Objective To investigate the diversity and composition of gut fungi microbiota in patients with polycystic kidney disease(PKD)and its correlation with clinical indicators.Methods A total of 44 PKD patients,44 patients with non-polycystic chronic kidney disease(NPCKD)and 22 healthy controls(HC)admitted to our hospital from February 2023 to February 2024 were recruited.ITS1 DNA sequencing was applied to analyze the gut fungal composition.Bioinformatics analysis was used to compare the diversity and structural differences of fungi among the 3 groups.Pearson correlation analysis was performed to analyze the relationship between gut fungi and clinical indicators.Results There were no significant differences in baseline characteristics(gender,age,body mass index,etc.)among the 3 groups,but statistical differences were seen in terms of serum indicators(such as serum creatinine,blood urea nitrogen,uric acid,estimated glomerular filtration rate,etc.)(P<0.01).Alpha diversity analysis showed no significant difference was seen between the PKD and HC groups,but the PKD group had significant differences to the NPCKD group(P<0.01).Beta diversity analysis revealed significant differences among the 3 groups and in pairwise comparisons(P=0.001).Fungi composition analysis found that the abundance of Candida was significantly higher in the PKD group than the other 2 groups(P<0.01),while the abundances of Aspergillus and Cladosporium were significantly lower in the PKD group than the HC group(P<0.05).Linear discriminant analysis(LEFSe)indicated that Candida was significantly enriched,while Aspergillus and Cladosporium were significantly reduced in the PKD group.Correlation analysis revealed that the abundance of Cladosporium was negatively correlated with cyst diameter and immunoglobulin light chain Kappa/Lambda ratio in the PKD group(P<0.05),while the abundance of Candida was positively correlated with liver/kidney cyst diameter(P<0.01).Conclusion PKD patients exhibit characteristic changes in gut fungi diversity and composition.The abundances of Cladosporium and Candida are closely associated with clinical indicators of PKD patients.

Objective To analyze the diversity and composition of gut microbiota in individuals with circadian rhythm disruption and their correlation with cognition.Methods Night shift workers and regular shift workers were subjected from our hospital during August 2022 and October 2024.The participants with circadian rhythm disorders were assigned into an experimental group(n=24),and those with normal circadian rhythms were into a control group(n=24).Their height,weight,age,gender,body mass index(BMI)and fresh fecal samples were collected,and Montreal Cognitive Assessment(MoCA)and Mini-Mental State Examination(MMSE)were used to evaluate their mental status.Metagenomics,Alpha and Beta diversity analyses,Linear Discriminant Analysis Effect Size(LEfSe),and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were employed to investigate the diversity and function characteristics of gut microbiota in the participants.Results There were no statistical differences between the 2 groups in baseline data,such as height,weight,gender,age,and BMI(P>0.05).Alpha diversity analysis indicated that no statistical differences were observed in the ACE,Chao1,Shannon,or Simpson indices between the 2 groups,while beta diversity analysis revealed significant differences(P<0.01),suggesting different structure of gut microbiota between them.In the experimental group,the abundance of Faecalibacterium prausnitzii and Agathobacter rectalis was decreased,while that of Escherichia coli and Phocaeicola vulgaratus was increased,with significant differences when compared with the control group(P<0.05).Additionally,KEGG functional analysis showed that the experimental group had obviously higher expression levels in Th17 cell differentiation and the IL-17 signaling pathway than the control group(P<0.05).Agathobacter rectalis and Faecalibacterium prausnitzii were positively correlated with MoCA score and MMSE score(P<0.05,P<0.01).Agathobacter rectalis was negatively correlated with the IL-17 signaling pathway and Th17 cell differentiation.Conclusion Individuals with circadian rhythm disorders have significant changes in the structure and function of gut microbiota when compared to those with normal circadian rhythms.Agathobacter rectalis may be involved in the regulation of the IL-17 signaling pathway and differentiation of Th17 cells,thereby possibly impacting the increases of cognitive score related to circadian rhythm disorders.

