1.Quercetin protects THP-1 macrophages against cellular stress response induced by Pseudomonas aeruginosa protease LasB
Yanying REN ; Xinwei LIU ; Rui ZHU ; Xiaojuan YOU ; Dengzhou LI ; Zhiqiang HE ; Chunxia WANG ; Ximing YANG ; Yongwei LI
Chinese Journal of Preventive Medicine 2024;58(3):337-346
Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.
2.Quercetin protects THP-1 macrophages against cellular stress response induced by Pseudomonas aeruginosa protease LasB
Yanying REN ; Xinwei LIU ; Rui ZHU ; Xiaojuan YOU ; Dengzhou LI ; Zhiqiang HE ; Chunxia WANG ; Ximing YANG ; Yongwei LI
Chinese Journal of Preventive Medicine 2024;58(3):337-346
Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.
3.The effect of pressure controlled ventilation-volume guaranteed combined with personalized positive end expiratory pressure ventilation on oxygenation function and postoperative pulmonary complications in patients undergoing brain tumor surgery
Jiakun LIU ; Pei SHI ; Fengzhi LIU ; Haiyan WU ; Yuelan WANG ; Ximing LI
Journal of Chinese Physician 2024;26(6):863-869
Objective:To explore the ventilation strategy of using pressure controlled ventilation-volume guaranteed (PCV-VG) mode combined with personalized positive end expiratory pressure (PEEP) during surgery, and its impact on oxygenation function and incidence of postoperative pulmonary complications (PPCs) in patients undergoing brain tumor surgery.Methods:Sixty patients who underwent elective brain tumor surgery at the Linyi People′s Hospital from January 2023 to June 2023 were selected. The patients were randomly divided into PCV-VG ventilation mode group (T group) and volume controlled ventilation (VCV) mode group (C group) using a random number table method, with 30 patients in each group. One patient was excluded from group T due to changes in the condition, and 29 patients were actually included. Group T adopted PCV-VG ventilation mode and searched for the maximum dynamic lung compliance (Cdyn) by titrating PEEP. The PEEP corresponding to the maximum Cdyn value was the optimal PEEP, which was maintained until the end of mechanical ventilation. Group C adopted a ventilation mode of VCV plus 5 cmH 2O fixed value PEEP. All patients underwent arterial blood gas analysis before anesthesia induction (T 1), 15 minutes after setting ventilation mode (T 2), and 15 minutes after extubation (T 3), recording arterial oxygen partial pressure (PaO 2), carbon dioxide partial pressure (PaCO 2), lactate (Lac), and blood glucose (Glu), and calculating oxygenation index (PaO 2/FiO 2). The levels of white blood cells (WBC), C-reactive protein (CRP), and procalcitonin (PCT) were recorded before and 72 hours after surgery. The incidence of postoperative PPCs at 72 hours was observed. Results:There was a statistically significant difference in the grouping effect and time effect of PaO 2 and PaCO 2 between group C and group T (all P<0.05), while there was no statistically significant difference in the interaction effect (all P>0.05); The differences in grouping effects, time effects, and interaction effects of PaO 2/FiO 2 between group C and group T were statistically significant (all P<0.05); The average value of the optimized PEEP obtained by T-group titration was 7.48 cmH 2O, corresponding to an average platform pressure (Pplat) of 14.90 cmH 2O, and an average value of 53.37 ml/cmH 2O corresponding to the maximum Cdyn value. Compared with the Pplat and Cdyn corresponding to the fixed value of 5 cmH 2O PEEP in Group C, the Pplat corresponding to the optimized PEEP in Group T was lower than that in Group C ( P<0.05), and the Cdyn was higher than that in group C ( P<0.05). The CRP level and incidence of PPCs in group T after 72 hours of surgery were significantly lower than those in group C (all P<0.05). Conclusions:Compared with VCV, the ventilation mode of PCV-VG combined with personalized PEEP can improve intraoperative lung ventilation and oxygenation function in patients with brain tumors, reduce the occurrence of pulmonary inflammation 72 hours after surgery, and lower the incidence of PPCs.
