Objective To investigate the characteristics and differences between two type 2 diabetic model mice,db/db and KK-Ay,as models of diabetic nephropathy(DN),and to verify them with irbesartan treatment.Methods A total of 102 db/db and 102 KK-Ay mice(12 weeks old)were screened based on the albumin-to-creatinine ratio(ACR)and 24-hour urinary total protein(UCFP),with 78 mice of each strain selected and divided into 6 groups.The db/db+irbesartan group and KK-Ay+irbesartan group were administered irbesartan(40 mg/kg),the db/db group and KK-Ay group received an equal volume of drinking water for experimental animals,and the db/db+test article A groups and KK-Ay+test article A groups were given different doses of test article A(only the result of mortality data in the test articles groups were presented in the study,while other result were not shown).Wild-type C57BL/6J(WTa/WTb group)served as the normal control and were given experimental animal drinking water.Daily clinical observations were made,body mass was measured weekly,and ACR and UCFP were monitored periodically.At the end of administration,the mice were dissected,and the brain,liver,lungs and kidneys were collected,and the organ coefficients(organ mass× 1000/body mass)were calculated.Blood was collected to measure serum total protein(TP),albumin(Alb),blood urea nitrogen(BUN),glucose(GLU),triglycerides(TG),and cholesterol(CHO).Kidney tissues were stained with HE,MASSON,and PAS to observe histopathological changes.Results Among the 78 KK-Ay mice,5 died over 8 weeks,while among the 78 db/db mice,21 died over 6 weeks.ACR increased with age in both the db/db and KK-Ay groups but decreased in the db/db+irbesartan and KK-Ay+irbesartan groups,though without statistical significance(P＞0.05).UCFP increased with age in the KK-Ay group but significantly decreased after irbesartan treatment(P＜0.05).Compared with the WT groups,the kidney coefficients in the db/db and KK-Ay groups showed no significant changes(P＞0.05),while the brain liver coefficients decreased significantly(P＜0.01)and the liver coefficients increased significantly(P＜0.01),but they did not change after irbesartan treatment(P＞0.05).Compared with the WT groups,GLU and TG levels were significantly increased in the db/db and KK-Ay groups(P＜0.01).Compared with the KK-Ay group,GLU and TG levels were significantly reduced in the KK-Ay+irbesartan group(P＜0.01).In the db/db and KK-Ay groups,kidney tissues showed thickening of the glomerular mesangial matrix and mesangial cell proliferation,which were alleviated after irbesartan treatment.Conclusions Both 12-week-old db/db and KK-Ay mice can serve as models for DN research.Compared with db/db mice,KK-Ay mice exhibited smaller individual differences in ACR,fewer animal deaths after urine collection,and significant improvements in GLU,TG and UCFP after irbesartan treatment.

Objective:To report the nationwide surveillance results of pathogenic profiles and antimicrobial resistance patterns of Gram-positive bloodstream infections in China in 2023.Methods:The clinical isolates of Gram-posttive bacteria from blood cultures were collected in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）during January to December 2023. Antimicrobial susceptibility testing was performed using the dilution method recommended by the Clinical and Laboratory Standards Institute（CLSI）. Statistical analyses were conducted using WHONET 5.6 and SPSS 25.0 software.Results:A total of 4 385 Gram-positive bacterial isolates were obtained from 60 participating center. The top five pathogens were Staphylococcus aureus（ n=1 544，35.2%），coagulase-negative Staphylococci（ n=1 441，32.9%）， Enterococcus faecium（ n=574，13.1%）， Enterococcus faecalis（ n=385，8.8%），and α-hemolytic Streptococci（ n=187，4.3%）. The prevalence of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）was 26.2%（405/1 544）and 69.8%（1 006/1 441），respectively. Notably，all Staphylococci remained susceptible to glycopeptide or daptomycin. Staphylococcus aureus demonstrated excellent susceptibility（>97.0%）to cephalobiol，rifampicin，trimethoprim-sulfamethoxazole，linezolid，minocycline，tigecycline，and eravacycline. No Enterococcus exhibiting resistance to linezolid were detected. Glycopeptide resistance was uncommon but more frequent in Enterococcus faecium（resistance to vancomycin and teicoplanin：both 1.7%）compared to Enterococcus faecalis（both 0.3%）. The detection rates of MRSA and MRCNS exhibited significant regional variations across the country（ χ2=17.674 and 148.650，respectively，both P<0.001）. No vancomycin-resistant Enterococci were detected in central China. Institutional comparison demonstrated higher prevalence of MRSA（ χ2=14.111， P<0.001）and MRCNS（ χ2=4.828， P=0.028）in provincial hospitals than that in municipal hospitals. Socioeconomic analysis identified elevated detection rates of both MRSA（ χ2=18.986， P<0.001）and MRCNS（ χ2=4.477， P=0.034）in less developed regions（per capita GDP

Objective:To report the results of bacterial resistant investigation collaborative system（BRICS）on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2023，and provide reference for clinical tretment of bloodstream infections and prevention and control of bacterial resistance.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of BRICS were collected during January 2023 to December 2023. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 were used to analyze the data.Results:During the study period，11 492 strains of Gram-negative bacteria were collected from 60 hospitals，of which 10 098（87.9%）were Enterobacterales and 1 394（12.1%）were non-fermentative bacteria. The top 5 bacterial species were Escherichia coli（50.0%）， Klebsiella pneumoniae（26.1%）， Pseudomonas aeruginosa（5.1%）， Acinetobacter baumannii complex（5.0%）and Enterobacter cloacae complex（4.1%）. The ESBL-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus mirablilis were 46.8%（2 685/5 741），18.3%（549/2 999）and 44.0%（77/175），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（76/5 741）and 15.0%（450/2 999）；32.9%（25/76）and 78.0%（351/450）of CREC and CRKP were sensitive to ceftazidime/avibactam combination，respectively. 94.7%（72/76）and 90.2%（406/450）of CREC and CRKP were sensitive to aztreonam/avibactam combination. Furthermore，57.9%（44/76）and 79.1%（356/450）were sensitive to imipenem/relebactam combination. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 64.6%（370/573），while more than 80.0% of CRAB complex was sensitive to tigecycline，eravacycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 17.0%（99/581）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of important Gram-negative bacteria resistance among different regions in China，with statistically significant differences in the prevalence of CREC，CRKP，CRPA and CRAB complex（ χ2=10.6，28.6，10.8 and 19.3， P<0.05）. The prevalence of ESBL-producing Escherichia coli， CREC，CRAB complex and CRKP were higher in provincial hospitals than those in municipal hospitals（ χ2=12.5，9.8，12.7 and 57.8，all P<0.01）. Conclusions:Gram-negative bacteria are the main pathogens causing bloodstream infections in China，and Escherichia coli is ranked in the top，while the trend of Klebsiella pneumoniae increases continuously with time. CRKP infection shows a slow upward trend，CREC infecton maintains a low prevalence level，and CRAB complex infection continues to exhibit a high prevalence rate. The composition and resistance patterns of pathogens causing bloodstream infections vary to some extent across different regions and levels of hospitals in China.

