1.Characterization and Application of Moisture Absorption Kinetics of Traditional Chinese Medicines Based on Double Exponential Model:A Review
Yanting YU ; Lei XIONG ; Yan HE ; Wei LIU ; Jing YANG ; Yao ZHANG ; Jiali CHEN ; Xiaojian LUO ; Xiaoyong RAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):340-346
Hygroscopicity research has long been a key focus and hot topic in Chinese materia medica(CMM). Elucidating hygroscopic mechanisms plays a vital role in formulation design, process optimization, and storage condition selection. Hygroscopic models serve as essential tools for characterizing CMM hygroscopic mechanisms, with various types available. The double exponential model is a kinetic mathematical model constructed based on the law of conservation of energy and Fick's first law of diffusion, tailored to the physical properties of CMM extracts. In recent years, this model has been extensively applied to simulate the dynamic moisture absorption behavior of CMM extracts and solid dosage forms under varying humidity conditions. It has revealed the correlation between moisture absorption kinetic parameters and material properties, offering a new perspective for characterizing the moisture uptake behavior of CMM. This paper systematically reviews the application progress of this model in the field of CMM, analyzes its advantages, disadvantages, and challenges in this domain, and explores its potential application trends in other fields. It aims to provide references for elucidating the moisture absorption mechanisms of CMM and researching moisture-proofing technologies, while also offering insights for its broader application in food and polymer materials.
2.Aging and perioperative brain health: Mechanisms, management, and future.
Peilin CONG ; Qian CHEN ; Qianqian WU ; Jing WANG ; Xinwei HUANG ; Qian ZHANG ; Zheping CHEN ; Huanghui WU ; Yuxin ZHANG ; Mengfan HE ; Zhouxiang LI ; Li TIAN ; Lize XIONG
Chinese Medical Journal 2025;138(19):2381-2398
Globally, over 300 million surgeries are performed each year, and more than 50% of surgeries involve patients aged 65 and older. Aging poses significant challenges to perioperative brain health, as the deterioration of brain structure and function increases susceptibility to postoperative neurological complications. Protecting perioperative brain health remains a worldwide clinical challenge. With senescence, the brain undergoes a progressive decline in homeostasis across various molecular, cellular, and regional functions. Anesthetics and surgical stimuli may accelerate the disruption of brain homeostasis and exacerbate age-related neurodegeneration. This review provides a framework for understanding how anesthesia and surgery can affect brain health in the aging population and contribute to postoperative neurological complications, with a particular focus on perioperative neurocognitive disorder.
3.Comparative study on the efficacy of Zero-PVA versus titanium plate in single-level ACDF treatment and analysis of radiographic influencing factors
Jing HUANG ; Yunyu XIONG ; Ziran ZHOU ; Hengtao HE ; Bing WANG
Chinese Journal of Orthopaedics 2025;45(4):205-214
Objective:To investigate the clinical and radiological differences in treating single-level cervical spondylosis using the Zero-profile interbody fusion cage (Zero-PVA) versus the titanium plate interbody fusion cage in anterior cervical discectomy and fusion (ACDF) and to analyze the correlation between clinical outcomes and radiological parameters.Methods:this retrospective study collected data from patients with cervical spondylosis who underwent single-level ACDF surgery at the First Affiliated Hospital of Kunming Medical University between January 2019 and June 2022. A total of 70 patients (32 males and 38 females) with an average age of 51.34 ± 8.62 years (range: 29 to 77 years) were included. Based on the type of interbody fusion device used, patients were divided into two groups: the cage-plate construct (CP) group (35 patients; average age: 51.51 ± 10.02 years, range: 29 to 77 years) and the zero-profile interbody fusion device with variable angle (ZP) group (35 patients; average age: 51.17±7.11 years, range: 38 to 68 years). Preoperative, one-week postoperative, and final follow-up (≥6 months postoperative) data were collected for visual analogue scale (VAS) scores, neck disability index (NDI) scores, and Japanese Orthopaedic Association (JOA) scores. Radiological parameters assessed included surgical segment intervertebral disc height, cervical lordosis angle, surgical segment Cobb angle, T 1 slope (T 1S), and C 2-C 7 sagittal vertical axis (C 2-C 7 SVA). Additional evaluations included Bazaz dysphagia ratings at one week and final follow-up, Odom surgical outcome ratings