4.Mechanism of the mechanosensitive channel Piezo1 regulating chondrocytes in post-traumatic osteoarthritis: a review
Gongwu YUAN ; Zhigang LI ; Xi KE ; Ximing LIU
Chinese Journal of Trauma 2024;40(11):1040-1048
Post-traumatic osteoarthritis (PTOA) is a chronic degenerative joint disease caused by joint injury and abnormal mechanical stress stimulation is one of its key mechanisms. Currently, it is difficult to repair the articular cartilage completely even with surgical treatment and there is a lack of effective treatments for PTOA. Chondrocytes, one of the primary cell types involved in the pathogenic process of PTOA, are highly sensitive to mechanical signals. The mechanosensitive ion channel Piezo1 serves the important functions of sensing external forces and mechanical stimuli and plays a crucial role in PTOA by regulating chondrocytes′ responses to mechanical stress. Therefore, its essential to understand the mechanisms of the Piezo1 channel in regulating PTOA chondrocytes in exploring new therapeutic strategies. To this end, the authors reviewed the research progress on the regulatory mechanisms of the Piezo1 channel on PTOA chondrocytes, aiming to provide a reference for future studies on PTOA mechanisms and treatment options.
5.Analysis of electrocardiographic features and in-hospital mortality in acute total left main artery occlusion and subtotal occlusion
Chunwei LIU ; Fan YANG ; Yuecheng HU ; Jingxia ZHANG ; Hongliang CONG ; Ximing LI
Tianjin Medical Journal 2024;52(7):755-761
Objective To investigate the difference of electrocardiographic(ECG)features between total left main artery(LM)occlusion and subtotal occlusion,and analyze risk factors of in-hospital mortality.Methods A total of 94 patients with left main complete occlusion and 99 patients with subtotal occlusion were included.ECG characteristics,coronary angiography and other clinical data were compared,and factors of hospital death were analyzed.The receiver operating characteristics(ROC)curve was used to analyze the predictive value of ECG characteristics in hospital death risk in patients with LM occlusion.The relationship between ECG characteristics,shock and collateral circulation were analyzed in patients with LM occlusion.Results Compared with the subtotal occlusion group,patients with LM occlusion presented with more ST-segment elevation(STE)in Ⅰ,avL,V2-V5,more STE in avR and avL,more left anterior fascicular block+right bundle branch block,prolonged QRS duration,less STE in avR and less STE in avR+V1.The in-hospital mortality was 46.8%(44/94)in LM occlusion and 14.1%(14/99)in LM subtotal occlusion.STE in avR+avL predicted total LM occlusion with a specificity of 0.97,and left anterior branch+right bundle branch block predicted total LM occlusion with a specificity of 1.00.In patients with total LM occlusion,STE in Ⅰ,avL,V2-V5,prolongation of QRS duration,shock,no collateral circulation,STE in Ⅰ,avL,V2-V5 combined with left anterior fascicular block+right bundle branch block,and STE in Ⅰ,avL,V2-V5 combined with shock predicted in-hospital mortality,with the area under the curve of 0.716,0.619,0.766,0.688,0.572,0.785,respectively.The diagnostic specificity of STE in Ⅰ,avL,V2-V5 combined with shock was 0.82,and the sensitivity was 0.75.STE in Ⅰ,avL,V2-V5 combined with left anterior fascicular block+right bundle branch block predicted in-hospital death in LM occlusion with a specificity of 0.94.The proportion of shock was higher in patients with STE in Ⅰ,avL,V2-V5,left anterior fascicular block+right bundle branch block and collateral flow absence(P<0.05).In patients with total occlusion,no collateral flow was observed in patients with STE in Ⅰ,avL,V2-V5.In patients with STE in avR(including avR+V1),82.4%of patients presented with right coronary collateral circulation supplying the left anterior descending coronary artery and left circumflex artery territory.In patients with STE in avR+avL,69.2%of patients presented with right coronary collateral circulation supplying left anterior descending coronary artery territory.Conclusion Total LM occlusion presents with different ECG features compared with subtotal occlusion.In LM total occlusion,the ECG features predict in-hospital mortality and are associated with different collateral circulation.
