In modern society, people are increasingly exposed to chronic stress, leading to various mental disorders. However, the activities of brain regions, especially neural firing patterns related to specific behaviors, remain unclear. In this study, we introduce a novel approach, NeuroSync, which integrates open-field behavioral testing with electrophysiological recordings from emotion-related brain regions, specifically the central amygdala and the paraventricular nucleus of the hypothalamus, to explore the mechanisms of negative emotions induced by chronic stress in mice. By applying machine vision techniques, we quantified behaviors in the open field, and signal processing algorithms elucidated the neural underpinnings of the observed behaviors. Synchronizing behavioral and electrophysiological data revealed significant correlations between neural firing patterns and stress-related behaviors, providing insights into real-time brain activity underlying behavioral responses. This research combines deep learning and machine learning to synchronize high-resolution video and electrophysiological data, offering new insights into neural-behavioral dynamics under chronic stress conditions.
Animals ; Mice ; Male ; Emotions/physiology* ; Stress, Psychological/physiopathology* ; Action Potentials/physiology* ; Mice, Inbred C57BL ; Behavior, Animal/physiology* ; Machine Learning ; Amygdala/physiopathology* ; Neurons/physiology* ; Paraventricular Hypothalamic Nucleus/physiopathology* ; Brain/physiology*

Circular RNA(circRNA),as a kind of non-coding RNA,regulates a variety of biological functions.To explore the effect of circRNA on PEDV replication in the host porcine intestinal epi-thelial cells,this study screened and analyzed the differentially expressed circRNAs by bioinforma-tic software in African Green Monkey renal cells(Vero-E6 cells)infected by porcine epidemic di-arrhea virus(PEDV),the differentially expressed circRNA ssc_circ_PIK3C2A was identified and the secondary structure was analyzed.PCR was used to identify the ssc_circ_PIK3C2A circRNA structure,the model of PEDV-infected IPEC-J2 cells was constructed,the TCID50 test was used to validate the viral titer of PEDV.The expression of circ_PIK3C2A was detected by qRT-PCR in IPEC-J2 infected by PEDV.circ_PIK3C2A qRT-PCR was performed to detect the expression of N gene of PEDV when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.The results showed that ssc_circ_PIK3C2 A is a porcine circular RNA with a typical circular structure,the virus titer of PEDV reached 10-6/mL after PEDV infected IPEC-J2 cells for 48 h,the expression of circ_PIK3C2A increased extremely(P＜0.01)at 6 h after PEDV-infection,with the extension of infec-tion time,its expression gradually decreased,and the expression was the lowest at 24 h,but there was no time-dependent trend.The expression of PEDV N gene decreased significantly when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.In conclusion,when PEDV infects IPEC-J2 cells,the expression of porcine circ_PIK3C2A decreases,and replication of PEDV increases signifi-cantly in IPEC-J2 cells.our result provides a basis for further study of the mechanism of circular RNA on PEDV replication and its physiological activities in host cells in the future.

OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.

BACKGROUND:Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries.NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development.However,the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.OBJECTIVE:To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.METHODS:Induced pluripotent stem cells were cultured in vitro.The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR.NKX2.1 was overexpressed in induced pluripotent stem cells,which were then induced to differentiate into lung stem cells.The expression of FoxA2,SOX9,and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation.The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.RESULTS AND CONCLUSION:(1)Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4,SOX2,and NANOG.(2)Compared with the non-transfected control group,the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(3)Seven days after induction of differentiation,compared with the non-transfected control group,the expression of lung stem cell-related markers FoxA2,SOX9,and P63 was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(4)Thirteen days after induction of differentiation,compared with the non-transfected control group,the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group.The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.

Transmissible gastroenteritis(TGE)is a highly contagious gastrointestinal disease of pigs caused by the transmissible gastroenteritis virus(TGEV).It results in severe diarrhea and high mortality in suckling piglets.As no specific treatment available,vaccination is considered to be one of the most cost-effective measures for preventing and controlling TGE.However,the immune protection provided by vaccines based on traditional TGEV strains is becoming inadequate due to the continuous emergence of strong and new strains of the virus,which poses a serious threat to the pig industry's health and development.Therefore,the development of new vaccines with im-proved efficiency and broad-spectrum coverage is urgently needed.This paper aims to summarize the pathogenic structural characteristics of TGEV,discuss the latest advancements in TGEV vac-cine research and development,and propose future strategies for the development of highly effec-tive TGEV vaccines.The findings of this paper can serve as a valuable reference for effectively pre-venting and controlling TGE in clinical practice.

