1.Astragalin Regulates Autophagy and Apoptosis of Astrocytes in L4-5 Spinal Dorsal Horn of Mouse Inflammatory Pain Model
Weishan ZHANG ; Jiahong LIN ; Can WANG ; Runheng ZHANG ; Junhua YANG ; Jing LIU ; Guoying LI ; Yuxin MA
Journal of Sun Yat-sen University(Medical Sciences) 2025;46(2):186-196
ObjectiveTo explore the effects of astragalin (AST) on autophagy and apoptosis of astrocytes in the L4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant (CFA). MethodsTwenty-four male C57BL/6 mice, aged six months, were randomly assigned to four groups: control group, saline group, CFA model group, and CFA+AST group, six mice in each group. The inflammatory pain model was established by injection of 10 µL CFA into the right lateral malleolus fossa. The saline group were injected with an equal amount of normal saline at the same site. The inflammatory pain mice in CFA+AST group were further treated with AST (60 mg/kg) intraperitoneally once a day for 21 consecutive days. Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1, apoptosis-related factors including Cleaved-Caspase3 and Caspase9, and the astrocyte marker such as GFAP in the L4-5 spinal dorsal horn of the mice in each group. Western blot was used to examine the protein expression levels of autophagy-related proteins(ATG12, Beclin-1) and apoptosis-related proteins(Caspase 3, Caspase 9) in the L4-5 spinal dorsal horn of mice. ResultsImmunofluorescent staining showed that in the L4-5 dorsal horn of the spinal cord, the fluorescence intensity of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) was significantly increased, while that of Cleaved-Caspase 3 (P<0.001) and Caspase 9 (P<0.000 1) was decreased in the CFA+AST group when compared to the CFA model group. Furthermore, AST could inhibit the activation of astrocytes. Western blot further confirmed that AST significantly upregulated the expression of ATG12 (P<0.000 1) and Beclin-1 (P<0.000 1) in the L4-5 spinal cord of CFA mice, and downregulated the expression of Caspase 3 (P<0.01) and Caspase 9 (P<0.001). ConclusionsAST promotes autophagy of astrocytes and inhibits their apoptosis in the L4-5 spinal dorsal horn of CFA mice.
2.Effects of Bushen Zhuyun Prescription on Endometrial Angiogenesis by Regulating Mitochondrial Function
Yuling ZHOU ; Jinglei ZHANG ; Can CAO ; Ying SUN ; Ruobing NIU ; Yingmei ZHANG ; Shuchan HU ; Lin LUO ; Ming HE
Chinese Journal of Information on Traditional Chinese Medicine 2024;31(1):91-96
Objective To observe the effects of Bushen Zhuyun Prescription on regulating mitochondrial function and endometrial angiogenesis;To explore its mechanism of improving endometrial receptivity.Methods The mouse model of controlled ovarian hyperstimulation(COH)was established,and the mice were randomly divided into normal group,model group,and Bushen group,with 20 mice in each group.Bushen group received Bushen Zhuyun Prescription for gavage for 11 d,and the normal group and model group received normal saline for gavage.The number of embryo implantation was counted,the endometrial morphology was observed by HE staining,α-smooth muscle actin(α-SMA)expression was observed by immunofluorescence staining.Human endometrial microvascular endothelial cells(HEMECs)were cultured in vitro,they were divided into control group,VEGFA group,Bushen group and VEGFA + Bushen group,and were intervened with VEGFA and/or Bushen Zhuyun Prescription medicated serum.The activities of mitochondrial respiratory chain complexes Ⅰ-Ⅳ,the content of ATP,the expression of PCNA and Caspase-3 were detected.Results Animal experiment showed that,compared with the normal group,the number of embryo implantation in model group significantly decreased(P<0.05),α-SMA protein expression in endometrial tissue significantly decreased(P<0.05);compared with the model group,the number of embryo implantation in Bushen group significantly increased(P<0.05),α-SMA protein expression in endometrial tissue significantly increased(P<0.05).Cell experiment showed that,Bushen Zhuyun Prescription medicated serum could increase the activity of mitochondrial respiratory chain complexes Ⅰ-Ⅳ and ATP content in HEMECs,promote PCNA protein expression,and inhibit Caspase-3 protein expression(P<0.05,P<0.01).Conclusion Bushen Zhuyun Prescription can promote endometrial angiogenesis through improving mitochondrial function.
