1.Identification and Analysis of bHLH Genes Related to Color Formation of Gastrodia elata Stem
Xue JIANG ; Dandan RAN ; Xiuwen WANG ; Xiaobo ZHANG ; Xiaohong OU ; Jie PAN ; Tao ZHOU ; Zhen OUYANG ; Jiao XU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):202-209
ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently, the gene family members were subject to analysis, including gene structure, chromosomal localization, cis-acting elements, gene synteny, and phylogeny. Combined with transcriptome data and quantitative Real-time PCR, the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally, correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca, unevenly distributed across 17 chromosomes and clustered into 16 subfamilies, with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes, 12 bHLH genes (such as bHLH62-3 and bHLH74) were associated with the bright yellow color of G elata f. elata stem, while 9 bHLH genes (such as PIL13, UNE12, and bHLH130) were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca, the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata, and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes, 21 bHLH genes participate in the coloration metabolic pathway regulation of stems, flowers, and fruits. Specifically, bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata, thus relevant to the color formation of stem. Additionally, GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.
2.Identification and Analysis of bHLH Genes Related to Color Formation of Gastrodia elata Stem
Xue JIANG ; Dandan RAN ; Xiuwen WANG ; Xiaobo ZHANG ; Xiaohong OU ; Jie PAN ; Tao ZHOU ; Zhen OUYANG ; Jiao XU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):202-209
ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently, the gene family members were subject to analysis, including gene structure, chromosomal localization, cis-acting elements, gene synteny, and phylogeny. Combined with transcriptome data and quantitative Real-time PCR, the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally, correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca, unevenly distributed across 17 chromosomes and clustered into 16 subfamilies, with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes, 12 bHLH genes (such as bHLH62-3 and bHLH74) were associated with the bright yellow color of G elata f. elata stem, while 9 bHLH genes (such as PIL13, UNE12, and bHLH130) were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca, the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata, and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes, 21 bHLH genes participate in the coloration metabolic pathway regulation of stems, flowers, and fruits. Specifically, bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata, thus relevant to the color formation of stem. Additionally, GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.
3.Evaluation of the accuracy of three-dimensional data acquisition from liquid- interference surfaces assisted by a scanner head with a compressed airflow system.
Xinkai XU ; Jianjiang ZHAO ; Sukun TIAN ; Zhongning LIU ; Xiaoyi ZHAO ; Xiaobo ZHAO ; Tengfei JIANG ; Xiaojun CHEN ; Chao MA ; Yuchun SUN
Journal of Peking University(Health Sciences) 2025;57(1):121-127
OBJECTIVE:
To quantitatively evaluate the accuracy of data obtained from liquid-interference surfaces using an intraoral 3D scanner (IOS) integrated with a compressed airflow system, so as to provide clinical proof of accuracy for the application of the compressed airflow system-based scanning head in improving data quality on liquid-interference surfaces.
METHODS:
The study selected a standard model as the scanning object, adhering to the "YY/T 1818-2022 Dental Science Intraoral Digital Impression Scanner" guidelines, a standard that defined parameters for intraoral scanning. To establish a baseline for accuracy, the ATOS Q 12M scanner, known for its high precision, was used to generate true reference values. These true values served as the benchmark for evaluating the IOS performance. Building on the design of an existing scanner, a new scanning head was developed to integrate with a compressed airflow system. This new design aimed to help the IOS capture high-precision data on surfaces where liquid-interference, such as saliva, might otherwise degrade scanning accuracy. The traditional scanning method, without airflow assistance, was employed as a control group for comparison. The study included five groups in total, one control group and four experimental groups, to investigate the effects of scanning lens obstruction, airflow presence, liquid media, and the use of the new scanning head on scanning process and accuracy. Each group underwent 15 scans, generating ample data for a robust statistical comparison. By evaluating trueness and precision in each group, the study assessed the impact of the compressed airflow system on the accuracy of IOS data collected from liquid-interference surfaces. Additionally, we selected Elite and Primescan scanners as references for numerical accuracy values.