Objective To explore the underlying mechanisms by which time-restricted feeding(TRF)attenuates dextran sodium sulfate(DSS)-induced colitis in mice via modulation of regenerating islet-derived protein 3 gamma(Reg3γ)expression and gut microbiota.Methods Six-week-old C57BL/6 mice were stratified by body weight and randomized into ad libitum feeding(AL)and TRF groups(n=7).The AL mice were given unrestricted food access,whereas the TRF mice were allowed feeding only during 00:00 and 08:00 daily,for totally 4 weeks.Mouse colitis model was induced at the fourth week by adding 2.5%dextran sodium sulfate(DSS)in drinking water for 6 d.Disease severity and the effects of TRF were assessed with disease activity index(DAI)scoring,colon length measurement,HE staining and histopathological scoring,and mRNA expression levels of regenerating islet-derived 3 gamma(Reg3g)and inflammatory cytokines in colonic tissues.Another 14 mice were randomized into AL plus antibiotic cocktail(AL+ABX)and TRF plus antibiotic cocktail(TRF+ABX)groups,with 7 animals in each group.ABX was administered to deplete gut microbiota and evaluate the microbiota dependence of TRF in attenuating colitis.Fecal samples from AL and TRF mice were analyzed by 16S ribosomal RNA sequencing(16S rRNA-seq),and serum lipopolysaccharide(LPS)level was measured.The colonic epithelial cells were collected for RNA-seq.Results After modeling,the AL mice exhibited typical colitis symptoms,such as weight loss,diarrhea,and hematochezia.TRF intervention significantly attenuated these above symptoms,with lower DAI scores from day 4 post-modeling(P<0.001),reduced colon shortening(P<0.01),preserved tissue architecture,and decreased inflammation.RT-qPCR analysis showed that TRF down-regulated colonic mRNA expression levels of Reg3g and pro-inflammatory cytokines(IL-1 β,IL-6,TNF-α)(P<0.05)while up-regulated that of anti-inflammatory factor IL-10(P<0.000 1)when compared with the corresponding levels in AL mice.ABX treatment led no significant differences between the AL+ABX and TRF+ABX groups in term of DAI score,colon length,or histopathology.Obviously down-regulated Reg3g was observed in the TRF+ABX group than the AL+ABX group(P<0.05),whereas L-1β,IL-6,TNF-α and IL-10 showed no notable changes.16S rRNA-seq revealed that TRF markedly reshaped gut microbiota composition,with increased Gram-positive bacterial abundance,reduced Gram-negative bacteria,with concomitant lower serum LPS level(P<0.001).RNA-seq also indicated significant suppression of NF-κB and other inflammation-related signaling pathways in the TRF group.Conclusion TRF attenuates DSS-induced colitis in mice by downr-egulating Reg3γ expression,reshaping gut microbiota,and reducing serum LPS level,and thereby suppressing NF-κB-mediated inflammatory signaling pathways.

Objective:To study the effects of Qingre Lishi Prescription on ATP-P2X7R-IL-1β signaling pathway in rats with acute gouty arthritis.Methods:Otally 60 SPF-grade SD rats were divided into normal group, model group, colchicine group, and Qingre Lishi Prescription low-, medium- and high-dosage groups according to random number table method; the normal group and model group were given distilled water at a dosage of 10 ml/kg by gavage, the colchicine group was given colchicine solution at a dosage of 0.6 mg/kg by gavage, and Qingre Lishi Prescription low-, medium- and high-dosage groups were given Qingre Lishi Prescription extract suspension at dosages of 11.76, 23.52, and 47.04 g/kg by gavage, once a day for 7 consecutive days. Except for the normal group, AGA models were prepared by gavage on the 4th day in all other groups, and the swelling index of the right posterior ankle joint of rats was measured at 6, 12, 24, 48, and 72 hours after modeling; after 7 days of administration, samples were taken and HE staining was used to observe the pathological changes in the synovial tissue of the right posterior ankle joint. ELISA was used to detect the levels of serum ATP, IL-1 β, IL-8, and IL-37, and immunohistochemistry was used to detect the positive rates of P2X7R and IL-1 β proteins in the synovial tissue of the right posterior ankle joint.Results:At 6, 12, 24, 48, and 72 hours after modeling, compared with the model group, the swelling index of the colchicine group and Qingre Lishi Prescription low-dosage group decreased ( P<0.01); the levels of serum ATP, IL - β, IL-8, and IL-37 decreased in the colchicine group and Qingre Lishi Prescription low-, medium-, and high-dosage groups ( P<0.01); the expressions of P2X7R and IL-1 β proteins in the synovial tissue of the right posterior ankle was reduced in the colchicine group and Qingre Lishi Prescription high-dosage group ( P<0.05). Conclusion:Qingre Lishi Prescription may improve joint swelling and alleviate synovial tissue inflammation in AGA model rats through ATP/P2X7R/IL-1β-mediated inflammatory factor pathway.

