the joint return period and the relationships among different types of them. The occurrence probability of a multi-characteristic drought event should be calculated by the integration of joint probability density function over the region corresponding to the event of interest, and the joint frequency of drought characteristics should not be used as the occurrence probability of the drought disaster losses greater (or less) than that caused by the drought with the same characteristics. In addition, drought processes identified by PDSI and standardized precipitation indices (SPI) from monthly and seasonal scales were compared, indicating the drought identified results through PDSI are almost consistent with the actual situations.

The Common and Different Neural Bases of Distraction and Reinterpretation: A Meta-Analysis of fMRI Studies.

Distraction and reinterpretation have been recognized as two different tactics of emotion regulation. As a tactic of attention deployment, distraction involves shifting attention to neutral information or performing a secondary task to distract attention from emotion stimuli of the primary task. Reinterpretation, a representative tactic of cognitive change, was defined as changing the meaning of a situation to enhance or reduce its emotional impact. Thus, there are significant differences between the two processes. We wondered if the neural mechanisms underlying distraction and reinterpretation are different. Even though their neural correlates have been widely studied with functional magnetic resonance imaging (fMRI), few studies were conducted to compare the two tactics directly. Here we conducted an activation likelihood estimation (ALE) meta-analyses to investigate the common or different neural bases of distraction and reinterpretation. Moreover, we also used the meta-analytic connectivity modeling (MACM) to identify the emotion regulation network of distraction and reinterpretation. Overall, we found that the left dorsal lateral prefrontal cortex (DLPFC) was consistently activated during distraction and reinterpretation, whereas the left amygdala and inferior frontal gyrus/ventrolateral prefrontal cortex (VLPFC) were specifically activated during reinterpretation alone. The results indicate that the neural basis of distraction and reinterpretation are similar but not identical. The MACM results showed that distraction and reinterpretation share a common emotion regulation network, including the bilateral DLPFC, the dorsal medial prefrontal cortex, the inferior parietal lobule, the insula, the left (pre) supplementary motor area, the left middle temporal gyrus, and the right superior temporal gyrus. However, that network may subserve different functions when adopting various emotion regulation strategies. In addition, we suggest that the emotion regulation network of the left VLPFC may be a specific regulatory network for reinterpretation.

Analysis of Pedestrian Fractures in Collisions Between Small Cars and Pedestrians Based on Surveillance Videos.

OBJECTIVE: To discuss the collision relationship and the cause of the fracture caused by traffic accidents in which the front of a small car collides with the side of a pedestrian while braking. METHODS: The surveillance videos of 42 traffic accidents involving the front of a small car colliding with the side of a pedestrian while braking were collected. By analyzing the surveillance videos and the paths, the speed of the collision, the relationship between the vehicle and the pedestrian upon collision, and the movement trajectory of the human body were clearly identified. The type and severity of the injuries were also determined through autopsy. The characteristics of the human injuries and vehicle paths were analyzed according to the collision speed (<40 km/h, 40-60 km/h, 60-90 km/h), and the correlations between the fracture and the height of the pedestrian, the height of the hood and the length of the hood were discussed. RESULTS: When a small car hits the side of a pedestrian, the front bumper first hits the lower limbs of the pedestrian, and then, the human body falls to the side of the vehicle, causing a secondary collision with the hood and front windshield; thus, the pedestrian is thrown at a speed similar to the speed of the vehicle, finally falling to the ground and sliding forward a certain distance. (1) When V is less than 40 km/h (n = 10), the pedestrian's head did not collide with the windshield, and the fatal injuries were caused by the individual striking the ground. (2) When V is greater than 40 km/h (n = 32), the majority (97%) of cases showed collision with the windshield. (3) When 40 to 60 km/h (n = 16), the pedestrian's head collided with the windshield, which can cause fatal injuries, and pelvic fractures and rib fractures occurred in 56.25% of patients. (4) When V is less than 60 km/h (n = 26), the ratio of the height of the pedestrian to the height of the hood was significantly smaller in the pelvic fracture group than in the nonpelvic fracture group (P < 0.01). (5) When 60 to 90 km/h (n = 16), there were holes in the windshield, and the pedestrians experienced severe head injuries, with cervical spine fracture occurring in 37.5% of patients, pelvic fractures occurring in 43.75% of patients, and rib fractures occurring in 31.25% of patients. CONCLUSIONS: When V is less than 40 km/h, the vehicle does not cause severe injuries in pedestrians; when V is greater than 40 km/h, the collisions of the pedestrian's head with the windshield lead to severe head injuries and the accident can cause severe pelvic and rib fractures; when V is greater than 60 km/h, the collisions of the pedestrian's head with the windshield can cause cervical spine fracture in addition to head injuries. The occurrence of human injuries is related to not only the vehicle speed but also factors such as the height of the pedestrian, the height of the hood and the length of the hood.