Objective:To explore the impact of the two-way referral model on compliance and prognosis in patients with heart failure.Methods:This bidirectional cohort study enrolled chronic heart failure (CHF) patients treated at Qilu Hospital of Shandong University or designated primary hospitals between March 2018 and March 2022. Patients were categorized into two groups based on referral status: two-way referral group (participating in the referral model with≥1 follow-up visit at primary hospitals) and the core hospital group (receiving treatment and follow-up exclusively at Qilu Hospital). Baseline clinical characteristics were collected and compared between groups. Patients underwent followed-up, with primary endpoints including follow-up rate, drug (β-blockers, angiotension converting enzyme inhibitor (ACEI)/angiotensin Ⅱ receptor blockers (ARB)/angiotensin receptor-neprilysin inhibitor (ARNI), sodium-glucose cotransporter 2 inhibitors and mineralocorticoid receptor antagonists) utilization rate and target dose achievement rate. Secondary endpoints encompassed changes from baseline in left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDd), and N-terminal pro-brain natriuretic peptide (NT-proBNP), plus cardiovascular mortality and heart failure rehospitalization. Generalized linear mixed models analyzed longitudinal trends in LVEF, LVEDd, and NT-proBNP levels. Kaplan-Meier curves and Cox regression evaluated LVEF recovery rates, supplemented by subgroup analyses. Multivariate logistic regression was used to identify factors influencing target dose achievement rate for β-blockers and ACEI/ARB/ARNI therapies in CHF patients.Results:A total of 357 patients were enrolled, aged 53 (41, 63) years, including 256 males (71.7%). 157 patients were in the two-way referral group and 200 patients in the core hospital-treated group. Compared with the core hospital-treated group, the two-way referral group had lower baseline LVEF (28 (22, 34)% vs. 31 (23, 36)%, P=0.021) and systolic blood pressure (116 (104, 125) mmHg vs. 121 (109, 134) mmHg (1 mmHg=0.133 kPa), P=0.010). The 12-month follow-up rate of the two-way referral group was higher than the core hospital-treated group (73.8% vs. 56.0%, P=0.004). No significant between-group differences were observed in drug utilization rate of β-blockers, ACEI/ARB/ARNI, or sodium-glucose cotransporter 2 inhibitors during follow-up (all P>0.05), while mineralocorticoid receptor antagonists use showed a declining trend in both groups. Although the core hospital-treated group had higher target dose achievement rates for β-blockers (65.4% vs. 49.3%, P=0.042) and ACEI/ARB/ARNI (79.8% vs. 65.8%, P=0.046) than the two-way referral group, multivariate logistic regression indicated that the two-way referral model was not a negative predictor for these outcomes (all P>0.05). Both groups showed improved NT-proBNP, LVEDd, and LVEF from baseline (all P<0.001) with no significant difference in trends between groups (all P>0.05). There was no significant difference in the composite incidence (7.6% vs. 6.5%, P=0.674) and cumulative incidence (log-rank P=0.684) of cardiovascular death and heart failure rehospitalization at 12 months between two groups. Conclusion:The two-way referral model demonstrates advantages in improving medication adherence, drug utilization rates, and targetdoseachievement rates among CHF patients. This model not only promotes cardiac functional recovery but also reduces risks of cardiovascular mortality and heart failure rehospitalization, achieving comparable therapeutic and management outcomes to those observed in core hospital-treated patients.

Objectives:To explore the urban-rural differences in the association between the Life's Essential 8(LE8)scores and the risk of new-onset cardiometabolic diseases(CMD)among adult Chinese.Methods:A total of 4 719 adults aged 18 years or older without CMD from the China Health and Nutrition Survey(CHNS)cohort from 2009 to 2015 were enrolled,including 1 909 in urban areas and 2 810 in rural areas.CMD included myocardial infarction,stroke,and diabetes.The LE8 score included diet,physical activity,sleep,nicotine exposure,body mass index(BMI),non-high-density lipoprotein cholesterol(non-HDL-C),blood glucose,and blood pressure.The score of each LE8 factor ranged from 0 to 100 points,and the average of the sum of the eight scores was the final score.The final LE8 score was divided into