Objective To investigate the effects of increasing the ejection seat backrest angle on the body pressure distribution of pilots of different body types,and to provide a basis for the design of ejection seats,flight training,and the development of strategies to alleviate muscle fatigue.Methods Male fighter pilots were divided into normal and overweight groups according to their body mass index(BMI),and a total of 40 pilots were tested for the distribution of sitting pressure under the two seat inclination angles of 20°and 33°by using the body pressure distribution measurement system.The effects of different seat inclination angles on sitting comfort were also analyzed.Results The pressure distributions of pilots with different body types were significantly different at different seat inclination angles.Compared with the 20°seat,the 33°seat condition had a larger cushion contact area[F(1,78)=40.281,P<0.001],a smaller average pressure and average pressure gradient[F(1,78)=32.030,P<0.001;F(1,78)=12.594,P<0.001],and significantly reduced average,maximum pressure,and maximal pressure gradients for the backrest[F(1,78)=10.516,P=0.002;F(1,78)=26.803,P<0.001;F(1,78)=4.918,P=0.029,respectively].In addition,overweight individuals with BMI had a notable increase in the cushion contact area[F(1,78)=21.038,P<0.001]and the backrest contact area[F(1,78)=8.301,P=0.005].No significant interaction was observed for angle and BMI.Conclusion An increased seat inclination angle results in a more uniform distribution of pressure across the human body,thereby increasing comfort.Moreover,the disparities in pressure and backrest distribution across disparate body types at varying seat angles provide a vital foundation for the optimized design of seating.

Objective To investigate the impact of blood sampling from different sites on coagula-tion results during systemic heparin anticoagulation period treated by continuous renal replacement therapy(CRRT).Methods Seventy-eight patients undergoing CRRT with systemic heparin anticoag-ulation were selected.Using a self-control method,blood samples were simultaneously collected from sampling port at input end of the CRRT extracorporeal circuit,the arterial pressure monitoring cathe-ter,and the peripheral vein of the patients.The four coagulation parameters were tested,and the differences in coagulation results among the blood sampling sites were compared.Results During CRRT with systemic heparin anticoagulation,the four coagulation parameters using blood samples col-lected from the sampling port at the input end of the CRRT extracorporeal circuit,the arterial pressure monitoring catheter,and the peripheral vein showed no statistically significant differences(P＞0.05).Subgroup analyses based on different hemofiltration machines and heparin doses also showed no statisti-cally significant differences(P＞0.05).Conclusion During CRRT,blood sampling from the sam-pling port at the input end of the circuit or the arterial pressure monitoring catheter can be used as an alternative to peripheral venous blood sampling,with no impact on coagulation results.

Objective To explore the relationship between the levels of serum dopamine,5-hydroxytryptamine,homovanillic acid and cognitive dysfunction in patients with Parkinson's disease.Methods A total of 118 Parkinson's disease patients admitted to Bazhong Central Hospital from February 2020 to February 2022 were selected as the disease group,and 106 healthy patients who underwent physical examination in the hospital during the same period were selected as the control group.The levels of serum do-pamine,5-hydroxytryptamine and homovanillic acid were measured in all subjects,and the cognitive function was evaluated with Montreal Cognitive Assessment Scale(MoCA).Parkinson's disease patients were divided into cognitive impairment group and non cognitive impairment group according to MoCA score.The serum do-pamine,5-hydroxytryptamine,homovanillic acid levels of the cognitive impairment group and the non cogni-tive impairment group were compared.Pearson correlation analysis was used to analyze the correlation be-tween serum dopamine,5-hydroxytryptamine and homovanillic acid levels and cognitive function of Parkinson's disease patients,and the receiver operating characteristic curve was drawn to analyze the evaluation value of serum dopamine,5-hydroxytryptamine and homovanillic acid levels on cognitive dysfunction of Parkinson's disease patients.Results The levels of serum dopamine,5-hydroxytryptamine,homovanillic acid and MoCA score in the disease group were lower than those in the control group,with statistical significance(P＜0.05).The levels of serum dopamine,5-hydroxytryptamine,homovanillic acid and MoCA score in advanced patients were lower than those in early patients,and the difference was statistically significant(P＜0.05).The levels of serum dopamine,serotonin and homovanillic acid in the cognitive dysfunction group were lower than those in the non-cognitive dysfunction group,and the difference was statistically significant(P＜0.05).The results of Pearson analysis showed that the levels of serum dopamine,5-hydroxytryptamine and homovanillic acid in Parkinson's disease patients were positively correlated with MoCA score(P＜0.05).The sensitivity of the combined assessment of serum dopamine,serotonin and homovanillic acid in Parkinson's disease patients was higher than that in Parkinson's disease patients alone(x2=7.413,P=0.006;x2=9.714,P=0.002;x2=8.541,P=0.003),the area under the curve(AUC)was higher than the AUC for assessing cognitive impair-ment in Parkinson's disease alone(Z=2.479,P=0.013;Z=2.271,P=0.023;Z=2.451,P=0.014).Conclusion There are differences in serum levels of dopamine,5-hydroxytryptamine,homovanillic acid and cognitive function among different stages of Parkinson's disease patients.Serum levels of dopamine,5-hydroxytryptamine,and homovanillic acid are closely related to cognitive function in Parkinson's disease pa-tients,all of which have evaluation value for the occurrence of cognitive dysfunction in Parkinson's disease pa-tients.However,the combination of the three serum indicators is more helpful for clinical evaluation and diag-nosis.