Pathological changes induced by phosphine poisoning: a study on 8 children.

Aluminum phosphide (ALP) has been extensively used as an economical and effective insecticide, rodenticide, and fumigant. The active ingredient of ALP is phosphine (PH3), the use of which can lead to accidental inhalation and mass poisoning with high mortality. Exposure to PH3 will give rise to global damage in the human body. This study reviewed 4 fatal accidents including 8 children with PH3 poisoning and aimed to determine the pathological changes that resulted from exposure to PH3 and, secondly, aimed to determine whether oxidative stress was involved in PH3-induced neurotoxicity using histopathological and immunohistochemistry (IHC) methods. After focusing on the pathological changes on the major organs, we found severe damage induced by PH3 in many systems, especially the neurological system, including neuronal, axonal, and vascular injuries as well as oxidative damage with increased expression of 4-hydroxy-2-trans-nonenal (4HNE), 8-hydroxy-2'-deoxyguanosine (8-OH-dG), and 3-nitrotyrosine (3-NT) in the brain, which indicated that oxidative stress was a crucial mechanism for neuronal death in PH3 toxicity. Moreover, we observed severe myocardial and hepatocellular fatty degeneration in the tissues of the heart and liver. We considered that these characteristic changes are a suggestive sign of PH3 poisoning and partly explained the toxic mechanism of PH3 (inhibition of mitochondrial oxidative phosphorylation). We hope that this research could improve the understanding of the toxicity of PH3 in both forensic and clinical practice.

Identifying novel microhaplotypes for ancestry inference.

The use of DNA to determine the ancestry of an individual is becoming more and more important in the areas of forensics. Kidd et al. (Forensic Sci Int Genet 12:215-224, 2014) have been the first to identify and catalog haplotypes, termed as minihaplotypes (1-10-kilobase spans) and microhaplotypes (≤ 200 bp), with potential use in forensic analysis. In the present study, we selected 10 short ancestry informative microhaplotypes by calculating the informativeness (In) according to Rosenberg et al. (Am J Hum Genet 73(6):1402-1422, 2003). In total, 2504 individuals from 26 populations in 1000 Genomes Project database Phase 3 were enrolled. Among the studied microhaplotypes, eight of them are comprised of 3 SNPs while two microhaplotypes are made up of 4 SNPs. The size (molecular extent) range of 10 microhaplotypes is 5 to 48 bp with an average of 31.4 bp. The heterozygosity value ranges from 0.2235 to 0.8958 with an average of 0.6593. The average power of discrimination (PD) values is 0.7944 and ranges from 0.3786 to 0.9242. Analyses of this dataset provided clear differentiation of the populations from the Africa, East Asia, South Asia, and Europe biogeographic regions. However, individuals from American ancestry were not well separated. To conclude, our results revealed the significance of using microhaplotypes as an ancestry informative marker. The present panel could offer a valid complementary tool in forensic applications.

Sudden death from an epileptic seizure due to capillary telangiectasias in the hippocampus.

Cerebral capillary telangiectasia (CCT) is a type of vascular malformation that is incidentally encountered in clinical practice. Diseased vessels are small and usually clinically benign over the course of a patient's life. Although most CCT patients are asymptomatic, the situation becomes complicated when trauma is encountered. A case of sudden death due to an epileptic episode after very mild head trauma is reported, including a retrospective study of 12 cases, to remind peers to pay close attention to CCT especially when located in important functional regions of the brain. After immunohistochemical staining and pathological examination, we speculated that the epileptogenic mechanism of CCT may be similar to that of hippocampal sclerosis. As the definite epileptogenic mechanism of CCT in the hippocampus is still elusive, we suggest that more research should be conducted on CCT.

