1.Safety and efficacy of acute stent implantation during endovascular treatment for patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis
Tian LIN ; Wanling WEN ; Juan DU ; Zheng WU ; Xiangkai KONG ; Wenbo DUAN ; Xiaoyun ZHANG ; Bin DU ; Yiling CAI ; Yongqiang CUI
Chinese Journal of Internal Medicine 2024;63(3):272-278
Objective:To investigate the efficacy and safety of acute stent implantation during endovascular treatment for patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis.Methods:A retrospective analysis was carried out on 46 patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis who received endovascular treatment at the Strategic Support Force Medical Center from January 2015 to August 2022. Twenty-seven patients underwent balloon angioplasty alone and 19 patients underwent acute stent implantation. The baseline characteristics, modified thrombolysis in cerebral infarction (mTICI) score of the responsible vessels, modified Rankin scale (mRS) score 90 days after operation, incidence of symptomatic intracranial hemorrhage and mortality of the two groups were evaluated.Results:The proportion of effective recanalization of the offending vessels (mTICI≥2b) in the acute stenting group was slightly higher than that in the balloon angioplasty group (16/19 vs. 81.5%), but the difference was not statistically significant ( P>0.05). Besides, there was no significant difference in the median of mRS between the acute stenting group [3.0(0, 4.0)] and the balloon angioplasty group [4.0(1.0, 5.0)] 90 days after operation ( P>0.05). In terms of safety, the incidence of symptomatic intracranial hemorrhage and mortality were comparable between the two groups ( P>0.05). Conclusions:The effect of acute stent implantation during endovascular treatment for patients with emergent large vessel occlusion due to intracranial atherosclerotic stenosis is not inferior to that of balloon angioplasty, and it does not increase the risk of intracranial bleeding complications.
2.Quercetin inhibits the activity of neuroendocrine tumor cells by regulating the GAS5/miR-18b-5p axis
Wen-Juan WU ; Bo LI ; Hai-Hong LÜ ; Jun CHEN ; Wen KOU
The Chinese Journal of Clinical Pharmacology 2024;40(10):1429-1433
Objective To investigate the inhibitory effect of quercetin on Gastro entero pancreatic NEN(GEP-NEN).Methods Human pancreatic neuroendocrine tumor BON-1 cells were randomly divided into control group,quercetin group(80 μmol·L-1 quercetin),quercetin+si-NC group(transfected with si-NC+80 μmol·L-1 quercetin),quercetin+si-growth arrest-specific+ranscript 5(GAS5)group(transfected with si-GAS5+80 μmol·L-1 quercetin).Dual luciferase reporter gene assay was used to verify the targeted binding of GASS5 to miR-18b-5p;real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the mRNA expression levels of B-cell lymphoma-2(Bcl-2)and Bel-2 associated X protein(Bax);positive expression of GAS5 and miR-18b-5p in cells was detected by fluorescence in situ hybridization(FISH)assay.Results Dual luciferase reporter gene results showed that GAS5 was targeted to miR-18b-5p.The GAS5 expression levels of control group,quercetin group,quercetin+si-NC group and quercetin+si-GAS5 group were 1.00±0.13,1.72±0.19,1.78±0.14 and 1.16±0.11,respectively;the expression levels of miR-18b-5p were 1.00±0.15,0.67±0.08,0.72±0.06 and 0.95±0.11 respectively;Bax mRNA expression levels were 1.00±0.12,2.17±0.25,2.32±0.28 and 1.37±0.15,respectively;Bcl-2 mRNA expression levels were 1.00±0.15,0.41±0.05,0.37±0.06 and 1.21±0.13,respectively.The above indexes were significantly different between quercetin group and control group(all P<0.05);the above indexes were significantly different between quercetin+si-NC group and quercetin+si-GAS5 group(all P<0.05).Conclusion Quercetin may slow down the development of GEP-NEN by targeting GAS5/miR-18b-5p molecular axis to inhibit cell growth.
