Objective To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. Methods B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Results Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ² = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. Conclusion The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system.

Objective:To explore the clinicopathological features of serum alpha-fetoprotein-producing gastric cancer (AFPGC) and its subtype hepatoid adenocarcinoma of the stomach (HAS), and to analyze the related factors affecting the prognosis of AFPGC and HAS patients.Methods:The clinical data of 91 patients with AFPGC who were admitted to the Second Affiliated Hospital of Dalian Medical University from February, 2010 to February, 2021 were retrospectively collected. According to the results of hepatoid differentiation, the patients were divided into HAS group (26 cases) and non-HAS group (65 cases). Meanwhile, 130 patients with alpha-fetoprotein (AFP) negative gastric cancer at the Second Affiliated Hospital of Dalian Medical University were selected by stratified sampling method as common gastric cancer group. The clinicopathological data of patients in the HAS group, non-HAS group and common gastric cancer group were compared and all the patients were followed up for 3 years. Kaplan-Meier survival curves were plotted, and log-rank test was used to analyze the median survival time. Multivariate Cox regression was performed to analyze the independent risk factors affecting the prognosis of patients. Chi-square test was used for statistical comparison.Results:The proportion of patients with poorly differentiated tumor in the HAS group was higher than that in the non-HAS group (84.6%(22/26) vs. 55.4%(36/65)), and the difference was statistically significant ( χ2=7.02, P=0.030). The proportions of patients with age < 60 years old, abnormal level of carcinoembryonic antigen (CEA), vascular tumor thrombus, liver metastasis, poor differentiated tumor, and postoperative chemotherapy in the HAS group were higher than those in the common gastric cancer group (34.6% (9/26) vs. 13.8% (18/130), 26.9% (7/26) vs. 7.7% (10/130), 73.1% (19/26) vs. 51.5% (67/130), 30.8% (8/26) vs. 11.5% (15/130), 84.6% (22/26) vs. 37.7% (49/130), and 69.2% (18/26) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=5.16, 6.39, 4.06, 4.94, 18.73, and 16.52; all P<0.05). The proportion of male patients in the non-HAS group was lower than that in the common gastric cancer group (58.5%(38/65) vs. 73.1%(95/130)), while the proportions of patients with age < 60 years old, abnormal levels of carbohydrate antigen (CA)15-3, CA19-9, CA242, and CEA in the non-HAS group were higher than those in the common gastric cancer group (40.0% (26/65) vs. 13.8%(18/130), 7.7%(5/65) vs. 0, 23.1%(15/65) vs. 10.0%(13/130), 18.5%(12/65) vs. 7.7%(10/130), and 23.1%(15/65) vs. 7.7%(10/130), respectively), and the differences were all statistically significant ( χ2=4.27, 16.69, 11.25, 6.03, 5.02, and 9.18; all P<0.05). The proportion of patients with maximum diameter of tumor ≥ 5 cm, clinical stage Ⅲ or Ⅳ, lymph node metastasis, extrahepatic metastasis, and postoperative chemotherapy in the non-HAS group were all higher than those in the common gastric cancer group (58.5% (38/65) vs. 42.3% (55/130), 64.6% (42/65) vs. 46.2% (60/130), 83.1% (54/65) vs. 54.6% (71/130), 47.7% (31/65) vs. 15.4% (20/130), and 75.4% (49/65) vs. 27.7% (36/130), respectively), and the differences were all statistically significant ( χ2=4.53, 10.80, 15.26, 23.41, and 40.08; all P<0.05). The incidence of liver metastasis in patients with AFP >100 μg/L was higher than that in patients with AFP ≤100 μg/L (47.6%(10/21) vs. 17.1%(12/70)), and the difference was statistically significant ( χ2=8.18, P=0.004). The results of Kaplan-Meier survival analysis showed that the median survival time of patients in the HAS group, non-HAS group and common gastric cancer group was 13, 28, and 54 months, respectively, and the difference was statistically significant (log-rank test, χ2=20.33, P<0.001). The results of multivariate Cox regression analysis revealed that CA19-9 ( HR=5.803, 95% confidence interval (95% CI): 1.545 to 21.794, P<0.001) and extrahepatic metastasis ( HR=2.747, 95% CI: 1.243 to 6.068, P=0.012) were independent risk factors affecting the prognosis of patients in the non-HAS group. CA15-3 ( HR=84.163, 95% CI: 5.085 to 1 392.920, P=0.002), CA125 ( HR=0.038, 95% CI: 0.006 to 0.257, P=0.001), and the degree of tumor differentiation ( HR=2.284, 95% CI: 1.101 to 36.677, P=0.039) were independent risk factors affecting the prognosis of patients in the HAS group. Conclusions:Compared to common gastric cancer, AFPGC is characterized by advanced clinical stages, with higher propensity for lymph node metastasis, liver metastasis, extrahepatic metastasis and poor prognosis. The higher the AFP level before surgery, the more possibility of liver metastasis after surgery. Serum CA15-3 and CA125 might be tumor markers in predicting the prognosis of HAS patients.