Assessment of ethylene glycol diacetate as an alternative carrier for use in agrochemical emulsifiable concentrate formulation.

The conventional emulsifiable concentrate (EC) formulation contains a large amount of aromatic solvents, which causes adverse effects to both the environment and human health due to the toxicity of the solvents. Here, we developed a 2.5% lambda-cyhalothrin EC formulation with ethylene glycol diacetate (EGDA) as the solvent, and the developed formulation serves as an environmental-friendly alternative to overcome the adverse effects of aromatic solvents. The physicochemical characterizations, wettability properties, phytotoxicity and bioassays of the EGDA-EC formulation were systematically investigated and compared with that of the EC formulation with xylene as the solvent. The results showed that both EC formulations had excellent emulsion properties and storage stabilities. Additionally, the EGDA-EC formulation possessed a higher flash point (96 °C), indicating safer production, storage and transport. The retentions of the EGDA-EC sample on leaves were 1.22-1.46-fold higher than that of the xylene-EC sample, and the EGDA-EC also exhibited lower surface tensions and contact angles, which would benefit decreasing drift-off and improving utilization. Furthermore, the bioassays demonstrated that the EGDA-EC formulation had lower acute toxicity to aquatic organisms and higher control efficacy to target insects compared with the xylene-EC formulation. Therefore, EGDA is a promising carrier for oil-soluble agrochemicals to improve their application performance and reduce their adverse effects.

Sudden death due to malignant hyperthermia with a mutation of RYR1: autopsy, morphology and genetic analysis.

Malignant hyperthermia (MH) is an autosomal dominant disorder characterized by abnormal calcium homeostasis in skeletal muscles in response to triggering agents. Autopsy, morphology, and genetic analysis were performed on a 19-year-old man who died rapidly after exposure to sevoflurane during maxillofacial surgery. Muscle spasm around the operation area and limb rigidity occurred and renal tubules full of myoglobin casts were observed by microscopy. Ultrastructural changes in the skeletal muscles and the myocardium were detected by transmission electron microscopy (TEM). Genetic analysis disclosed a ryanodine receptor type 1 (RYR1) gene mutation and a nucleoide mutation in chromosome 19q (G1021A) in the deceased and his father. According to the fore mentioned results, the relationship between the cause of death and MH was confirmed. Thus, genetic analysis can be an important procedure in diagnosing MH.

[Nasal resorption of prim-O-glucosylcimifugin and 5-O-methylvisammioside in rats].

In this experiment, rat nasal mucosa absorption characteristics of prim-O-glucosylcimifugin and 5-O-methylvisammioside were studied to provide a basis for drug delivery of Toutongning nasal spray. The nasal mucosa absorption test in rats was conducted with in situ nasal perfusion method after pH 6 buffer solution was used to prepare high, medium and low concentrations of prim-O-glucosylcimifugin, 5-O-methylvisammioside mixed solution as liquid circulation in nasal cavity. Then the concentrations of the circulating liquid compositions to be measured were determined by HPLC, and the absorption rates of prim-O-glucosylcimifugin and 5-O-methylvisammioside under different pH conditions were also investigated. According to the results, the absorption rate constant was (0.588±0.041)×10⁻³, (0.547±0.023)×10⁻³, (0.592±0.063)×10⁻³ min⁻¹ for prim-O-glucosylcimifugin high, middle and low concentrations, and (0.438±0.041)×10⁻³, (0.407±0.023)×10⁻³, and (0.412±0.063)×10⁻³ min⁻¹ for 5-O-methylvisammioside high, middle and low concentrations. There was no significant difference among high, middle and low concentration groups, and the absorption under pH 6 was better than that under other pH conditions. Therefore, we can get the conclusion that the main active ingredient of Toutongning nasal sprays can be absorbed through the nasal mucosa, and it is feasible to make nasal spray; in addition, pH 6 of nasal spray is scientific and reasonable.

Expression patterns of SH3BGR family members in zebrafish development.

