Efferocytosis refers to the process of phagocytes engulfing and clearing the cells after programmed cell death. In recent years, an increasing number of studies have shown that the mechanisms of efferocytosis are closely related to drug-induced liver injury, hepatic ischemia-reperfusion injury, viral hepatitis, cholestatic liver diseases, metabolic-associated fatty liver disease, alcoholic liver disease, and other liver disorders. This review summarized the research progress on the role of efferocytosis in liver diseases, with the hope of providing new targets for the prevention and treatment of liver diseases.
Humans ; Liver Diseases/metabolism* ; Animals ; Phagocytosis/physiology* ; Phagocytes ; Efferocytosis

Objective:To analyze pancreatic cancer incidence and mortality data in China and worldwide and to provide data for pancreatic cancer prevention and control efforts.Methods:Data of pancreatic cancer incidence and mortality rates, along with historical and predictive data, were obtained from the GLOBOCAN 2022 database. Epidemiological characteristics of pancreatic cancer was analyzed by region, sex, age and Human Development Index (HDI). Spearman's correlation coefficient test was used to assess the relationship between HDI and age-standardized incidence rate (ASIR) and age-standardized mortality rate (ASMR).Results:In 2022, the global number of new cases and deaths of pancreatic cancer will be 511 thousand and 467 thousand, respectively, with an ASIR and ASMR of 4.7/10 5 and 4.2/10 5, respectively. North America and Europe had the highest pancreatic cancer incidence and mortality rates of 8.5/10 5 and 7.3/10 5, respectively. Global ASIR and ASMR in men were both 1.4 times higher than those in women. HDI levels were positively correlated with ASIR ( r=0.79, P＜0.001) and ASMR ( r=0.78, P＜0.001) of pancreatic cancer in all regions. The number of pancreatic cancer cases and deaths in China were 119 thousand and 106 thousand, respectively, while the ASIR and ASMR of pancreatic cancer were 4.4/10 5 and 3.9/10 5, respectively. Both ASIR and ASMR in men were both 1.5 times higher than those in women in China. The number of pancreatic cancer incidence and death cases in China in 2050 is predicted to be 216 thousand and 204 thousand cases, with an increase of 81.5% and 92.5% compared with 2022, respectively. Conclusions:The disease burden of pancreatic cancer varies significantly among different regions, genders and ages. Pancreatic cancer incidence and mortality are positively correlated with HDI. The incidence and mortality rates of pancreatic cancer in China are close to the global average, but the number of new cases and deaths is high. Prevention and control should be strengthened to improve the survival of pancreatic cancer patients.

