Objective:Investigating the association of the systemic immune-inflammation index and the pan-inflammation index with incident non-alcoholic fatty liver disease (NAFLD).Methods:This retrospective cohort study included 42 891 participants who underwent at least two health examinations at the Second Affiliated Hospital of Dalian Medical University between 2014 and 2023. Based on their levels of the systemic immune inflammation index (SII) and the pan immune inflammation value (PIV), participants were respectively divided into four quartile groups (Q1 to Q4). Cox proportional hazards models were employed to analyze the association of different SII and PIV levels, as well as their quartile groups, with new onset NAFLD in the total population and across various subgroups. Restricted cubic splines were used to examine the dose response relationship between these inflammatory indices and incident NAFLD. Additionally, sensitivity analyses were conducted to confirm the robustness of the findings.Results:After adjusting for confounding factors, the natural logarithm-transformed lnSII ( HR=1.247, 95% CI: 1.184-1.314, P<0.001) and lnPIV ( HR=1.192, 95% CI: 1.148-1.238, P<0.001) were significantly positively associated with the risk of NAFLD. When the subjects were grouped by SII quartiles (Q1-Q4), compared with those in Q1, participants in Q2, Q3, and Q4 exhibited progressively higher risks of incident NAFLD:11.9% ( HR=1.119, 95% CI: 1.051-1.192, P<0.001), 17.1% ( HR=1.171, 95% CI: 1.100-1.248, P<0.001), and 29.1% ( HR=1.291, 95% CI: 1.211-1.377, P<0.001), respectively. Quartile analysis of PIV yielded similar trends: the risk of incident NAFLD increased for 10.4% ( HR=1.104, 95% CI: 1.034-1.179, P=0.003), 18.7% ( HR=1.187, 95% CI: 1.112-1.266, P<0.001), and 30.5% ( HR=1.305, 95% CI: 1.223-1.393, P<0.001) in Q2, Q3, and Q4 group respectively when compared to that in Q1 group. Subgroup analysis confirmed consistent associations of SII and PIV with elevated NAFLD risk across all subgroups. Conclusion:Elevated levels of SII and PIV are significantly associated with increased risk of NAFLD.

Objective:To investigate transition pattern of health status among middle-aged and elderly population in China based on frailty index.Methods:In this retrospective cohort study, middle-aged and elderly people were selected from the China Health and Retirement Longitudinal Study (CHARLS) in 2011; and 1 434 subjects were followed up to 2015. The frailty index was calculated from the prevalence of chronic diseases, daily activity ability and blood biomarkers, and the frailty state was divided by quartiles of the frailty index. Markov models were constructed to determine the transition probabilities of different frailty states.Results:The mean age of the 1 434 subjects was (59.0±9.4) years and the mean frailty index was 0.11±0.05. In the healthy individuals, 63.0% remained healthy after a four-year follow-up; during the same follow-up period, 40.9% of the mildly frail individuals and 23.0% of the moderately frail individuals remained in their baseline frailty status. Increasing age leaded to a gradual increase in the probability of the population shifting to a severely frailty state. Women were more likely to shift to severe frailty status than men (0.029 vs 0.019, Z=3.03, P=0.002). Conclusion:Among middle-aged and elderly population in China, the transition of health states follows a pattern where higher frailty levels are associated with lower stability. Advanced age and female gender are identified as risk factors for progression to severe frailty.

Objective:To explore the correlation of cumulative atherogenic index of plasma (cumAIP) and cumulative atherosclerosis index (cumAI) with new-onset non-alcoholic fatty liver disease (NAFLD).Methods:From January 2017 to December 2023, 2 472 subjects who underwent health checkups at the Second Affiliated Hospital of Dalian Medical University for 3 consecutive years were enrolled. Triglyceride, total cholesterol, high density lipoprotein cholesterol and their measurement time intervals were used to calculate cumAIP and cumAI. The subjects were divided into Q1, Q2, Q3 and Q4 groups with the threshold values of 25th percentile, median and 75th percentile of the baseline atherogenic index of plasma (AIP) and atherosclerotic index (AI) subjects. Cox regression model was used to analyze the effects of cumAIP and cumAI on the new-onset NAFLD, restricted cubic spline was performed to analyze the nonlinear association between cumAIP and cumAI and new-onset NAFLD, and the clinical decision curve was used to compare the decision value of different indicators for NAFLD. Results:The risk of NAFLD gradually increased along with the increasing of cumAIP and cumAI. In the quartile groups of cumAIP, the incidence of Q1 to Q4 groups was 6.15%, 8.74%, 15.05%, and 25.08%, respectively. In the quartile groups of cumAI, the incidence of Q1 to Q4 groups was 5.99%, 11.17%, 15.21%, and 22.65%, respectively. After adjusting the confounding factors, the risk of new-onset NAFLD in the high-level group ( Q4) was higher than that in the low-level cumAIP group ( Q1) ( HR=3.15, 95% confidence interval (95% CI): 2.15 to 4.63, P<0.001) and the high-level cumAI group ( Q4) ( HR=2.74, 95% CI: 1.82 to 4.10, P<0.001). cumAIP and cumAI showed a significant nonlinear association with new-onset NAFLD ( χ2=119.15, 94.53; both P<0.001). The cumAIP had higher predictive value for NAFLD than the other cumulative lipid metrics and baseline AIP or AI. Conclusion:CumAIP and cumAI can be served as new predictive indicators of NAFLD, with a particular focus on the dynamic cumulative changes of AIP, which can achieve effective early screening for NAFLD.

