1.The regulatory role of the RUS family in plant growth and development.
Yao HU ; Sirui LI ; Xinxin ZHANG ; Qinglin TANG ; Dayong WEI ; Shibing TIAN ; Yang YANG ; Zhimin WANG
Chinese Journal of Biotechnology 2024;40(1):81-93
The chloroplast genome encodes many key proteins involved in photosynthesis and other metabolic processes, and metabolites synthesized in chloroplasts are essential for normal plant growth and development. Root-UVB (ultraviolet radiation B)-sensitive (RUS) family proteins composed of highly conserved DUF647 domain belong to chloroplast proteins. They play an important role in the regulation of various life activities such as plant morphogenesis, material transport and energy metabolism. This article summarizes the recent advances of the RUS family proteins in the growth and development of plants such as embryonic development, photomorphological construction, VB6 homeostasis, auxin transport and anther development, with the aim to facilitate further study of its molecular regulation mechanism in plant growth and development.
Female
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Pregnancy
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Humans
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Ultraviolet Rays
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Biological Transport
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Chloroplasts/genetics*
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Embryonic Development
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Plant Development/genetics*
2.Integrating proteomics and targeted metabolomics to reveal the material basis of liver-gallbladder damp-heat syndrome in chronic hepatitis B
LI Ni&rsquo ; ao ; GONG Yuefeng ; WANG Jia ; CHEN Qingqing ; SU Shibing ; ZHANG Hua ; LU Yiyu
Digital Chinese Medicine 2024;7(4):320-331
Methods:
CHB patients and healthy volunteers were enrolled from Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine between August 21, 2018 and December 31, 2020. They were divided into three groups: healthy group, LGDHS group, and latent syndrome (LP) group. Proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ) was performed to identify differentially expressed proteins (DEPs). Metabolomic profiling via ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was applied to serum samples to detect differentially regulated metabolites (DMs). Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment were employed to explore dysregulated pathways. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were utilized to visualize group separation and identify key metabolites and proteins contributing to LGDHS differentiation. Receiver operating characteristic (ROC) curve analysis evaluated the diagnostic performance of key biomarkers, while logistic regression models assessed their predictive accuracy. P values were corrected for multiple tests using the Benjamini-Hochberg method to control the false discovery rate (FDR). Validation of potential biomarkers was conducted using independent microarray data and real-time quantitative polymerase chain reaction (RT-qPCR).
Results:
A total of 150 participants were enrolled, including healthy group (n = 45), LGDHS group (n = 60), and LP group (n = 45). 254 DEPs from proteomics data and 72 DMs from metabolomic profiling were identified by PCA and OPLS-DA. DEPs were mainly enriched in immune and complement pathways, while DMs involved in amino acid and energy metabolism. The integrated analysis identified seven key biomarkers: α1-acid glycoprotein (ORM1), asparagine synthetase (ASNS), solute carrier family 27 member 5 (SLC27A5), glucosidase II alpha subunit (GANAB), hexokinase 2 (HK2), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and maltase-glucoamylase (MGAM). Microarray validation confirmed the diagnostic potential of these genes, with area under the curve (AUC) values for ROC analysis ranging from 0.536 to 0.759. Among these, ORM1, ASNS, and SLC27A5 showed significant differential ability in differentiating LGDHS patients (P = 0.016, P = 0.035, and P < 0.001, respectively), with corresponding AUC of 0.749, 0.743, and 0.759, respectively. A logistic regression model incorporating these three genes demonstrated an AUC of 0.939, indicating a high discriminatory power for LGDHS. RT-qPCR further validated the differential expression of ORM1 and SLC27A5 between LGDHS and LP groups (P = 0.011 and P = 0.034, respectively), with ASNS showing a consistent trend in expression (P = 0.928).
Conclusion
This study integrates multi-omics approaches to uncover the molecular mechanisms underlying LGDHS in CHB. The identification of biomarkers ORM1, ASNS, and SLC27A5 offers a solid basis for the objective diagnosis of LGDHS, contributing to the standardization and modernization of TCM diagnostic practices.