Objective To construct a non-trace deletion mutant of exlA in Pseudomonas aeruginosa strain NY8755(NY8755ΔexlA)and investigate the basic characteristics of pore-forming toxin ExlA.Methods The NY8755ΔexlA was constructed using the secondary homologous recombination method.C57BL/6J female mice ages 6 to 8 weeks were infected with NY8755 and NY8755 ΔexlA via aerosolized intratraheal inoculation respectively.Within 7 days of infection,the survival and weight changes of the mice were observed and recorded before the proinflammatory cytokines in the bronchoal-veolar lavage fluid(BALF)of the infected mice in the two groups were detected.Results The sequencing results showed that NY8755 ΔexlA was constructed.After 1×107 CFU NY8755 and NY8755 ΔexlA were infected,all the mice in the wild-type strain group died within 48 hours,while those in the mutant strain group began to die after 48 hours,and 40%of them remained alive 7 days later.The weight of surviving mice in the mutant strain group decreased but recovered gradually.After 12 hours of infection,there were more bloody exudates(redder in color)in the BALF of the wild-type strain group than in the mutant strain group,and the contents of proinflammatory cytokines interleukin-1β(IL-1β)and interleukin-17A (IL-17A)were significantly different. Conclusion Pseudomonas aeruginosa pore-forming toxin ExlA is the key pathogenic virulence factor of the exlA-positive Pseudomonas aeruginosa,which can significantly affect the survival status of mice and cause obvious inflammation in mice. Very little information is available on the action mechanisms of ExlA. In this study, The NY8755ΔexlA and the C57BL/6J mouse models infected with NY8755 and NY8755ΔexlA have been constructed that may be used for the investigation of pathogenesis of exlA-positive Pseudomonas aeruginosa.

Objective To construct a regulatory network of competing endogenous RNA(ceRNA)with prognostic value for bladder urothelial carcinoma(BLCA),and analyze the relationship between key messenger RNA(mRNA)and immune function.Methods The UCSC Xena database was used to download mRNA expression data from 404 BLCA patients and 28 normal individuals and key mRNAs were screened by differential analysis.ENCORI database was utilized to search microRNAs(miRNAs)that bind to key mRNAs and all long non-coding RNAs(LncRNAs)that bind to miRNAs.The expression data of miRNA and LncRNA were downloaded from TCGA database,co-expression analysis was performed to identify key mRNA with all miRNAs and miRNA with all LncRNAs,and thus key miRNAs and LncRNAs were screened out.Survival analysis was conducted based on the differences in expression levels of these key mRNAs,miRNAs,and LncRNAs between tumor patients and normal individuals,and finally a ceRNA regulatory network was constructed.The correlation between key mRNAs and immune cells,immune checkpoints(CD274,PDCD1 and CTLA4),and immune cell marker genes(IG)was analyzed using the TIMER 2.0 database.Results A total of 22 key mRNAs were screened,with the most significant difference being proline 3-hydroxylase 4(P3H4).The expression of P3H4 in patients with BLCA was high,and survival time was shorter in patients with high expression.A sum of 33 miRNAs and 14 LncRNAs were screened using the key mRNAs as the central link.Through co-expression analysis and survival analysis,hsa-miR-151a-3p and MIR100 HG were identified as the key miRNA and key LncRNA with prognostic value.The differences in the above analysis results were statistically significant(all P＜0.05).Based on these findings,a ceRNA regulatory network consisting of 1 mRNA,1 miRNA,and 1 LncRNA was constructed.Immunoassay firstly revealed a significant positive correlation between double positive T cells and P3H4 expression in the tumor microenvironment of BLCA.Moreover,there were 3 types of immune cells(tumor-associated neutrophils,and tumor-associated macrophages,dendritic cells),3 immune checkpoints(CD274,PDCD1,CTLA4),and 15 IGs with significant correlation with P3H4.These differences were statistically significant(all P＜0.01).Conclusion This study could help to reveal the progression mechanism of BLCA.The constructed ceRNA network and immune analysis can offer new insights into potential biological targets and immunotherapy directions for the diagnosis,treatment,and prediction of BLCA patients.