1.Research progress on calcium activities in astrocyte microdomains.
Fu-Sheng DING ; Si-Si YANG ; Liang ZHENG ; Dan MU ; Zhu HUANG ; Jian-Xiong ZHANG
Acta Physiologica Sinica 2025;77(3):534-544
Astrocytes are a crucial type of glial cells in the central nervous system, not only maintaining brain homeostasis, but also actively participating in the transmission of information within the brain. Astrocytes have a complex structure that includes the soma, various levels of processes, and end-feet. With the advancement of genetically encoded calcium indicators and imaging technologies, researchers have discovered numerous localized and small calcium activities in the fine processes and end-feet. These calcium activities were termed as microdomain calcium activities, which significantly differ from the calcium activities in the soma and can influence the activity of local neurons, synapses, and blood vessels. This article elaborates the detection and analysis, characteristics, sources, and functions of microdomain calcium activities, and discusses the impact of aging and neurodegenerative diseases on these activities, aiming to enhance the understanding of the role of astrocytes in the brain and to provide new insights for the treatment of brain disorders.
Astrocytes/cytology*
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Humans
;
Animals
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Calcium/metabolism*
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Calcium Signaling/physiology*
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Brain/physiology*
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Aging/physiology*
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Membrane Microdomains/physiology*
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Neurodegenerative Diseases/physiopathology*
2.Functional characterization of flavonoid glycosyltransferase AmGT90 in Astragalus membranaceus.
Guo-Qing PENG ; Bing-Yan XU ; Jian-Ping HUANG ; Zhi-Yin YU ; Sheng-Xiong HUANG
China Journal of Chinese Materia Medica 2025;50(6):1534-1543
Astragalus membranaceus(A. membranaceus), a traditional tonic, contains flavonoids as one of its main bioactive components and key indicators for quality standard detection. These compounds predominantly exist in glycosylated forms after glycosylation modification within the plant. The catalytic products of flavonoid glycosyltransferases in A. membranaceus have been reported to be mostly monoglycosides, and only AmUGT28 catalyzes luteolin to form diglycosides. In this study, we cloned a glycosyltransferase gene, AmGT90, from A. membranaceus, with an ORF length of 1 335 bp, encoding 444 amino acids, and the protein had a relative molecular mass of 50.5 kDa. Phylogenetic tree analysis indicated that AmGT90 belongs to the UGT74 family. In vitro enzymatic reaction showed that AmGT90 had broad substrate specificity and could catalyze the glycosylation of various flavonoids, including isoflavones, flavones, flavanones, and chalcones. AmGT90 not only catalyzed the formation of monoglycosides but also diglycosides. In addition, the mechanism of AmGT90 catalyzing the formation of diglycosides from luteolin was preliminarily explored. The experimental results showed that AmGT90 may preferentially recognize C4'-OH of luteolin and then recognize C7-OH to form diglycosides. This study reported a glycosyltransferase from A. membranaceus capable of converting flavonoids into monoglycosides and diglycosides. This finding not only enhances our understanding of the biosynthetic pathways of flavonoid glycosides in A. membranaceus but also introduces a new component for glycoside production through synthetic biology.
Glycosyltransferases/chemistry*
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Flavonoids/chemistry*
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Astragalus propinquus/classification*
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Phylogeny
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Glycosylation
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Plant Proteins/chemistry*
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Substrate Specificity
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Cloning, Molecular
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Amino Acid Sequence
3.Molecular mechanism of magnesium alloy promoting macrophage M2 polarization through modulation of PI3K/AKT signaling pathway for tendon-bone healing in rotator cuff injury repair.
Xianhao SHENG ; Wen ZHANG ; Shoulong SONG ; Fei ZHANG ; Baoxiang ZHANG ; Xiaoying TIAN ; Wentao XIONG ; Yingguang ZHU ; Yuxin XIE ; Zi'ang LI ; Lili TAN ; Qiang ZHANG ; Yan WANG
Chinese Journal of Reparative and Reconstructive Surgery 2025;39(2):174-186
OBJECTIVE:
To evaluate the effect of biodegradable magnesium alloy materials in promoting tendon-bone healing during rotator cuff tear repair and to investigate their potential underlying biological mechanisms.