Objective To explore the diagnostic value of mediastinal lymph nodes CT image features in pulmonary sarcoidosis.Methods The imageologic data of the patients with pulmonary sarcoidosis(pulmonary sarcoidosis group)and lung cancer complicating mediastinal lymph node metastasis(lung cancer group)con-firmed by EBUS-TBNA in this hospital from June 2015 to November 2023 were analyzed retrospectively.The chest plain scan CT and enhanced scan CT were performed in all cases,and the imaging characteristics were compared between the two groups.The receiver operating characteristic(ROC)curve was drawn and the area under the curve(AUC)was calculated.The diagnostic value of CT imaging characteristics in pulmonary sar-coidosis was analyzed.Results The proportion of female patients in the lung sarcoidosis group was signifi-cantly higher than that in the lung cancer group,while the age was smaller than that in the lung cancer group,and the differences were statistically significant(P＜0.05).The number of mediastinal lymph nodes,location,short diameter and CT enhancement value in the pulmonary sarcoidosis group had statistical difference(P＜0.05).The ROC curve analysis showed that AUC of lymph node short diameter reciprocal,CT enhancement value and position in detection alone were 0.586,0.785 and 0.505 respectively,and AUC of the three combined reciprocal was the highest(0.789).Conclusion The lymph node short diameter,CT enhancement value and po-sition have certain value in clinical diagnosis of pulmonary sarcoidosis and the 3-indicator combination could increase the diagnostic efficiency of pulmonary sarcoidosis.

Liquiritigenin(LG)is a flavone of pharmacological importance,however,its application potential is severely limited due to its poor water solubility.LG could be disassociated slightly in water to form phenolate anion,therefore,better solubilization effect is expected by inclusion with cationic cyclodextrins(CCDs).In this work,four kinds of CCDs modified with amino groups at the primary face were synthesized,and their solid inclusion complexes with LG were successfully prepared by preparing their saturated solutions.The formation of the solid inclusion complexes was confirmed by scanning electron microscopy(SEM)and powder X-ray diffraction(PXRD),and their supramolecular binding behavior in solution was studied using multiple techniques.A 1∶1 inclusion stoichiometry of inclusion complexation was defined using Job plot by ultraviolet-visible(UV-vis)spectroscopy,and their binding stability constants(Ks)were determined as 2862.77,3494.70,6521.85 and 9599.48 L/mol using UV-vis spectroscopic titration,far more superior to that of nativeβ-CD(Ks=236.79 L/mol).This indicated that the amino side chains on CCDs could actively participate in the inclusion complexation through anion-cation interactions,significantly strengthening the host-guest binding between CCDs and LG.The inclusion modes were further elucidated based on proton and two-dimensional rotating-frame overhauser enhancement spectroscopy(2D-ROESY)nuclear magnetic resonance(NMR)experiments and molecular docking.Water solubility of LG was dramatically promoted up to 4.9 mg/mL,which was 70-fold higher than that of native LG.This study could draw inspiration for the binding and solubilization of phenols such as flavones by design of cationic macrocyclic molecules.

Hepatic stellate cells (HSCs) are the primary fibrogenic cells in the liver, and their activation plays a crucial role in the development and progression of hepatic fibrosis. Here, we report that retinoid X receptor-alpha (RXRα), a unique member of the nuclear receptor superfamily, is a key modulator of HSC activation and liver fibrosis. RXRα exerts its effects by modulating calcium/calmodulin-dependent protein kinase kinase β (CaMKKβ)-mediated activation of AMP-activated protein kinase-alpha (AMPKα). In addition, we demonstrate that K-80003, which binds RXRα by a unique mechanism, effectively suppresses HSC activation, proliferation, and migration, thereby inhibiting liver fibrosis in the CCl₄ and amylin liver NASH (AMLN) diet animal models. The effect is mediated by AMPKα activation, promoting mitophagy in HSCs. Mechanistically, K-80003 activates AMPKα by inducing RXRα to form condensates with CaMKKβ and AMPKα via a two-phase process. The formation of RXRα condensates is driven by its N-terminal intrinsic disorder region and requires phosphorylation by CaMKKβ. Our results reveal a crucial role of RXRα in liver fibrosis regulation through modulating mitochondrial activities in HSCs. Furthermore, they suggest that K-80003 and related RXRα modulators hold promise as therapeutic agents for fibrosis-related diseases.

