Objective To explore the regulatory mechanism of NFE2L2/KEAP1 in alveolar bone repair induced by periodontitis.Methods A rat periodontitis model was established and divided into four groups:Control group(n=6);Periodontitis model group(n=6);Periodontitis+lentivirus empty vector group(n=6);Periodontitis+NFE2L2 overexpression plasmid group(n=6).Histopathological changes in each group were observed using HE staining.TRAP staining was used to detect osteoclast positivity,while ELISA was employed to measure inflammatory cytokine levels in tissues.Immunofluorescence and qPCR were used to detect NFE2L2 expression,and western blot was used to assess the expression of osteogenic proteins ALPL2,RUNX2,and COL1.Primary periodontal ligament cells(hPDLCs)were cultured,and cells were transfected to overexpress NFE2L2 and KEAP1.The cells were divided into six groups:Normal group;Model group;pcDNA-NC group;pcDNA-NFE2L2 group;pc-NFE2L2+pcDNA-NC group;pc-NFE2L2+pcDNA-KEAP1 group.A cellular model was established,and the morphology of primary hPDLCs was observed under a microscope.Cell proliferation was assessed using CCK-8.Osteogenic mineralization was observed using alizarin red staining,and western blot was used to detect osteogenic proteins and autophagy markers.Cell migration was observed using a scratch assay.Results(1)After model induction,redness,swelling of the gums,extensive inflammatory infiltration,and alveolar bone resorption were observed,confirming successful model establishment.Partial tissue recovery occurred after NFE2L2 overexpression via lentivirus.(2)After model induction,osteoclast positivity increased,confirming successful model establishment.Overexpression of NFE2L2 reduced osteoclast positivity(P<0.001).(3)After model induction,levels of IL-1β,IL-10,and TNF-α were significantly higher than in the normal group(P<0.05),confirming successful model establishment.Transfection with NFE2L2 lentivirus reduced inflammatory cytokine levels(P<0.0001).After model induction,osteogenic protein expression decreased compared to the normal group,but overexpression of NFE2L2 increased osteogenic protein expression(P<0.05).(5)LPS treatment significantly reduced cell viability,while NFE2L2 overexpression enhanced it(P<0.0001).(6)LPS treatment reduced calcified nodules,while NFE2L2 overexpression increased them.Addition of pcDNA-KEAP1 reduced mineralized nodules.(7)LPS treatment decreased osteogenic protein expression,while NFE2L2 overexpression increased it.However,addition of pcDNA-KEAP1 reduced osteogenic protein expression(P<0.05).(8)LPS treatment reduced cell migration,whereas NFE2L2 overexpression enhanced it(P<0.0001).(9)Expression of autophagy markers decreased after LPS treatment,but increased after transfection with NFE2L2 plasmid.However,addition of pcDNA-KEAP1 reduced the expression of autophagy markers(P<0.05).Conclusion This study identified the regulatory role of NFE2L2/KEAP1 in periodontitis,providing a scientific basis for the treatment of periodontitis.