Objective:To explore the clinical features and genetic mutation sites of 28 patients with Congenital fibrinogen disorders (CFDs).Methods:A total of 28 unrelated CFDs patients admitted to Wenzhou People′s Hospital from June 2018 to April 2023 were enrolled into this research. A total of 2.7 mL of peripheral blood was collected from each patient for coagulation function tests, which included thrombin time (TT), fibrinogen activity (Fg: C), fibrinogen antigen (Fg: Ag), and gene detection. The Sanger sequencing method was employed to verify variations in the fibrinogen (Fg) protein-coding gene across 28 patients. Bioinformatics analyses, including harmfulness analysis, conservation analysis across different species, and spatial simulation predictions of variant proteins, were conducted by PolyPhen-2, PROVEAN, SnapGene, and Pymol softwares on the variant sites of these patients. Pathogenicity ratings for the detected variant sites were performed in accordance with the Standards and Guidelines for the Interpretation of Sequence variants by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG Guidelines). This study received approval from the Ethics Committee of Wenzhou People′s Hospital (Approval No. KY-2023-269), and informed consent was obtained from all participants before enrollment.Results:The clinical and genetic characteristics of 28 patients with CFDs in this study were as follows. ①Clinical data: Among the 28 patients, 2 cases were diagnosed with type I CFDs, while 26 cases were diagnosed with type II CFDs. And 50.0% (14/28) of the patients exhibited no clinical manifestations, while 28.6% (8/28) presented with bleeding manifestations, and 7.1% (2/28) exhibited thrombus manifestations, 3.6% (1/28) experienced both bleeding and thrombosis. Among female patients, 13.0% (3/23) exhibited a history of habitual abortion. All patients demonstrated TT and a significant decrease in Fg: C. ②Sanger sequencing revealed a total of 10 types of heterozygous variations in the FGA, FGB, and FGG genes across 28 patients, distributed among 9 loci. The variation at the γ c. 902G>A/c.901C>T accounted for the highest proportion (35.7%, 10/28), followed by the Bβ c. 569 A>G (28.6%, 8/28). ③Biological informatics analysis: the Aα c. 180+ 1G>T mutation was predicted to be highly deleterious. And the Aα c. 104G>A, Bβ c. 425T>G, Bβ c. 586C>T, and γ c. 902G>A/c.901C>T variations were also predicted to be harmful. Conservation analysis indicates that the 9 variant sites were highly conserved among homo sapiens, musculus, ovis aries, scrofa, and rattus. Spatial conformation analysis revealed that some variations lead to an increase or decrease in the number of hydrogen bonds. ④ACMG guideline rating analysis: Among the ten variations in the Fg protein-coding genes FGA, FGB, and FGG identified in 28 patients, 9 variations (Aα c. 104G>A, Aα c. 180+ 1G>T, Bβ c. 425T>G, Bβ c.569A>G, Bβ c. 586C>T, Bβ c. 643G>A, γ c. 901C>T, γ c. 902G>A, γ c. 1001A>C) were classified as pathogenic, while one variation (γ c. 908C>G) was classified as likely pathogenic. Conclusion:In this study, the majority of CFDs patients are diagnosed with type II CFDs, with 50% presenting clinical symptoms predominantly manifesting as bleeding, thrombosis, and recurrent miscarriage. The mutation hotspots are mainly located in exon 2 of FGA, exon 4 of FGB, and exon 8 of FGG.

BACKGROUND:We have successfully isolated and cultured neonatal mouse cerebrospinal fluid-contacting neurons in vitro,but there is no study that reports an effective method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons.There is no study on whether the self-renewal ability of cerebrospinal fluid-contacting neurons changes with age.OBJECTIVE:To establish a method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons in vitro,and to characterize the self-renewal ability of adult mouse cerebrospinal fluid-contacting neurons and neonatal mouse cerebrospinal fluid-contacting neurons in vitro.METHODS:Primary cells containing cerebrospinal fluid-contacting neurons were isolated from the cervical medulla of adult mouse (3 months of age) in adherent culture and transfected with lentivirus fused with multimodal imaging genes.High-purity adult mouse cerebrospinal fluid-contacting neurons were obtained by puromycin screening in suspension culture in complete medium.The expression of neural stem cell markers Nestin and SOX2 was detected by immunofluorescence in adult mouse cerebrospinal fluid-contacting neurons,and the ability of adult mouse cerebrospinal fluid-contacting neurons to form spheres and pass on in vitro was observed.An equal number (5×103/mL) of passage 3 adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons were divided into two groups under the same conditions and inoculated into ultra-low adhesion plates containing complete medium in suspension culture at 5％ CO2,37℃ thermostat,respectively.The self-renewal capacity of adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons was characterized by in vitro spheroid formation,CCK8 assay,qPCR,and western blot assay.RESULTS AND CONCLUSION:(1) High-purity cerebrospinal fluid-contacting neurons were successfully isolated from adult mouse,which expressed Nestin and SOX2 in vitro,and were able to form neurospheres and pass on continuously.(2) The in vitro self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse,and the neurospheres formed by day 4 of cell culture in neonatal mouse were about 150 μm in diameter,whereas the neurospheres formed by adult mouse tactile neurons were only 40 μm in diameter (P<0.0001).(3) CCK8 proliferation assay showed that the proliferative activity of adult mouse cerebrospinal fluid-contacting neurons was significantly weaker than that of neonatal mouse at all time points after culture (P<0.0001).(4) qPCR and western blot assay revealed that the mRNA (P<0.0001) and protein expression levels (P<0.01) of Nestin and SOX2 in cerebrospinal fluid-contacting neurons of adult mouse were significantly decreased compared with those of neonatal mouse.(5) The above results indicated that the self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse in vitro.