METHODS:
Forty-eight 8-week-old Sprague Dawley rats were taken and randomly divided into groups A, B, and C. Rotator cuff tear models were created and repaired using magnesium alloy sutures in group A and Vicryl Plus 4-0 absorbable sutures in group B, while only subcutaneous incisions and sutures were performed in group C. Organ samples of groups A and B were taken for HE staining at 1 and 2 weeks after operation to evaluate the safety of magnesium alloy, and specimens from the supraspinatus tendon and proximal humerus were harvested at 2, 4, 8, and 12 weeks after operation. The specimens were observed macroscopically at 4 and 12 weeks after operation. Biomechanical tests were performed at 4, 8, and 12 weeks to test the ultimate load and stiffness of the healing sites in groups A and B. At 2, 4, and 12 weeks, the specimens were subjected to the following tests: Micro-CT to evaluate the formation of bone tunnels in groups A and B, HE staining and Masson staining to observe the regeneration of fibrocartilage at the tendon-bone interface after decalcification and sectioning, and Goldner trichrome staining to evaluate the calcification. Immunohistochemical staining was performed to detect the expressions of angiogenic factors, including vascular endothelial growth factor (VEGF) and bone morphogenetic protein 2 (BMP-2), as well as osteogenic factors at the tendon-bone interface. Additionally, immunofluorescence staining was used to examine the expressions of Arginase 1 and Integrin beta-2 to assess M1 and M2 macrophage polarization at the tendon-bone interface. The role of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway in tendon-bone healing was further analyzed using real-time fluorescence quantitative PCR.
RESULTS:
Analysis of visceral sections revealed that magnesium ions released during the degradation of magnesium alloys did not cause significant toxic effects on organs such as the heart, liver, spleen, lungs, and kidneys, indicating good biosafety. Histological analysis further demonstrated that fibrocartilage regeneration at the tendon-bone interface in group A occurred earlier, and the amount of fibrocartilage was significantly greater compared to group B, suggesting a positive effect of magnesium alloy material on tendon-bone interface repair. Additionally, Micro-CT analysis results revealed that bone tunnel formation occurred more rapidly in group A compared to group B, further supporting the beneficial effect of magnesium alloy on bone healing. Biomechanical testing showed that the ultimate load in group A was consistently higher than in group B, and the stiffness of group A was also greater than that of group B at 4 weeks, indicating stronger tissue-carrying capacity following tendon-bone interface repair and highlighting the potential of magnesium alloy in enhancing tendon-bone healing. Immunohistochemical staining results indicated that the expressions of VEGF and BMP-2 were significantly upregulated during the early stages of healing, suggesting that magnesium alloy effectively promoted angiogenesis and bone formation, thereby accelerating the tendon-bone healing process. Immunofluorescence staining further revealed that magnesium ions exerted significant anti-inflammatory effects by regulating macrophage polarization, promoting their shift toward the M2 phenotype. Real-time fluorescence quantitative PCR results demonstrated that magnesium ions could facilitate tendon-bone healing by modulating the PI3K/AKT signaling pathway.
CONCLUSION
Biodegradable magnesium alloy material accelerated fibrocartilage regeneration and calcification at the tendon-bone interface in rat rotator cuff tear repair by regulating the PI3K/AKT signaling pathway, thereby significantly enhancing tendon-bone healing.
Animals
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Rotator Cuff Injuries/metabolism*
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Rats, Sprague-Dawley
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Signal Transduction
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Wound Healing/drug effects*
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Alloys/pharmacology*
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Rats
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Proto-Oncogene Proteins c-akt/metabolism*
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Rotator Cuff/metabolism*
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Macrophages/metabolism*
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Magnesium/pharmacology*
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Phosphatidylinositol 3-Kinases/metabolism*
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Vascular Endothelial Growth Factor A/metabolism*
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Male
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Biocompatible Materials
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Bone Morphogenetic Protein 2/metabolism*
4.Imaging changes of the intervertebral disc after posterior cervical single door enlarged laminoplasty for cervical spinal stenosis with disc herniation.
Yan-Dong ZHANG ; Xu-Hong XUE ; Sheng ZHAO ; Gui-Xuan GE ; Xiao-Hua ZHANG ; Shi-Xiong WANG ; Ze GAO
China Journal of Orthopaedics and Traumatology 2025;38(6):572-580
OBJECTIVE:
To explore prevalence, incidence and possible factors of immediate herniated discs after posterior cervical expansive open-door laminoplasty (EODL).