Neuropathic pain is a chronic condition caused by damage or dysfunction in the nervous system. While the spleen may influence neuropathic pain, its role has been poorly understood. This study demonstrates that the spleen plays a crucial role in regulating neuropathic pain through the bed nucleus of the stria terminalis (BNST) - paraventricular nucleus of the hypothalamus (PVN) neural circuit in a chronic constriction injury (CCI) mouse model. Splenectomy, splenic denervation, or splenic sympathectomy significantly increased the mechanical withdrawal threshold (MWT) and reduced macrophage infiltration in the dorsal root ganglia (DRG) of CCI mice. Pseudorabies virus injections into the spleen revealed connections to the BNST and PVN in the brain. Chemogenetic inhibition of the BNST-PVN circuit increased macrophage infiltration in the DRG and decreased the MWT; these effects were reversed by splenectomy, splenic denervation, or sympathectomy. These findings underscore the critical role of the spleen, regulated by the BNST-PVN circuit, in neuropathic pain.
Animals ; Neuralgia/pathology* ; Septal Nuclei/physiopathology* ; Male ; Spleen/physiopathology* ; Paraventricular Hypothalamic Nucleus/physiopathology* ; Mice, Inbred C57BL ; Splenectomy ; Mice ; Neural Pathways/physiopathology* ; Disease Models, Animal ; Ganglia, Spinal/physiopathology* ; Sympathectomy ; Macrophages

The study aimed to investigate the effect of the CD200R1 gene deletion on microglia activation and nigrostriatal dopamine neuron loss in the Parkinson's disease (PD) process. The CRISPR-Cas9 technology was applied to construct the CD200R1^-/- mice. The primary microglia cells of wild-type and CD200R1^-/- mice were cultured and treated with bacterial lipopolysaccharide (LPS). Microglia phagocytosis level was assessed by a fluorescent microsphere phagocytosis assay. PD mouse model was prepared by nigral stereotaxic injection of recombinant adeno-associated virus vector carrying human α-synuclein (α-syn). The changes in the motor behavior of the mice with both genotypes were evaluated by cylinder test, open field test, and rotarod test. Immunohistochemical staining was used to assess the loss of dopamine neurons in substantia nigra. Immunofluorescence staining was used to detect the expression level of CD68 (a key molecule involved in phagocytosis) in microglia. The results showed that CD200R1 deletion markedly enhanced LPS-induced phagocytosis in vitro by the microglial cells. In the mouse model of PD, CD200R1 deletion exacerbated motor behavior impairment and dopamine neuron loss in substantia nigra. Fluorescence intensity analysis results revealed a significant increase in CD68 expression in microglia located in the substantia nigra of CD200R1^-/- mice. The above results suggest that CD200R1 deletion may further activates microglia by promoting microglial phagocytosis, leading to increased loss of the nigrostriatal dopamine neurons in the PD model mice. Therefore, targeting CD200R1 could potentially serve as a novel therapeutic target for the treatment of early-stage PD.
Animals ; Microglia/physiology* ; Mice ; Phagocytosis ; Parkinson Disease/genetics* ; Disease Models, Animal ; Receptors, Cell Surface/physiology* ; Dopaminergic Neurons/pathology* ; Antigens, CD/metabolism* ; Gene Deletion ; Substantia Nigra ; Mice, Inbred C57BL ; Mice, Knockout ; Cells, Cultured ; Male ; alpha-Synuclein ; CD68 Molecule ; Orexin Receptors

Resveratrol(Res) is a kind of polyphenolic compound, possessing multiple biological activities such as antioxidant, anti-inflammatory, cardioprotective, and anticancer effects. In recent years, the cardiovascular protective mechanism of Res has become a research hotspot. Studies have shown that Res has a protective effect on the cardiovascular system through various pathways, such as inhibiting oxidative stress, regulating ferroptosis of cells, improving ischemia-reperfusion(I/R) injury, regulating lipid metabolism, suppressing inflammatory responses, and enhancing endothelial function. It can also alleviate cardiotoxicity caused by drugs and chemicals. In terms of oxidative stress, Res reduces the level of intracellular reactive oxygen species(ROS) by enhancing the expression of proteins such as silent information regulator 1(SIRT1) and regulating mitochondrial function, thereby alleviating myocardial cell damage. Regarding ferroptosis, Res inhibits the occurrence of ferroptosis by regulating the expression of proteins related to iron metabolism. Res can also improve I/R injury through mechanisms such as activating autophagy and the mitochondrial quality control network. In regard to improving endothelial function, Res protects the function of endothelial cells by regulating multiple signaling pathways, such as downregulating the PREP1-mediated pathway. Res can also regulate lipid metabolism and inhibit the progression of atherosclerosis. In terms of inflammatory responses, Res exerts anti-inflammatory effects through mechanisms such as inhibiting the nuclear factor-kappa B(NF-κB) signaling pathway. In addition, Res has an improving effect on cardiotoxicity caused by different drugs or environmental factors. However, the clinical application of Res still faces limitations such as poor pharmacokinetic properties. In the future, in-depth exploration is needed at multiple levels from basic research to clinical application to clarify the dose-response relationship and standardize the standards of medication regimens with the expectation of providing more effective strategies for the prevention and treatment of cardiovascular diseases.
Humans ; Resveratrol/pharmacology* ; Animals ; Cardiotonic Agents/pharmacology* ; Oxidative Stress/drug effects* ; Cardiovascular Diseases/genetics* ; Cardiovascular System/metabolism* ; Signal Transduction/drug effects*