Objective To investigate the protective effects of Kaempferol(KAE)against hepatic lipid de-position induced by a high-fat diet,as well as the underlying mechanisms.Methods C57BL/6J male mice were fed a high-fat diet for 22 weeks to establish a chronic nonalcoholic steatohepatitis(NASH)model.KAE was admin-istered during the last 8 weeks as an interventional agent to evaluate its effects.Liver lipid deposition was assessed,and the expression levels of endoplasmic reticulum(ER)stress-related proteins,activation of the Farnesoid X Re-ceptor(FXR)signaling pathway,and the expression of lipid synthesis-related genes were analyzed.In vitro,pal-mitic acid(PA)was used to stimulate AML-12 cells to induce lipid accumulation.Additionally,siRNA targeting FXR was transfected into AML-12 cells to investigate the role of the ER stress-FXR signaling pathway in mediating the effects of KAE.Results The intervention of kaempferol inhibited the rapid weight gain induced by a high-fat diet,reduced serum total cholesterol,triglyceride levels,and ALT activity,effectively alleviated large-scale lipid aggregation in the liver,thereby exerting a protective effect against hepatic lipid deposition in NASH.Mechanisti-cally,KAE decreased hepatic ER stress,promoted the expression of FXR and its activation marker SHP,thereby suppressing the expression of FASN and reducing hepatic lipid synthesis.In vitro,KAE treatment significantly re-versed the inhibitory effect of excessive ER stress on FXR activity,as evidenced by the upregulation of FXR activ-ity leading to decreased FASN expression and reduced steatosis in AML-12 cells.Moreover,FXR knockdown mark-edly abolished the protective effects of KAE on lipid deposition in AML-12 cells exposed to PA,by eliminating the promoting effect of KAE on SHP expression and the SHP-mediated suppression of SREBP1c.Conclusions KAE treatment alleviated ER stress,thereby enhancing FXR/SHP signaling and subsequently suppressing lipid synthesis to reduce hepatic lipid accumulation.These findings suggest that KAE holds therapeutic potential for the manage-ment of hepatic steatosis in NASH.

The interstitial protein UL14 of pseudorabies virus(PRV)constitutes a crucial compo-nent for viral replication and virulence invasion.It can interact with other viral proteins to complete the infection of the host,rendering it one of the potentially important antiviral targets.In this stud-y,the PRV DX strain isolated in the laboratory was used as the parental strain.The recombinant UL14 protein was expressed and purified through the prokaryotic expression system.BALB/c mice were immunized with this protein as an antigen.After three rounds of subcloning screening,three hybridoma cell lines against the UL14 protein were obtained,named 1B4,1B5,and 1F2,respective-ly.The immunoreactivity of the antibodies was detected by immunofluorescence assay(IFA).The results showed that the antibody titers of the 1B4 and 1B5 strains were not lower than 1∶3 200,and that of the 1F2 strain was not lower than 1∶2 000.The results of western blot(WB)detection showed that the antibody titers of the three strains were all not lower than 1∶8 000.The results of indirect ELISA detection showed that the antibody titers in ascites were all not lower than 112 800.The results of the identification of the antigenic epitopes of the UL14 protein showed that the antigenic epitope recognized by 1B4 was 1 MFASDRRERRVRLAEAFQRE20,and the antigenic epitopes recognized by 1B5 and 1F2 were speculated to be 33GRADKKNPEFVRAFMAAKQAR53.After PRV infected susceptible cells,the temporal expression of the UL14 protein was detected by IF A using the prepared monoclonal antibodies.It was found that the temporal expression of UL14 was similar to that of gC,and the result of RT-qPCR of UL14 gene was consistent with IF A,indi-cating that UL14 might be a late gene during the PRV replication.The subcellular localization of UL14 after virus infection was detected by IFA,and it was found that the UL14 protein could be transferred from the cytoplasm to the nucleus and the expression of UL14 protein increased with the extension of infection time.This study provides a good research tool for further investigating on the function of the PRV UL14 protein and might contribute to the further study of the PRV replication mechanism mediated by the PRV UL14 protein.