METHODS:
Totally 29 patients with cervical spinal stenosis and intervertebral disc herniation who underwent EODL from October 2020 to December 2021 were collected, including 24 males and 5 females, aged from 43 to 81 years old with an average of (61.3±9.0) years old;the courses of disease ranged from 1 to 120 months with an average of (36.4±37.0) months. Three or more intervertebral discs on C3-C7 were observed. The clinical efficacy was evaluated according to Japanese Orthopaedic Association (JOA) score before operation, 3 days and 1, 3, 6 and 12 months after operation, respectively. The changes of herniated disc before and after operation were measured by multipoint area method and two-dimensional distance method, and incidence and percentage of herniated disc regression were further calculated. Cervical imaging parameters such as Cobb angle (C3-C7), intervertebral angle, T1 slope (T1S), spinal canal sagittal diameter, K-line angle, dural sac sagittal diameter were measured and compared before and after operation. Pearson correlation was used to analyze correlation between cervical sagittal imaging parameters and disc herniation changes before and after operation.
RESULTS:
All patients obtained grade A wound healing, and 14 of them were followed up for 3(1.00, 5.25) months. There were no immediate or long-term postoperative complications. Totally 101 herniated intervertebral discs were measured, of which 79 regression numbers were obtained by area measurement. The number of intervertebral disc regressions by distance measurement was 77. There was no statistically significant difference in Cobb angle, intervertebral angle, T1S and K-line angle of C3-C7 (P>0.05), however, there were statistically significant differences in sagittal diameter of spinal canal, sagittal diameter of dural sac, and JOA score before and after operation(P<0.05). The regression ratio of disc herniation ranged from 5% to 50%, and regression ratio of disc herniation was greater than 25% in 45.57%(36/79). Disc herniation in C4,5 was positively correlated with sagittal plane diameter in C5(r=0.423, P=0.028). There was a negative correlation between changes of C3,4 and C3,4 intervertebral angle (r=-0.450, P=0.041). The improvement rate of cervical JOA score immediately after operation was (59.54±15.07) %, and postoperative follow-up improved to (76.57±14.66) %.
CONCLUSION
Herniated disc regression immediately after EODL is a common occurrence, and EODL should be selected as far as possible under the premise of satisfying surgical indications. The regression of disc herniation is positively correlated with spinal canal sagittal diameter, and spinal canal should be enlarged as far as possible in the appropriate scope during EODL, so as to create more opportunities and conditions for disc regression and achieve better clinical results.
Humans
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Female
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Male
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Intervertebral Disc Displacement/diagnostic imaging*
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Spinal Stenosis/diagnostic imaging*
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Laminoplasty/methods*
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Middle Aged
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Aged
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Cervical Vertebrae/diagnostic imaging*
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Adult
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Aged, 80 and over
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Intervertebral Disc/surgery*
5.Clinical efficacy of bone cement filling combined with lower extremity arterial balloon dilation in the treatment of Wagner Ⅳ grade diabetic foot.
Jia-Min HOU ; Sheng-Gang WU ; Feng WEI ; Xiong-Feng LI
China Journal of Orthopaedics and Traumatology 2025;38(9):955-959
OBJECTIVE:
To explore clinical efficacy of bone cement filling combined with lower extremity arterial balloon dilation in treating Wagner grade Ⅳ diabetic foot (DF).
METHODS:
From January to October 2024, 9 Wagner grade Ⅳ DF patients with lower extremity vascular occlusion were admitted, including 7 males and 2 females, aged from 51 to 87 years old;5 patients on the left side and 4 patients on the right side. All patients were underwent stageⅠdebridement of the affected foot and bone cement filling, and treated with lower extremity arterial balloon dilation after operation, they were. After the formation of the induced membrane, stageⅡwound repair was performed. The wound healing time and condition were observed. Ankle-brachial index (ABI) was used to evaluate the lower extremity vascular perfusion before operation and 3 months after operation, respectively.