at final follow-up, fusion grades, adjacent vertebral ectopic ossification grades, and adjacent segment degeneration. Patients were further classified into a symptom relief group and an incomplete symptom relief group based on Odom ratings. Logistic regression analysis was performed to examine the correlation between radiological parameters and postoperative outcomes. Statistical methods such as ROC curves, Youden index, product index, and Euclidean index were used to determine optimal cut-off values for radiological parameters affecting postoperative outcomes. Results:There were no statistically significant differences between the two groups in terms of gender, age, follow-up duration, surgical segment, or classification ( P>0.05), confirming comparability. At both one-week post-operation and final follow-up, VAS, NDI, and JOA scores showed no significant differences between the groups ( P>0.05). However, the incidence of dysphagia in the CP group was 49% at one week post-operation and 20% at final follow-up, significantly higher than that in the ZP group (14% at one week and 0 at final follow-up; P<0.05). The intervertebral space height in the CP group was 9.16±0.84 mm at one week post-operation and 8.19±1.11 mm at final follow-up, which was significantly higher than in the ZP group (8.22±1.23 mm at one week and 7.57±1.14 mm at final follow-up; P<0.05). The surgical segment Cobb angle in the CP group at final follow-up (7.34°±4.21°) was significantly greater than in the ZP group (4.63°±4.32°; t=2.661, P=0.010). Regarding heterotopic ossification (HO) of the superior vertebral body, in the CP group, 27 cases were grade 0, 7 cases were grade 1, and 1 case was grade 2. In the ZP group, 34 cases were grade 0 and 1 case was grade 1, with a statistically significant difference between the groups (χ 2=6.128, P=0.028). Binary logistic regression analysis revealed that a reduction in intervertebral space height of ≥0.63 mm at one week post-operation and final follow-up ( OR=2.913, P=0.021) and a heterotopic ossification grade of ≥1 ( OR=5.178, P=0.042) were correlated with incomplete symptom relief after single-level ACDF surgery. Conclusions:Both Zero-PVA and titanium plate interbody fusion devices demonstrated favorable clinical outcomes in single-level ACDF surgery. However, a reduction in intervertebral space height and heterotopic ossification of the superior vertebral body may be risk factors for incomplete symptom relief following single-level ACDF.
4.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
5.Research and application of a new deep learning based strategy for platelet histogram review
Enming ZHANG ; Chao YANG ; Xianchun CHEN ; Yan LIN ; Taixue AN ; Haixia LI ; Yongjian HE ; Zhiwei LIU ; Limei FENG ; Wanying LIN ; Tie XIONG ; Kai QIU ; Ya GAO ; Lizhu HUANG ; Jing HE ; Chunyan WANG ; Dehua SUN ; Bo SITU ; Lei ZHENG
Chinese Journal of Laboratory Medicine 2025;48(9):1201-1206
Objective:To develop an artificial intelligence (AI)-based platelet review strategy to identify abnormal platelet histograms with no significant difference between initial impedance platelet count (PLT-I) and PLT-F results.Methods:This study included 5 119 routine blood analysis in Nanfang Hospital of Southern Medical University and its Ganzhou branch from July 2023 and March 2024. Specimens exhibiting abnormal platelet histograms and an initial platelet count >40×10?/L underwent review using the fluorescent platelet count (PLT-F) channel. Consistency of the results was defined as a difference between impedance platelet count (PLT-I) and PLT-F less than ±20% of the PLT-F results. A deep learning model was developed using platelet and red blood cell histogram data from a training set of 3 807 specimens. The model′s diagnostic performance was evaluated on an independent external validation set ( n=805) using receiver operating characteristic (ROC) curve analysis. Changes in the number of reviewed samples and sample turnaround time were analyzed to assess its clinical utility. Results:The deep learning model based on platelet and red blood cell histograms achieved an area under the ROC curve (AUC) of 0.854 in the training set. At a cutoff value of 0.1, the sensitivity was 0.954 and specificity was 0.358. The model could reduce review by 16.80% (190/1 131). In the validation set, the AUC was 0.805, with a sensitivity of 0.955 and specificity of 0.307, corresponding to a reduction of 17.41% (47/270) in reviewed specimens.Conclusion:The platelet review prediction model developed based on deep learning technology can efficiently identify samples with consistent results before and after review, reducing unnecessary reviews and shortening specimen testing time, thereby improving the efficiency of platelet test.