The interstitial protein UL14 of pseudorabies virus(PRV)constitutes a crucial compo-nent for viral replication and virulence invasion.It can interact with other viral proteins to complete the infection of the host,rendering it one of the potentially important antiviral targets.In this stud-y,the PRV DX strain isolated in the laboratory was used as the parental strain.The recombinant UL14 protein was expressed and purified through the prokaryotic expression system.BALB/c mice were immunized with this protein as an antigen.After three rounds of subcloning screening,three hybridoma cell lines against the UL14 protein were obtained,named 1B4,1B5,and 1F2,respective-ly.The immunoreactivity of the antibodies was detected by immunofluorescence assay(IFA).The results showed that the antibody titers of the 1B4 and 1B5 strains were not lower than 1∶3 200,and that of the 1F2 strain was not lower than 1∶2 000.The results of western blot(WB)detection showed that the antibody titers of the three strains were all not lower than 1∶8 000.The results of indirect ELISA detection showed that the antibody titers in ascites were all not lower than 112 800.The results of the identification of the antigenic epitopes of the UL14 protein showed that the antigenic epitope recognized by 1B4 was 1 MFASDRRERRVRLAEAFQRE20,and the antigenic epitopes recognized by 1B5 and 1F2 were speculated to be 33GRADKKNPEFVRAFMAAKQAR53.After PRV infected susceptible cells,the temporal expression of the UL14 protein was detected by IF A using the prepared monoclonal antibodies.It was found that the temporal expression of UL14 was similar to that of gC,and the result of RT-qPCR of UL14 gene was consistent with IF A,indi-cating that UL14 might be a late gene during the PRV replication.The subcellular localization of UL14 after virus infection was detected by IFA,and it was found that the UL14 protein could be transferred from the cytoplasm to the nucleus and the expression of UL14 protein increased with the extension of infection time.This study provides a good research tool for further investigating on the function of the PRV UL14 protein and might contribute to the further study of the PRV replication mechanism mediated by the PRV UL14 protein.

Objective To investigate the protective effects of Kaempferol(KAE)against hepatic lipid de-position induced by a high-fat diet,as well as the underlying mechanisms.Methods C57BL/6J male mice were fed a high-fat diet for 22 weeks to establish a chronic nonalcoholic steatohepatitis(NASH)model.KAE was admin-istered during the last 8 weeks as an interventional agent to evaluate its effects.Liver lipid deposition was assessed,and the expression levels of endoplasmic reticulum(ER)stress-related proteins,activation of the Farnesoid X Re-ceptor(FXR)signaling pathway,and the expression of lipid synthesis-related genes were analyzed.In vitro,pal-mitic acid(PA)was used to stimulate AML-12 cells to induce lipid accumulation.Additionally,siRNA targeting FXR was transfected into AML-12 cells to investigate the role of the ER stress-FXR signaling pathway in mediating the effects of KAE.Results The intervention of kaempferol inhibited the rapid weight gain induced by a high-fat diet,reduced serum total cholesterol,triglyceride levels,and ALT activity,effectively alleviated large-scale lipid aggregation in the liver,thereby exerting a protective effect against hepatic lipid deposition in NASH.Mechanisti-cally,KAE decreased hepatic ER stress,promoted the expression of FXR and its activation marker SHP,thereby suppressing the expression of FASN and reducing hepatic lipid synthesis.In vitro,KAE treatment significantly re-versed the inhibitory effect of excessive ER stress on FXR activity,as evidenced by the upregulation of FXR activ-ity leading to decreased FASN expression and reduced steatosis in AML-12 cells.Moreover,FXR knockdown mark-edly abolished the protective effects of KAE on lipid deposition in AML-12 cells exposed to PA,by eliminating the promoting effect of KAE on SHP expression and the SHP-mediated suppression of SREBP1c.Conclusions KAE treatment alleviated ER stress,thereby enhancing FXR/SHP signaling and subsequently suppressing lipid synthesis to reduce hepatic lipid accumulation.These findings suggest that KAE holds therapeutic potential for the manage-ment of hepatic steatosis in NASH.