This study aims to investigate the effect of Lactiplantibacillus plantarum LP12 on obe-sity prevention.In our study,Lactiplantibacillus plantarum LP12 was added to the diet for feed-ing,and the blood biochemistry status of rabbit,as well as the antioxidant effect of serum and liver samples were analyzed by determining the body weight change and feed intake of Japanese White rabbits.The changes in colony structure and abundance were also analyzed by 16S rDNA sequen-cing.The results showed that supplementation of Lactiplantibacillus plantarum LP12 inhibits weight gain,decreases serum glucose and ALT levels,and increases SOD activity in the liver.16S RNA gene sequencing analysis showed that the addition of Lactiplantibacillus plantarum LP12 increases the abundance of Bacteroidetes and Desulfovibrioides at the phylum level,and the supple-mentation of Lactiplantibacillus plantarum LP12 increases the abundance of Muribaculaceae at the genus level.Predictive analysis of microbiota function revealed that the supplementation of Lactiplantibacillus plantarum LP12 positively regulated iron-sulfur clusters and Zn-dependent proteases.In conclusion,the addition of Lactiplantibacillus plantarum effectively inhibits weight gain in Japanese White rabbits,enhances the antioxidative activity of the liver,and induces altera-tions in the gut microbiota composition of these rabbits.These findings lay an experimental foun-dation for further exploring the mechanisms by which Lactobacillus plantarum LP12 exerts its preventive effects against obesity and promotes metabolic health.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

The synthetic PDCoV N protein gene was optimized and cloned into the pET-30a vector to obtain the pET-30a-N plasmid.Thenthe recombinant plasmid was transformed into three strains of BL21 E.coli using heat-shock to explore protein expression conditions.The expressed proteins was purified using Ni Focurose 6FF(IMAC)and used as antigen to immunize the New Zealand White rabbit to prepare the polyclonal antibody against the PDCoV N protein.The antibody titer was measured by indirect ELISA method.The specificity for the antibody was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the pET-30a-N plasmid showed high expression level in BL21 StarTM(DE 3).The optimal expression condition was 37 ℃ 4 h.The purity of the target protein could reach 90.3％after purification.Indirect ELISA showed that the antibody titers was up to 1∶204 800.Western blot and IFA showed that the produced rabbit polyclonal antibody exhibited good specificity.In conclusion,the polyclonal antibody was prepared which specifically recognized the PDCoV N proteins.The results provided some references for the subsequent exploration of PDCoV N protein function and laid a foundation for establishing a diag-nostic method for PDCoV.

This study aims to investigate the effect of Lactiplantibacillus plantarum LP12 on obe-sity prevention.In our study,Lactiplantibacillus plantarum LP12 was added to the diet for feed-ing,and the blood biochemistry status of rabbit,as well as the antioxidant effect of serum and liver samples were analyzed by determining the body weight change and feed intake of Japanese White rabbits.The changes in colony structure and abundance were also analyzed by 16S rDNA sequen-cing.The results showed that supplementation of Lactiplantibacillus plantarum LP12 inhibits weight gain,decreases serum glucose and ALT levels,and increases SOD activity in the liver.16S RNA gene sequencing analysis showed that the addition of Lactiplantibacillus plantarum LP12 increases the abundance of Bacteroidetes and Desulfovibrioides at the phylum level,and the supple-mentation of Lactiplantibacillus plantarum LP12 increases the abundance of Muribaculaceae at the genus level.Predictive analysis of microbiota function revealed that the supplementation of Lactiplantibacillus plantarum LP12 positively regulated iron-sulfur clusters and Zn-dependent proteases.In conclusion,the addition of Lactiplantibacillus plantarum effectively inhibits weight gain in Japanese White rabbits,enhances the antioxidative activity of the liver,and induces altera-tions in the gut microbiota composition of these rabbits.These findings lay an experimental foun-dation for further exploring the mechanisms by which Lactobacillus plantarum LP12 exerts its preventive effects against obesity and promotes metabolic health.

