1.Mechanism of Neochlorogenic Acid in Ameliorating Psoriatic Keratinocyte Proliferation and Inflammation by Targeting HSP90 to Modulate NF-κB/NLRP3 Signaling Pathway
Mengyao JIANG ; Xinwei ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):89-98
ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid (NA) and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe (NA-P) and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8(CCK-8). The potency of NA and NA-P was evaluated in the safe concentration range, and the effects of 0-100 μmol·L-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-23 (IL-23), and interleukin-17A (IL-17A) inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA), and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the effects of NA on the mRNA expression of keratin 16 (K16) in HaCaT cells, S100 calcium-binding protein A9 (S100A9), S100 calcium-binding protein A7 (S100A7), IL-6, IL-17A, and chemokine 1 (CXCL1). In vitro fluorescence labeling and competition experiments using NA-P were performed, and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry (Pull-down/LC-MS/MS) were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting (Pull down-WB), cellular heat transfer analysis combined with protein immunoblot (CETSA-WB) experiments, and molecular docking. Finally, Real-time PCR was utilized to detect the effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the mRNA expression of IL-1β, nucleotide-binding oligomeric structural domain-like receptor protein 3 (NLRP3), apoptosis-associated speckled-like protein (ASC), and cysteine protease-1 (Caspase-1) in HaCaT cells. Protein immunoblot (Western blot) was used to detect the effects of phosphorylated p65 (p-p65), p65, phosphorylated human nuclear factor-κB inhibitory protein α (p-IκBα), human nuclear factor κB inhibitory protein α (IκBα), and heat shock protein 90 (HSP90) expression. ResultsIn the 200 μmol·L-1 safe concentration range, HaCaT cell proliferation, increased expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors, and increased mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L-1 NA and NA-P groups was inhibited, and the expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors was decreased in the NA-L, NA-M, NA-H, and NA-P-H groups. The mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 was decreased (P<0.05). High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L-1 NA competing with 100 μmol·L-1 NA-P. Compared with that in the blank group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1, as well as the expression of p-p65/p65, p-IκBα/IκBα, and HSP90 protein, were increased in HaCaT cells in the M5 group (P<0.05). Compared with that in the M5 group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1 of cells in the NA-L group, the NA-M group, the NA-H group, and the NA-P-H group was decreased (P<0.05). p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups (P<0.05), and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein, and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1, HSP90B1, and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.
2.Mechanism of Neochlorogenic Acid in Ameliorating Psoriatic Keratinocyte Proliferation and Inflammation by Targeting HSP90 to Modulate NF-κB/NLRP3 Signaling Pathway
Mengyao JIANG ; Xinwei ZHANG ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):89-98
ObjectiveTo investigate the target proteins directly bound by neochlorogenic acid (NA) and the molecular mechanisms that ameliorate the proliferation and inflammatory response of psoriatic keratinocytes. MethodsM5-induced HaCaT cells were used as a psoriatic keratinocyte proliferation and inflammatory cell model. The synthesized NA probe (NA-P) and NA prodrug were first evaluated for cell viability using a cell proliferation/cell counting kit-8(CCK-8). The potency of NA and NA-P was evaluated in the safe concentration range, and the effects of 0-100 μmol·L-1 NA and probe on M5-induced proliferation of HaCaT cells were detected using CCK-8. The effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-23 (IL-23), and interleukin-17A (IL-17A) inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA), and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to measure the effects of NA on the mRNA expression of keratin 16 (K16) in HaCaT cells, S100 calcium-binding protein A9 (S100A9), S100 calcium-binding protein A7 (S100A7), IL-6, IL-17A, and chemokine 1 (CXCL1). In vitro fluorescence labeling and competition experiments using NA-P were performed, and target protein angling and analysis using pull-down experiments combined with liquid chromatography-mass spectrometry (Pull-down/LC-MS/MS) were conducted. Target validation was performed using pull-down experiments combined with protein immunoblotting (Pull down-WB), cellular heat transfer analysis combined with protein immunoblot (CETSA-WB) experiments, and molecular docking. Finally, Real-time PCR