a low group(<67.5 points),a medium group(67.5-77.5 points),and a high group(>77.5 points)according to the tertile of the final LE8 score.There were 780,564,and 565 cases in the low,medium,and high LE8 score groups in urban areas,respectively.There were 838,934,and 1 038 cases in the low,medium,and high LE8 score groups in rural areas.The primary outcome was the number of participants newly diagnosed with diabetes,stroke,and/or myocardial infarction during the follow-up period.The Cox proportional hazards regression model was used to analyze the urban-rural difference in the association between LE8 and the risk of new-onset CMD.Results:The total cardiovascular health LE8 scores in rural areas was higher than that in urban residents(72.9 points vs.70.2 points,P<0.01).During the average follow-up period([5.86±0.74]years),there were 176 new cases of CMD,including 67 cases in urban areas and 109 cases in rural areas.Multivariate Cox analysis showed that in the fully adjusted model,compared with the urban-higher group,the rural-lower group(HR=1.65,95%CI:1.27-2.15,P=0.001)had the greatest increased risk of new CMD,followed by the urban-lower group(HR=1.49,95%CI:1.01-2.21,P=0.046),the rural-middle group(HR=1.42,95%CI:1.06-1.90,P=0.020),and the rural-higher group(HR=1.16,95%CI:1.01-1.33,P=0.032).Multivariable Cox regression results showed that in urban areas,compared with those with LE8 scores≥70 points,those with LE8 scores<70 points and with unfavorable BMI score(HR=2.70,95%CI:1.51～4.84,P<0.01),blood glucose scores(HR=2.09,95%CI:1.24-3.51,P<0.01),and blood pressure score(HR=3.73,95%CI:1.69-8.24,P<0.01)faced increased risk of CMD;in rural areas,compared with≥70 points group,<70 points group with unfavorable BMI score(HR=3.92,95%CI:2.43-6.30,P<0.01),non-HDL-C score(HR=2.96,95%CI:1.94-4.52,P<0.01),blood glucose score(HR=2.59,95%CI:1.71-3.94,P<0.01)and blood pressure score(HR=1.85,95%CI:1.18-2.92,P<0.01)faced increased risk of CMD.The risk of CMD associated with BMI,blood glucose,and non-HDL-C scores<70 points was higher in rural areas than in urban areas.Conclusions:Although rural residents have slightly higher LE8 scores than urban residents,the risk of CMD associated with lower LE8 scores is particularly significant in rural areas.Strengthening the monitoring and primary prevention of cardiovascular health among rural residents is of great significance to reduce the burden of CMD in China.

Objective:To describe the trend of changes in the disease burden of age-related hearing loss in China and globally from 1990 to 2021,to forecast the prevalence and years lived with disability(YLD)rates of age-related hearing loss from 2022 to 2036,and to provide a reference for the prevention and control of the disease burden associated with age-related hearing loss.Methods:Using the Global Burden of Disease 2021(GBD2021)data,this study selected age-standardized prevalence rate(ASPR)and YLD as indicators.The disease burden and long-term trends of age-related hearing loss in China and globally from 1990 to 2021 were described by different socio-demographic index(SDI)and gender.Joinpoint regression was used to calculate the average annual percent change(AAPC)to assess the trend changes in the disease burden.Decomposition analysis was applied to explore the relative im-pacts of aging,population growth,and epidemiological changes on the variation in disease burden.An autoregressive integrated moving average(ARIMA)model was used to forecast the age-standardized pre-valence rate and YLD rates from 2022 to 2036.Results:The prevalence of age-related hearing loss in China in 2021 was 82 162.49(73 288.08-89 187.21)per 100 000,higher than the global SDI level of 66 238.16(59 982.54-72 669.82)per 100 000,the high SDI region's level of 57 650.42(52 059.12-63 889.02)per 100 000,the upper-middle SDI region's level of 69 115.59(62 494.18-75 340.64)per 100 000,the middle SDI region's level of 72 365.56(65 181.43-78 912.01)per 100 000,the lower-middle SDI region's level of 64 439.66(58 368.22-71 468.27)per 100 000,and the low SDI region's level of 61 725.25(55 749.18-68 477.67)per 100 000.The age-related hearing loss YLD rate in China was 2 762.98[95％uncertainty interval(UI):1 855.28-3 880.68]per100000,higher than the global SDI level of 2 236.75(95％UI:1 511.56-3 155.88)per 100 000,the high SDI region's level of 1 805.79(95％UI:1 212.69-2 577.17)per 100 000,the upper-middle SDI re-gion's level