3.Accuracy and clinical efficacy of robot-assisted transiliac-transsacral screw placement for posterior pelvic ring frac-tures
Xue-Lin WANG ; Zheng-Jie WU ; Yan-Hui ZENG ; Can-Hui LI ; Jian-Sheng ZHOU ; Shi HONG
China Journal of Orthopaedics and Traumatology 2024;37(6):605-608
Objective To explore accuracy and clinical effect of robot-assisted implantation of sacroiliac penetrating screw in orthopedic surgery for posterior pelvic ring fracture.Methods The clinical data of 24 patients with posterior pelvic ring frac-ture treated with robot-assisted sacroiliac penetration screws from August 2022 to August 2023 were retrospectively analyzed,including 10 males and 14 females;aged from 21 to 73 years old with an average of(49.29±14.48)years old;according to Tile pelvic fractures,13 patients were type B and 11 were type C.The effect of screw placement was evaluated according to Gras criteria based on postoperative CT scan results.At the final follow-up,fracture healing was evaluated according to Matta score,and functional recovery was evaluated by Majeed score.Results All patients were followed up for 3 to 13 months with an aver-age of(6.00±3.28)months.Totally 36 sacroiliac penetrating screws,18 S1 penetrating screws,18 S2 penetrating screws were inserted,a total of 29 were excellent and 7 good according to Gras standard.Screw adjustment times was 0.00(0.00,0.75)times.At the final follow-up,Matta score was excellent in 18 patients,5 good and 1 moderate,and the maximum displacement distance was 2.55(0.00,5.65)mm.Majeed score was 84.37±8.38,15 patients were excellent,7 good and 2 moderate.Con-clusion Robot could accurately and safely assist in the placement of sacroiliac joint screws for the treatment of posterior pelvic ring fractures,and promote postoperative functional recovery of patients.
4.Development of pre-deployment base-based training course for medical units in peacekeeping level-II hospitals
Can CHEN ; Wenqiong DU ; Renqing JIANG ; Xin ZHONG ; Wei ZHANG ; Lin CHEN ; Zhaowen ZONG
Chinese Journal of Medical Education Research 2024;23(4):482-486
Objective:To establish a practical and systematic pre-deployment base-based training course for medical units in peacekeeping level-II hospitals.Methods:We surveyed the current status of pre-deployment training of medical teams in peacekeeping level-II hospitals, and organized expert discussions to preliminarily construct centralized training course contents. At the same time, the Delphi method was used to conduct two rounds of questionnaire-based consultation to evaluate the overall framework design of the course and the rationality and importance scores of each module. The contents of the pre-deployment training course were further selected and optimized based on the positive coefficient (%), authority coefficient (Cr), coefficient of concordance ( W), and coefficient of variation (Cv). Results:The contents of the pre-deployment training course for the medical units of peacekeeping level-II hospitals were preliminarily established. The experts scored high for the combat onsite first-aid skills module [rationality score, (9.32±0.75) points; importance score, (9.45±0.65) points] and the combat wound treatment module [rationality score, (9.33±0.75) points; importance score, (9.28±0.74) points], and scored lowest for the field survival module [rationality score, (7.95±2.28) points; importance score, (7.87±2.16) points].Conclusions:The established contents of the pre-deployment base-based training course for the medical units of peacekeeping level-II hospitals are reasonable and practical, which lays the foundation for perfecting the pre-deployment training system of peacekeeping medical teams.
5.Ku70 Functions as an RNA Helicase to Regulate miR-124 Maturation and Neuronal Cell Differentiation
Ai-Xue HUANG ; Rui-Ting LI ; Yue-Chao ZHAO ; Jie LI ; Hui LI ; Xue-Feng DING ; Lin WANG ; Can XIAO ; Xue-Mei LIU ; Cheng-Feng QIN ; Ning-Sheng SHAO
Progress in Biochemistry and Biophysics 2024;51(6):1418-1433
ObjectiveHuman Ku70 protein mainly involves the non-homologous end joining (NHEJ) repair of double-stranded DNA breaks (DSB) through its DNA-binding properties, and it is recently reported having an RNA-binding ability. This paper is to explore whether Ku70 has RNA helicase activity and affects miRNA maturation. MethodsRNAs bound to Ku protein were analyzed by RNA immunoprecipitation sequencing (RIP-seq) and bioinfomatic anaylsis. The expression relationship between Ku protein and miRNAs was verified by Western blot (WB) and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assays. Binding ability of Ku protein to the RNAs was tested by biolayer interferometry (BLI) assay. RNA helicase activity of Ku protein was identified with EMSA assay. The effect of Ku70 regulated miR-124 on neuronal differentiation was performed by morphology analysis, WB and immunofluorescence assays with or without Zika virus (ZIKV) infection. ResultsWe revealed that the Ku70 protein had RNA helicase activity and affected miRNA maturation. Deficiency of Ku70 led to the up-regulation of a large number of mature miRNAs, especially neuronal specific miRNAs like miR-124. The knockdown of Ku70 promoted neuronal differentiation in human neural progenitor cells (hNPCs) and SH-SY5Y cells by boosting miR-124 maturation. Importantly, ZIKV infection reduced the expression of Ku70 whereas increased expression of miR-124 in hNPCs, and led to morphologically neuronal differentiation. ConclusionOur study revealed a novel function of Ku70 as an RNA helicase and regulating miRNA maturation. The reduced expression of Ku70 with ZIKV infection increased the expression of miR-124 and led to the premature differentiation of embryonic neural progenitor cells, which might be one of the causes of microcephaly.
