Objective To investigate the expression of exosomal α-synuclein(α-syn),serum lipocalcitin-2(LCN-2)levels and the relationship with disease severity in Parkinson's disease(PD)patients.Methods 106 PD patients admitted to Yulin Hospital,the First Affiliated Hospital of Xi'an Jiaotong University,from June 2020 to June 2023 were selected.The PD patients were divided into the early group(n=31),the middle group(n=45)and the late group(n=30)according to their conditions,and 92 healthy volunteers who had outpatient medical checkups during the same period were selected as the control group.The exosome α-syn and serum LCN-2 levels were detected,and the differences in exosome α-syn and serum LCN-2 levels were compared in PD patients with different disease severity levels.Factors affecting the prevalence of PD and the value of exosomal α-syn and serum LCN-2 in diagnosing PD were analyzed.Results The levels of exosomal α-syn(3 086.23±359.46 pg/ml)and serum LCN-2(4.92±1.32ng/ml)in the PD group were higher than those in the control group(1 993.24±244.35 pg/ml,1.96±0.34 ng/ml),and the differences were statistically significant(t=24.636,20.909,all P<0.05).The levels of exosomal α-syn(3 312.72±53.46pg/ml)and serum LCN-2(5.76±0.28ng/ml)in the late-stage group were higher than those in the early-stage(2 843.65±65.29pg/ml,4.02±0.32ng/ml)and the middle-stage group(3 102.35±105.48pg/ml,4.98±0.41ng/ml),and the differences were statistically significant(t=9.092～30.644,all P<0.05).Family history of PD,high level of exosomal α-syn and high level of LCN-2 were the risk factors for the development of PD(Wald χ2=8.121,7.002,4.805,all P<0.05).The AUC(95%CI)of diagnosing PD with serum LCN-2 in combination with exosome α-syn was higher than that of exosome α-syn,while serum LCN-2 alone was used for diagnosis(Z=4.869,5.103,all P<0.05).Conclusion Serum exosomal α-syn and serum LCN-2 levels are significantly higher in PD patients,which can be associated with the exacerbation of PD,and combined exosomal α-syn and serum LCN-2 can assess the risk of developing PD.

Objective To investigate the expression of exosomal α-synuclein(α-syn),serum lipocalcitin-2(LCN-2)levels and the relationship with disease severity in Parkinson's disease(PD)patients.Methods 106 PD patients admitted to Yulin Hospital,the First Affiliated Hospital of Xi'an Jiaotong University,from June 2020 to June 2023 were selected.The PD patients were divided into the early group(n=31),the middle group(n=45)and the late group(n=30)according to their conditions,and 92 healthy volunteers who had outpatient medical checkups during the same period were selected as the control group.The exosome α-syn and serum LCN-2 levels were detected,and the differences in exosome α-syn and serum LCN-2 levels were compared in PD patients with different disease severity levels.Factors affecting the prevalence of PD and the value of exosomal α-syn and serum LCN-2 in diagnosing PD were analyzed.Results The levels of exosomal α-syn(3 086.23±359.46 pg/ml)and serum LCN-2(4.92±1.32ng/ml)in the PD group were higher than those in the control group(1 993.24±244.35 pg/ml,1.96±0.34 ng/ml),and the differences were statistically significant(t=24.636,20.909,all P<0.05).The levels of exosomal α-syn(3 312.72±53.46pg/ml)and serum LCN-2(5.76±0.28ng/ml)in the late-stage group were higher than those in the early-stage(2 843.65±65.29pg/ml,4.02±0.32ng/ml)and the middle-stage group(3 102.35±105.48pg/ml,4.98±0.41ng/ml),and the differences were statistically significant(t=9.092～30.644,all P<0.05).Family history of PD,high level of exosomal α-syn and high level of LCN-2 were the risk factors for the development of PD(Wald χ2=8.121,7.002,4.805,all P<0.05).The AUC(95%CI)of diagnosing PD with serum LCN-2 in combination with exosome α-syn was higher than that of exosome α-syn,while serum LCN-2 alone was used for diagnosis(Z=4.869,5.103,all P<0.05).Conclusion Serum exosomal α-syn and serum LCN-2 levels are significantly higher in PD patients,which can be associated with the exacerbation of PD,and combined exosomal α-syn and serum LCN-2 can assess the risk of developing PD.