6.Comparative study on the efficacy of Zero-PVA versus titanium plate in single-level ACDF treatment and analysis of radiographic influencing factors
Jing HUANG ; Yunyu XIONG ; Ziran ZHOU ; Hengtao HE ; Bing WANG
Chinese Journal of Orthopaedics 2025;45(4):205-214
Objective:To investigate the clinical and radiological differences in treating single-level cervical spondylosis using the Zero-profile interbody fusion cage (Zero-PVA) versus the titanium plate interbody fusion cage in anterior cervical discectomy and fusion (ACDF) and to analyze the correlation between clinical outcomes and radiological parameters.Methods:this retrospective study collected data from patients with cervical spondylosis who underwent single-level ACDF surgery at the First Affiliated Hospital of Kunming Medical University between January 2019 and June 2022. A total of 70 patients (32 males and 38 females) with an average age of 51.34 ± 8.62 years (range: 29 to 77 years) were included. Based on the type of interbody fusion device used, patients were divided into two groups: the cage-plate construct (CP) group (35 patients; average age: 51.51 ± 10.02 years, range: 29 to 77 years) and the zero-profile interbody fusion device with variable angle (ZP) group (35 patients; average age: 51.17±7.11 years, range: 38 to 68 years). Preoperative, one-week postoperative, and final follow-up (≥6 months postoperative) data were collected for visual analogue scale (VAS) scores, neck disability index (NDI) scores, and Japanese Orthopaedic Association (JOA) scores. Radiological parameters assessed included surgical segment intervertebral disc height, cervical lordosis angle, surgical segment Cobb angle, T 1 slope (T 1S), and C 2-C 7 sagittal vertical axis (C 2-C 7 SVA). Additional evaluations included Bazaz dysphagia ratings at one week and final follow-up, Odom surgical outcome ratings at final follow-up, fusion grades, adjacent vertebral ectopic ossification grades, and adjacent segment degeneration. Patients were further classified into a symptom relief group and an incomplete symptom relief group based on Odom ratings. Logistic regression analysis was performed to examine the correlation between radiological parameters and postoperative outcomes. Statistical methods such as ROC curves, Youden index, product index, and Euclidean index were used to determine optimal cut-off values for radiological parameters affecting postoperative outcomes. Results:There were no statistically significant differences between the two groups in terms of gender, age, follow-up duration, surgical segment, or classification ( P>0.05), confirming comparability. At both one-week post-operation and final follow-up, VAS, NDI, and JOA scores showed no significant differences between the groups ( P>0.05). However, the incidence of dysphagia in the CP group was 49% at one week post-operation and 20% at final follow-up, significantly higher than that in the ZP group (14% at one week and 0 at final follow-up; P<0.05). The intervertebral space height in the CP group was 9.16±0.84 mm at one week post-operation and 8.19±1.11 mm at final follow-up, which was significantly higher than in the ZP group (8.22±1.23 mm at one week and 7.57±1.14 mm at final follow-up; P<0.05). The surgical segment Cobb angle in the CP group at final follow-up (7.34°±4.21°) was significantly greater than in the ZP group (4.63°±4.32°; t=2.661, P=0.010). Regarding heterotopic ossification (HO) of the superior vertebral body, in the CP group, 27 cases were grade 0, 7 cases were grade 1, and 1 case was grade 2. In the ZP group, 34 cases were grade 0 and 1 case was grade 1, with a statistically significant difference between the groups (χ 2=6.128, P=0.028). Binary logistic regression analysis revealed that a reduction in intervertebral space height of ≥0.63 mm at one week post-operation and final follow-up ( OR=2.913, P=0.021) and a heterotopic ossification grade of ≥1 ( OR=5.178, P=0.042) were correlated with incomplete symptom relief after single-level ACDF surgery. Conclusions:Both Zero-PVA and titanium plate interbody fusion devices demonstrated favorable clinical outcomes in single-level ACDF surgery. However, a reduction in intervertebral space height and heterotopic ossification of the superior vertebral body may be risk factors for incomplete symptom relief following single-level ACDF.