3.Chinese Translation of the Stanford Expectations of Treatment Scale and Its Application Evaluation on Traditional Chinese Medicine for Diarrhea-Predominant Irritable Bowel Syndrome with Liver-Constraint and Spleen-Deficiency Syndrome
Shibing LIANG ; Yingying ZHANG ; Zhijie WANG ; Zeyu YU ; Mei HAN ; Huijuan CAO ; Guoyan YANG ; Shihuan CAO ; Hongjie CHENG ; Qiaoyan ZHANG ; Youzhu SU ; Yufei LI ; Jianping LIU
Journal of Traditional Chinese Medicine 2024;65(19):1994-2001
ObjectiveTo adapt the Stanford Expectations of Treatment Scale(SETS) into Chinese(C-SETS) and test the feasibility, validity and reliability of its application in patients with diarrhea-predominant irritable bowel syndrome(IBS-D) with liver-constraint and spleen-deficiency syndrome treated with traditional Chinese medicine(TCM). MethodsWe obtained authorisation from the developer of the SETS, and followed the principle of "two-way translation" to translate the SETS by literal translation and back translation to form the C-SETS. Ninety-six IBS-D patients with liver-constraint and spleen-deficiency syndrome were enrolled as respondents and filled out C-SETS before receiving treatment; the feasibility was assessed by the recall rate, completion rate and the duration of filling out the scale; the reliability was assessed by Cronbach's α; the structural validity was assessed by exploratory and confirmatory factor analysis, and the content validity was assessed by correlation analysis. ResultsThe C-SETS consists of 10 items, with the 1st, 3rd, and 5th rating items constituting the Positive Expectations subscale, and the 2nd, 4th, and 6th rating items constituting the Negative Expectations subscale, each of which is rated on a 7-point Likert Scale. The recall of C-SETS was 100%(96/96), the completion rate was 89.58%(86/96); Cronbach's α for the Positive and Negative Treatment Expectations subscales were 0.845 and 0.854, respectively; exploratory factor analysis showed that the coefficient of commonality for all six entries was larger than 0.4, and that the six entries could be used by both factors to explain 77.092% of the total variance; validation factor analysis showed that the goodness-of-fit index, comparative fit index, root mean square of approximation error, canonical fit coefficient, and chi-square degrees of freedom ratio took the values of 0.943, 1.003, 0, 0.943, and 0.626, respectively; and the results of Spearman's analysis suggested that the C-SETS had good content validity. ConclusionThe C-SETS has well feasibility, reliability, and validity, which initially proves that it can be used as a tool to assess the treatment expectation of patients with IBS-D with liver-constraint and spleen-deficiency syndrome before receiving TCM treatment.
4.Methodology for Developing Patient Guideline (2):Process and Methodology
Lijiao YAN ; Ning LIANG ; Nannan SHI ; Sihong YANG ; Ziyu TIAN ; Dan YANG ; Xiaojia NI ; Yufang HAO ; Wei CHEN ; Ruixiang WANG ; Yingfeng ZHOU ; Shibing LIANG ; Shuyu YANG ; Yujing ZHANG ; Ziteng HU ; Jianping LIU
Journal of Traditional Chinese Medicine 2024;65(21):2194-2198
At present, the process and methodology of patient guidelines (PGs) development varies greatly and lacks systematic and standardised guidance. In addition to the interviews with PG developers, we have sorted out the relevant methodology for the adaptation and development of existing clinical practice guideline recommendations and facilitated expert deliberations to achieve a consensus, so as to finally put forward a proposal for guidance on the process and methodology for the development of PGs. The development of PGs can be divided into the preparation stage, the construction stage, and the completion stage in general, but the specific steps vary according to the different modes of development of PGs. The development process of Model 1 is basically the same as the patient version of the guideline development process provided by the International Guidelines Network, i.e., team formation, screening of recommendations, guideline drafing, user testing and feedback, approval and dissemination. The developer should also first determine the need for and scope of translating the clinical practice guideline into a patient version during the preparation phase. Model 2 adds user experience and feedback to the conventional clinical practice guideline development process (forming a team, determining the scope of the PG, searching, evaluating and integrating evidence, forming recommendations, writing the guideline, and expert review). Based on the different models, we sort out the process and methods of PG development and introduce the specific methods of PG development, including how to identify the clinical problem and how to form recommendations based on the existing clinical practice guidelines, with a view to providing reference for guideline developers and related researchers.