RESULTS:
The scanning accuracy on liquid-interference surfaces was significantly reduced in terms of both trueness and precision [Trueness: 18.5 (6.5) vs. 38.0 (6.7), P < 0.05; Precision: 19.1 (8.5) vs. 31.7 (15.0), P < 0.05]. The use of the new scanning head assisted by the compressed airflow system significantly improved the scanning accuracy [Trueness: 22.3(7.6) vs. 38.0 (6.7), P < 0.05; Precision: 25.8 (9.6) vs. 31.7 (15.0), P < 0.05].
CONCLUSION
The scanning head based on the compressed airflow system can assist in improving the accuracy of data obtained from liquid-interference surfaces by the IOS.
Imaging, Three-Dimensional/methods*
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Humans
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Dental Impression Technique/instrumentation*
4.Veronica anagallis-aquatica L. iridoid glycosides alleviate heart failure via metabolites homoveratrumic acid and 2-hydroxy-3,4-dimethoxybenzoic acid mediated by the gut microbiota.
Manjiong WANG ; Xiaobo GUO ; Hanfang LIU ; Xiao LI ; Yue YAO ; Qing FU ; Yu JIN ; Shuaishuai NI ; Xiaokang LI ; Chaojiang XIAO ; Bei JIANG ; Conglong XIA ; Jian LI ; Yixiang XU
Acta Pharmaceutica Sinica B 2025;15(6):3338-3342
The iridoid glycosides from Veronica anagallis-aquatica L. alleviate heart failure by modulating the gut microbiota and influencing the production of two metabolites with potential antihypertrophic effects, HVA and 2OH-VA.Image 1.
5.MAUP Effect on Spatial Pattern of Pseudostellaria heterophylla Production Regions in China
Leting ZHANG ; Tao ZHOU ; Chengdong XU ; Zhixian JING ; Chenghong XIAO ; Hui WANG ; Tingting SHI ; Jiawei HUANG ; Xiaobo ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):183-191
ObjectiveTo investigate the modifiable areal unit problem (MAUP) in the spatial pattern of Pseudostellaria heterophylla production regions and reveal the impact of statistical scales on the spatial distribution characteristics of this medicinal plant species. MethodsUsing multi-source data (literature records, field surveys, and statistical data), we systematically analyzed the spatial patterns across three administrative levels (provincial, prefectural, and county scales). Spatial autocorrelation (Moran's I) analysis, high-low clustering (Getis-Ord General G), and hot/cold spot analysis (Getis-Ord Gi*) were employed. ResultsThe literature-based analysis showed that the production regions of P. heterophylla presented random distribution on the provincial scale and significant aggregation on the prefectural scale. The field survey data showed that the production regions displayed random distribution on the provincial scale but significant aggregation on both prefectural and county scales. The statistical data revealed that the production regions lacked spatial autocorrelation on the provincial scale but demonstrated significant aggregation on prefectural and county scales. ConclusionMAUP effects have substantive implications for understanding and decision-making in the arrangement of medicinal plant production regions. The county scale proves to be the most sensitive and explanatory level for analyzing the spatial pattern of P. heterophylla production regions, providing a critical foundation for habitat modeling, suitability evaluation, and ecological cultivation planning of medicinal plants.