6.A Study on the Anti-Angiogenic Activity of an RG-I Polysaccharide from Lonicera macranthoides Flowers
Can JIN ; Liyan LIN ; Huixian CHEN ; Kan DING
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(5):1172-1181
Objective This study aims to investigate the structure and anti-angiogenic activity of homogeneous polysaccharides in Honeysuckle flowers,constructing the theoretical basis for its widespread application.Methods Homogeneous Honeysuckle polysaccharide LF-02-2 was obtained through water extraction,alcohol precipitation,anion exchange chromatography,and gel permeation chromatography.The structure of LF-02-2 was deduced through molecular weight determination,monosaccharide composition analysis,sugar residue linkage analysis,partial acid hydrolysis,and sugar aldonic acid reduction combined with NMR data.Additionally,in vitro tube formation experiments using human microvascular endothelial cells(HMEC-1)were conducted to assess its anti-angiogenic activity.Results Structural analysis of LF-02-2 revealed a homogeneous polysaccharide with a weight-average molecular weight of 74.1 kDa.The monosaccharide composition included rhamnose(Rha),galactose(Gal),galacturonic acid(GalA),and arabinose(Ara)in molar ratios of 10.43:14.94:6.66:67.97.The main chain consisted of 1,4-linked α-Galp A,1,2-linked α-Rhap,and 1,2,4-linked α-Rhap.Branches were connected to O-4 of 1,2,4-linked α-Rhap and included β-Galp with terminal connections and 1,4,6-linked β-Galp and α-Araf with terminal and 1,5-linked connections.Tube formation experiments demonstrated that LF-02-2 significantly inhibited tube formation in human microvascular endothelial cells(HMEC).Conclusion LF-02-2,an RG-I pectic polysaccharide from Honeysuckle flowers,exhibited significant anti-angiogenic activity,suggesting its potential for development as an anti-angiogenic drug.
7.Scutellarin inhibits proliferation and migration of human prostate cancer cell line PC-3
Yanhong XIAO ; Mingdong JIANG ; Yeyuan LIN ; Can RAN ; Bo LIANG
Basic & Clinical Medicine 2024;44(9):1229-1235
Objective To investigate the effect of scutellarin(STR)on the proliferation and migration of human prostate cancer cell line(PC-3)and its underlying mechanism.Methods PC-3 cells were divided into low-dose STR group,medium-dose STR group,high-dose STR group,colivelin(STAT3 activator)group,high-dose STR+colivelin group and control group.CCK-8 assay and colony formation experiments were applied to detect cell prolif-eration;Scratch experiment was applied to detect cell migration;Transmission electron microscopy was applied to observe the mitochondrial ultrastructure of PC-3 cells.The intracellular free Fe2+,malondialdehyde(MDA)content and reactive oxygen species(ROS)levels were detected by colorimetric method;RT-qPCR was applied to detect the mRNA expression of member 11 of solute vector family(SLC7A11),proliferating cell nuclear antigen(PCNA)and matrix metalloproteinase-9(MMP-9)in cells;Western blot was used to detected p-STAT3 and GPX4 proteins in cells.Results Compared to control group,the mitochondrial structure of PC-3 cells in the low-dose STR group,medium-dose STR group and high-dose STR group was significantly disrupted.The A450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression,and p-STAT3,GPX4 protein all reduced.While Fe2+,MDA content,and that ROS level increased with dose-dependent way(P<0.05).Compared with control group,the destruction of mitochondrial structure in cells from colivelin group improved;The A450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression and p-STAT3,GPX4 protein all increased,while Fe2+MDA content,and ROS level decreased(P<0.05).Compared with high-dose STR group,the damage of mitochondrial structure in PC-3 cells in the high-dose STR+colivelin group was re-duced.The A450 value,colony formation rate,scratch healing rate,PCNA,SLC7A11,MMP-9 mRNA expression,and p-STAT3,GPX4 protein increased,while Fe2+,MDA content and ROS level decreased(P<0.05).Conclusions The mechanism by of STR reducing proliferation and migration of prostate cancer cells is potentially related to the inhibition of STAT3/GPX4 pathway.