7.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
8.Research and application of a new deep learning based strategy for platelet histogram review
Enming ZHANG ; Chao YANG ; Xianchun CHEN ; Yan LIN ; Taixue AN ; Haixia LI ; Yongjian HE ; Zhiwei LIU ; Limei FENG ; Wanying LIN ; Tie XIONG ; Kai QIU ; Ya GAO ; Lizhu HUANG ; Jing HE ; Chunyan WANG ; Dehua SUN ; Bo SITU ; Lei ZHENG
Chinese Journal of Laboratory Medicine 2025;48(9):1201-1206
Objective:To develop an artificial intelligence (AI)-based platelet review strategy to identify abnormal platelet histograms with no significant difference between initial impedance platelet count (PLT-I) and PLT-F results.Methods:This study included 5 119 routine blood analysis in Nanfang Hospital of Southern Medical University and its Ganzhou branch from July 2023 and March 2024. Specimens exhibiting abnormal platelet histograms and an initial platelet count >40×10?/L underwent review using the fluorescent platelet count (PLT-F) channel. Consistency of the results was defined as a difference between impedance platelet count (PLT-I) and PLT-F less than ±20% of the PLT-F results. A deep learning model was developed using platelet and red blood cell histogram data from a training set of 3 807 specimens. The model′s diagnostic performance was evaluated on an independent external validation set ( n=805) using receiver operating characteristic (ROC) curve analysis. Changes in the number of reviewed samples and sample turnaround time were analyzed to assess its clinical utility. Results:The deep learning model based on platelet and red blood cell histograms achieved an area under the ROC curve (AUC) of 0.854 in the training set. At a cutoff value of 0.1, the sensitivity was 0.954 and specificity was 0.358. The model could reduce review by 16.80% (190/1 131). In the validation set, the AUC was 0.805, with a sensitivity of 0.955 and specificity of 0.307, corresponding to a reduction of 17.41% (47/270) in reviewed specimens.Conclusion:The platelet review prediction model developed based on deep learning technology can efficiently identify samples with consistent results before and after review, reducing unnecessary reviews and shortening specimen testing time, thereby improving the efficiency of platelet test.