of 2 316.58(95％UI:1 557.53-3 274.87)per 100 000,the middle SDI region's level of 2 480.99(95％UI:1 678.17-3 489.24)per 100 000,the lower-middle SDI region's level of 2 313.28(95％UI:1 578.35-3 271.50)per 100 000,and the low SDI region's level of 2 383.55(95％UI:1 623.66-3 365.68)per 100 000.From 1990 to 2021,both the prevalence and YLD rate of age-related hearing loss in China showed an increasing trend,rising by an average of 0.18％(95％CI:0.16％-0.19％)and 0.29％(95％CI:0.27％-0.30％)per year,respectively.The rates of in-crease in prevalence were the same for both men and women,with men showing a 0.18％increase(95％CI:0.17％-0.19％,P＜0.001)and women showing a 0.18％increase(95％CI:0.16％-0.19％,P＜0.001).However,the YLD rate increase was faster in men than in women,with men experiencing a 0.32％increase(95％CI:0.27％-0.37％,P＜0.001)and women experiencing a 0.27％increase(95％CI:0.26％-0.28％,P＜0.001).Decomposition analysis showed that population growth was the main factor driving the increase in prevalence and YLD rate globally and across different SDI regions.However,aging was the primary factor contributing to the increase in prevalence and YLD rate in China.ARIMA model predictions suggested that the prevalence and YLD rate of age-related hearing loss would continue to rise from 2022 to 2036,with the predicted prevalence and YLD rate in 2036 reaching 89 723.99 per 100 000 and 2 872.47 per 100 000,respectively.Conclusion:The prevalence and disease burden of age-related hearing loss in individuals aged 60 and above in China rank first globally.From 1990 to 2021,both the prevalence and YLD rate of age-related hearing loss have shown a continuous upward trend,consistently surpassing the levels observed in various SDI regions worldwide.The prevalence and disease burden of age-related hearing loss are particularly significant among elderly men.Moreover,projections indicate that the disease burden of age-related hearing loss will continue to rise over the next 15 years.Therefore,it is urgent to pay close attention to age-related hearing loss in this specific population,and early intervention measures are crucial to reduce the disease burden associated with age-related hearing loss.

Objective To analyze the mutation status of Kirsten rat sarcoma viral oncogene homolog(KRAS),phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA),v-raf murine sarcoma viral oncogene homolog B1(BRAF)genes,and the expression of mismatch repair(MMR)and human epidermal growth factor receptor 2(HER-2)proteins in tumor tissues of patients with colorectal cancer(CRC)harboring neuroblastoma rat sarcoma viral oncogene homolog(NRAS)gene mutations,and explore their relationships with the clinicopathological characteristics of CRC patients.Methods The clinicopathological data of 546 patients with NRAS mutation CRC were retrospectively analyzed.The mutation status of NRAS,KRAS,PIK3CA,and BRAF genes was detected by AmoyDx amplification refractory mutation system(ARMS)-polymerase chain reaction(PCR)kit(fluorescent PCR method),the expression levels of MMR and HER-2 proteins were detected by immunohistochemical staining EnVision method,and the relationship between them and the clinicopathological characteristics of patients were analyzed.Results The mutation rate of single-point mutations in the NRAS gene was 98.35%(537/546),double-point mutations in the NRAS gene were 1.65%(9/546),and double mutations in the NRAS and KRAS genes were 1.47%(8/546).No patients were found to harbor mutations in the PIK3CA or BRAF genes.The types of NRAS mutations included Q61R(or Q61K,Q61L,Q61H)mutations(266/546,48.72%),G12D(or G12S)mutations(154/546,28.21%),G13R(or G12C,G12V,G12A,G13V)mutations(134/546,24.54%),and A146T mutation(1/546,0.18%).G13R(or G12C,G12V,G12A,G13V)mutations in the NRAS gene were more likely to occur in the rectum cancer patients(P=0.035);although the tumors had a larger diameter(P=0.029),the patients had a longer progression-free survival after surgery(P=0.028).Among patients with NRAS gene mutations,HER-2 positive expression was associated with perineural invasion(P=0.003),and the patients with deficient MMR were younger on average(P=0.041)and were associated with double-point mutations in the NRAS gene(P=0.018).Conclusion CRC harboring NRAS mutations may have unique clinicopathological characteristics and molecular phenotypes,providing possibilities for individualized treatment and prognosis evaluation of CRC.