OBJECTIVE To compare the short-term efficacy of autologous frozen tragus perichondrium and fresh perichondrium in repairing tympanic membrane perforations,and to explore the clinical application value of autologous frozen tragus perichondrium.METHODS Twenty-five patients with bilateral tympanic membrane perforations from March 2021 to October 2023 were selected,including 7 males and 18 females.Bilateral tympanoplasty was completed in stages.The initial operation was set as the control group,in which the ventral perichondrium of the tragus was used for tympanoplasty,and the dorsal perichondrium of the tragus was reserved and stored at-80℃ultra-low temperature in a sterile container.The second operation was set as the observation group,in which the thawed frozen perichondrium was used for contralateral tympanoplasty.The differences in the healing rate of tympanic membrane,postoperative hearing,operation time and surgical bleeding volume were compared between the two groups.RESULTS All patients were followed up for three months.The success rate of tympanic membrane healing in the observation group was 96%(24/25),and that in the control group was 92%(23/25).There was no statistically significant difference between the two groups(χ2=0.36,P>0.05).The operation time and surgical bleeding volume of patients in the observation group were lower than those in the control group[(48.64±4.64)min vs.(67.92±5.69)min,(5.32±1.54)ml vs.(9.65±1.73)ml],and the differences were statistically significant(t=13.93,t=12.09,P all<0.05).The postoperative air conduction hearing thresholds and air-bone conduction gap of the two groups of patients were lower than those before operation(all P<0.05).There was no difference in air conduction hearing threshold,bone conduction hearing threshold and air-bone conduction difference between the groups(all P>0.05).CONCLUSION The application of autologous frozen tragus perichondrium has effectively shortened the operation time of the contralateral ear,avoided the trauma caused by taking materials again,and has the characteristics of minimally invasive and high efficiency.The method is feasible and the curative effect is accurate.

Animal quarantine is essential for controlling the spread of animal diseases,ensuring the healthy development of the livestock industry,and safeguarding public health and food safety.This paper reviews and organizes the development history,achievements,and current dilemmas of China's animal quarantine.It proposes further policy reflections and suggestions from the aspects of man-agement concepts,system design,and specific implementation to adapt to the requirements of the new era.

Animal quarantine is essential for controlling the spread of animal diseases,ensuring the healthy development of the livestock industry,and safeguarding public health and food safety.This paper reviews and organizes the development history,achievements,and current dilemmas of China's animal quarantine.It proposes further policy reflections and suggestions from the aspects of man-agement concepts,system design,and specific implementation to adapt to the requirements of the new era.

Objective To explore the effect and mechanism of uric acid(UA)in regulating the larval growth and development of Drosophila melanogaster.Methods A total of 1350 newly hatched first-instar larvae of wild-type Drosophila melanogaster(W11 18)were collected,and the Drosophila melanogaster model of hyperurice-mia was constructed with a high purine diet.The larvae were assigned into three groups(n=150):control(stand-ard corn meal medium),low-dose adenine(corn meal medium containing 0.05％adenine),and high-dose ad-enine(corn meal medium containing 0.10％adenine),and two parallel groups were set up.The growth and de-velopment of larvae in each group was observed,and the UA and hormone levels were measured.In addition,the expression levels of genes involved in growth and development were determined.Results Compared with the con-trol group,the low-and high-dose adenine groups showed elevated UA levels(both P＜0.001)and prolonged de-velopmental period(P=0.024,P＜0.001).The high-dose adenine group showed decreased survival rate,pupa-tion rate,and eclosion rate and elevated levels of juvenile hormone(JH)and 20-hydroxyecdysone(20E)(all P＜0.001).The PCR results showed that compared with the control group,high-dose adenine upregulated the mRNA levels of reactive oxygen species(ROS),forkhead box O(FOXO),and mammalian target of rapamycin(mTOR)while downregulating the mRNA levels of Sestrin,mTOR complex 1(mTORC1),and AMP-activated protein kinase(all P＜0.001).Conclusion High concentrations of UA may promote the expression of ROS/FOXO/mTORC1/mTOR signaling pathway by regulating the levels of JH and 20E,thereby inhibiting the larval growth and development of Drosophila melanogaster.