OBJECTIVE: To explore the clinical features and genetic mutation sites of 28 patients with Congenital fibrinogen disorders (CFDs). METHODS: A total of 28 unrelated CFDs patients admitted to Wenzhou People's Hospital from June 2018 to April 2023 were enrolled into this research. A total of 2.7 mL of peripheral blood was collected from each patient for coagulation function tests, which included thrombin time (TT), fibrinogen activity (Fg:C), fibrinogen antigen (Fg:Ag), and gene detection. The Sanger sequencing method was employed to verify variations in the fibrinogen (Fg) protein-coding gene across 28 patients. Bioinformatics analyses, including harmfulness analysis, conservation analysis across different species, and spatial simulation predictions of variant proteins, were conducted byPolyPhen-2, PROVEAN, SnapGene, and Pymol softwares on the variant sites of these patients. Pathogenicity ratings for the detected variant sites were performed in accordance with the Standards and Guidelines for the Interpretation of Sequence variants by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG Guidelines). This study received approval from the Ethics Committee of Wenzhou People's Hospital (Approval No. KY-2023-269), and informed consent was obtained from all participants before enrollment. RESULTS: The clinical and genetic characteristics of 28 patients with CFDs in this study were as follows. CLINICAL DATA: Among the 28 patients, 2 cases were diagnosed with type I CFDs, while 26 cases were diagnosed with type II CFDs. And 50.0% (14/28) of the patients exhibited no clinical manifestations, while 28.6% (8/28) presented with bleeding manifestations, and 7.1% (2/28) exhibited thrombus manifestations, 3.6% (1/28) experienced both bleeding and thrombosis. Among female patients, 13.0% (3/23) exhibited a history of habitual abortion. All patients demonstrated TT and a significant decrease in Fg:C. Sanger sequencing revealed a total of 10 types of heterozygous variations in the FGA, FGB, and FGG genes across 28 patients, distributed among 9 loci. The variation at the γ c.902G>A/c.901C>T accounted for the highest proportion (35.7%, 10/28), followed by the Bβ c.569 A>G (28.6%, 8/28). Biological informatics analysis: the Aα c.180+1G>T mutation was predicted to be highly deleterious. And the Aα c.104G>A, Bβ c.425T>G, Bβ c.586C>T, and γ c.902G>A/c.901C>T variations were also predicted to be harmful. Conservation analysis indicates that the 9 variant sites were highly conserved among homo sapiens, musculus, ovis aries, scrofa, and rattus. Spatial conformation analysis revealed that some variations lead to an increase or decrease in the number of hydrogen bonds. ACMG guideline rating analysis: Among the ten variations in the Fg protein-coding genes FGA, FGB, and FGG identified in 28 patients, 9 variations (Aα c.104G>A, Aα c.180+1G>T, Bβ c.425T>G, Bβ c.569A>G, Bβ c.586C>T, Bβ c.643G>A, γ c.901C>T, γ c.902G>A, γ c.1001A>C) were classified as pathogenic, while one variation (γ c.908C>G) was classified as likely pathogenic. CONCLUSION In this study, the majority of CFDs patients are diagnosed with type II CFDs, with 50% presenting clinical symptoms predominantly manifesting as bleeding, thrombosis, and recurrent miscarriage. The mutation hotspots are mainly located in exon 2 of FGA, exon 4 of FGB, and exon 8 of FGG.
Humans ; Female ; Male ; Afibrinogenemia/congenital* ; Fibrinogen/metabolism* ; Mutation ; Genotype ; Adult ; Child ; Adolescent ; Child, Preschool ; Infant