6.Correlation between pulmonary cavitary tuberculosis and immune and nutritional status and prediction model construction in the Kangbei Plateau
Jing FENG ; Juan HAN ; Jiqiong LI ; Jiangcun NIMA ; Lamu ZHAXI ; Zhongbi XU ; XiaoBo HU
Chongqing Medicine 2025;54(11):2497-2502
Objective To investigate the association between cavitary pulmonary tuberculosis and im-munonutritional status in the Kangbei Plateau and construct a prediction model.Methods A total of 121 pa-tients with active pulmonary tuberculosis treated at Ganzi County People's Hospital from June 2023 to June 2024 were retrospectively enrolled and divided into cavitary group(n=53)and non-cavitary group(n=68)based on imaging findings.Clinical and laboratory data were collected.Influencing factors for pulmonary cavi-tation in tuberculosis were screened using univariate and multivariate logistic regression analysis to construct a prediction model.The model's performance was evaluated using receiver operating characteristic(ROC)curve analysis and the Hosmer-Lemeshow test.Results The cavitary group had higher proportions of smokers,spu-tum-positive cases,patients with disease duration>6 months,and patients with respiratory symptoms at ini-tial diagnosis,as well as higher neutrophil-to-lymphocyte ratio(NLR),platelet-to-lymphocyte ratio(PLR),neutrophil count,and platelet count than the non-cavitary group(P<0.05).Conversely,the prognostic nutri-tional index(PNI),lymphocyte count,and albumin(ALB)level were lower in the cavitary group(P<0.05).Multivariate analysis identified disease duration,NLR,and ALB as influencing factors for pulmonary cavitation in tuberculosis patients(P<0.05).The prediction model yielded an area under the curve(AUC)of 0.909,and the Hosmer-Lemeshow test indicated good model fit(χ2=10.740,P=0.217).Conclusion Pulmonary cavitation in high-altitude tuberculosis patients is associated with disease duration,ALB,and NLR.The com-bined predictive value of these three indicators is high.
7.UNOS based study for long-term outcomes and risk factors of liver transplantation recipients receiving grafts from donor aged 80 years and over
Libin DONG ; Xiaobo WANG ; Shuqi CAI ; Kai WANG ; Xiao XU
Chinese Journal of Organ Transplantation 2024;45(6):382-390
Objective:To investigate the long-term prognosis of liver transplantation (LT) recipients who received grafts from donors aged ≥80 years and the associated risk factors.Methods:Clinical and follow-up data of LT recipients from January 2002 to June 2023 were retrospectively analyzed using the United Network for Organ Sharing (UNOS) database. Donors were categorized into three groups : non-elderly donors (NED, age < 60 years), elderly donors (ED, age 60-79 years), and very elderly donors (VED, age ≥80 years). Propensity score matching (PSM) was used to reduce baseline selection bias among the groups. Survival rates were calculated using the Kaplan-Meier method, and differences among the groups were compared using the Log-Rank test. Univariate and multivariate Cox proportional hazards regression analyses were performed to identify independent risk factors for overall survival (OS) in the VED group. Recipients were further subdivided into three age groups (<50 years, 50-69 years, and ≥70 years) to compare survival outcomes among NED, ED, and VED groups.Results:A total of 115, 089 LT recipients were included, comprising 95, 973 (83.4%) in the NED group, 18, 520 (16.1 %) in the ED group, and 596 (0.5 %) in the VED group. After 3∶3∶1 PSM, each group included 1, 623 recipients for NED and ED, and 541 for VED, with no significant differences in baseline data. The 10-year OS rates for NED, ED, and VED groups were 61.8%, 55.9%, and 47.8%, respectively, and the 10-year graft survival (GS) rates were 61.3 %, 53.8%, and 45.9 %, respectively, with all comparisons showing statistical significance ( P< 0.001). In recipients aged <70 years, the VED group had significantly lower OS and GS rates (49.0% and 47.1 %, respectively ) compared to the NED (63.7 % and 61.8%) and ED (57.7% and 55.2 %) groups ( P< 0.001 ). For recipients aged ≥70 years, there were no significant differences in 10-year OS and GS among the NED (47.2% and 48.7 %), ED (47.0 % and 48.7 %), and VED (40.0 % and 39.2 %) groups ( P= 0.992 and P= 0.996, respectively). Cox regression analysis identified cold ischemia time ≥8 hours ( HR=1.447, 95% CI: 1.088-1.923, P=0.011), pre-transplant ICU dependence ( HR=1.803, 95% CI: 1.176–2.765, P=0.007), and hepatitis B/C virus infection ( HR =1.432, 95% CI: 1.057-1.941, P=0.020) as independent risk factors for OS in the VED group. Conclusions:Liver grafts from VED grafts significantly reduce long-term OS and GS in recipients, except in those aged ≥70 years where prognosis is comparable to recipients of NED and ED grafts.. For the VED group, factors such as cold ischemia time ≥8 hours, pre-transplant ICU dependence, and hepatitis B/C virus infection markedly influence the prognosis.