8.Standardized operational protocol for the China Human Brain Bank Consortium(2nd edition)
Xue WANG ; Zhen CHEN ; Juan-Li WU ; Nai-Li WANG ; Di ZHANG ; Juan DU ; Liang YU ; Wan-Ru DUAN ; Peng-Hao LIU ; Han-Lin ZHANG ; Can HUANG ; Yue-Shan PIAO ; Ke-Qing ZHU ; Ai-Min BAO ; Jing ZHANG ; Yi SHEN ; Chao MA ; Wen-Ying QIU ; Xiao-Jing QIAN
Acta Anatomica Sinica 2024;55(6):734-745
Human brain banks use a standardized protocol to collect,process and store post-mortem human brains and related tissues,along with relevant clinical information,and to provide the tissue samples and data as a resource to foster neuroscience research according to a standardized operating protocols(SOP).Human brain bank serves as the foundation for neuroscience research and the diagnosis of neurological disorders,highlighting the crucial rule of ensuring the consistency of standardized quality for brain tissue samples.The first version of SOP in 2017 was published by the China Human Brain Bank Consortium.As members increases from different regions in China,a revised SOP was drafted by experts from the China Human Brain Bank Consortium to meet the growing demands for neuroscience research.The revised SOP places a strong emphasis on ethical standards,incorporates neuropathological evaluation of brain regions,and provides clarity on spinal cord sampling and pathological assessment.Notable enhancements in this updated version of the SOP include reinforced ethical guidelines,inclusion of matching controls in recruitment,and expansion of brain regions to be sampled for neuropathological evaluation.
9.Efficacy and safety of remimazolam tosylate for general anesthesia in morbidly obese patients
Gong CHEN ; Yan-Xi LU ; Jin LI ; Fan ZHANG ; Can-Can CHENG ; Xin-Lin YIN ; Sai-Ying WANG ; Huan CHANG
Chinese Pharmacological Bulletin 2024;40(5):859-864
Aim To evaluate the effectiveness and safety of remimazolam tosylate for administering general anesthesia in morbidly obese patients.Methods This clinical trial was conducted at a single center from De-cember 2021 to October 2023.It assessed 108 morbid-ly obese patients(body mass index,BMI≥40)who underwent laparoscopic sleeve gastrectomy.Patients were randomly assigned to either the remimazaolam group(Group R)or the propofol group(Group P)for general anesthesia induction and maintenance.The primary outcome was to compare the incidence of ad-verse events and postoperative recovery characteristics between the two groups.Results During induction pe-riod,the incidence of adverse events was higher in group P,including hypotension(P<0.01),hypox-emia(P<0.05),bradycardia(P<0.01),and in-creased vasopressor requirement(P<0.05).The time to loss of consciousness and BIS falling to 60 was shor-ter in group P than in group R(P<0.01).There were no statistically significant differences between the two groups in terms of postoperative quality of recovery(QoR-40 score),24-hour postoperative pain visual an-alogue scale(VAS)scores and morphine consump-tion.In conclusion,remimazolam tosylate,utilized for anesthesia induction in morbidly obese patients,signif-icantly reduced hypotension and hypoxemia compared to propofol,while it could also maintain similar postop-erative recovery quality.Conclusions Remimazolam is effective in reducing the incidence of hypotension and hypoxaemia during the induction period of general anaesthesia in morbidly obese patients and it is compa-rable to propofol in terms of quality of postoperative re-covery.
10.Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Analysis and Imaging of Porcine Brain Tissue Section Using Precoated Nanocarbon-Sodium Dodecyl Sulfate Composite Matrix
Si-Jia ZHUANG ; Yu-Lin SHEN ; Fan YANG ; Can GONG ; Xu XU
Chinese Journal of Analytical Chemistry 2024;52(7):985-994,中插22-中插29
Nanomaterial matrices have unique advantages in small molecule analysis by matrix-assisted laser desorption ionization mass spectrometry(MALDI-MS).On the basis of comparing different surfactants,a method for pre-coating nanocarbon sodium dodecyl sulfate(SDS)composite matrix was established,which could alleviate the issue of aggregation of nanocarbon particles as a matrix.The experimental results showed that the nanocarbon-SDS composite pre-coated matrix effectively suppressed the background ion peak of the nano matrix,significantly improved the MS peak intensity and signal-to-noise ratio of the tested substance.Under the optimal conditions,cholesterol,1,2-dipalmitoyl-sn-glyceryl-3-phosphorylcholine(DPPC),and triglyceride standards were analyzed,with intra-point repeatability(RSD)of MS peak intensity≤5%and inter-point repeatability≤7%.As to measurement of cyclic adenosine monophosphate and DPPC samples in the range of 0.05?1.0 mg/mL,linear correlation coefficient(R2)between peak intensity and concentration was greater than 0.993,indicating good quantitative analysis potential.The nanocarbon-SDS composite pre-coating matrix was used for MALDI-MS imaging of pig brain tissue,and the differences in component distribution between pig brain white matter and gray matter were observed,demonstrating the potential of nanocarbon-SDS composite pre-coating matrix for MALDI-MS imaging.

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