6.Preliminary research on clinical application of a new flexible patch ECG
Yibei LU ; Dongxia JIN ; Zhenchun SONG ; Haoyuan MA ; Yan LI ; Tianxu HAO ; Ximing LI
Tianjin Medical Journal 2024;52(10):1095-1099
Objective To explore the diagnostic value of a wearable flexible patch ECG instrument in arrhythmia,the alarm situation during clinical application,patient satisfaction and safety.Methods A total of 1 443 subjects wore flexible patch ECG and conventional dynamic ECG(control)for 24 h to test the validity and consistency of arrhythmia diagnosis,and counted the alarm of remote ECG and the occurrence of related adverse events during the wearing of the instrument.Results There were 987 cases of arrhythmia detected by flexible patch ECG and 992 cases by conventional dynamic ECG.The total coincidence rate of arrhythmia diagnosis was 98.7%.The mean heart rate was measured by flexible patch ECG(75.4±11.4)times/min,conventional dynamic heart rate(71.5±12.1)times/min,the intra-group correlation coefficient(ICC)of 2 instruments was 0.892(95%CI:0.537-0.956),with good repeatability.The correct alarm rate of flexible patch ECG was 100%.The incidences of skin pruritus(0.28%vs.1.32%),skin allergy,redness and swelling(0.14%vs.0.69%)and electrode strip shedding(0 vs.0.28%)during wearing the flexible patch electrocardiogram were lower than those of the conventional holter electrocardiogram(P<0.05).Conclusion The flexible patch ECG has few adverse reactions,high comfort,good safety and clinical applicability.
7.Effects of Jianpi Huatan Fang on lipid metabolism and FOXO1/PDK4 expression in the ovary in rats with polycystic ovary syndrome and insulin resistance
Luyu YANG ; Anying LIN ; Zhengwang YANG ; Lan LI ; Ximing YU
Acta Laboratorium Animalis Scientia Sinica 2024;32(4):451-460
Objective To investigate the effects of Jianpi Huatan Fang on lipid metabolism and expression levels of FOXO1 and PDK4 in rats with polycystic ovary syndrome(PCOS)and insulin resistance(IR).Methods Forty female rats were divided randomly into a blank control group(n=10)and a model group(n=30).A PCOS-IR rat model was established using a high-fat diet(8 weeks)combined with letrozole(added at weeks 4~8).Thirty successfully modeled rats were then divided randomly into a model control group,a Jianpi Huatan Fang group(11.07 g/kg),and a metformin group(0.2 g/kg)(n=10 rats per group).Drug intervention was given for 4 weeks.The general status,Traditional Chinese medicine syndrome score,and weight changes were observed in each group.Histopathological changes in the ovary were detected by hematoxylin and eosin staining.Fasting blood glucose and blood lipids were measured using a blood biochemical analyzer.Levels of sex hormones including follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),estradiol(E2),and insulin were determined by enzyme linked immunosorbent assay and changes in the expression of FOXO1 and PDK4 in the ovaries were determined by Western Blot and real-time fluorescence quantitative polymerase chain reaction.Results Rats in the model control group exhibited symptoms of spleen deficiency and phlegm-dampness and showed significant weight gain,sex hormone disorders,polycystic ovarian changes,and dysregulation of glucose and lipid metabolism,compared with rats in the blank control group(P<0.01).In addition,weight increase was slower in rats in the Jianpi Huatan Fang group and the metformin group compared with that in the model control group,and follicle development was improved.Serum LH,T,LH/FSH,fasting blood glucose,insulin,homeostatic model assessment for IR,cholesterol,triglycerides,and low-density lipoprotein cholesterol decreased,estradiol and FSH increased,and ovarian FOXO1 and PDK4 mRNA and protein expression levels were down regulated in the Jianpi Huatan Fang group compared with the findings in the model control group(P<0.05,P<0.01).Serum high-density lipoprotein cholesterol content was also increased,but the difference was not significant(P>0.05).Conclusions Jianpi Huatan Fang can effectively regulate sex hormone secretion,improve ovarian reproductive function,and regulate glucose and lipid metabolism in obese PCOS-IR rats,possibly via inhibiting the FOXO1/PDK4 pathway.