Objective:Investigate the characteristics of hospitalized patients infected with parvovirus B19 (B19V) and the potential impact of B19V on disease progression and clinical outcomes in patients.Methods:The subjects of this observational retrospective study were 62 hospitalized patients who tested positive for B19V among 3 129 inpatients who visited Peking University People′s Hospital between April 1, 2022, and August 31, 2023, and underwent metagenomic next-generation sequencing (mNGS) for pathogens. Patient data regarding pathogen detection, clinical characteristics, and outcomes were collected, along with other laboratory test results, to assess hospitalization duration, incidence of mixed infections during hospitalization, and clinical outcomes within 30 days. The chi-square test was utilized to analyze differences in patient mortality rates and other relevant factors.Results:B19V infection waspobserved in patients with hematological disorders and solid organ transplant recipients (79.03%, 49/62). Additionally, B19V frequently coexists with other pathogens (40/62), with the highest rates of mixed infections involving human herpesvirus 6, cytomegalovirus, and Epstein-Barr virus, having 20, 9, and 8 cases respectively. The most common site of B19V detection is peripheral blood (χ 2=0.000 3, P<0.001), although it can also be detected in other bodily fluid samples. The 30 day mortality rate among B19V infected patients was 19.35% (12/62) 10 out of 12 deceased patients had concurrent mixed infections with other pathogens. Moreover, patients with mixed B19V infections exhibited a higher mortality rate compared to those with B19V infection alone (χ 2=4.546, P=0.033). Comparative analysis of 24 patients who underwent multiple mNGS tests revealed that patients with chronic B19V infection (12 cases) had a higher mortality rate compared to those with transient B19V infection (12 cases) (χ 2=12, P<0.001). Conclusions:Parvovirus B19 (B19V) infection is more commonly observed in patients with hematological disorders and solid organ transplant recipients, often coexisting with other pathogens. The mortality rate within 30 days of B19V infection is elevated, particularly when accompanied by concurrent mixed infections with other pathogens. Moreover, patients with chronic and persistent B19V infection face an increased risk of mortality.

Approximately 140 million people worldwide are homozygous carriers of APOE4 (ε4), a strong genetic risk factor for late onset familial and sporadic Alzheimer's disease (AD), 91% of whom will develop AD at earlier age than heterozygous carriers and noncarriers. Susceptibility to AD could be reduced by targeted editing of APOE4, but a technical basis for controlling the off-target effects of base editors is necessary to develop low-risk personalized gene therapies. Here, we first screened eight cytosine base editor variants at four injection stages (from 1- to 8-cell stage), and found that FNLS-YE1 variant in 8-cell embryos achieved the comparable base conversion rate (up to 100%) with the lowest bystander effects. In particular, 80% of AD-susceptible ε4 allele copies were converted to the AD-neutral ε3 allele in human ε4-carrying embryos. Stringent control measures combined with targeted deep sequencing, whole genome sequencing, and RNA sequencing showed no DNA or RNA off-target events in FNLS-YE1-treated human embryos or their derived stem cells. Furthermore, base editing with FNLS-YE1 showed no effects on embryo development to the blastocyst stage. Finally, we also demonstrated FNLS-YE1 could introduce known protective variants in human embryos to potentially reduce human susceptivity to systemic lupus erythematosus and familial hypercholesterolemia. Our study therefore suggests that base editing with FNLS-YE1 can efficiently and safely introduce known preventive variants in 8-cell human embryos, a potential approach for reducing human susceptibility to AD or other genetic diseases.
Humans ; Apolipoprotein E4/genetics* ; Cytosine ; Mutation ; Blastocyst ; Heterozygote ; Gene Editing ; CRISPR-Cas Systems

Objective To explore the effect of eversion carotid endarterectomy(eCEA)on the cog-nitive function in elderly carotid artery stenosis patients with different ages.Methods A total of 56 elderly patients undergoing eCEA in Department of Vascular Surgery of No.920 Hospital of Joint Logistics Support Force from May 2019 to May 2022 were enrolled and divided into a 60-69 year old group(31 cases)and a 70-80 year old group(25 cases).Mini-mental state examination(MMSE)Scale was used to analyze the patients within 1 week before surgery and 1 and 6 months after surgery.Results In the 60-69 years old group,their MMSE scores in 1 and 6 months after surgery were significantly higher than those before surgery(24.71±3.67 vs 23.52±3.70,P＜0.05;25.48±3.19 vs 23.52±3.70,P＜0.01).For the 70-80 year old group,the MMSE score in 6 months after surgery was obviously higher than that before surgery and that in 1 month after sur-gery(25.44±3.42 vs 23.76±3.81,P＜0.01;25.44±3.42 vs 23.90±3.65,P＜0.01).The improve-ment of MMSE score between 1 month after surgery and before surgery was notably more obvi-ous in the 60-69 year old group than the 70-80 year old group,but the improvement between 6 months and 1 month after surgery was statistically declined in the 60-69 year old group than the 70-80 year old group(P＜0.05).Conclusion eCEA can significantly improve cognitive function in elderly patients with carotid artery stenosis,and obvious efficacy is observed in the early stage in the 60-69 years old patients.