was utilized to detect the effects of 20, 40, 80 μmol·L-1 NA and 80 μmol·L-1 NA-P on the mRNA expression of IL-1β, nucleotide-binding oligomeric structural domain-like receptor protein 3 (NLRP3), apoptosis-associated speckled-like protein (ASC), and cysteine protease-1 (Caspase-1) in HaCaT cells. Protein immunoblot (Western blot) was used to detect the effects of phosphorylated p65 (p-p65), p65, phosphorylated human nuclear factor-κB inhibitory protein α (p-IκBα), human nuclear factor κB inhibitory protein α (IκBα), and heat shock protein 90 (HSP90) expression. ResultsIn the 200 μmol·L-1 safe concentration range, HaCaT cell proliferation, increased expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors, and increased mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 were observed in the M5 group compared with the blank group. Cell proliferation in 5-100 μmol·L-1 NA and NA-P groups was inhibited, and the expression of TNF-α, IL-1β, IL-23, and IL-17A inflammatory factors was decreased in the NA-L, NA-M, NA-H, and NA-P-H groups. The mRNA expression of K16, S100A9, S100A7, IL-6, IL-17A, and CXCL1 was decreased (P<0.05). High-confidence targets were screened for HSP90 protein by Pull-down/LC-MS/MS using 200 μmol·L-1 NA competing with 100 μmol·L-1 NA-P. Compared with that in the blank group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1, as well as the expression of p-p65/p65, p-IκBα/IκBα, and HSP90 protein, were increased in HaCaT cells in the M5 group (P<0.05). Compared with that in the M5 group, the mRNA expression of NLRP3, IL-1β, ASC, and Caspase-1 of cells in the NA-L group, the NA-M group, the NA-H group, and the NA-P-H group was decreased (P<0.05). p-p65/p65 and p-IκBα/IκBα were decreased in the NA-M and NA-H groups (P<0.05), and there was no change in HSP90 protein. Pull down-WB showed that NA could directly target HSP90 protein, and NA binding to HSP90 protein enhanced its thermal stability. Molecular docking of NA with HSP90 family proteins HSP90AA1, HSP90B1, and HSP90AB1 all resulted in highly stable binding. ConclusionNA can inhibit the proliferation and inflammatory response of psoriatic keratinocytes by a mechanism that may be achieved by targeting HSP90 to modulate the NF-κB/NLRP3 signaling pathway.
3.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
4.Mechanism of Tibetan Medicine Sanwei Doukoutang to Improve Cognitive Dysfunction in 5×FAD Mice Based on Wnt/β-catenin Signaling Pathway
Shuran LI ; Yaxin WANG ; Jing SUN ; Lei BAO ; Zihan GENG ; Dan XIE ; Ronghua ZHAO ; Yanyan BAO ; Qiyue SUN ; Jingsheng ZHANG ; Xinwei WANG ; Xinying LI ; Xihe CUI ; Xiaowei YANG ; LIUXIAN ; Mengyao CUI ; Qingshan LIU ; Shanshan GUO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):54-60
ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang (SWDK) on cognitive dysfunction in mice suffering from Alzheimer's disease (AD) and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group, total ginsenoside group(0.04 g·kg-1), high-, medium-, and low-dose groups of SWDK (32.60, 16.30, 8.15 g·kg-1), with 10 mice in each group, and ten wild-type mice of the same age were used as the normal group, male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin (HE) staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin (SYP) and postsynaptic dense substance 95 (PSD95) in mice serum were detected by enzyme-linked immunosorbent assay (ELISA). The positive expression of brain-derived neurotrophic factor(BDNF) in the dentate gyrus (DG) region of mouse brain tissue was observed by immunohistochemistry (IHC). The protein levels of BDNF, Wnt family member 3A(Wnt3a), and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice, the model group of mice had significantly more complex swimming routes and lower swimming speed (P<0.01), significantly lower percentage of time spent in the target quadrant (P<0.01), and a significantly lower percentage of freezing time (P<0.05). The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95(P<0.01) in the serum of mice were reduced, and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice were obviously reduced (P<0.05, P<0.01). Compared with the model group, the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes, the high- and medium- dose SWDK groups significantly higher swimming speeds (P<0.01), significantly higher percentage of time spent in the target quadrant (P<0.01), obviously higher percentage of Freezing time (P<0.05), and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP (P<0.05, P<0.01), PSD95 (P<0.01), increased BDNF-positive cells in the DG region of brain tissue, and obviously elevated levels of BDNF, Wnt3a, and β-catenin proteins in the hippocampus of mice (P<0.05, P<0.01). ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice, and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway, which promotes BDNF expression and thereby enhances synaptic plasticity, allowing neuronal signaling to be restored.