The stimulator of interferon genes(STING),an integral adaptor protein in the DNA-sensing pathway,plays a pivotal role in the innate immune response against infections.Additionally,it presents a valuable therapeutic target for infectious diseases and cancer.We observed that fangchinoline(Fan),a bis-benzylisoquinoline alkaloid(BBA),effectively impedes the replication of vesicular stomatitis virus(VSV),encephalomyocarditis virus(EMCV),influenza A virus(H1 N1),and herpes simplex virus-1(HSV-1)in vitro.Fan treatment significantly reduced the viral load,attenuated tissue inflammation,and improved survival in a viral sepsis mouse model.Mechanistically,Fan activates the antiviral response in a STING-dependent manner,leading to increased expression of interferon(1FN)and interferon-stimulated genes(ISGs)for potent antiviral effects in vivo and in vitro.Notably,Fan interacts with STING,preventing its degradation and thereby extending the activation of IFN-based antiviral responses.Collectively,our findings highlight the potential of Fan,which elicits antiviral immunity by suppressing STING degra-dation,as a promising candidate for antiviral therapy.

Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.

Objectives:To discuss the inhibitory effect of Lactobacillus reuteri on the replication of rotavirus(RV)strain SA11 in vivo and in vitro,and to clarify its effect on the expression of related immune factors.Methods:For in vitro experiments,Lactobacillus reuteri was cultured and identified,and the standard curve and growth curve were plotted to screen the optimal time and concentration for Lactobacillus reuteri cultivation.The cells were infected with Lactobacillus reuteri at the concentrations of 5×108,10×108,50×108,100×108,200×108,and 500×108 CFU·mL-1,and the surival rates of Caco-2 cells were detected by trypan blue staining method.Various concentrations of Lactobacillus reuteri were co-incubated with RV in vitro and applied to the Caco-2 cells.The cells were divided into negative control group(NC group),positive control group(PC group),and 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups.Immunofluorescence focus method was used to detect the viral titers in the Caco-2 cells after treated with Lactobacillus reuteri and real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the copy numbers of RV VP6 gene in the Caco-2 cells after treated with various concentrations of Lactobacillus reuteri.In in vivo experiments,25 litters of SPF suckling mice were divided into control group,RV group(infected with SA11 strain),Ab-NC group(treated with antibiotic to deplete gut microbiota),Ab-RV group(depleting gut microbiota and then infected with SA11 strain),and Ab-Lac-RV group(depleting gut microbiota,treated with Lactobacillus reuteri,and then infected with SA11 strain).The fecal samples were collected on days 2,4,6,8,and 10 gavage,colon tissue sample were collected on day 4 of and RT-qPCR method was used to detect the copy numbers of RV VP6 gene in feces and the mRNA expression levels of interleukin(IL)-1β,IL-8,IL-10,interferon-γ(IFN-γ),and tumor necrosis factor-α(TNF-α)in colon tissue of the suckling mice in vartious groups.Results:The Lactobacillus reuteri grew well,with round,smooth,and milky white convex colonies and neat edges.After Gram staining,the bacteria appeared purple,irregular,and square-shaped rods.16SrDNA sequencing showed 99%sequence homology,indicating successful activation of Lactobacillus reuteri.The number of live Lactobacillus reuteri was linearly related to the absorbance(A)value,and the standard curve for regression analysis was Y=0.437 5X+0.000 6,R2=0.999 4.During the 0-2 h cultivation period,the bacteria were at the logarithmic growth phase with slow growth;from 2-14 h,the bacteria grew rapidly and stabilized at 14-16 h,reaching the growth rate peak at 16 h,after which they entered the decline phase.Infection with Lactobacillus reuteri at concentrations of 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 resulted in the survival rates of Caco-2 cells were all>90%,so these concentrations were selected for the further experiments.Compared with PC group,the copy numbers of RV VP6 gene in the Caco-2 cells in 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with PC group,the viral titers in the Caco-2 cells in 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with control group,the numbers of gut microbiota colonies in Ab-NC,Ab-RV,and Ab-Lac-RV groups were significantly decreased,indicating successful depletion of gut microbiota in the suckling mice.On days 2 and 4 after gavage,the RV VP6 gene copy number in the feces in Ab-RV group was significantly lower than that in RV group(P<0.05).On days 4,6,8,and 10 after gavage,the RV VP6 gene copy number in the feces in Ab-Lac-RV group was significantly lower than that in Ab-RV group(P<0.05).Compared with control group,the expression levels of IL-1β,IL-10,IFN-γ,and TNF-α mRNA in colon tissue in Ab-RV and Ab-Lac-RV groups were significantly increased(P<0.05 or P<0.01),while the expression level of IL-8 mRNA was significantly decreased(P<0.05),and the expression level of IL-10 mRNA in colon tissue in Ab-LAC-RV group was significantly increased(P<0.01).Conclusion:Lactobacillus reuteri may inhibit the RV replication by upregulating the expressions of IL-1β,IL-10,IFN-γ,and TNF-α mRNA and downregulating the expression of IL-8 mRNA.