5.Pathological characteristics of heart and spleen in a kawasaki disease model induced by candida albicans watersoluble fraction
Jiaxin ZHANG ; Zongli ZHANG ; Yifan LUO ; Tao LI ; Shibing XI
Chinese Journal of Clinical and Experimental Pathology 2024;40(7):719-724
Purpose To investigating the pathological characteristics of the heart and spleen in a Kawasaki disease(KD)model induced by candida albicans watersoluble fraction(CAWS)and to provide a research basis for the pathological mechanisms of KD.Methods Intraperitoneal injection of CAWS was used to establish a KD model in C57BL/6 mice.Specimens of the heart,aorta,and spleen were collected to measure body weight and the weights of the heart and spleen.HE staining was utilized to examine the morphological alterations in the heart,aorta,and spleen.The expression levels of the ma-ture macrophage marker(F4/80),TNF-α,and IL-1β in the spleen were measured using qRT-PCR and immunofluorescence.its clinical pathological characteristics were analyze and relevant literatures were reviewed.Results Compared to the control group,there were no significant differences in the heart and body mass of the model group mice,but their spleen mass signif-icantly increased(P<0.05).In the myocardial interstitium of model group mice,there was infiltration of mononuclear cells,disorder of the elastic fibers in the aortic wall along with mucoid degeneration,disruption of the splenic red and white pulp struc-ture,and significant macrophage infiltration.In the model group mice,the mRNA levels and immunofluorescence staining inten-sity of spleen F4/80(2.58-fold,P<0.000 1),TNF-α(1.43-fold,P<0.001),and IL-1β(3.62-fold,P<0.000 1)were significantly higher than those in the control group.Conclusion Mice with KD models induced by CAWS can exhibit patholog-ical alterations akin to KD in the heart and aorta,significant en-largement of the spleen,infiltration by macrophages,with ele-vated expressions of TNF-α and IL-1β.
6.Clinical features and risk factors of pulmonary hemorrhage in extremely preterm infants
Min QI ; Ye ZHANG ; Shibing YANG ; Xing LI ; Zhou JIANG
Chinese Journal of Neonatology 2023;38(4):200-204
Objective:To study the clinical features and risk factors of pulmonary hemorrhage in extremely preterm (EPT) infants.Methods:From February 2018 to January 2022, EPT infants admitted to NICU of our hospital and diagnosed with pulmonary hemorrhage were retrospectively assigned into the observation group and those without pulmonary hemorrhage were assigned into the control group. Univariate analysis and multivariate logistic regression analysis were used to compare the clinical features and determine risk factors of pulmonary hemorrhage in EPT infants.Results:A total of 114 EPT infants were included, including 28 cases (24.6%) in the observation group with pulmonary hemorrhage and 86 cases in the control group. Pulmonary hemorrhage mainly occurred within the first week after birth. Univariate analysis showed that the observation group had higher incidences of following events than the control group: birth asphyxia, delivery room intubation, severe respiratory distress syndrome, hyperglycemia, thrombocytopenia, severe acidosis, shock, score for neonatal acute physiology with perinatal extension-Ⅱ (SNAPPE-Ⅱ) ≥37 and the highest lactate level. Birth weight was lower in the observation group than the control group ( P<0.05). Logistic regression analysis showed that SNAPPE-Ⅱ≥37, shock and hyperglycemia were risk factors of pulmonary hemorrhage ( OR=4.081, 4.610 and 3.355, respectively, all P<0.05). The incidences of mortality and intracranial hemorrhage in the observation group were higher than the control group. The duration of mechanical ventilation in the observation group was longer than the control group ( P<0.05). No significant differences existed in the duration of nasal continuous positive airway pressure, assist mechanical ventilation and total oxygen use, the incidences of grade Ⅱ-Ⅲ bronchopulmonary dysplasia, retinopathy of prematurity and the length of hospital stay ( P>0.05). Conclusions:SNAPPE-Ⅱ≥37, shock and hyperglycemia are early risk factors for pulmonary hemorrhage in EPT infants. EPT infants with pulmonary hemorrhage have higher incidences of mortality and intracranial hemorrhage, requiring longer periods of mechanical ventilation.