5.Interventional revascularization combined with perforator composite flap for staged treatment of peripheral arterial disease with ankle soft tissue defects.
Xiaoguang GUO ; Zhiguo WANG ; Zheng KANG ; Yanzhou LI ; Junxian YANG ; Weihua FENG ; Honglüe TAN ; Guoqiang JIN ; Xinwei WANG
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(12):1580-1585
OBJECTIVE:
To explore the effectiveness of primary interventional revascularization combined with secondary perforator composite flap in the treatment of peripheral arterial disease (PAD) accompanied by soft tissue defects around the ankle.
METHODS:
Between January 2022 and January 2025, 12 patients with PAD and soft tissue defects around the ankle were admitted. Among them, there were 9 males and 3 females; their ages ranged from 52 to 82 years, with an average of 68.9 years. The causes of injury included 4 cases of traffic accident, 5 cases of falls, 1 case of falling from height, 1 case of foreign body puncture injury, and 1 case of electric shock injury. The infection duration ranged from 1 month to 35 years, with a median duration of 3.5 months. The wound size ranged from 5.5 cm×3.0 cm to 15.0 cm×9.0 cm. The ankle-brachial index (ABI) was 0.32±0.12. The visual analogue scale (VAS) score for pain was 3.3±0.5. Preoperative vascular stenosis assessment was performed in all patients, with primary intervention to dredge large and medium-sized arteries, followed by secondary repair of the wound using a perforator composite flap. The flap size ranged from 6.5 cm×4.0 cm to 16.0 cm×10.0 cm. The donor sites were sutured directly or repaired with skin grafts. After two stages of treatment, the effectiveness was evaluated by measuring ABI, observing flap survival and wound healing, assessing VAS scores, and American Orthopedic Foot and Ankle Society (AOFAS) scores.
RESULTS:
All 12 cases completed two stages of treatment; all patients were followed up after the second-stage treatment, with a follow-up period ranging from 7 to 28 months, with an average of 16.8 months. After the first-stage treatment, the skin temperature around the ankle was significantly higher than that before treatment, and the ABI increased to 0.71±0.07, with a significant difference ( t=9.918, P<0.001). After the second-stage treatment, the blisters on the distal end of the skin flap occurred in 3 cases. The flaps survived and the wounds healed, with a healing time ranging from 10 to 14 days (mean, 11.8 days). The incisions at the donor site healed by first intention, and the skin grafts survived. The VAS score was 0.5±0.5 at 3 weeks, which was significantly lower than that before treatment ( t=13.675, P<0.001). No infection recurrence occurred during follow-up. At 6 months after the second-stage treatment, the AOFAS score of the ankle joint ranged from 92 to 97, with an average of 94.7, all reaching excellent.
CONCLUSION
Interventional revascularization combined with perforator composite flap for staged treatment of PAD with ankle soft tissue defects can obtain good effectiveness, by unclogging the main blood vessels, improving lower limb blood supply, and improving the survival rate of the skin flap.