Sarin(GB)is a typical representative of nerve agents with high toxicity,and very low amount can cause death.GB can cause water and atmospheric environment poisoning,so the detection of GB in water and air is of great significance.In this work,a colorimetric sensor array(CSA)based on GB inhibition of cholinesterase activity was constructed to detect GB with high selectivity.A 4×4 colorimetric array was constructed using acetylcholinesterase(AChE),butyryl cholinesterase(BuChE)and the corresponding substrate acetylthiocholine iodide(S-ACh),butyryl thiocholine iodide(S-BCh),acetylcholine chloride(ACh),butyryl choline chloride(BCh)and 2,6-dichloroindophenol ethyl ester(DCIE).The linear curve of the sensor was Y=131.3×lgC+271.6(R2=0.997),where Y was the array response Euclidean distance,C was the concentration of GB(mg/L),the linear range was 0.03?0.32 mg/L,and the detection limit was 27.6 μg/L.The method could effectively distinguish chemical warfare agents(CWA)such as VX,Soman(GD),mustard gas(HD),Louie reagent(L),and had high anti-interference ability,sensitivity and good repeatability.It was successfully applied to the detection of GB in simulated water and simulated air samples,and the sample recovery rate was 97.2% ?100.9%.This method would be potentially applied to the field rapid detection of nerve agents.

Lentiviral vectors(LVs)are key gene delivery tools for integrating target genes into the host genome,but they may also pose risks of insertional mutagenesis.The vector copy number(VCN)in cells is critical for determining the safety of gene modification.However,the reliability and accuracy of its quantification process are influenced by multiple factors.Developing cell reference materials with specific vector copy numbers represents a viable approach to enhance the reliability and consistency of measurement results,enabling quality control of the quantification process and traceability of outcomes.However,the preparation of such reference materials faces challenges in cell sample design,preparation protocols,and advanced quantification techniques.In this study,T lymphocyte cell line Jurkat-based reference materials with LV gene copy numbers of 1 and 2 copy/cell were developed.A high-precision duplex digital polymerase chain reaction(dPCR)method was established to quantify the LV gene and endogenous genes simultaneously.Additionally,the results of dPCR were cross-validated through next-generation sequencing and flow cytometric analysis.Ultimately,confocal microscopy characterization results showed that the developed cell reference materials had intact morphology.The quantification result of VCN-1 was(1.07±0.11)copy/cell,and that of VCN-2 was(2.09±0.21)copy/cell.These cell reference materials demonstrated compliance with stability and homogeneity requirements,and could be applied for quality control throughout the VCN measurement workflow and metrological traceability,improving the accuracy,comparability,and validity of copy number measurements.

Objective To establish a rapid screening and analysis method for etomidate and its ana-logues using a portable mass spectrometer equipped with a thermal desorption-atmospheric pressure chemical ionization source-linear ion trap.Methods A 10 μL aliquot of a standard solution at a con-centration of 1 μg/mL was taken,and after the solvent evaporated,the sample was inserted into the in-let of the portable mass spectrometer for detection.By adjusting the collision-induced dissociation pa-rameters,the molecular ion peak and fragment ion peak information of the standard were obtained and used to establish a reference database.In addition,the method was applied to 29 seized liquid and plant samples.Results A screening system for etomidate and its analogues was established based on the portable mass spectrometer and the corresponding mass spectrometry library.The system enables qualitative screening analysis by identifying primary protonated molecular ions and secondary product ions of etomidate and its analogues.The limits of detection for etomidate and its 12 analogues ranged from 0.1 to 10 μg/mL.Etomidate and its analogues were detected in all 29 liquid and plant samples.However,this method could not distinguish between isomeric imidazole esters,such as isopropoxate and propoxate.Additionally,when testing 2-SH-etomidate,there was a false positive for the detection of etomidate.Conclusion This study established a rapid screening method for etomidate and its ana-logues using a portable mass spectrometer.The method combines the high sensitivity of mass spectrome-try with the on-site applicability of portable devices,significantly improving detection efficiency and meeting the on-site detection needs of etomidate and its analogues.