Objective:To study and establish the quality standard of Polyoxyl 15 hydroxystearate(HS15),a phar-maceutical excipient,and systematically evaluate its functionality-related characteristics and safety.HS15 was applied to the preparation of docetaxel(DTX)injection to further investigate the safety and pharmacokinetic char-acteristics of the injection in vitro and in vivo.Methods:Based on the general USP-NF2024,EP11.0 and the fourth general rules of the Chinese Pharmacopoeia 2020 edition,the quality standards of HS15 were studied.Combined with the functional properties of surfactants,the critical micelle concentration of HS15 was investigated,and its safety was investigated by hemolysis test and vascular irritation test in vitro.HS15 was further applied to the preparation of DTX injection,and the safety and efficacy of the preparation were comprehensively evaluated by in vitro cytotoxicity test and in vivo pharmacokinetic study.Results:According to the experimental results and the pharmacopoeia of various countries,the quality standard of HS15 was preliminarily formulated.When the concen-tration of HS15 was 1 mg·mL-1,the hemolysis rate was about0.2%,the vascular irritation was small,and the DTX injection was safe in vitro and in vivo.The pharmacokinetic behavior was in line with expectations.Conclusion:This study successfully established the quality standard of HS15,and its functional correlation index research and safety evaluation strategy can provide reference for the quality control of similar excipients.The appli-cation of HS15 in the preparation of DTX injection provides a theoretical and experimental basis for its application in the development of insoluble antitumor drug injection.

Objective:To study and establish the quality standard of Polyoxyl 15 hydroxystearate(HS15),a phar-maceutical excipient,and systematically evaluate its functionality-related characteristics and safety.HS15 was applied to the preparation of docetaxel(DTX)injection to further investigate the safety and pharmacokinetic char-acteristics of the injection in vitro and in vivo.Methods:Based on the general USP-NF2024,EP11.0 and the fourth general rules of the Chinese Pharmacopoeia 2020 edition,the quality standards of HS15 were studied.Combined with the functional properties of surfactants,the critical micelle concentration of HS15 was investigated,and its safety was investigated by hemolysis test and vascular irritation test in vitro.HS15 was further applied to the preparation of DTX injection,and the safety and efficacy of the preparation were comprehensively evaluated by in vitro cytotoxicity test and in vivo pharmacokinetic study.Results:According to the experimental results and the pharmacopoeia of various countries,the quality standard of HS15 was preliminarily formulated.When the concen-tration of HS15 was 1 mg·mL-1,the hemolysis rate was about0.2%,the vascular irritation was small,and the DTX injection was safe in vitro and in vivo.The pharmacokinetic behavior was in line with expectations.Conclusion:This study successfully established the quality standard of HS15,and its functional correlation index research and safety evaluation strategy can provide reference for the quality control of similar excipients.The appli-cation of HS15 in the preparation of DTX injection provides a theoretical and experimental basis for its application in the development of insoluble antitumor drug injection.

OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.

BACKGROUND:Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries.NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development.However,the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.OBJECTIVE:To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.METHODS:Induced pluripotent stem cells were cultured in vitro.The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR.NKX2.1 was overexpressed in induced pluripotent stem cells,which were then induced to differentiate into lung stem cells.The expression of FoxA2,SOX9,and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation.The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.RESULTS AND CONCLUSION:(1)Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4,SOX2,and NANOG.(2)Compared with the non-transfected control group,the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(3)Seven days after induction of differentiation,compared with the non-transfected control group,the expression of lung stem cell-related markers FoxA2,SOX9,and P63 was significantly increased in the NKX2.1 overexpression group(P＜0.000 1).(4)Thirteen days after induction of differentiation,compared with the non-transfected control group,the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group.The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.

Circular RNA(circRNA),as a kind of non-coding RNA,regulates a variety of biological functions.To explore the effect of circRNA on PEDV replication in the host porcine intestinal epi-thelial cells,this study screened and analyzed the differentially expressed circRNAs by bioinforma-tic software in African Green Monkey renal cells(Vero-E6 cells)infected by porcine epidemic di-arrhea virus(PEDV),the differentially expressed circRNA ssc_circ_PIK3C2A was identified and the secondary structure was analyzed.PCR was used to identify the ssc_circ_PIK3C2A circRNA structure,the model of PEDV-infected IPEC-J2 cells was constructed,the TCID50 test was used to validate the viral titer of PEDV.The expression of circ_PIK3C2A was detected by qRT-PCR in IPEC-J2 infected by PEDV.circ_PIK3C2A qRT-PCR was performed to detect the expression of N gene of PEDV when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.The results showed that ssc_circ_PIK3C2 A is a porcine circular RNA with a typical circular structure,the virus titer of PEDV reached 10-6/mL after PEDV infected IPEC-J2 cells for 48 h,the expression of circ_PIK3C2A increased extremely(P＜0.01)at 6 h after PEDV-infection,with the extension of infec-tion time,its expression gradually decreased,and the expression was the lowest at 24 h,but there was no time-dependent trend.The expression of PEDV N gene decreased significantly when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.In conclusion,when PEDV infects IPEC-J2 cells,the expression of porcine circ_PIK3C2A decreases,and replication of PEDV increases signifi-cantly in IPEC-J2 cells.our result provides a basis for further study of the mechanism of circular RNA on PEDV replication and its physiological activities in host cells in the future.