RESULTS:
The wounds of all 9 patients healed completely, and the healing time ranged from 45 to 65 days. All patients were followed up for at least 6 months without recurrence. The skin of the affected foot wound healed with keratinization, and there was mild scar hyperplasia locally (1 patient had necrosis of the adjacent toe after stageⅠsurgery and was debridement and toe amputation again). The narrowed or occluded blood vessels of the lower extremities were all recanalized. ABI recovered from 0.3 to 0.5 before operation to 1.0 to 1.1 at 3 months after operation.
CONCLUSION
Bone cement filling combined with lower extremity arterial balloon dilation for the treatment of grade Wagner Ⅳ DF is conducive to promoting healing of the affected foot, effectively preventing secondary ulceration of the affected foot, and clinical therapeutic effect is satisfactory.
Humans
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Male
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Female
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Middle Aged
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Diabetic Foot/surgery*
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Aged
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Bone Cements/therapeutic use*
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Aged, 80 and over
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Lower Extremity/blood supply*
6.Surgical approaches to varicocele: a systematic review and network meta-analysis.
Lin-Jie LU ; Kai XIONG ; Sheng-Lan YUAN ; Bang-Wei CHE ; Jian-Cheng ZHAI ; Chuan-Chuan WU ; Yang ZHANG ; Hong-Yan ZHANG ; Kai-Fa TANG
Asian Journal of Andrology 2025;27(6):728-737
Surgical methods for varicocele remain controversial. This study intends to evaluate the efficacy and safety of different surgical approaches for treating varicocele through a network meta-analysis (NMA). PubMed, Embase, Cochrane, and Web of Science databases were thoroughly searched. In total, 13 randomized controlled trials (RCTs) and 24 cohort studies were included, covering 9 different surgical methods. Pairwise meta-analysis and NMA were performed by means of random-effects models, and interventions were ranked based on the surface under the cumulative ranking curve (SUCRA). According to the SUCRA, microsurgical subinguinal varicocelectomy (MSV; 91.6%), microsurgical retroperitoneal varicocelectomy (MRV; 78.2%), and microsurgical inguinal varicocelectomy (MIV; 76.7%) demonstrated the highest effectiveness in reducing postoperative recurrence rates. In this study, sclerotherapy embolization (SE; 87.2%), MSV (77.9%), and MIV (67.7%) showed the best results in lowering the risk of hydrocele occurrence. MIV (82.9%), MSV (75.9%), and coil embolization (CE; 58.7%) were notably effective in increasing sperm motility. Moreover, CE (76.7%), subinguinal approach varicocelectomy (SV; 69.2%), and SE (55.7%) were the most effective in increasing sperm count. SE (82.5%), transabdominal laparoscopic varicocelectomy (TLV; 76.5%), and MRV (52.7%) were superior in shortening the length of hospital stay. The incidence rates of adverse events for MRV (0), SE (3.3%), and MIV (4.1%) were notably low. Cluster analyses indicated that MSV was the most effective in the treatment of varicocele. Based on the existing evidence, MSV may represent the optimal choice for varicocele surgery. However, selecting clinical surgical strategies requires consideration of various factors, including patient needs, surgeon experience, and the learning curve.
Humans
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Male
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Embolization, Therapeutic/methods*
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Microsurgery/methods*
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Randomized Controlled Trials as Topic
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Sclerotherapy/methods*
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Treatment Outcome
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Urologic Surgical Procedures, Male/methods*
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Varicocele/surgery*
7.Lactobacillus plantarum ZG03 alleviates oxidative stress via its metabolites short-chain fatty acids.
Shuxian LIN ; Lina GUO ; Yan MA ; Yao XIONG ; Yingxi HE ; Xinzhu XU ; Wen SHENG ; Suhua XU ; Feng QIU
Journal of Southern Medical University 2025;45(10):2223-2230
OBJECTIVES:
To investigate the efficacy of Lactobacillus plantarum ZG03 (L. plantarum ZG03) for ameliorating oxidative stress in zebrafish.