8.Redo-Bentall surgery for aortic root lesions:a report of case series
Xiaobo PENG ; Fan LI ; Tianbo LI ; Chencheng LIU ; Bo XU ; Han XIA ; Yingbin XIAO ; Yong WANG
Journal of Army Medical University 2024;46(10):1158-1163
Objective To observe the clinical efficacy of Redo-Bentall surgery in the reoperation of aortic root lesions.Methods A retrospective analysis was performed on 46 patients who underwent Redo-Bentall surgery for aortic root lesions in our department from June 2010 to April 2022.They were 35 males and 11 females,at a mean age of 43.37±12.79 years,in 4.96±6.76 years since the last operation.General clinical data in perioperative period and during follow-up were collected and analyzed.Kaplan-Meier survival analysis was used to compare the survival rates of each etiological group.Results There were 9 cases of central end otitis,12 cases of Behset's disease,and 25 cases of other causes.After operation,4 cases(8.70%)experienced cardiac arrest,4 cases(8.70%)renal failure,2 cases(4.35%)gastrointestinal bleeding,2 cases(4.35%)new third-degree atrioventricular block and 2 cases(4.35%)permanent pacemaker placement.In perioperative period,3 cases(6.52%)died in hospital.During a mean follow-up of 5.03±3.27 years after discharge,5 cases(11.63%)were lost to follow-up,1 case died(2.33%),1 case had lacunar infarction(2.33%),and no severe bleeding or embolism complications was observed in the rest patients.The long-term survival rate was significantly lower in the endocarditis group(62.3%)and the Behcet's disease group(70%)than the other etiological groups(80%,P<0.05).Conclusion The application of Redo-Bentall in the reoperation of aortic root lesions is safe and effective,but the survival rate is quite lower in the patients with infective endocarditis and Behcet's disease.
9.PIM1 mediates oxidized low-density lipoprotein-induced phenotypic switching of vascular smooth muscle cells in ApoE-/-mice
Mengmeng FU ; Gengrui ZHONG ; Mengqi XU ; Xiaobo WANG ; Hanqin WANG
Chinese Journal of Pathophysiology 2024;40(10):1854-1863
AIM:To investigate the role of proviral integration site for Moloney murine leukemia virus 1(PIM1)in the phenotypic switching of vascular smooth muscle cells(VSMCs)induced by oxidized low-density lipoprotein(oxLDL),and to explore the underlying mechanisms.METHODS:Eighteen male ApoE-/-mice(8 weeks old)were ran-domly divided into general diet group and high-fat diet group,with 9 mice per group.After 16 weeks,aortic samples were analyzed using HE staining to observe plaque formation.In vitro,VSMCs were exposed to oxLDL to induce phenotypic transformation.Western blot and immunofluorescence were used to measure the protein expression levels of PIM1 and phe-notypic markers including α-smooth muscle actin(α-SMA),smooth muscle protein 22α(SM22α),osteopontin(OPN),and CD68.Glycolysis levels were assessed by detecting the expression of glycolytic enzymes 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase(PFKFB3)and hexokinase 2(HK2)by Western blot,and lactate secretion was measured using a lactate test kit.The effects of SMI-4a(a specific inhibitor of PIM1)and PIM1 small interfering RNA on oxLDL-in-duced phenotypic markers in VSMCs were evaluated.Moreover,the impact of 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one(3PO;a glycolysis inhibitor)on oxLDL-induced phenotypic switching and glycolysis in VSMCs was investigated.RESULTS:HE staining revealed atherosclerotic plaque formation in the aortas of ApoE-/-mice fed with high-fat diet.Im-munofluorescence showed high accumulation of PIM1 and OPN in the tunica intima of atherosclerotic plaques.Compared with control group,aortic plaques exhibited significantly elevated levels of PIM1,OPN and CD68 proteins(P<0.01),ac-companied by reduced expression of contractile phenotype markers α-SMA and SM22α(P<0.01).In vitro,oxLDL treat-ment led to gradual decrease in α-SMA and SM22α expression(P<0.05 or P<0.01),while OPN and CD68 expression in-creased(P<0.05 or P<0.01).Moreover,oxLDL significantly up-regulated the protein expression of PIM1,PFKFB3 and HK2,and increased lactate secretion in VSMCs(P<0.05 or P<0.01).Knockdown of PIM1 or treatment with SMI-4a markedly attenuated these oxLDL-induced effects on VSMCs(P<0.05 or P<0.01).Treatment with 3PO also abolished ox-LDL-induced phenotypic transformation and glycolysis in VSMCs(P<0.05 or P<0.01).CONCLUSION:PIM1 highly accumulates in the atherosclerotic plaques of ApoE-/-mice.The phenotypic transformation of VSMCs was correlated with the expression of PIM1.PIM1 can regulate the phenotypic transformation of oxLDL-treated VSMCs by inducing glycolysis.