SH3 domain-binding glutamic acid-rich (SH3BGR) gene family is composed of SH3BGR, SH3BGRL, SH3BGRL2, and SH3BGRL3 which encodes a cluster of small thioredoxin-like proteins and shares a Src homology 3 (SH3) domain. However, biological functions of SH3BGR family members are largely elusive. Given that zebrafish (Danio rerio) sh3bgrl, sh3bgrl2, sh3bgrl3, and sh3bgr are evolutionally identical to their corresponding human orthologues, we analyzed the spatiotemporal expression of SH3BGR family members in zebrafish embryonic development stages by in situ hybridization. Our results revealed that except sh3bgrl, other members are all maternally expressed, especially for sh3bgrl3 that is strongly expressed from one-cell stage to juvenile fishes. In situ expression patterns of SH3BGR members are similar in the very early developmental stages, including with commonly strong expression in intestines, olfactory bulbs, and neuromasts for neural system building up. Organ-specific expressions are also demonstrated, of which sh3bgr is uniquely expressed in sarcomere, and sh3bgrl3 in liver. sh3bgrl and sh3bgrl2 are similarly expressed in intestines, notochords, and neuromasts after 12-h post-fertilization of embryos. Eventually, messenger RNAs (mRNAs) of all sh3bgr members are mainly constrained into intestines of juvenile fishes. Collectively, our study clarified the expression patterns of sh3bgr family members in diverse organogenesis in embryonic development and indicates that SH3BGR members may play predominant roles in neural system development and in maintenance of normal function of digestive organs, especially for intestine homeostasis. However, their expression patterns are varied with the development stages and organ types, suggesting that the aberrant expression of these members would result in multiple diseases.

Three homicides with darts tainted with succinylcholine: autopsy and toxicology.

In emergency departments, intoxication with the muscle relaxant succinylcholine (SUX) often leads to a potentially lethal respiratory paralysis or other deleterious side effects. However, homicide cases with SUX poisoning are very rare because the toxic or lethal concentration ranges of SUX have not yet been determined. We described three uncommon homicide cases due to acute poisoning by darts contaminated with SUX. All the victims died quickly (less than 30 min) after being shot by an especially designed dart gun. Succinylmonocholine (SMC), a metabolite of SUX, was used as a marker to detect the latter. HPLC-MS/MS analysis demonstrated the presence of SUX in the droplet residues of the darts and SMC in the blood and urine in all cases. SMC concentrations of 0.45, 14.0, and 17.9 ng/ml were detected in the victims' blood and 259.0 ng/ml in the urine from the third case. The main pathological changes consisted of hemorrhage of the injured soft tissues, visceral congestion, severe pulmonary edema, and multifocal petechial hemorrhage of the heart and lungs. Taken together, the findings supported a diagnosis of fatal SUX poisoning. Futhermore, our study provided a reference for the lethal concentrations of SUX poisoning.

Screening and Identification of High-Yielding Strains of Conjugated Linoleic Acid and Optimization of Conditions for the Conversion of CLA.

Conjugated linoleic acid (CLA) is a class of naturally occurring octadecadienoic acid in humans and animals and is a general term for a group of conformational and positional isomers of linoleic acid. In order to obtain the development of excellent lactic acid strains with a high production of conjugated linoleic acid, 32 strains with a possible CLA conversion ability were obtained by initial screening using UV spectrophotometry, and then the strains were re-screened by gas chromatography, and finally, the strain with the highest CLA content was obtained. The strains were optimized for cultivation by changing the amount of substrate addition, inoculum amount, and fermentation time. The results showed that the yield of the experimentally optimized strain for the conversion of conjugated linoleic acid could reach 94.68 ± 3.57 µg/mL, which was 74.4% higher than the initial yield of 54.28 ± 2.12 µg/mL of the strain. The results of this study can provide some basis for the application of conjugated linoleic acid production by Lactobacillus paracasei in the fermentation of lactic acid bacteria.

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.

Ginseng aconitum decoction (Shenfu Tang) provides neuroprotection by ameliorating impairment of blood-brain barrier in cerebral ischemia-reperfusion injury.