METHODS:
We evaluated the growth pattern of L. plantarum ZG03, observed its morphology using field emission scanning electron microscopy, and assessed its safety and potential efficacy with whole-genome sequencing for genetic analysis. FITC-labeled ZG03 was used to observe its intestinal colonization in zebrafish. In a zebrafish model of 2% glucose-induced oxidative stress, the effect of ZG03 was evaluated by assessing the changes in neutrophils in the caudal hematopoietic tissue (CHT), superoxide dismutase (SOD) activity, reactive oxygen species (ROS) levels, and malondialdehyde (MDA) content. Liquid chromatography-mass spectrometry-based targeted metabolomics was used for analyzing short-chain fatty acids (SCFAs) in the zebrafish, and the antioxidant effects of the key metabolites (acetate, propionate, and caproate) were tested.
RESULTS:
On MRS agar, L. plantarum ZG03 formed circular, smooth, moist, and milky-white colonies with a rod-shaped cell morphology. Genomic analysis revealed abundant sugar metabolism gene clusters. After inoculation of FITC-labeled L. plantarum ZG03 in zebrafish, green fluorescence was clearly observed in the intestinal bulb, mid-intestine, and hind intestine. In zebrafish with glucose-induced oxidative stress, L. plantarum ZG03 significantly reduced ROS levels and the number of neutrophils in the CHT with increased SOD activity. L.plantarum ZG03 significantly increased the content of SCFAs including acetic acid, propionic acid, and caproic acid in zebrafish metabolites. In addition, sodium acetate, sodium propionate, and sodium caproate in the SCFAs significantly increased SOD activity in the zebrafish models.
CONCLUSIONS
L. plantarum ZG03 ameliorates oxidative stress in a glucose-induced zebrafish model through its metabolites, particularly the SCFAs including acetic acid, propionic acid and caproic acid.
Animals
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Zebrafish/metabolism*
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Oxidative Stress
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Lactobacillus plantarum/metabolism*
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Fatty Acids, Volatile/metabolism*
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Probiotics
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Reactive Oxygen Species/metabolism*
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Superoxide Dismutase/metabolism*
8.Effect of Ophiopogonin D on lipopolysaccharide-induced apoptosis of alveolar epithelial cells
Qing-Xin KANG ; Shen-Shan JIAO ; Zheng XIONG ; Hui-Ming XI ; Xun-Sheng JIANG ; Zi-Long ZHANG
The Chinese Journal of Clinical Pharmacology 2024;40(12):1744-1748
Objective To investigate the effect of Ophiopogonin D on lipopolysaccharide(LPS)-induced apoptosis of alveolar epithelial cells by regulating the interleukin-6(IL-6)/Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods A549 AT Ⅱ cells cultured in vitro were randomly divided into four groups:control group,LPS group,LPS+Ophiopogonin D group,LPS+Ophiopogonin D+colivelin(JAK2/STAT3 signal activator)group,except for the control group,and cells in all other groups were established injury models while being grouped with Ophiopogonin D and colivelin for treatment.Cell counting kit-8(CCK-8)experiment and flow cytometry were applied to detect cell proliferation and apoptosis in each group;Western blotting was applied to detect the expression of IL-6/JAK2/STAT3 signaling pathway proteins of cells in each group.Results The apoptosis rates of A549 cells in control group,LPS group,LPS+Ophiopogonin D group and LPS+Ophiopogonin D+colivelin group were(2.52±0.73)%,(52.43±4.14)%,(1.67±0.52)%and(47.94±3.43)%;IL-6 protein levels were 0.14±0.03,0.49±0.05,0.17±0.04 and 0.45±0.06,and p-JAK2/JAK2 protein levels were 0.17±0.04,0.64±0.08,0.19±0.06 and 0.61±0.07;p-STAT3/STAT3 protein levels were 0.20±0.06,0.69±0.10,0.22±0.07 and 0.65±0.09;the apoptosis rates of AT Ⅱ cells were(3.01±0.69)%,(55.16±3.94)%,(2.35±0.71)%and(50.28±3.78)%;the levels of IL-6 protein were 0.11±0.03,0.87±0.13,0.19±0.04 and 0.84±0.12;the p-JAK2/JAK2 protein levels were 0.13±0.04,0.56±0.08,0.15±0.03 and 0.53±0.07;p-STAT3/STAT3 protein levels were 0.30±0.08,0.79±0.14,0.33±0.09 and 0.75±0.13.The above indexes:control group,LPS+Ophiopogonin D group compared with LPS group,LPS+Ophiopogonin D+colivelin group compared with LPS+Ophiopogonin D group,the differences were statistically significant(all P<0.05).Conclusion Ophiopogonin D can reduce LPS induced inflammation and oxidative stress levels by inhibiting the activation of IL-6/JAK2/STAT3 signaling pathway,ultimately reducing LPS-induced apoptosis of alveolar epithelial cells.
