Different projection neurons of basolateral amygdala participate in the retrieval of morphine withdrawal memory with diverse molecular pathways.

Context-induced retrieval of drug withdrawal memory is one of the important reasons for drug relapses. Previous studies have shown that different projection neurons in different brain regions or in the same brain region such as the basolateral amygdala (BLA) participate in context-induced retrieval of drug withdrawal memory. However, whether these different projection neurons participate in the retrieval of drug withdrawal memory with same or different molecular pathways remains a topic for research. The present results showed that (1) BLA neurons projecting to the prelimbic cortex (BLA-PrL) and BLA neurons projecting to the nucleus accumbens (BLA-NAc) participated in context-induced retrieval of morphine withdrawal memory; (2) there was an increase in the expression of Arc and pERK in BLA-NAc neurons, but not in BLA-PrL neurons during context-induced retrieval of morphine withdrawal memory; (3) pERK was the upstream molecule of Arc, whereas D1 receptor was the upstream molecule of pERK in BLA-NAc neurons during context-induced retrieval of morphine withdrawal memory; (4) D1 receptors also strengthened AMPA receptors, but not NMDA receptors, -mediated glutamatergic input to BLA-NAc neurons via pERK during context-induced retrieval of morphine withdrawal memory. These results suggest that different projection neurons of the BLA participate in the retrieval of morphine withdrawal memory with diverse molecular pathways.

Formation of chronic morphine withdrawal memories requires C1QL3-mediated regulation of PSD95 in the mouse basolateral amygdala.

Chronic morphine withdrawal memory formation is a complex process influenced by various molecular mechanisms. In this study, we aimed to investigate the contributions of the basolateral amygdala (BLA) and complement component 1, q subcomponent-like 3 (C1QL3), a secreted and presynaptically targeted protein, to the formation of chronic morphine (repeat dosing of morphine) withdrawal memory using conditioned place aversion (CPA) and chemogenetic methods. We conducted experiments involving the inhibition of the BLA during naloxone-induced withdrawal to assess its impact on CPA scores, providing insights into the significance of the BLA in the chronic morphine memory formation process. We also examined changes in C1ql3/C1QL3 expression within the BLA following conditioning. Immunofluorescence analysis revealed the colocalization of C1QL3 and the G protein-coupled receptor, brain-specific angiogenesis inhibitor 3 (BAI3) in the BLA, supporting their involvement in synaptic development. Moreover, we downregulated C1QL3 expression in the BLA to investigate its role in chronic morphine withdrawal memory formation. Our findings revealed that BLA inhibition during naloxone-induced withdrawal led to a significant reduction in CPA scores, confirming the critical role of the BLA in this memory process. Additionally, the upregulation of C1ql3 expression within the BLA postconditioning suggested its participation in withdrawal memory formation. The colocalization of C1QL3 and BAI3 in the BLA further supported their involvement in synaptic development. Furthermore, downregulation of C1QL3 in the BLA effectively hindered chronic morphine withdrawal memory formation, emphasizing its pivotal role in this process. Notably, we identified postsynaptic density protein 95 (PSD95) as a potential downstream effector of C1QL3 during chronic morphine withdrawal memory formation. Blocking PSD95 led to a significant reduction in the CPA score, and it appeared that C1QL3 modulated the ubiquitination-mediated degradation of PSD95, resulting in decreased PSD95 protein levels. This study underscores the importance of the BLA, C1QL3 and PSD95 in chronic morphine withdrawal memory formation. It provides valuable insights into the underlying molecular mechanisms, emphasizing their significance in this intricate process.

Novel insights into the effects of 5-hydroxymethfurural on genomic instability and phenotypic evolution using a yeast model.