Ischemic stroke (IS) remains one of the most serious threats to human life. Early blood-brain barrier damage (BBB) is the cause of parenchymal cell damage. Repair of the structure and function of the BBB is beneficial for the treatment of IS. The traditional prescription ginseng aconitum decoction (GAD) has a long history in the treatment of cardiovascular and cerebrovascular diseases, however, the effect of GAD on the BBB disruption and underlying mechanisms remains largely unknown. To address these issues, in vitro models of BBB were established with brain endothelial cells (bEnd.3). We found that GAD reduced the leakage of the fluorescent probe FITC-dextran (P < 0.01) and increased the expression of tight junction proteins (Claudin-5, ZO-1) (P < 0.05) in the BBB model in vitro. Furthermore, to investigate the BBB protective effects of GAD in vivo. A total of 25 male C57/BL6 mice (20 - 22 g) were randomly divided into 5 groups (n = 5 per group): (1) Sham group (saline), (2) MCAO group (saline), (3) MCAO + CG group (Chinese ginseng 8 mg/kg/day), (4) MCAO + AC group (aconite 8 mg/kg/day), (5) MCAO + GAD group (GAD 8 mg/kg/day).We constructed IS model in mice and found that GAD treatment reduced IgG leakage (P < 0.05), up-regulated the expression of tight junction proteins Claudin-5, Occludin, and ZO-1 (P < 0.05). Further mechanism study showed that fatty acid oxidation (FAO) of vascular endothelial cells is involved in the protection of the BBB after IS, and GAD regulates FAO (P < 0.05) to protect BBB. In addition, we found the effect of GAD was stronger than that of Chinese ginseng (CG) (P < 0.05) and aconite (AC) (P < 0.01) alone. We concluded that GAD ameliorated the BBB dysfunction by regulating FAO involving vascular endothelial cells after IS. At the same time, the prescription is more effective than single traditional Chinese medicine.

Predictive value of MRI-based deep learning model for lymphovascular invasion status in node-negative invasive breast cancer.

To retrospectively assess the effectiveness of deep learning (DL) model, based on breast magnetic resonance imaging (MRI), in predicting preoperative lymphovascular invasion (LVI) status in patients diagnosed with invasive breast cancer who have negative axillary lymph nodes (LNs). Data was gathered from 280 patients, including 148 with LVI-positive and 141 with LVI-negative lesions. These patients had undergone preoperative breast MRI and were histopathologically confirmed to have invasive breast cancer without axillary LN metastasis. The cohort was randomly split into training and validation groups in a 7:3 ratio. Radiomics features for each lesion were extracted from the first post-contrast dynamic contrast-enhanced (DCE)-MRI. The Least Absolute Shrinkage and Selection Operator (LASSO) regression method and logistic regression analyses were employed to identify significant radiomic features and clinicoradiological variables. These models were established using four machine learning (ML) algorithms and one DL algorithm. The predictive performance of the models (radiomics, clinicoradiological, and combination) was assessed through discrimination and compared using the DeLong test. Four clinicoradiological parameters and 10 radiomic features were selected by LASSO for model development. The Multilayer Perceptron (MLP) model, constructed using both radiomic and clinicoradiological features, demonstrated excellent performance in predicting LVI, achieving a high area under the curve (AUC) of 0.835 for validation. The DL model (MLP-radiomic) achieved the highest accuracy (AUC = 0.896), followed by DL model (MLP-combination) with an AUC of 0.835. Both DL models were significantly superior to the ML model (RF-clinical) with an AUC of 0.720. The DL model (MLP), which integrates radiomic features with clinicoradiological information, effectively aids in the preoperative determination of LVI status in patients with invasive breast cancer and negative axillary LNs. This is beneficial for making informed clinical decisions.

Regulation of Erythroid Differentiation via the HIF1α-NFIL3-PIM1 Signaling Axis Under Hypoxia.