7.Effects of adriamycin resistance cell-derived exosomes on the proliferation and migration of osteosarcoma cells through drug resistance transmission
Chao SUN ; Wei FENG ; Lihua ZHANG ; Chenyang MENG ; Huiqin XUE ; Wei ZHAO ; Yuxin WANG ; Ziheng WANG ; Liang SUN ; Shibing GUO
Chinese Journal of Orthopaedics 2023;43(10):645-658
Objective:To explore the relationship and underlying mechanism between exosomes derived from doxorubicin-resistant osteosarcoma cells and MDR1 and miRNAs. Methods:MG63 and U2OS cell lines were selected to construct doxorubicin-resistant strains, and the 50% inhibitory concentration (half maximal inhibitory concentration, IC 50) of drug-resistant and sensitive strains was detected by MTT, and fluorescence staining was performed at intervals of 15 min between 15 and 120 min to detect the change of fluorescence intensity. RT-PCR and Western Blot were used to detect the expression levels of MDR1 P-gp to verify the drug resistance of osteosarcoma cells. Exosomes were identified by particle size analysis and Western Bolt detection. The endocytosis of PKH26-labeled exosomes from doxorubicin-resistant cells was observed, and the proliferation level and migration of exosomes from doxorubicin-resistant cells co-cultured with osteosarcoma cells were detected by MTT assay and cell scratch assay. The differential expression levels of miRNAs in osteosarcoma-sensitive and drug-resistant cells were verified by sequencing and bioinformatics analysis and RT-PCR assay. Tumor growth, serum exosome identification and mRNA expression level of miR-21-5p in tumor-bearing nude mice between normal osteosarcoma cell group and drug-resistant group, drug-resistant+normal exosome group, drug-resistant+drug-resistant+drug-resistant exosome group were observed. MDR1 expression level in tumor tissue was detected by RT-PCR, Western Blot and immunohistochemistry. Results:The IC 50 of two adriamycin resistant strains were 2.21 vs. 11.81 μg/ml and 0.93 vs. 11.81 μg/ml, respectively, and the fluorescence intensity decreased faster than that of normal strains. The relative mRNA expression levels of MDR1 in two cell lines were normal 1.12±0.16, 1.02±0.11 and drug-resistant 2.15±0.10, 2.127±0.12, respectively. The relative protein expression of P-gp was normal 0.92±0.11, 0.73±0.10 and drug-resistant 0.46±0.03, 0.30±0.04, the differences were statistically significant ( P<0.05). Drug-resistant exosomes can enter osteosarcoma cells through endocytosis and concentrate in the cytoplasm when co-cultured with normal strains. Osteosarcoma cells were co-cultured with drug-resistant exosomes at 2, 4, 6, and 8 μg/ml adriamycin, respectively. Compared with normal group, the proliferation level in drug-resistant group was significantly increased. Compared with the normal cell group 35.95±3.92, 6.72±3.55 and the normal exosome group 51.22±5.55, 19.31±1.93, the drug-resistant cell group 54.20±9.32, 19.24±2.88 and drug-resistant exosome group 76.40±5.41, 30.26±4.87, all had significantly higher cell mobility, the difference was statistically significant ( P<0.05). Exosome sequencing and biogenic analysis of 10 highly upregated miRNAs to validate mRNA expression differences between normal and drug-resistant strains by RT-PCR, showing a significant increase in miR-21-5p expression level of drug-resistant strains (5.89±0.26 vs. 0.99±0.06; 1.05±0.07 vs. 8.80±0.93, P<0.05), the difference was statistically significant ( P<0.05). In MG63 and U2OS, the normal cell group and drug-resistant cell group, and the normal exosome group and drug-resistant exosome group were compared, the tumor volume and the terminal tumor weight of nude mice were increased to varying degrees. MRNA relative expression levels of miR-21-5p in serum exosomes of nude mice after drug intervention were 0.86±0.07 and 0.86±0.05 in normal cell group, respectively. The values were 1.13±0.12, 1.14±0.12 in drug-resistant cell group, 0.71±0.05, 0.75±0.03 in normal exosome group, and 0.90±0.07, 0.93±0.04 in drug-resistant exosome group. Compared with normal and drug-resistant strains, the expression levels of normal and drug-resistant exosome groups were increased, with statistical significance ( P<0.05). Conclusion:The exosomes of drug-resistant cells in osteosarcoma could enhance the proliferation level and migration ability of cells through intercellular transfer of MDR1 and miRNAs. The expression of MDR1 and miR-21-5p in drug-resistant cells and tumor-forming nude mouse serum and tumor tissues were up-regulated which suggested that it might be involved in regulating the drug resistance process of osteosarcoma.