Objective To explore and analyze the characteristics and transmission routes of carbapenem-resistant Klebsiella pneumoniae(CRKP)strains in intensive care unit(ICU).Methods From January to October 2023,17 clinical infection isolates(clinical infection group),5 active screening isolates(active screening group),and 7 envi-ronmental isolates(environmental group)of CRKP in the liver surgery ICU of a hospital were selected and analyzed by whole-genome sequencing.The differences in resistance genes,virulence genes,and sequence typing(ST)were compared,and transmission routes were analyzed based on the phylogenetic tree.Results 29 strains of CRKP car-ried 4-18 resistance genes and 52-98 virulence genes,respectively.There were no statistically significant diffe-rences in genotype distribution of resistance genes,the number of virulence genes,and gene types among three groups of CRKP(all P＞0.05).ST showed that 29 CRKP strains mainly consisted of two categories:ST11 and ST15.Based on the phylogenetic tree constructed from the core genome,there were 7 highly homologous groups of CRKP,among which 4 groups had clear epidemiological associations.Conclusion CRKP in ICU carries more re-sistance and virulence genes,and some strains are highly homologous in ST and phylogenetic tree,which may lead to cross transmission.In the future,prevention and control measures should be strengthened to reduce the trans-mission of CRKP.

The Keap1-Nrf2 pathway has been shown to be an important defense mechanism against oxidative stress, which may be an effective therapeutic strategy for many diseases. The research progresses on Keap1-Nrf2 pathway in inflammatory diseases were mainly reviewed. The basic components and activation mechanism of Keap1-Nrf2 pathway were introduced. The relationship between Keap1-Nrf2 pathway and the crosstalk between NF-κB pathway and HO-1 pathway, the expression of inflammatory mediators and enzymes, and inflammatory bodies were expounded. Natural product-derived inhibitors, small molecule inhibitors targeting Keap1-Nrf2 pathway and their clinical progress were introduced, and the potential application value of Keap1-Nrf2 pathway in the treatment of inflammation was discussed.

Objective:To compare the efficacy of low-dose febuxostat and low-dose benzbromarone in lowering serum uric acid and their impact on liver function in male patients with renal underexcretion gout.Methods:This prospective cohort study enrolled 303 patients with renal underexcretion gout and normal baseline liver function. Participants were assigned to either the low-dose febuxostat group(20 mg qd) or the low-dose benzbromarone group(25 mg qd). A linear mixed-effects model was used to compare the uric acid target achievement rate(<360 μmol/L) and changes in liver enzyme levels between the two groups.Results:At week 4, the proportion of patients achieving the serum uric acid target(<360 μmol/L) was significantly higher in the benzbromarone group than that in the febuxostat group(64.2% vs 42.3%, P<0.001), with a trend toward greater efficacy at weeks 8 and 12. The incidence of alanine aminotransferase(ALT) or aspartate aminotransferase(AST) elevation above the upper limit was significantly higher in the febuxostat group compared to the benzbromarone group(35.2% vs 13.85%, P<0.001). After adjusting for baseline liver enzyme levels in the mixed-effects model, mean ALT and AST levels remained significantly higher in the febuxostat group than those in the benzbromarone group at weeks 4, 8, and 12( P<0.05). In the febuxostat group, ALT and AST levels significantly increased over time during weeks 0-4 and 4-8 ( P<0.001), peaking at week 8 followed by a decreasing trend. By week 12, the levels were not significantly different from baseline ( P>0.05). Whereas there was no significant difference at each follow-up time point in the benzbromarone group( P>0.05). Conclusions:In male patients with renal underexcretion gout, low-dose benzbromarone demonstrated superior urate-lowering efficacy and better hepatic safety compared to low-dose febuxostat.