10.Surveillance of bacterial resistance in tertiary hospitals across China:results of CHINET Antimicrobial Resistance Surveillance Program in 2022
Yan GUO ; Fupin HU ; Demei ZHU ; Fu WANG ; Xiaofei JIANG ; Yingchun XU ; Xiaojiang ZHANG ; Fengbo ZHANG ; Ping JI ; Yi XIE ; Yuling XIAO ; Chuanqing WANG ; Pan FU ; Yuanhong XU ; Ying HUANG ; Ziyong SUN ; Zhongju CHEN ; Jingyong SUN ; Qing CHEN ; Yunzhuo CHU ; Sufei TIAN ; Zhidong HU ; Jin LI ; Yunsong YU ; Jie LIN ; Bin SHAN ; Yunmin XU ; Sufang GUO ; Yanyan WANG ; Lianhua WEI ; Keke LI ; Hong ZHANG ; Fen PAN ; Yunjian HU ; Xiaoman AI ; Chao ZHUO ; Danhong SU ; Dawen GUO ; Jinying ZHAO ; Hua YU ; Xiangning HUANG ; Wen'en LIU ; Yanming LI ; Yan JIN ; Chunhong SHAO ; Xuesong XU ; Wei LI ; Shanmei WANG ; Yafei CHU ; Lixia ZHANG ; Juan MA ; Shuping ZHOU ; Yan ZHOU ; Lei ZHU ; Jinhua MENG ; Fang DONG ; Zhiyong LÜ ; Fangfang HU ; Han SHEN ; Wanqing ZHOU ; Wei JIA ; Gang LI ; Jinsong WU ; Yuemei LU ; Jihong LI ; Qian SUN ; Jinju DUAN ; Jianbang KANG ; Xiaobo MA ; Yanqing ZHENG ; Ruyi GUO ; Yan ZHU ; Yunsheng CHEN ; Qing MENG ; Shifu WANG ; Xuefei HU ; Wenhui HUANG ; Juan LI ; Quangui SHI ; Juan YANG ; Abulimiti REZIWAGULI ; Lili HUANG ; Xuejun SHAO ; Xiaoyan REN ; Dong LI ; Qun ZHANG ; Xue CHEN ; Rihai LI ; Jieli XU ; Kaijie GAO ; Lu XU ; Lin LIN ; Zhuo ZHANG ; Jianlong LIU ; Min FU ; Yinghui GUO ; Wenchao ZHANG ; Zengguo WANG ; Kai JIA ; Yun XIA ; Shan SUN ; Huimin YANG ; Yan MIAO ; Mingming ZHOU ; Shihai ZHANG ; Hongjuan LIU ; Nan CHEN ; Chan LI ; Jilu SHEN ; Wanqi MEN ; Peng WANG ; Xiaowei ZHANG ; Yanyan LIU ; Yong AN
Chinese Journal of Infection and Chemotherapy 2024;24(3):277-286
Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5%were gram-positive and 70.5%were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3%,76.7%and 77.9%,respectively.Overall,94.0%of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8%of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2%in the isolates from children and 95.7%in the isolates from adults.The resistance rate to carbapenems was lower than 13.1%in most Enterobacterales species except for Klebsiella,21.7%-23.1%of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1%to 13.3%.The prevalence of meropenem-resistant strains decreased from 23.5%in 2019 to 18.0%in 2022 in Pseudomonas aeruginosa,and decreased from 79.0%in 2019 to 72.5%in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

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