Objective To investigate the effect of ring finger protein 130(RNF130)on myocardial ischemia-reperfusion injury(MI/RI)and its potential mechanism.Methods Male C57BL/6J mice were divided into four groups(n=6):Sham,MI/RI,MI/RI+Vector,and MI/RI+RNF130 overexpression(MI/RI+RNF130OE).Cardiac function was evaluated by echocardiography 24 hours after ischemia-reperfusion.Pathological changes,oxidative damage,and apoptosis in myocardial tissues were observed via IHC,DHE,and TUNEL staining.Protein expression was detected using Western blot,immunofluorescence,and immunohistochemistry.Proteomic analysis was performed to identify downstream proteins regulated by RNF130,and protein-protein interactions were validated by immunoprecipitation(IP)assay.Results Compared with the MI/RI+Vector group,RNF130 overexpression significantly improved cardiac function,as indicated by increased left ventricular ejection fraction(EF)and fractional shortening(FS),reduced myocardial infarction area,and decreased expression of NOX-2 and BAX proteins(P＜0.05).DHE and TUNEL staining showed that RNF130 overexpression alleviated myocardial oxidative damage and apoptosis(P＜0.05).Proteomic analysis and IP assays revealed a significant interaction between RNF130 and PARP1,with PARP1 expression inversely correlated with RNF130.Conclusions RNF130 may mitigate MI/RI injury by regulating the PARP1 ubiquitination pathway,providing a new target for therapeutic intervention.

Coronavirus is the largest RNA virus with known genome so far,among which porcine coronavirus is an important pathogen causing acute diarrhea in piglets.Due to its highly mutable RNA nature,this virus poses a substantial challenge for disease prevention and control,leading to substantial annual economic losses within China's swine industry.The analysis of the mechanism of RNA synthesis of coronaviruses is helpful for understanding the genetic variation of coronaviruses and screening antiviral drugs.At present,more in-depth studies mainly focus on Murine coronavir-us(MHV)and human Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).In order to investigate the replication mechanism of porcine coronaviruses,this article takes MHV,SARS-CoV-2 and other coronaviruses as references to summarises the mechanism of regulating viral rep-lication in the non-coding region of porcine coronaviruses such as Porcine epidemic diarrhea virus(PEDV),Transmissible gastroenteritis virus(TGEV),Porcine deltacoronavirus(PDCoV),and Swine acute diarrhea syndrome coronavirus(SADS-CoV).Placed a specific emphasis on the func-tional roles of conserved motifs and secondary structures within the 5'UTR in facilitating viral rep-lication,elucidated how conserved structures in the 3'UTR regulate the same process.The signifi-cance of the interaction between the non-coding region of the virus and the host were discussed,and variations in the viral 5'UTR and 3'UTR were scrutinized,thereby establishing a solid theoret-ical foundation for the development of antiviral drugs that target UTRs and high titer vaccine can-didate strains.

Objective To investigate the effect of ring finger protein 130(RNF130)on myocardial ischemia-reperfusion injury(MI/RI)and its potential mechanism.Methods Male C57BL/6J mice were divided into four groups(n=6):Sham,MI/RI,MI/RI+Vector,and MI/RI+RNF130 overexpression(MI/RI+RNF130OE).Cardiac function was evaluated by echocardiography 24 hours after ischemia-reperfusion.Pathological changes,oxidative damage,and apoptosis in myocardial tissues were observed via IHC,DHE,and TUNEL staining.Protein expression was detected using Western blot,immunofluorescence,and immunohistochemistry.Proteomic analysis was performed to identify downstream proteins regulated by RNF130,and protein-protein interactions were validated by immunoprecipitation(IP)assay.Results Compared with the MI/RI+Vector group,RNF130 overexpression significantly improved cardiac function,as indicated by increased left ventricular ejection fraction(EF)and fractional shortening(FS),reduced myocardial infarction area,and decreased expression of NOX-2 and BAX proteins(P＜0.05).DHE and TUNEL staining showed that RNF130 overexpression alleviated myocardial oxidative damage and apoptosis(P＜0.05).Proteomic analysis and IP assays revealed a significant interaction between RNF130 and PARP1,with PARP1 expression inversely correlated with RNF130.Conclusions RNF130 may mitigate MI/RI injury by regulating the PARP1 ubiquitination pathway,providing a new target for therapeutic intervention.

