ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

ObjectiveTo investigate the functional role and underlying molecular mechanisms of fumarylacetoacetate hydrolase （FAH） in the progression of glioblastoma （GBM）. MethodsDifferential expression analysis was performed on the TCGA-GBM， GSE4290， and GSE116520 datasets. Weighted gene co-expression network analysis （WGCNA） was used to identify key modules， and Cox regression and risk modeling were used to screen prognostic genes. Immune infiltration analysis of prognostic genes was carried out by using single-cell RNA sequencing panels. The clinical expression signature of FAH in GBM was analyzed in the TCGA and HPA databases. The functional role of FAH was validated by in vitro and in vivo experiments， and pathway analysis was performed to explore the underlying mechanisms. ResultsA total of 152 overlapping genes were identified across the three GBM datasets （P<0.05）. WGCNA revealed that the turquoise module was most strongly associated with tumor purity， stromal score， immune score， and ESTIMATE score （P<0.001）. Compared with normal tissues， three prognostic genes （CTSD， FAH， and THBD） were upregulated in GBM and correlated with immune infiltration （P<0.05）. FAH mRNA and protein levels were elevated in GBM tissues relative to normal tissues， and its expression was significantly associated with age stratification and TP53 mutation （P<0.05）. CCK-8 assay results showed that， compared with the shNC group， the proliferative activity of GBM cells in the shFAH group was reduced （P<0.001）. Transwell migration and invasion assays demonstrated that， relative to the shNC group， the numbers of migrated and invaded cells in the shFAH group decreased （P<0.05）. Western blot analysis revealed that the protein expression levels of PI3K， p-AKT， and p-mTOR in the shFAH group decreased compared with those in the shNC group （P<0.05）. In vivo subcutaneous xenograft experiments further confirmed that tumor volume and weight significantly decreased in the shFAH group compared with the shNC group （P<0.001）. ConclusionFAH promotes GBM progression by activating the PI3K/AKT/mTOR signaling pathway and may serve as a potential therapeutic target for GBM.

This study investigated the types and distribution of rodents,and the infection status of eight pathogens in the high-altitude areas of western Sichuan,to provide a basis for control of rodent-borne diseases and pathogen surveillance in rodents in the area.From August to November of 2023,rodents were captured through the night method in high-altitude areas of western Sichuan.Nucleic acids were collected from the rodents'livers and lungs,and eight important pathogens were detected:Dabie bandavirus,Han-tavirus,Bartonella,Francisella tularensis,Anaplasma phagocytophilum,Rickettsia mooseri,Orientia tsutsugamushi,and Leptospira interrogans.The chi-square test was used to compare the composition ratios of rodent species and the difference in pathogen positivity rates among groups.A total of 114 rodents of nine species were captured.The dominant species in this area were Apodemus agrarius(22.81%),Apodemus chevrieri(18.42%),Niviventer confucianus(17.54%),Apodemus latronum(16.67%),and Apodemus peninsu-lae(13.16%).Other rodent species included Rattus nitidus(4.39%),Neodon irene(4.39%),Chodsigoa hypsibia(1.75%),and Nivi-venter excelsior(0.88%).Statistically significant differences in rodent species composition were observed among regions,altitudes,and habitats(χ2Region=112.358,P<0.05;χ2Altitude=96.843,P<0.05;χ2Habitat=48.842,P<0.05).The liver and lung pathogen results showed that the positivity rate of Bartonella was highest(29/114),whereas those of the other seven pathogens were 0%-4.39%.Five rodents were co-infected with two or more pathogens,and the composite positivity rate was 4.36%(5/114).Statistically significant differences in the positivity rates were observed for Leptospira interrogans among species(χ2=6.568,P=0.028)and Anaplasma phagocytophilum among habitats(χ2=7.596,P=0.027);however,no significant differences in the positivity rates of other pathogens were found among rodent species,regions,altitudes,habitats,and sexes(P>0.05).Thus,rodent species were abundant in the high-altitude areas of western Sichuan and carried a variety of pathogens.Multiple pathogens showed compound infections,among which the positivity rate of Bartonella was relatively high.The total infection rate of pathogens in living areas was relatively high,and the risk of pathogenesis to the population is greater.Therefore,rodent control and disease monitoring efforts should be strengthened.