ObjectiveTo investigate the antiviral effect of Menispermi Rhizoma total alkaloids and its relationship with the type Ⅰ interferon （IFN-Ⅰ） signaling pathway. MethodThe effects of Menispermi Rhizoma total alkaloids on the intracellular replication of influenza A virus （H1N1）， vesicular stomatitis virus （VSV）， and cerebral myocarditis virus （EMCV） were detected by fluorescent inverted microscope， flow cytometry， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and Western blot. A mouse model infected with H1N1 was constructed， and the mice were divided into a control group， H1N1 model group， Menispermi Rhizoma total alkaloids groups （10， 20， 30 mg·kg^-1）， and oseltamivir group （40 mg·kg^-1）， so as to study the effects on the weight and survival rate of infected mice. Real-time PCR was used to detect the activation effect of Menispermi Rhizoma total alkaloids on the IFN-Ⅰ pathway in cells， and the relationship between the antiviral effect of Menispermi Rhizoma total alkaloids in IFNAR1 knockout A549 cells （IFNAR1^-/--A549） and IFN-Ⅰ pathway was detected. ResultCompared with the control group， the virus proliferated significantly in the model group （P<0.01）. Compared with the model group， Menispermi Rhizoma total alkaloids could significantly inhibit the replication of H1N1， VSV， and EMCV in vitro （P<0.01）， inhibit the weight loss of the mice infected with the H1N1 in vivo， and improve the survival rate of mice （P<0.05）. In addition， Menispermi Rhizoma total alkaloids activated the IFN-I pathway and relied on this pathway to exert the function of antiviral infection. ConclusionMenispermi Rhizoma total alkaloids exert antiviral effects in vivo and in vitro by activating the IFN-Ⅰ pathway.

ObjectiveTo investigate the situation and development trend of the disease burden of acute hepatitis B in China in 1990 — 2019. MethodsThe Global Burden of Disease 2019 was used to analyze the incidence rate， mortality rate， and disability-adjusted life year （DALY） rate of acute hepatitis B in different sex and age groups and predict the trend of the incidence rate of acute hepatitis B. ResultsIn 2019， the incidence rate， mortality rate， and DALY rate of acute hepatitis B in China were 1 623.71/100 000， 0.20/100 000， and 10.04/100 000 respectively， which were reduced by 42.03%， 79.38%， and 80.21%， respectively， compared with the data in 1990， and women showed lower incidence rate， mortality rate， and DALY rate of acute hepatitis B than men. In 2019， the 20～<54 years group had the highest incidence rate （2 285.85/100 000） and DALY rate （10.53/100 000）， and the ≥55 years group had the highest mortality rate of 0.52/100 000. The Joinpoint regression model analysis showed that the incidence rate， mortality rate， and DALY rate of acute hepatitis B in China tended to decrease from 1990 to 2019， with an average annual percent change of -1.9%， -5.2%， and -5.5%， respectively （P<0.05）. The grey prediction model GM （1，1） showed that the incidence rate of acute hepatitis B will decrease from 2020 to 2030 in China. ConclusionThe disease burden of acute hepatitis B tended to decrease from 1990 to 2019 in China， indicating that the prevention and treatment measures for acute hepatitis B have achieved a marked effect in China； however， due to the large population base of China， active preventive measures should be further adopted to reduce the disease burden of acute hepatitis B.