Humans
;
Male
;
Female
;
Middle Aged
;
Aged
;
Peripheral Arterial Disease/surgery*
;
Soft Tissue Injuries/surgery*
;
Perforator Flap/blood supply*
;
Plastic Surgery Procedures/methods*
;
Aged, 80 and over
;
Ankle/blood supply*
;
Treatment Outcome
;
Ankle Brachial Index
;
Skin Transplantation/methods*
6.The association between body mass index and in-hospital major adverse cardiovascular and cerebral events in patients with acute coronary syndrome
Qing ZHOU ; Dan ZHU ; Yiting WANG ; Wenyue DONG ; Jie YANG ; Jun WEN ; Jun LIU ; Na YANG ; Dong ZHAO ; Xinwei HUA ; Yida TANG
Chinese Journal of Cardiology 2024;52(1):42-48
Objective:To assess the association between body mass index (BMI) and major adverse cardiovascular and cerebrovascular events (MACCE) among patients with acute coronary syndrome (ACS).Methods:This was a multicenter prospective cohort study, which was based on the Improving Care for Cardiovascular Disease in China (CCC) project. The hospitalized patients with ACS aged between 18 and 80 years, registered in CCC project from November 1, 2014 to December 31, 2019 were included. The included patients were categorized into four groups based on their BMI at the time of admission: underweight (BMI<18.5 kg/m 2), normal weight (BMI between 18.5 and 24.9 kg/m 2), overweight (BMI between 25.0 and 29.9 kg/m 2), and obese (BMI≥30.0 kg/m 2). Multivariate logistic regression models was used to analyze the relationship between BMI and the risk of in-hospital MACCE. Results:A total of 71 681 ACS inpatients were included in the study. The age was (63.4±14.7) years, and 26.5% (18 979/71 681) were female. And the incidence of MACCE for the underweight, normal weight, overweight, and obese groups were 14.9% (322/2 154), 9.5% (3 997/41 960), 7.9% (1 908/24 140) and 7.0% (240/3 427), respectively ( P<0.001). Multivariate logistic regression analysis showed a higher incidence of MACCE in the underweight group compared to the normal weight group ( OR=1.30, 95% CI 1.13-1.49, P<0.001), while the overweight and obese groups exhibited no statistically significant difference in the incidence of MACCE compared to the normal weight group (both P>0.05). Conclusion:ACS patients with BMI below normal have a higher risk of in-hospital MACCE, suggesting that BMI may be an indicator for evaluating short-term prognosis in ACS patients.
7.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.
8.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.
9.Correlation analysis on serum proline dehydrogenase levels and left ventricular systolic function in patients with chronic heart failure
Kaitong YANG ; Lili HE ; Qingjuan ZUO ; Xinwei YU ; Yifang GUO
Journal of Jilin University(Medicine Edition) 2024;50(6):1719-1727
Objective:To discuss the differences in serum proline dehydrogenase(ProDH)levels among chronic heart failure(CHF)patients with different ejection fraction types,and to clarify the effect of ProDH levels on cardiac function.Methods:A retrospective analysis of clinical data of 118 CHF patients was conducted.These patients were divided into heart failure with reduced ejection fraction(HFrEF)group(n=39),heart failure with mid-range ejection fraction group(HFmrEF)(n=42),and heart failure with preserved ejection fraction(HFpEF)group(n=37).A total of 45 non-CHF patients hospitalized during the same period were collected as control group.The general data of all the subjects in various groups were collected,and the levels of biochemical indicators and cardiac structure indicators in serum of all the subjects were detected.Spearman correlation analysis and point-biserial correlation analysis were used to analyze the correlation between serum ProDH levels and various biochemical indicators;multivariate Logistic regression analysis was used to analyze the factors influencing HFrEF and HFmrEF.Results:Compared with control group,the usage rate of beta-blockers of the patients in HFpEF group was significantly increased(P<0.05);in HFmrEF group,the percentage of male patients,the usage rate of statins,and the usage rate of beta-blockers were all significantly increased(P<0.05);in HFrEF group,the age and systolic blood pressure(SBP)of the patients were significantly decreased(P<0.05),while the usage rates of statins and beta-blockers of the patients were significantly increased(P<0.05).Compared with HFpEF group,the age of the patients in HFmrEF group was significantly decreased(P<0.05),and the percentage of male patients and the usage rate of statins were significantly increased(P<0.05);the age of the patients in the HFrEF