9.Multiparametric CT features for prediction of the risk classification of gastric stromal tumor
Chengyao XIE ; Zhiqi YANG ; Xiaofeng CHEN ; Zhiqiang ZHAGN ; Sheng ZHANG ; Xiong ZHANG ; Wenfeng LING
Journal of Practical Radiology 2024;40(3):394-397
Objective To investigate the value of multiparametric CT features for predicting the risk classification of gastric stro-mal tumor(GST).Methods The clinical data from 139 patients with GST were retrospectively collected.According to the patho-logical risk results,the patients were divided into two groups:a low-risk GST group(including very low-and low-risk)with 75 patients and a high-risk GST group(including medium and high-risk)with 64 patients.The CT features between low-risk GST group and high-risk GST group were compared using chi-squared test or t-test.The risk factors of high-risk GST were identified by univariate analysis.The prediction models were built by multivariate logistic regression.The performance of models were evaluated by receiver oper-ating characteristic(ROC)curve.Results There were significant differences in the maximum tumor diameter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate,venous phase enhance-ment degree rate,cystic,and necrosis between low-risk GST group and high-risk GST group,which were associated with the risk classification of GST.The area under the curve(AUC)of the quantitative features-based model that combined maximum tumor diam-eter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate and venous phase enhancement degree rate,showed a significantly higher performance than the qualitative features-based model that incorporated cystic and necrosis(0.981 vs 0.850,P<0.001).Conclusion Maximum tumor diameter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate,venous phase enhance-ment degree rate,as well as cystic and necrosis,are associated with the risk classification of GST and can predict the high-risk GST.
10.Association between physical activity and anxiety/depression symptoms among college students in Jiangxi Province
ZHOU Jianfeng, MA Sheng, XIONG Jianping, SONG Yongjing, ZOU Zhiyong
Chinese Journal of School Health 2024;45(12):1722-1726
Objective:
To investigate the association between physical activity frequency and anxiety/depression symptoms among college students in Jiangxi Province, so as to provide a basis for the physical and mental health development of adolescents.
Methods:
From July to October 2023, 5 761 college students from 11 colleges and universities in Jiangxi Province were selected by convenience sampling to conduct an online questionnaire survey regarding physical activity and mental health. Anxiety symptoms and depressive symptoms were assessed by General Anxiety Disorder Scale-7 and Patient Health Questionnaires-9, respectively. Contingency table was used to analyze the distribution of different anxiety and depression symptoms by physical activity frequency, and ordered Logistic regression was used to analyze the association between physical activity frequency with anxiety and depressive symptoms, and stratified analysis was performed based on gender.
Results:
The detection rate of anxiety symptoms of college students was 43.6% ( n =2 513), and the detection rate of depression symptoms was 63.9% ( n =3 682). There were significant differences in the detection rate of anxiety and depression symptoms among different gender and physical activity frequency groups ( χ 2=15.98, 106.32; 30.65, 150.88, P <0.05). The detection rates of severe anxiety and depression symptoms of male and female who never exercise were higher (severe anxiety: 6.1% for male and 5.2% for female, severe depression: 8.7% for male and 7.4% for female).The results of ordered Logistic regression analysis showed that compared with college students who never exercised, male college students who were physically active almost every day were negatively correlated with mild anxiety and moderate depression symptoms ( OR=0.53,0.33,P <0.05). In addition, male college students who do physical activity 3-4 times a week were negatively correlated with moderate depressive symptoms ( OR =0.43), while male college students who do physical activity 1-2 times a week were negatively correlated with severe depressive symptoms ( OR =0.21) ( P <0.05). Physical activity was negatively correlated with different degress of anxiety and depression symptoms in female college students (anxiety: OR =0.27-0.74, depression: OR =0.18-0.75, P < 0.05).
Conclusions
The frequency of physical activity in college students (especially female college students) is negatively correlated with anxiety and depressive symptoms. It is suggested to improve the frequency of physical activity to promote physical and mental health.


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