Objective To investigate the expressions of IL-18,IL-18 binding protein isoform a(IL-18BPa)and IL-18Rα in blood CD4+Th1 cells of patients with allergic asthma(AA),and with allergic rhinitis and asthma syndrome(ARA),and the effects of allergens on their expressions.Methods Blood samples were collected from patients with AA,ARA and healthy control(HC)subjects.Flow cytometry was used to evaluate the actions of allergens on the expressions of IL-18,IL-18BPa and IL-18Rα in CD4+Th1 cells.Ovalbumin(OVA)-induced AA mouse models were established,and the expression level of IL-18Rα along with the influence of IL-18 on its expression in blood and lung Th1 cells was also explored by flow cytometry.Results Compared with HC,we observed increased IL-18 while decreased IL-18BPa expression in blood Th1 cells of AA and ARA patients.Moreover,allergens upregulated the expression levels of IL-18,IL-18BPa and IL-18Rα in Th1 cells of patients with AA and ARA.Additionally,intratracheal challenge with OVA plus IL-18 increased the proportions of blood and lung Th1 cells of HC and AA mice,and upregulated IL-18Rα expression on blood and lung Th1 cells of HC mice.Conclusion Allergens may be involved in the pathogenesis of airway allergic diseases by inducing the expressions of IL-18 and IL-18Rα in CD4+Th1 cells.

Objective:To evaluate the technical feasibility and perioperative safety of pyeloplasty assis-ted by the CarinaTM modular laparoscopic surgical robotic system in patients with ureteropelvic junction obstruction(UPJO).Methods:From November to December 2024,five consecutive patients diagnosed with UPJO underwent robot-assisted pyeloplasty using the CarinaTM modular laparoscopic surgical system at Peking University First Hospital.Data on patient demographics,intraoperative parameters(including docking time,console time,and estimated blood loss),perioperative outcomes,follow-up results,and surgeons' subjective evaluations of system performance were prospectively collected.Descriptive statistics were used;continuous variables were presented as median(range),and categorical variables as frequen-cy and percentage.Results:The cohort included four females and one male.All the patients successfully completed the robotic procedure without conversion to open or conventional laparoscopic surgery.The me-dian age was 32 years(24-37 years),and the median body mass index was 21.6 kg/m2(15.8-27.3 kg/m2).The median docking time was 8 min(3-12 min),and the median console time was 91 min(71-125 min).Intraoperative blood loss was uniformly 20 mL.The median postoperative drainage du-ration was 3 d(0-4 d),and the median length of hospital stay was 4 d(4-9 d).No Clavien-Dindo grade Ⅲ or higher complications occurred.All the patients had their double-J stents removed at 2 months postoperatively,and pain in the ipsilateral flank,reported preoperatively by all the five patients,was al-leviated.The subjective surgical success rate was 100％.Surgeons reported stable system performance throughout all the procedures,with no instances of mechanical arm interference or visual drift affecting surgical fluency.Conclusion:Preliminary findings indicate that pyeloplasty using the domestically deve-loped CarinaTM modular laparoscopic robotic system is technically feasible and perioperatively safe for the treatment of UPJO.

Objective The aim of this study is to discuss the causal relationship between periodontal disease(PD)and prostate cancer(PCa).Methods A two-sample bidirectional Mendelian randomization(MR)analysis based on publicly statistical data from genome-wide association studies(GWAS)was conducted.MR Egger,weighted medium,simple mode and weighted mode were supplemented,while inverse variance weighted analysis(IVW)was the main method of analysis.Heterogeneity testing,pleiotropy testing and leave-one-out testing were used to assess the sensitivity and stabili-ty.Results The results of MR analysis showed that PD had no significant impact on the occurrence of PCa:East Asian(IVW,PD:OR=1.07,P=0.48);European(IVW,PD:OR=1.00,P=0.37,periodontitis:OR=1.03,P=0.14,chronic gingivitis:OR=0.99,P=0.37,chronic periodontitis:OR=1.03,P=0.22).The reverse MR analysis also did not show a causal relationship between PCa and PD:East Asian(IVW,PD:OR=0.97,P=0.22);European(IVW,PD:OR=0.84,P=0.44,periodontitis:OR=1.01,P=0.75,chronic gingivitis:OR=0.93,P=0.23,chronic periodontitis:OR=0.99,P=0.80).The results of other analysis were consistent with those of IVW analysis.Conclusions The results of our two-sample bidirectional MR analysis do not support a causal relationship between PD and PCa.

Objective To analyze the disability level and long-term care needs assessment policies issued by 49 pilot cities in China's long-term care insurance system,providing insights for further improvement of the system.Methods Policy documents related to long-term care insurance,published between June 2016 and December 2024 on official platforms of local governments,the Ministry of Human Resources and Social Security,the National Healthcare Security Administration,and the National Public Service Platform for Standards Information,were retrieved.A comparative analysis was used to summarize and compare the content of these policy texts.Results Currently,there are 49 pilot cities implementing the long-term care insurance system in China.47 cities have issued disability level assessment standards,with 38 cities adopting the trial standards issued by the National Healthcare Security Administration.In 42 pilot cities,the disability level assessment includes indicators such as activities of daily living,cognitive function,and sensory and communication abilities.22 cities have issued policies related to care needs assessment.In most cities,the care needs assessment partially or fully includes indicators from the disability level assessment.Conclusion There is no national uniformity in disability level assessment standards,and the care needs assessment framework needs further improvement.It is recommended to standardize disability level assessment criteria,clearly delineate the boundaries and content of disability level and care needs assessments,and enhance the development of the care needs assessment system.