8.Study on mechanism of Jianpi Huatan Prescription in insulin resistance of polycystic ovary syndrome through AMPK/LKB1 signaling pathway
Tian LUO ; Xiao LI ; Anying LIN ; Luyu YANG ; Lan LI ; Ximing YU
International Journal of Traditional Chinese Medicine 2024;46(10):1303-1309
Objective:To investigate the effects of Jianpi Huatan Prescription on a rat model of polycystic ovary syndrome (PCOS) with insulin resistance (IR) induced by letrozole combined with high fat diet based on the liver kinase B1 (LKB1)/AMP activated protein kinase (AMPK) signaling pathway; To discuss its mechanism.Methods:A total of 40 SD female rats were divided into model group (30 rats) and blank group (10 rats). The model group rats were fed with high-fat feed from the first week of modeling for 8 consecutive weeks, and from the fourth week onwards, were given a daily gavage of letrozole solution for 5 consecutive weeks; the blank group was fed normally, and from the 4th week onwards, an equal amount of sodium carboxymethyl cellulose solution was administered. After the completion of modeling, the modeling group was divided into Jianpi Huatan Prescription group, Metformin group, and model group according to random number table method. The Jianpi Huatan Prescription group and the metformin group were given corresponding drug interventions, while the other two groups were simultaneously administered an equal amount of physiological saline for a total of 4 weeks. Ovarian morphological changes were observed using HE staining; intraperitoneal glucose tolerance test (IPGTT) was used to detect glucose tolerance in rats; Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estrogen (E 2), total testosterone (T), fasting blood glucose (FBG), and fasting insulin (FINS) in rats; Real time fluorescence quantitative PCR and Western blot techniques were used to detect changes in LKB1, AMPK mRNA and protein expression in the LKB1/AMPK signaling pathway of rat ovarian tissue. Results:Compared with the model group, the polycystic changes in the ovaries of rats in the Jianpi Huatan Prescription group were improved, and the growth rate of body weight decreased. The levels of FBG, FINS, HOMA-IR, and IPGTT decreased ( P<0.05 or P<0.01), while serum FSH and HDL-C levels were significantly up-regulated ( P<0.05 or P<0.01). The levels of T, LH, LH/FSH, TC, TG, and LDL-C decreased ( P<0.05 or P<0.01); The mRNA and protein expression levels of LKB1 and AMPK in ovarian tissue increased ( P<0.05). Conclusion:Jianpi Huatan Prescription can improve PCOS-IR in rats caused by the combination of letrozole and high-fat diet, regulate various levels of sex hormones, and improve glucose and lipid metabolism disorders by regulating the LKB1/AMPK signaling pathway, which has a repairing effect on polycystic ovary disease in rats.
9.Research Progress of Artificial Intelligence in the Diagnosis and Treatment of Anorectal Diseases
Yiwen ZHANG ; Ximing WANG ; Zilong LI ; Xinzhang ZHANG ; Changxian CHEN ; Weijun LIU ; Zhenyong ZHANG
Journal of Kunming Medical University 2024;45(2):1-6
In the past 20 years,the development of artificial intelligence has made rapid progress,and it is increasingly applied in the medical field,including medical image-assisted diagnosis and treatment,health management,disease risk prediction and so on.In this paper,the application status of artificial intelligence-assisted detection and diagnosis system based on deep learning in anorectal diseases is summarized,and the new methods related to the diagnosis and treatment of anorectal diseases at home and abroad are summarized.It mainly reviews the research progress of artificial intelligence technology in the diagnosis and treatment of anal fistula,perianal abscess,hemorrhoids and other anorectal diseases.
10.Research progress in the prevention of contrast-related acute kidney injury
Tianjin Medical Journal 2024;52(3):327-331
Contrast-associated acute kidney injury(CA-AKI)is an important complication caused by the use of contrast medium(CM)in diagnostic or interventional surgery.At present,it has become one of the major causes of acute renal insufficiency in hospitalized patients.Choosing a relatively low toxic CM and reducing the exposure time and dose of CM can prevent CA-AKI occurrence to some extent.Drugs such as statins and postoperative hydration can reduce the risk of CA-AKI.In addition,nanomedicine has shown a benefit in animal models.This paper reviews the current prevention and treatment of CA-AKI to lay the foundation for further study of new interventions and provide a theoretical basis for clinical treatment.

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