OBJECTIVE To sum marize the pro cedure and informatization construction of centralized volume-based procurement（VBP）in our hospital ，in order to give references for normal development of centralized VBP. METHODS The standardized workflow system was established ，including using flow chart method to establish standardized workflow ，carrying out procedure training and inspection of procedure implementation ， and continuously conducting procedure optimization. The information system was developed for the task links that needed a lot of calculation to improve the automation level of information processing. RESULTS & CONCLUSIONS Eight specific work procedures were established in our hospital ，including the work procedures of submitting the demand data of centralized VBP and the implementation of centralized VBP ，and has taken measures such as procedure training ，establishing supervision and inspection system and using auxiliary means to promote the implementation of the procedure ，so as to optimize the procedure and work form. An informatization platform for the clinical task allocation of the agreed purchase quantity of centralized VBP and a supervision platform for the daily use of VBP were also established in our hospital，then the two tasks with a large amount of calculation could be finished. Standardized workflow system and informatization platform construction has improved the operation and supervision efficiency of centralized VBP in our hospital ，ensured the completion of centralized purchase tasks and saved human resources ，which has a certain promotion value.

Objective:To predict the pathogens of bloodstream infection (BSI) in hematopoietic stem cell transplantation (HSCT) patients by plasma microbial cell-free DNA (mcfDNA) sequencing with and without additional amplification.Methods:A total of 978 HSCT patients were enrolled in Peking University People′s Hospital from March to July 2021, and the 7 428 blood samples were prospectively collected from pretransplant conditioning period to 4 months after transplantation. The plasma samples were separated and then cryopreserved. According to blood culture results and whether there were plasma samples before BSI onset, twenty-eight HSCT patients with positive blood culture (39 plasma samples within 1-8 days before BSI onset) and 9 HSCT patients with negative blood culture (9 plasma samples) were filtered. The 39 samples were performed with mcfDNA additional and non-additional amplification sequencing, and the 9 samples were only performed with additional amplification sequencing. With the blood culture results as the gold standard, the consistency between the sequencing and the blood culture results was observed. Student t test and Wilcoxon test were used for statistical analysis. Results:Without additional amplification sequencing, only 7 samples sequencing results were consistent with the blood culture results, and the total pathogen detection rate was 17.95% (7/39). The rates within 3 days and 4-8 days were 23.81% (5/21) and 2/18, respectively. The main pathogenic type detected was gram-negative bacteria (5/7). With additional amplification sequencing, the total pathogen detection rate was 59.26% (16/27) and the rate within 3 days was 8/13. The number of gram-positive bacteria detected was elevated (13/16) and the number of additional microorganisms in additional amplification sequencing was increased significantly ( P=0.001 0), compared with non-additional amplification sequencing. Moreover, additional sequencing analysis of 9 samples from patients with negative culture result showed that no pathogen was detected in six samples, and the common Torque teno virus in HSCT patients was detected in only three samples. Conclusion:The pathogen detection rate of plasma mcfDNA additional amplification sequencing was better than that of non-additional amplification sequencing in HSCT patients before BSI onset, especially in the first three days, which has the potential to predict BSI pathogens.