5-Hydroxymethfurural (5-HMF) is naturally found in a variety of foods and beverages and represents a main inhibitor in the lignocellulosic hydrolysates used for fermentation. This study investigated the impact of 5-HMF on the genomic stability and phenotypic plasticity of the yeast Saccharomyces cerevisiae. Using next-generation sequencing technology, we examined the genomic alterations of diploid S. cerevisiae isolates that were subcultured on a medium containing 1.2 g/L 5-HMF. We found that in 5-HMF-treated cells, the rates of chromosome aneuploidy, large deletions/duplications, and loss of heterozygosity were elevated compared with that in untreated cells. 5-HMF exposure had a mild impact on the rate of point mutations but altered the mutation spectrum. Contrary to what was observed in untreated cells, more monosomy than trisomy occurred in 5-HMF-treated cells. The aneuploidy mutant with monosomic chromosome IX was more resistant to 5-HMF than the diploid parent strain because of the enhanced activity of alcohol dehydrogenase. Finally, we found that overexpression of ADH6 and ZWF1 effectively stabilized the yeast genome under 5-HMF stress. Our findings not only elucidated the global effect of 5-HMF on the genomic integrity of yeast but also provided novel insights into how chromosomal instability drives the environmental adaptability of eukaryotic cells.IMPORTANCESingle-cell microorganisms are exposed to a range of stressors in both natural and industrial settings. This study investigated the effects of 5-hydroxymethfurural (5-HMF), a major inhibitor found in baked foods and lignocellulosic hydrolysates, on the chromosomal instability of yeast. We examined the mechanisms leading to the distinct patterns of 5-HMF-induced genomic alterations and discovered that chromosomal loss, typically viewed as detrimental to cell growth under most conditions, can contribute to yeast tolerance to 5-HMF. Our results increased the understanding of how specific stressors stimulate genomic plasticity and environmental adaptation in yeast.

Projection neurons from medial entorhinal cortex to basolateral amygdala are critical for the retrieval of morphine withdrawal memory.

The medial entorhinal cortex (MEC) is crucial for contextual memory, yet its role in context-induced retrieval of morphine withdrawal memory remains unclear. This study investigated the role of the MEC and its projection neurons from MEC layer 5 to the basolateral amygdala (BLA) (MEC-BLA neurons) in context-induced retrieval of morphine withdrawal memory. Results show that context activates the MEC in morphine withdrawal mice, and the inactivation of the MEC inhibits context-induced retrieval of morphine withdrawal memory. At neural circuits, context activates MEC-BLA neurons in morphine withdrawal mice, and the inactivation of MEC-BLA neurons inhibits context-induced retrieval of morphine withdrawal memory. But MEC-BLA neurons are not activated by conditioning of context and morphine withdrawal, and the inhibition of MEC-BLA neurons do not influence the coupling of context and morphine withdrawal memory. These results suggest that MEC-BLA neurons are critical for the retrieval, but not for the formation, of morphine withdrawal memory.

Baseline Blood Levels of Mucin-1 Are Associated with Crucial On-Treatment Adverse Outcomes in Patients with Idiopathic Pulmonary Fibrosis Receiving Antifibrotic Pirfenidone.

Mucin-1 is a multi-functional glycoprotein expressed by type II alveolocytes and may be detectable in the circulation following pulmonary fibrosis. The prognostic utility of baseline pre-treatment blood levels of mucin-1 in patients with idiopathic pulmonary fibrosis (IPF) receiving antifibrotics has not yet been fully established. We retrospectively studied a cohort of patients (from two hospitals) with IPF who were receiving pirfenidone for >12 weeks. Baseline blood mucin-1 levels were measured via sandwich enzyme-linked immunosorbent assays. We investigated the performance of mucin-1 levels in longitudinally predicting the risks of acute exacerbation of IPF (AE-IPF) and severe adverse outcomes (SAO), including lung transplantation and death. Seventy patients were included; 20 developed AE-IPF; and 31 had SAO during the follow-up period. Patients with baseline mucin-1 levels ≥2.5 ng/mL had enhanced risks of AE-IPF (adjusted hazard ratio [aHR], 14.07; 95% confidence interval [CI], 4.26-46.49) and SAO within 2 years (aHR, 7.87; 95% CI, 2.86-21.70) and anytime during the follow-up (aHR, 4.68; 95% CI, 2.11-10.39). The risks increased across subgroups with increasing mucin-1 levels. Patients in the "mucin-1 ≥ 2.5" group also exhibited an accelerated decline in DLCO. This study supports baseline blood mucin-1 levels as a biomarker for IPF that predicts adverse outcomes during pirfenidone treatment.