group was significantly decreased(P<0.05),and the usage rate of statins was significantly increased(P<0.05).Compared with HFmrEF group,the SBP of the patients in HFrEF group was significantly decreased(P<0.05).Compared with control group,the serum levels of low-density lipoprotein cholesterol(LDL-c)of the patients in HFpEF and HFmrEF groups were significantly decreased(P<0.05),while the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP)were significantly increased(P<0.05);the serum levels of glomerular filtration rate(GFR)and ProDH of the patients in HFrEF group were significantly decreased(P<0.05),and the levels of fasting blood glucose(FBG)and NT-proBNP were significantly increased(P<0.05).Compared with HFpEF group,the serum hemoglobin(Hb)level of the patients in HFmrEF group was significantly increased(P<0.05);the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05),while the ProDH level was significantly decreased(P<0.05).Compared with HFmrEF group,the serum NT-proBNP level of the patients in HFrEF group was significantly increased(P<0.05).Compared with control group,the left atrial diameter(LAD)and the ratio of early diastolic mitral inflow velocity to early diastolic mitral annular velocity(E/Em)of the patients in HFpEF,HFmrEF,and HFrEF groups were significantly increased(P<0.05);the left ventricular end-diastolic diameter(LVEDD)of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the left ventricular ejection fraction(LVEF)were significantly decreased(P<0.05).Compared with HFpEF group,the LVEDD of the patients in HFmrEF and HFrEF groups were significantly increased(P<0.05),and the LVEF were significantly decreased(P<0.05);the LAD of the patients In HFrEF group was significantly increased(P<0.05).Compared with HFmrEF group,the E/Em ratio of the patients in HFrEF group was significantly increased(P<0.05),and the LVEF was significantly decreased(P<0.05).The serum ProDH levels of the patients were negatively correlated with LVEDD(r=-0.210,P=0.007)and positively correlated with LVEF(r=0.220,P=0.005).Male and elevated FBG levels were the risk factors for cardiac function,while the increasing serum GFR and ProDH levels were the protective factors for cardiac function.Conclusion:There are differences in ProDH levels among the CHF patients with different ejection fraction types.The patients with poorer cardiac function have lower serum ProDH levels,and higher ProDH levels may be beneficial for improving the left ventricular systolic function in the CHF patients.
10.Dual-source CT virtual monoenergetic imaging for differentiating prostate cancer and benign prostatic hyperplasia
Xinwei ZHONG ; Yi CHEN ; Bowen YUE ; Jianhui LI ; Hao ZHANG ; Xiaohong CHEN ; Xiaofeng CHEN ; Junliang DENG ; Wenzhong HOU ; Zhiqi YANG
Chinese Journal of Medical Imaging Technology 2024;40(11):1749-1753
Objective To observe the value of dual-source CT(DECT)virtual monoenergetic imaging(VMI)for differentiating prostate cancer(PC)and benign prostatic hyperplasia(BPH).Methods Thirty-three patients with PC(PC group)and 44 patients with BPH(BPH group)were retrospectively enrolled,and 40-100 keV(with 10 keV interval)VMI were reconstructed based on arterial phase DECT images,respectively.Clinical data,focal CT value and contrast-to-noise ratio(CNR)of VMI with different energy levels and conventional linear fusion images of arterial phase were compared between groups.Binary logistic regression models were constructed based on clinical data being significantly different between groups,focal CT value of VMI with energy level with the highest CNR,also focal CT value of conventional linear fusion images of arterial phase,respectively.The area under the receiver operating characteristic curve(AUC)was calculated to evaluate the efficacy of each model for differentiating PC and BPH.Results Free prostate-specific antigen(f-PSA),total prostate-specific antigen(t-PSA),focal CT value of 40-100 keV VMI and conventional linear fusion images of arterial phase in PC group were all higher,while short diameter in PC group was smaller than those in BPH group(all P<0.05).Logistic regression models were constructed based on f-PSA,t-PSA,short diameter,CT value at 40 keV VMI and CT value at conventional linear fusion images of arterial phase,respectively,with AUC for differentiating PC and BPH of 0.879,0.902,0.701,0.911 and 0.857,respectively.Conclusion DECT VMI could be used as a supplementary examination for prostate diseases,and 40 keV VMI had the best efficacy for differentiating PC and BPH.

Result Analysis
Print
Save
E-mail