8.Analysis of Imaging Performance Standards of CBCT X-IGRT System Used in Radiotherapy.
Shibing XIE ; Peichen WANG ; Chunying JIAO ; Chengxin LIANG ; Xintao ZHANG ; Jiajie XIE
Chinese Journal of Medical Instrumentation 2023;47(6):608-611
This article briefly describes the imaging performance standards of the kilovolt X-ray image guidance system used in radiotherapy, analyzes the main aspects that should be considered in the image quality of X-IGRT system, and focuses on parameters that should be considered in the imaging performance evaluation criteria of the CBCT X-IGRT. The purpose is to sort out the imaging performance evaluation standards of kilovolt X-IGRT system, clarify the image quality requirements of X-IGRT equipment, and reach a consensus when evaluating the imaging performance of X-IGRT system.
Radiotherapy Planning, Computer-Assisted/methods*
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Cone-Beam Computed Tomography/methods*
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Spiral Cone-Beam Computed Tomography
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Radiotherapy, Intensity-Modulated/methods*
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Radiotherapy, Image-Guided/methods*
9.Therapeutic Effect and Mechanism of Flavonoids from Sophora Flavescens Ait. on Heat Stress-induced Reproductive Dysfunction in Mice
Yongwei LAI ; Qian LU ; Chao YE ; Yanchun WANG ; Yizhong ZHANG ; Shibing LIU ; Kuang REN ; Hongyan FAN
Chinese Journal of Modern Applied Pharmacy 2023;40(24):3368-3376
Abstract
OBJECTIVE To investigate the effect and mechanism of flavonoids from Sophora flavescens Ait. on testicular tissue damage in male mice induced by local heat stress in the scrotum. METHODS TCMSP database was used to screen the targets of flavonoids in Sophora flavescens Ait., and the bioinformatics analysis was performed on the target. The mouse model of scrotal heat stress was used and the flavonoids of Sophora flavescens Ait. was used for intervention. The sperm density and sperm aberration rate of mice in each group were measured, and the morphological changes of testicular tissue were observed. Tumor necrosis factor-α (TNF-α) and interleukin 17(IL-17) mRNA and protein levels in testicalar were detected of by q-PCR and Western blotting. Na+-K+-ATPase, Mg2+-ATPase, Ca2+-ATPase, lactate dehydrogenase(LDH), sorbitol dehydrogenase(SDH) activity, malondialdehyde (MDA), NO, TNF-α level and the content of testosterone in serum were detected in tissue homogenate. RESULTS Heat stress could lead to the decrease of sperm density and increase of aberrant sperm, the obvious thinning of testicular spermatogenic epithelium, the decrease of cell level and quantity, the significant decrease of ATPase, LDH, SDH activities, and the increase of MDA, NO content, TNF-α and IL-17 expression in testicular tissue. After the intervention with 250, 500 mg·kg-1·d-1 flavonoids of Sophora flavescens Ait., the quality of sperm and the damage of testicular tissue morphology were improved. The level of TNF-α and IL-17 in serum and testicular tissue were decreased, and the activity of ATPase, SDH and the level of testosterone were increased. CONCLUSION The mechanism of flavonoids of Sophora flavescens Ait. is through inhibiting the inflammatory factor TNF-α and IL-17 levels, improve the anti-lipid peroxidation ability and inhibite the role of NO, enhance the activity of energy enzymes in spermatogenesis, improve the level of serum testosterone, and improve the reproductive disorders caused by heat stress.