BACKGROUND:We have successfully isolated and cultured neonatal mouse cerebrospinal fluid-contacting neurons in vitro,but there is no study that reports an effective method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons.There is no study on whether the self-renewal ability of cerebrospinal fluid-contacting neurons changes with age.OBJECTIVE:To establish a method for isolating and culturing high-purity adult mouse cerebrospinal fluid-contacting neurons in vitro,and to characterize the self-renewal ability of adult mouse cerebrospinal fluid-contacting neurons and neonatal mouse cerebrospinal fluid-contacting neurons in vitro.METHODS:Primary cells containing cerebrospinal fluid-contacting neurons were isolated from the cervical medulla of adult mouse (3 months of age) in adherent culture and transfected with lentivirus fused with multimodal imaging genes.High-purity adult mouse cerebrospinal fluid-contacting neurons were obtained by puromycin screening in suspension culture in complete medium.The expression of neural stem cell markers Nestin and SOX2 was detected by immunofluorescence in adult mouse cerebrospinal fluid-contacting neurons,and the ability of adult mouse cerebrospinal fluid-contacting neurons to form spheres and pass on in vitro was observed.An equal number (5×103/mL) of passage 3 adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons were divided into two groups under the same conditions and inoculated into ultra-low adhesion plates containing complete medium in suspension culture at 5％ CO2,37℃ thermostat,respectively.The self-renewal capacity of adult mouse and neonatal mouse cerebrospinal fluid-contacting neurons was characterized by in vitro spheroid formation,CCK8 assay,qPCR,and western blot assay.RESULTS AND CONCLUSION:(1) High-purity cerebrospinal fluid-contacting neurons were successfully isolated from adult mouse,which expressed Nestin and SOX2 in vitro,and were able to form neurospheres and pass on continuously.(2) The in vitro self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse,and the neurospheres formed by day 4 of cell culture in neonatal mouse were about 150 μm in diameter,whereas the neurospheres formed by adult mouse tactile neurons were only 40 μm in diameter (P<0.0001).(3) CCK8 proliferation assay showed that the proliferative activity of adult mouse cerebrospinal fluid-contacting neurons was significantly weaker than that of neonatal mouse at all time points after culture (P<0.0001).(4) qPCR and western blot assay revealed that the mRNA (P<0.0001) and protein expression levels (P<0.01) of Nestin and SOX2 in cerebrospinal fluid-contacting neurons of adult mouse were significantly decreased compared with those of neonatal mouse.(5) The above results indicated that the self-renewal ability of cerebrospinal fluid-contacting neurons in adult mouse was significantly weaker than that of neonatal mouse in vitro.

Coronavirus is the largest RNA virus with known genome so far,among which porcine coronavirus is an important pathogen causing acute diarrhea in piglets.Due to its highly mutable RNA nature,this virus poses a substantial challenge for disease prevention and control,leading to substantial annual economic losses within China's swine industry.The analysis of the mechanism of RNA synthesis of coronaviruses is helpful for understanding the genetic variation of coronaviruses and screening antiviral drugs.At present,more in-depth studies mainly focus on Murine coronavir-us(MHV)and human Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).In order to investigate the replication mechanism of porcine coronaviruses,this article takes MHV,SARS-CoV-2 and other coronaviruses as references to summarises the mechanism of regulating viral rep-lication in the non-coding region of porcine coronaviruses such as Porcine epidemic diarrhea virus(PEDV),Transmissible gastroenteritis virus(TGEV),Porcine deltacoronavirus(PDCoV),and Swine acute diarrhea syndrome coronavirus(SADS-CoV).Placed a specific emphasis on the func-tional roles of conserved motifs and secondary structures within the 5'UTR in facilitating viral rep-lication,elucidated how conserved structures in the 3'UTR regulate the same process.The signifi-cance of the interaction between the non-coding region of the virus and the host were discussed,and variations in the viral 5'UTR and 3'UTR were scrutinized,thereby establishing a solid theoret-ical foundation for the development of antiviral drugs that target UTRs and high titer vaccine can-didate strains.