Objective To provide methodological examples for related research,the influencing factors of the evolution of Qi deficiency syndrome in chronic obstructive pulmonary disease(COPD)based on a nonlinear mixed effects model was explored.Methods A research questionnaire on the influencing factors of the evolution of Qi deficiency syndrome in chronic obstructive pulmonary disease was developed,and clinical data of 650 COPD patients on the 1st and 14th day of acute exacerbation,the 1st and 28th day of risk window,the first day of stable period,and the 90th day were dynamically collected from 10 tertiary hospitals across the country.8 baseline data including gender and age were collected through the PROC NLMIXED process by SAS 9.4 software.Coronary heart disease,diabetes and hypertension accounted for the highest proportion.Nine concurrent syndromes including wind cold syndrome and phlegm heat syndrome were used as fixed effects,and individual level was used as random effects to gradually fit the model and screen the influencing factors of Qi deficiency syndrome in the entire process of disease occurrence and development.Results A total of 637 eligible cases were included,and clinical datas were dynamically collected on the 1st and 14th day of acute exacerbation,the 1st and 28th day of the risk window,the 1st and 90th day of the stable period.It was found that the number of acute exacerbations,alcohol consumption,concomitant hypertension,coronary heart disease,blood stasis syndrome,yin deficiency syndrome,yang deficiency syndrome,6-minute walking distance,and the modified Medical Research Council Dyspnea Questionnaire(mMRC)had an impact on the evolution of Qi deficiency syndrome in the previous year(P<0.05).Conclusion The use of a nonlinear mixed effects model revealed the relevant factors affecting the evolution of Qi deficiency syndrome from complex multi temporal dynamic data,providing methodological references for other related studies.

This study investigated the regulatory effects of carvacrol on lipopolysaccharide(LPS)-induced inflammatory response and endoplasmic reticulum(ER)stress in bovine mammary epithe-lial cells using in vitro cell culture techniques.Western blot analysis revealed significantly elevated expression levels of NF-κB pathway-related proteins and ER stress marker proteins in mastitis samples compared to healthy mammary tissues(P＜0.05).Cells were divided into a blank control group and carvacrol(CAV)treatment groups with varying concentrations(50,100,250,500,750,1 000 μmol/L).After 24 hours of culture,cell proliferation was assessed using the CCK-8 assay.An inflammatory model was established by stimulating MAC-T cells(a bovine mammary epithelial cell line)with LPS(10 mg/L)for 12 h,followed by measurement of the transcriptional levels of inflammatory-related genes(IL-6,IL-1β,and TNFα).MAC-T cells were pretreated with low,medi-um,and high doses of CAV for 12 h before LPS stimulation.Molecular docking analysis was per-formed to examine the interaction between CAV and GRP78,a key ER stress protein.Real-time quantitative PCR(qPCR)was used to analyze the mRNA expression levels of inflammatory cyto-kines(IL-6,IL-1β,TNFα),while Western blot was employed to assess the expression levels of NF-κB pathway proteins(p-IκB,p-NF-κB p65)and ER stress-related proteins(p-PERK,p-IRE1α,ATF6,GRP78,CHOP).The results from Western blot and qPCR demonstrated that CAV alleviated LPS-induced inflammatory response and cellular damage by inhibiting the NF-κB signa-ling pathway and ER stress.

Objective:To investigate repeated dose toxicity of CpG 2395 and summarize common safety characteristics of CpG ODNs.Methods:Rats were randomly divided into 7 groups and injected with different doses of CpG 2395,CpG-Alum adjuvant system and normal saline into hindlimb muscles(4 administrations,2 weeks apart).After administration,clinical observations were conducted at different time points,accompanied by other examinations,such as body weight,food consumption,body temperature,hematology,coagulation,clinical chemistry,urinalysis,cytokines,gross pathology and histopathology.Additionally,repeated dose toxicity results of other CpG ODNs were summarized and analyzed.Results:CpG 2395 possessed relevant attributes to induce significant immunostimu-latory effects,and addition of aluminum adjuvants could enhance these processes.Different CpG ODNs had common safety characteris-tics.Relevant symptoms would be mainly alleviated or eliminated without delayed toxicity after end of administrations,and dose-toxicity relationship had been confirmed.Conclusion:CpG 2395 is considered to be safe and controllable at proposed clinical dose,and based on common safety characteristics,possible local and systemic adverse reactions can be predicted in advance before clinical application of CpG 2395.