10.Design and application of Checklist for quality control in intensive care unit
Shibing ZHAO ; Qi ZOU ; Chao ZHANG ; Lunjun ZHANG ; Qiang WU ; Ximing DENG ; Huaxue WANG
Chinese Critical Care Medicine 2021;33(4):466-471
Objective:To design a Checklist for quality control in intensive care unit and observe the effect of clinical application.Methods:By consulting guidelines and literature, such as Critical care medicine professional medical quality control index (2015 edition), the quality control Checklist of intensive care unit was designed. It included four parts: quality control data collection, medical record quality verification, special diagnosis and treatment, and hospital infection prevention and control supervision. Every month, a doctor with a senior professional title served as the quality control director, and was responsible for the quality control of the department's medical care, including collecting data of the past 24 hours during the morning handover, discussing and registering special diagnosis and treatment behaviors that would be performed on the day, and coordinating with the nursing team leader, controlling the quality of the whole department throughout the day, such as supervising each medical staff if they had unreasonable behaviors, checking the running and discharge medical records, and inspecting the status of the staff on duty. The data in 2018, 2019 (Checklist implemented) and 2017 (Checklist not implemented) were retrospectively analyzed, including the status of admitted patients, department management information, length of intensive care unit (ICU) stay, and the incidence of three-tube infection [ventilator-associated pneumonia (VAP), catheter-related bloodstream infection (CRBSI), catheter-associated urinary tract infection (CAUTI)], and standardized mortality, etc. Results:From 2017 to 2019, the number of patients admitted was 373, 446, and 480, with annual growth of 19.57% and 7.62% in 2018 and 2019, respectively, and an increase of 28.69% in 2019 compared with 2017. There was no statistically significant difference in the average age and acute physiology and chronic health evaluationⅡ (APACHEⅡ) of patients in the three years. Compared with 2017, the length of ICU stay of patients in 2018 and 2019 were significantly shortened (days: 8.99±6.12, 9.14±7.02 vs. 10.20±7.21), and the incidence of VAP, CRBSI and CAUTI were significantly reduced [VAP (cases/1 000 ventilation days): 12.97±3.60, 9.62±3.14 vs. 17.48±4.89, CRBSI (cases/1 000 catheter days): 3.75±2.19, 3.87±1.87 vs. 6.19±3.13, CAUTI (cases/1 000 catheter days): 3.29±2.18, 3.28±1.87 vs. 5.61±3.18]. The standardized mortality were also significantly reduced [(77.27±7.24)%, (70.61±7.49)% vs. (84.41±9.05)%], the number of non-compliance with hospital infection prevention per month decreased significantly (person times: 54.00±6.30, 41.08±10.76 vs. 72.08±19.68), and the number of special diagnosis and treatment per month increased significantly (person times: 1 056.67±235.27, 1 361.75±278.48 vs. 722.25±145.96), the rate of etiology submission before antimicrobial treatment [(93.21±3.68)%, (96.59±2.49)% vs. (87.86±5.28)%] and deep vein thrombosis (DVT) prevention rate [(91.13±6.36)%, (96.23±2.99)% vs. (85.58±7.68)%] were significantly improved, and all the differences were statistically significant (all P < 0.05). All medical records in the three years were Grade A, but the average scores in 2018 and 2019 were higher than those in 2017 (96.82±2.84, 96.73±2.94 vs. 93.70±3.33, both P < 0.01). Compared with 2018, the incidence of VAP, the rate of etiology submission before antimicrobial treatment, the DVT prevention rate, and the standardized mortality rate in 2019 were further improved, and the number of non-compliance with hospital infection prevention per month decreased and the number of special diagnosis and treatment per month increased, and the differences were statistically significant (all P < 0.05). Conclusion:The application of quality control Checklist in intensive care unit can build an effective quality control system, reduce the incidence of three-tube infection, standardized mortality and length of ICU stay, improve the quality control awareness and execution of medical staff, and promote the improvement of medical quality.


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