Childhood primary renal tubular diseases are chronic kidney diseases characterized by impaired renal tubular reabsorption. Primary renal tubular disease has diverse clinical manifestations and lacks of specificity. Laboratory tests are limited，making it prone to missed diagnosis and misdiagnosis. Based on the current knowledge of renal tubular diseases，authors propose early warning signals of renal tubular diseases such as family history of primary tubular diseases，unexplained polyhydramnios during pregnancy，polydipsia，polyuria，delayed growth and development or rickets，decreased muscle strength and tone，unexplained electrolyte disturbance，hyperuricemia，acid-base disturbance，positive urine sugar test，renal tubular proteinuria，urinary imaging examination suggesting kidney stones，calcium deposition，renal cysts and early onset of eye，ear，joint and neuron injury.Meanwhile，some universal management strategies for primary renal tubular disease are proposed，emphasizing the importance of multidisciplinary collaboration，genetic testing and individualized intervention to improve the long-term prognosis of childhood primary renal tubular diseases.

Objective:To evaluate the potential of IgG N-glycans as diagnostic biomarker for ankylosing spondylitis (AS) by comparing and analyzing the IgG N-glycan profiles with AS and healthy controls.Methods:A 1∶1 matched case-control study design was adopted, 81 AS patients who visited the Department of Rheumatology and Immunology at Taian City Central Hospital and the Second Affiliated Hospital of Shandong First Medical University between July 2020 and June 2021 were recruited. These patients were matched with 81 healthy individuals undergoing routine physical checkup. The levels of IgG N-glycosylation in human plasma were quantitatively measured using ultrahigh-performance liquid chromatography. Binomial logistic regression analysis was performed to identify IgG N-glycan biomarkers associated with AS.Results:A total of 14 primary glycans and 13 derived traits showed statistically significant differences between the AS case group and the control group. Binomial logistic regression analysis showed that glycan peak 4, agalactosylated glycans, fucosylated glycans, and fucosylated agalactosylated glycans were positively associated with AS[ OR(95% CI)=1.12(1.01, 1.42), 1.21(1.03, 1.43), 1.48(1.08, 2.03), and 1.27(1.04, 1.55); P=0.036, 0.022, 0.039, 0.020, respectively]. In terms of diagnostic performance, the single glycan GP4 exhibited the largest area under the ROC curve, with an AUC (95% CI) 0.751 (0.677, 0.826), while the combined glycan indicators (GP4+G0+F+FG0) achieved an AUC (95% CI) 0.768(0.697, 0.840). Conclusion:IgG N-glycans have the potentials to serve as candidate biomarkers for AS, and warrants further investigation.

Objective To analyze the influencing factors and prognosis of failed initial invasive mechani-cal ventilation weaning in extremely premature infants.Methods A retrospective analysis was conducted on the clinical data of 143 extremely premature infants who were delivered at Jiangxi Maternal and Child Health Hospital and treated in the neonatal intensive care unit(NICU)from July 2021 to June 2024 and received in-vasive mechanical ventilation within 72 hours after birth.According to whether re-intubation was required within 72 hours after the initial weaning,they were divided into the successful weaning group(n=110)and the failed weaning group(n=33).Stepwise logistic regression was used to analyze the influencing factors and prognosis of failed initial invasive mechanical ventilation weaning.Results There were statistically significant differences between the two groups with different gestational ages at birth,birth weights,tracheal intubation in the delivery room or operating room,abnormal C reactive protein at admission,fraction of inspiration O2(FiO2)at admission,gestational age before weaning from the ventilator,weight before weaning from the vent-ilator,patent ductus arteriosus(PDA,≥2.5 mm),proportion of≥3 tracheal intubation times,invasive me-chanical ventilation time,oxygen supply time,and hospitalization expenses(P<0.05).The results of multiva-riate logistic regression analysis showed that gestational age at birth,abnormal C reactive protein at admis-sion,FiO2 at admission,gestational age before weaning from the ventilator,PDA(≥2.5 mm),duration of in-vasive mechanical ventilation,pulmonary hemorrhage,feeding intolerance,time to total enteral feeding,shock,and length of hospital stay were independent influencing factors for failed initial invasive mechanical ventila-tion weaning(P<0.05).Conclusion Early prevention and early treatment of risk factors are the keys to the successful weaning of extremely premature infants.