Pseudolaric acid B triggers ferritinophagy and ferroptosis via upregulating NCOA4 in lung adenocarcinoma cells.

BACKGROUND: Ferroptosis is a novel subtype of programmed cell death caused by iron-dependent lipid peroxidation and excessive reactive oxygen species (ROS) production. Small-molecule ferroptotic drugs have the probability of selectively targeting the specific features of aggressive tumor cells. In particular, pseudolaric acid B (PAB) triggered ferroptosisin breast cancer cells. The aim of this study is to explore the antitumor effect of PAB on A549 cells and provide a theoretical basis for the further development and clinical application of PAB. METHODS: First, relevant databases were used to predict of target genes related to PAB, Then, EdU proliferation assay, colony formation and wound-healing assays were applied to calculate A549 cells proliferative abilities. Measurement of ferrous iron, lipid peroxidation, ROS, malondialdehyde (MDA) and glutathione (GSH) were utilized to explore the relevant mechanism. RESULTS: We showed that PAB decreased the viability of lung adenocarcinoma cells in vitro, which was accompanied by abnormally elevated levels of intracellular ferrous iron and overproduction of lipid reactive oxidate species (L-ROS). In turn, deferoxamine (DFO) significantly rescued PAB-induced lipid peroxidation. PAB also improved the intracellular labile iron pool by promoting ferritin autophagy via the upregulation of the nuclear receptor coactivator 4 (NCOA4). Moreover, silencing of NCOA4 alleviated PAB-inducedferroptotic death and reduced the levels of intracellular ferrous iron. CONCLUSIONS: In summary, PAB-triggered ferroptosis in lung adenocarcinoma cells by enhancing ferritinophagy. thus, PAB is a potential therapeutic agent for lung adenocarcinoma.

Transcriptional co-activator p300 participates in atrial fibrosis induced by high hydrostatic pressure by regulating TGF-β/Smad3 signaling pathway / 中国药理学通报

Aim To investigate the role and potential mechanism of transcriptional co-activator p300 in atrial fibrosis caused by high hydrostatic pressure. Methods The left atrial appendage tissues of humans in three groups of sinus rhythm, atrial fibrillation (AF), hypertension and AF were collected. The expressions of p300 protein and TGF-β/Smad3 signaling pathway and fibrotic factors as type I/III collagen Alphal chain (Col-lAl/Col-3Al), matrix metalloproteinase 2/9 (MMP-2/9) were tested by Western blot. Mouse atrial appendage fibroblasts were cultured under hydrostatic pressures of 0, 20 and 40 mmHg. The fibroblasts cultured under 40 mmHg pressure were treated with curcumin and p300 interference RNA. Western blot was used to test changes in the expression of p300 and the above fibrosis indicators. CCK-8 method was used to test changes of cell proliferation. Results The expressions of p300 and TGF-β/Smad3 signaling pathway proteins and fibrotic factors in AF group and hypertension combined with AF group were significantly higher than those in sinus rhythm group (P < 0. 05). 40 mmHg high hydrostatic pressure stimulation in vitro could increase the expression of p300 and fibrotic factors in fibroblasts (P < 0. 0 5) and enhance the proliferation ability (P < 0. 05). Both curcumin and p300 interfering RNA could reverse the increased expression of p300 and fibrotic factors (P < 0. 05) and decrease cell proliferation (P < 0. 05) induced by hydrostatic pressure. Conclusions High hydrostatic pressure can induce atrial fibrosis, which involves the participation of p300 in this process by regulating the TGF-β/Smad3 signaling pathway.