9.Mortality, morbidity, and care practices for 1750 very low birth weight infants, 2016-2021
Yang HE ; Meng ZHANG ; Jun TANG ; Wanxiu LIU ; Yong HU ; Jing SHI ; Hua WANG ; Tao XIONG ; Li ZHANG ; Junjie YING ; Dezhi MU
Chinese Medical Journal 2024;137(20):2452-2460
Background::Very low birth weight (VLBW) infants are the key populations in neonatology, wherein morbidity and mortality remain major challenges. The study aimed to analyze the clinical characteristics of VLBW infants.Methods::A retrospective cohort study was conducted in West China Second Hospital between January 2016 and December 2021. Neonates with a birth weight of <1500 g were included. Mortality, care practices, and major morbidities were analyzed, and compared with those of previous 7 years (2009-2015).Results::Of the total 1750 VLBW, 1386 were infants born with birth weight between 1000-1499 g and 364 infants were born with weight below 1000 g; 42.9% (751/1750) required delivery room resuscitation; 53.9% (943/1750) received non-invasive ventilation only; 38.2% (669/1750) received invasive ventilation; 1517 VLBW infants received complete treatment. Among them, 60.1% (912/1517) of neonates had neonatal respiratory distress syndrome (NRDS), 28.7% (436/1517) had bronchopulmonary dysplasia (BPD), 22.0% (334/1517) had apnea, 11.1% (169/1517) had culture-confirmed sepsis, 8.4% (128/1517) had pulmonary hemorrhage, 7.6% (116/1517) had severe intraventricular hemorrhage (IVH)/periventricular leukomalacia (PVL), 5.7% (87/1517) had necrotizing enterocolitis (NEC), and 2.0% (31/1517) had severe retinopathy of prematurity. The total and in-hospital mortality rates were 9.7% (169/1750) and 3.0% (45/1517), respectively. The top three diagnoses of death among those who had received complete treatment were sepsis, NRDS, and NEC. In 2009-2015, 1146 VLBW were enrolled and 895 infants received complete treatment. The proportions of apnea, IVH, and IVH stage ≥3/PVL, were higher in 2009-2015 compared with those in 2016-2021, while the proportions of NRDS and BPD were characterized by significant increases in 2016-2021. The total and in-hospital mortality rates were 16.7% (191/1146) and 5.6% (50/895) respectively in 2009-2015.Conclusion::Among VLBW infants born in 2016-2021, the total and in-hospital mortality rates were lower than those of neonates born in 2009-2015. Incidences of NRDS and BPD increased in 2016-2021, which affected the survival rates and long-term prognosis of VLBW.
10.Role of T3SS in promoting Pseudomonas aeruginosa internalization in pulmonary epithelial cells via ERK/ROS signaling pathway
Junzhi XIONG ; Hua YU ; Xingmin WANG ; Xiaomei HE ; Qian DAI ; Jing QIU
Journal of Army Medical University 2024;46(22):2493-2504
Objective To explore the role and underlying mechanism of type Ⅲ secretion system(T3SS)in regulating the internalization of Pseudomonas aeruginosa(PA)into pulmonary epithelial cells.Methods The human non-small cell lung cancer A549 cells were infected with or without PA strains,including wild-type PAO1(a standard experimental PA strain),△exsA(knockout of the critical activator for T3SS genes),△pscJ(T3SS secretion-defective strain)and PAO1-E(EGTA-induced high expression of T3SS genes).The A549 cells pretreated with ERK inhibitor U0126 or reactive oxygen species(ROS)inhibitor apocynin(APO)/N-acetyl-L-cysteine(NAC)were infected with PAO1 or PAO1-E strain.Thus,the experiment was grouped as follows:the mock-treated group,PAO1-or PAO1-E-infected group,inhibitor-treated group,and PAO1/PAO1-E plus inhibitor-treated group.Extracellular bacteria were killed by gentamicin,and the cell lysates were diluted and then plated on PA screening plates.Bacterial amounts were detected by counting colony-forming units(CFUs).The production of ROS was analyzed using fluorescent probe labeling and flow cytometry.The activation of the ERK pathway was detected by Western blotting.Results Compared with the PAO1-infected group,the intracellular bacteria and ROS level in △exsA-or△pscJ-infected cells were lower(P<0.05,P<0.01),so was the generation of ROS(P<0.01);In contrast,those of the PAO1-E strain-infected cells displayed an opposite trend(P<0.01).Compared with the PAO1-or PAO1-E-infected group,the cells pretreated with APO/NAC followed by PAO1 or PAO1-E infection showed reduced intracellular bacterial amounts(P<0.01).Compared to the PAO1-infected A549 cells,the phosphorylation level of ERK was increased in the △exsA-or △pscJ-infected cells(P<0.01),while that level was suppressed in the PAO1-E-treated cells(P<0.01).Compared with the PAO1-infected group,the PAO1-infected cells pretreated with U0126 displayed reduced ERK activation,elevated ROS production,and increased intracellular counts of PAO1(P<0.01).Conclusion T3SS-mediated inhibition of the ERK pathway promotes the production of ROS and the internalization of PA in lung epithelial cells.

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