Aims: Erythropoiesis is controlled by several factors, including oxygen level under different circumstances. However, the role of hypoxia in erythroid differentiation and the underlying mechanisms are poorly understood. We studied the effect and mechanism of hypoxia on erythroid differentiation of K562 cells and observed the effect of hypoxia on early erythropoiesis of zebrafish. Results: Compared with normal oxygen culture, both hemin-induced erythroid differentiation of K562 cells and the early erythropoiesis of zebrafish were inhibited under hypoxic treatment conditions. Hypoxia-inducible factor 1 alpha (HIF1α) plays a major role in the response to hypoxia. Here, we obtained a stable HIF1α knockout K562 cell line using the CRISPR-Cas9 technology and further demonstrated that HIF1α knockout promoted hemin-induced erythroid differentiation of K562 cells under hypoxia. We demonstrated an HIF1-mediated induction of the nuclear factor interleukin-3 (NFIL3) regulated in K562 cells under hypoxia. Interestingly, a gradual decrease in NFIL3 expression was detected during erythroid differentiation of erythropoietin-induced CD34+ hematopoietic stem/progenitor cells (HSPCs) and hemin-induced K562 cells. Notably, erythroid differentiation was inhibited by enforced expression of NFIL3 under normoxia and was promoted by the knockdown of NFIL3 under hypoxia in hemin-treated K562 cells. In addition, a target of NFIL3, pim-1 proto-oncogene, serine/threonine kinase (PIM1), was obtained by RNA microarray after NFIL3 knockdown. PIM1 can rescue the inhibitory effect of NFIL3 on hemin-induced erythroid differentiation of K562 cells. Innovation and Conclusion: Our findings demonstrate that the HIF1α-NFIL3-PIM1 signaling axis plays an important role in erythroid differentiation under hypoxia. These results will provide useful clues for preventing the damage of acute hypoxia to erythropoiesis.

Ash1L ameliorates psoriasis via limiting neuronal activity-dependent release of miR-let-7b.

BACKGROUND AND PURPOSE: Psoriasis is a common autoimmune skin disease that significantly diminishes patients' quality of life. Interactions between primary afferents of the somatosensory system and the cutaneous immune system mediate the pathogenesis of psoriasis. This study aims to elucidate the molecular mechanisms of how primary sensory neurons regulate psoriasis formation. EXPERIMENTAL APPROACH: Skin and total RNA were extracted from wild-type (WT) and ASH1-like histone lysine methyltransferase (Ash1l+/- ) mice in both naive and imiquimod (IMQ)-induced psoriasis models. Immunohistochemistry, quantitative real-time polymerase chain reaction (qRT-PCR) and fluorescence-activated cell sorting (FACS) were then performed. Microfluidic chamber coculture was used to investigate the interaction between somatosensory neurons and bone marrow dendritic cells (BMDCs) ex vivo. Whole-cell patch clamp recordings were used to evaluate neuronal excitability after Ash1L haploinsufficiency in primary sensory neurons. KEY RESULTS: The haploinsufficiency of ASH1L, a histone methyltransferase, in primary sensory neurons causes both neurite hyperinnervation and increased neuronal excitability, which promote miR-let-7b release from primary afferents in the skin in a neuronal activity-dependent manner. With a 'GUUGUGU' core sequence, miR-let-7b functions as an endogenous ligand of toll-like receptor 7 (TLR7) and stimulates the activation of dermal dendritic cells (DCs) and interleukin (IL)-23/IL-17 axis, ultimately exacerbating the symptoms of psoriasis. Thus, by limiting miR-let-7b release from primary afferents, ASH1L prevents dermal DC activation and ameliorates psoriasis. CONCLUSION AND IMPLICATIONS: Somatosensory neuron ASH1L modulates the cutaneous immune system by limiting neuronal activity-dependent release of miR-let-7b, which can directly activate dermal DCs via TLR7 and ultimately lead to aggravated psoriatic lesion.

Methylation changes of liver DNA during the formation of gallstones.

Aim: To explore the overall methylation changes in liver tissues during the formation of gallstones, as well as the key pathways and genes involved in the process. Methods: Reduced-representation bisulfite sequencing and RNA sequencing were conducted on the liver tissues of mice with gallstones and control normal mice. Results: A total of 8705 differentially methylated regions in CpG and 1410 differentially expressed genes were identified. The joint analysis indicated that aberrant DNA methylation may be associated with dysregulated gene expression in key pathways such as cholesterol metabolism and bile secretion. Conclusion: We propose for the first time that methylation changes in some key pathway genes in liver tissue may be involved in the formation of gallstones.