Objective:Investigating the association of the systemic immune-inflammation index and the pan-inflammation index with incident non-alcoholic fatty liver disease (NAFLD).Methods:This retrospective cohort study included 42 891 participants who underwent at least two health examinations at the Second Affiliated Hospital of Dalian Medical University between 2014 and 2023. Based on their levels of the systemic immune inflammation index (SII) and the pan immune inflammation value (PIV), participants were respectively divided into four quartile groups (Q1 to Q4). Cox proportional hazards models were employed to analyze the association of different SII and PIV levels, as well as their quartile groups, with new onset NAFLD in the total population and across various subgroups. Restricted cubic splines were used to examine the dose response relationship between these inflammatory indices and incident NAFLD. Additionally, sensitivity analyses were conducted to confirm the robustness of the findings.Results:After adjusting for confounding factors, the natural logarithm-transformed lnSII ( HR=1.247, 95% CI: 1.184-1.314, P<0.001) and lnPIV ( HR=1.192, 95% CI: 1.148-1.238, P<0.001) were significantly positively associated with the risk of NAFLD. When the subjects were grouped by SII quartiles (Q1-Q4), compared with those in Q1, participants in Q2, Q3, and Q4 exhibited progressively higher risks of incident NAFLD:11.9% ( HR=1.119, 95% CI: 1.051-1.192, P<0.001), 17.1% ( HR=1.171, 95% CI: 1.100-1.248, P<0.001), and 29.1% ( HR=1.291, 95% CI: 1.211-1.377, P<0.001), respectively. Quartile analysis of PIV yielded similar trends: the risk of incident NAFLD increased for 10.4% ( HR=1.104, 95% CI: 1.034-1.179, P=0.003), 18.7% ( HR=1.187, 95% CI: 1.112-1.266, P<0.001), and 30.5% ( HR=1.305, 95% CI: 1.223-1.393, P<0.001) in Q2, Q3, and Q4 group respectively when compared to that in Q1 group. Subgroup analysis confirmed consistent associations of SII and PIV with elevated NAFLD risk across all subgroups. Conclusion:Elevated levels of SII and PIV are significantly associated with increased risk of NAFLD.

Objective:To investigate transition pattern of health status among middle-aged and elderly population in China based on frailty index.Methods:In this retrospective cohort study, middle-aged and elderly people were selected from the China Health and Retirement Longitudinal Study (CHARLS) in 2011; and 1 434 subjects were followed up to 2015. The frailty index was calculated from the prevalence of chronic diseases, daily activity ability and blood biomarkers, and the frailty state was divided by quartiles of the frailty index. Markov models were constructed to determine the transition probabilities of different frailty states.Results:The mean age of the 1 434 subjects was (59.0±9.4) years and the mean frailty index was 0.11±0.05. In the healthy individuals, 63.0% remained healthy after a four-year follow-up; during the same follow-up period, 40.9% of the mildly frail individuals and 23.0% of the moderately frail individuals remained in their baseline frailty status. Increasing age leaded to a gradual increase in the probability of the population shifting to a severely frailty state. Women were more likely to shift to severe frailty status than men (0.029 vs 0.019, Z=3.03, P=0.002). Conclusion:Among middle-aged and elderly population in China, the transition of health states follows a pattern where higher frailty levels are associated with lower stability. Advanced age and female gender are identified as risk factors for progression to severe frailty.

Objective:To explore the correlation of cumulative atherogenic index of plasma (cumAIP) and cumulative atherosclerosis index (cumAI) with new-onset non-alcoholic fatty liver disease (NAFLD).Methods:From January 2017 to December 2023, 2 472 subjects who underwent health checkups at the Second Affiliated Hospital of Dalian Medical University for 3 consecutive years were enrolled. Triglyceride, total cholesterol, high density lipoprotein cholesterol and their measurement time intervals were used to calculate cumAIP and cumAI. The subjects were divided into Q1, Q2, Q3 and Q4 groups with the threshold values of 25th percentile, median and 75th percentile of the baseline atherogenic index of plasma (AIP) and atherosclerotic index (AI) subjects. Cox regression model was used to analyze the effects of cumAIP and cumAI on the new-onset NAFLD, restricted cubic spline was performed to analyze the nonlinear association between cumAIP and cumAI and new-onset NAFLD, and the clinical decision curve was used to compare the decision value of different indicators for NAFLD. Results:The risk of NAFLD gradually increased along with the increasing of cumAIP and cumAI. In the quartile groups of cumAIP, the incidence of Q1 to Q4 groups was 6.15%, 8.74%, 15.05%, and 25.08%, respectively. In the quartile groups of cumAI, the incidence of Q1 to Q4 groups was 5.99%, 11.17%, 15.21%, and 22.65%, respectively. After adjusting the confounding factors, the risk of new-onset NAFLD in the high-level group ( Q4) was higher than that in the low-level cumAIP group ( Q1) ( HR=3.15, 95% confidence interval (95% CI): 2.15 to 4.63, P<0.001) and the high-level cumAI group ( Q4) ( HR=2.74, 95% CI: 1.82 to 4.10, P<0.001). cumAIP and cumAI showed a significant nonlinear association with new-onset NAFLD ( χ2=119.15, 94.53; both P<0.001). The cumAIP had higher predictive value for NAFLD than the other cumulative lipid metrics and baseline AIP or AI. Conclusion:CumAIP and cumAI can be served as new predictive indicators of NAFLD, with a particular focus on the dynamic cumulative changes of AIP, which can achieve effective early screening for NAFLD.