ObjectiveTo study the possible mechanism of Chaihu Shugan Powder （柴胡疏肝散， CSP） in the treatment of functional dyspepsia （FD）. MethodsTwenty-four SD rats were randomly divided into a normal group， a model group， a CSP group and a probiotic group， with six rats in each group.The tail-clamping provocation method was used in all groups except for the normal group to replicate the FD rat model. Simultaneously， the normal group and the model group were given 10 ml/（kg·d） of saline by gavage， while the CSP group and the probiotic group were given 9.6 g/（kg·d） of CSP aqueous decoction and 0.945 g/（kg·d） of probiotic aqueous solution by gavage， respectively， twice daily for four weeks. After four weeks， the gastric emptying and small intestinal propulsion rates were detected in each group of rats. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the gastric sinusoids and duodenum of the rats. The changes in the intestinal flora were analyzed by 16s rDNA high-throughput gene sequencing， and the expressions of the duodenal zona occludin 1 （ZO-1） and Occludin were detected by immunohistochemistry and western blotting. Pearson correlation analysis was performed on intestinal flora and ZO-1 and Occludin protein expression. ResultsThe gastric antrum tissue structure was clear in all groups， and the gland structure was regular， with smooth gastric tissue mucosa and no pathological changes such as erosion and ulcer. Compared to those in the normal group， the intestinal villi in the duodenal tissue in the model group were significantly reduced or atrophied， and the goblet cells were arranged in disorder， with eosinophilic infiltration； the gastric emptying rate and small intestinal propulsion rate， as well as ZO-1 and Occludin protein expression in duodenal tissue significantly decreased （P＜0.01）. Compared to those in the model group， the duodenal tissue structure was clear， and the length intestinal villi was longer， with goblet cells neatly arranged in the CSP group and the probiotic group； no obvious eosinophil infiltration was found， and the gastric emptying rate and small intestinal propulsion rate as well as ZO-1 and Occludin protein expression significantly increased in the CSP group； a small amount of eosinophil infiltration was found， and the gastric emptying rate and Occludin protein expression significantly increased in the probiotic group （P＜0.05 or P＜0.01）. Beta diversity analysis of intestinal flora showed that the overall structure of intestinal flora in the model group changed significantly compared to that in the normal group （P＜0.01）. The overall structure of the intestinal flora in the CSP group and the probiotic group was closer to the normal group than the model group. Species composition analysis showed that the relative abundance of the Firmicutes decreased， while the relative abundance of the Bacteroidetes and norank_f_Muribaculaceae increased， and the Bacteroidetes/Firmicutes value increased in the model group than those in the normal group （P＜0.05 or P＜0.01）. Compared to those in the model group， the relative abundance of the Firmicutes increased， while the relative abundance of the Bacteroidetes and norank_f_Muribaculaceae， as well as the Bacteroidetes/Firmicutes value decreased in the CSP group and the probiotic group （P＜0.05 or P＜0.01）. There was no statistically significant difference in each indicator between the probiotic group and the CSP group （P＞0.05）. Pearson correlation analysis showed that at the phylum level， Firmicutes was positively correlated with ZO-1 （r=0.610， P=0.016） and Occludin （r=0.694， P=0.004） protein expression. Bacteroidetes was negatively correlated with ZO-1 （r=-0.557， P=0.031） and Occludin （r=-0.662， P=0.007） protein expression. At the genus level， norank_f_Muribaculaceae was negatively correlated with ZO-1 （r=-0.727， P=0.002） and Occludin （r=-0.760，P=0.001） protein expression. ConclusionCSP can restore the structure of intestinal flora， regulate the abundance levels of Firmicutes， Bacteroidetes and norank_f_Muribaculaceae， up-regulate ZO-1 and Occludin proteins， and thus repairing the duodenal mucosal barrier， and playing a therapeutic role in FD rats.

Emerging evidence suggests that intron-detaining transcripts (IDTs) are a nucleus-detained and polyadenylated mRNA pool for cell to quickly and effectively respond to environmental stimuli and stress. However, the underlying mechanisms of detained intron (DI) splicing are still largely unknown. Here, we suggest that post-transcriptional DI splicing is paused at the Bact state, an active spliceosome but not catalytically primed, which depends on Smad Nuclear Interacting Protein 1 (SNIP1) and RNPS1 (a serine-rich RNA binding protein) interaction. RNPS1 and Bact components preferentially dock at DIs and the RNPS1 docking is sufficient to trigger spliceosome pausing. Haploinsufficiency of Snip1 attenuates neurodegeneration and globally rescues IDT accumulation caused by a previously reported mutant U2 snRNA, a basal spliceosomal component. Snip1 conditional knockout in the cerebellum decreases DI splicing efficiency and causes neurodegeneration. Therefore, we suggest that SNIP1 and RNPS1 form a molecular brake to promote spliceosome pausing, and that its misregulation contributes to neurodegeneration.
Spliceosomes/metabolism* ; Introns/genetics* ; RNA Splicing ; RNA, Messenger/genetics* ; Cell Nucleus/metabolism*

Objective : To screen the RNA binding proteins interacting with NBV nonstructural protein σNS in host cells and to analyze their bioinformatics functions. Methods : In this study , the eukaryotic expression vector pEF⁃HA⁃MB⁃S3 of NBV σ NS was constructed and transfected into HEK293T cells after verification. After RNase A treatment , the obtained protein lysate was enriched by immunoprecipitation to enrich σNS binding proteins , identified and analyzed by LC⁃MS/MS mass spectrometry , and the properties and specific functions of proteins were discovered with the help of related Bioinformatics tools. Results : In this study , 32 candidate RNA binding proteins interacting with NBV σNS proteins were successfully screened , and the results of bioanalysis showed that these proteins were mainly located in cytoplasm and nucleus , and were mainly involved in biological processes such as cell metabolism , biological regulation , virus translation and transcription. Conclusion This study preliminarily analyzes the function of RNA binding proteins interacting with NBV σNS , which lays a foundation for further study on the mechanism of σNS protein in NBV life cycle.