Surveillance and analysis of Haemaphysalis longicornis with SFTS virus in Tumen river basin located in the frontiers of Russia, Korea and Northeast China / 中华疾病控制杂志

Objective To mastered the distribution characteristics of Haemaphysalis longicornis in the Tumen River basin along the border between Russia, Korea and Northeast China, and understand the status of Haemaphysalis longicornis carrying the virus of fever with thrombocytopenia syndrome (SFTS), then, isolate the virus and analyze its genetic characteristics. Method Ticks were collected from Hunchun, Tumen, Helong and Longjing cities in the Tumen River basin of Jilin Province from April to September, 2017. Haemaphysalis longicornis was selected and grouped. SFTS virus was detected by real-time quantitative polymerase chain reaction (RT-qPCR). Virus isolation was carried out in Vero cells, besides, S,M,L gene segments were amplified . The homology of S, M and L gene segments was compared, phylogenetic tree was established, and their gene characteristics were analyzed. Result Haemaphysalis longicornis mainly distributed in Hunchun and Tumen City in the lower reaches of Tumen River. It was the dominant species in the two counties, reaching 71.85% and 87.62% respectively. A virus named YBHC-TICK2-2017/CHINA was isolated from Haemaphysalis longicornis collected in Hunchun. The sequences of S segment (1 746 bp), M segment (3 336 bp) and L segment (6 376 bp) of the virus were 98.00%-99.00%, homologous to those of SFTS virus isolates from China and Korea recorded in National Center for Biotechnology Information (NCBI) Gen Bank. Phylogenetic tree analysis showed that the S segment gene sequence of the virus strain was divided into a cluster with Jilin strain (KT890282) in China, and M segment and L segment gene sequence with Jiangsu strain (KR230781) in China. Conclusions Haemaphysalis longicornis are widely distributed in the lower reaches of the Tumen river. It was the first time that SFTS virus was isolated from Haemaphysalis longicornis in this area, suggesting that this area is important for SFTS prevention.

Investigation of free ticks carrying CRT and compound infection with SFTSV in Yanbian area of Jilin Province / 中华疾病控制杂志

Objective To understand the condition of tick carrying CRT(Candidatus rickettsia tarasevichiae) and compound infection with SFTSV(Sever fever thrombocytopenia syndrome virus) in Yanbian area of Jilin province. Methods Free ticks were collected from 6 counties including Yanji, Wangqing, Hunchun, Dunhua, Antu and Helong in the Korean Autonomous Prefecture of Jilin Province(Yanbian Prefecture) by using the flagging, and they were classified morphologically. The CRT and SFTSV in ticks were detected by Nest PCR(Nested polymerase chain reaction) and Real Time RT-PCR(Real-time Quantitative polymerase chain reaction) methods. Target DNA sequencing was performed for CRT positive products, and the homology of gene sequence and phylogenetic relationship were analyzed. Results A total of 1032 ticks were collected in this study, including ixodes persulcatus (35.56%) and dermacenter silvarum (20.64%). haemaphysalis japonica (20.45%), haemaphysalis longicornis (10.47%), haemaphysalis concinna (8.33%),others( 4.55%). The CRT was detected from the Ixodes persulcatus, Haemaphysalis longicornis, Haemaphysalis japonica Dermacentor silvarum. The MIR(Minimum infection rate per 100 ticks) of CRT was 10.47%.The SFTSV was detected from the Haemaphysalis-concinna, Haemaphysalis-japonica,Ixodes-persulcatus,Haemaphysalis-longicornis,Dermacentor-silvarum. The MIR of SFTSV was 2.52 %. Three species of ticks, including Ixodes persulcatus(2.45%), Haemaphysalis japonica(1.42%), and dermacentor silvarum(0.47%), had CRT and SFTSV compound infections, and the MIR of two pathogens compound infections was 1.26 %. In this study, the gene sequence of CRT positive PCR products ompA and 17kDa with nucleotide sequence of Xinyang plant of HeNan XinYang strain (KX365196.1),had homology of 100%. Phylogenetic analysis showed that CRT ompA and the HeNan XinYang strain (KX365196.1) gene sequences formed a cluster in Yanbian, while the 17 kDa gene formed an independent branch. Conclusions For the first time, CRT was detected from free ticks in Yanbian area of Jilin Province, and it was found that Ixodes persulcatus may be the main medium of transmission of the pathogen. At the same time, it was found that CRT and SFTSV have compound infection in ticks of Yanbian area. Therefore, it can be clearly identified that Yanbian area in Jilin Province is the natural source of CRT, and there are two pathogenic compound infections in the local ticks.