3.Pharmacokinetics of JS026 and JS026-JS016 for single intravenous administration in healthy volunteers
Yan TIAN ; Hui-Jing YE ; Jing-Jing WANG ; Nan-Yang LI ; Juan MA ; Xi TAN ; Fan WU ; Jie WANG ; Shu-Yan YU ; Xiao-Jie WU ; Jin-Jie HE ; Jing ZHANG ; Wen-Hong ZHANG
The Chinese Journal of Clinical Pharmacology 2024;40(15):2251-2255
Objective To evaluate tolerability,safety and pharmacokinetics of JS026 and JS026-JS016 single dose intravenous infusion in healthy adults.Methods This phase 1,randomized,double-blind,placebo-controlled,dose-escalation study totally included 48 participants:32 healthy subjects were enrolled in JS026 single intravenous infusion groups and 16 healthy subjects were enrolled in JS026-JS016 groups.JS026 was sequentially administered from low dose to high dose(30-1 000 mg),with intravenous infusion of JS026 or placebo in JS026 single-dose groups,and intravenous infusion of JS026-JS016 or placebo in the combination drug groups.Blood was collected according to the time point designed for trial.Serum concentrations of JS026 and JS016 were determined by enzyme linked immunosorbnent assay(ELISA),and pharmacokinetics parameters were calculated by WinNonlin 8.2.The power model method was used to evaluate the linear analysis of dose and drug exposure.Results 47 subjects completed trial and 1 subject lost to follow-up.After a single intravenous injection of JS026 of 30 mg,100 mg,300 mg,600 mg,and 1 000 mg,mean Cmax were(9.47±1.53),(33.20±4.95),(96.10±13.70),(177.00±22.20)and(353.00±56.70)μg·mL-1,respectively;mean AUC0-∞ were(4 225.00±607.00),(1.78 × 104±3 268.00),(5.83 × 104±1 038.00),(1.07 × 105±152.00),(1.66 × 105±327.00)μg·h·mL-1,respectively;mean t1/2 of JS026 were 563-709 h.The Cmax and AUC0-∞ of JS026 were basically similar alone or in combination with JS016.The results of Power model showed that Cmax and AUC0-∞ increased approximately linearly with the increasing dose of JS026.Treatment emergent adverse event was not increasing when dose increased and most of adverse event associated with drugs were abnormal on laboratory tests and haematuria,thus JS026 and JS016 was well tolerated in all groups.Conclusion The single intravenous infusion of JS026 can almost be thought to be a linear relationship between the doses and drug serum exposure.JS016 had no significant effect on serum concentration of JS026 and JS026 was well tolerated and safe in healthy subjects within 30-1 000 mg.
4.Clinical trial on rituximab in the treatment of patients with neuromyelitis optica spectrum disorder
Xue-Juan LIN ; Wen-Jing WU ; Jing-Yi TONG
The Chinese Journal of Clinical Pharmacology 2024;40(16):2311-2315
Objective To explore the changes in peripheral blood interleukin-23(IL-23)/IL-17A axis and microRNA-365-5p(miR-363-5p)in patients with neuromyelitis optica spectrum disorder(NMOSD)before and after treatment with rituximab.Methods NMOSD patients were divided into aquaporin-4 immunoglobulin G(AQP-4IgG)positive group and AQP-4IgG negative group based on the presence of AQP-4IgG antibodies.Both groups received continuous treatment with rituximab(PTX)for over one year.The therapeutic effect of the two groups was compared,and the annual recurrence times,ADL scores and peripheral blood expression levels of IL-23,IL-17A and miR-363-5p were compared before treatment and 12 months after treatment,and the occurrence of adverse drug reactions were recorded.Results AQP-4IgG positive group and AQP-4IgG negative group included 73 cases and 27 cases respectively.The clinical therapeutic effect of AQP-4IgG positive group and AQP-4IgG negative group was 84.93%and 70.37%,with no statistical significance(P>0.05).Before treatment,the annual recurrence times of AQP-4IgG positive group and AQP-4IgG negative group were(1.36±0.32)and(1.33±0.41)times per year,the ADL scores were(62.36±5.76)and(63.27±5.38)points,IL-23 were(325.23±116.35)and(328.79±120.38)pg·mL-1,IL-17A were(68.46±12.38)and(69.61±13.41)pg·mL-1,miR-363-5p were 1.76±0.10 and 1.77±0.19,respectively.After 12 months of treatment,the annual recurrence times of AQP-4IgG positive group and AQP-4IgG negative group were(0.28±0.16)and(0.31±0.12)times per year,ADL scores were(85.10±10.36)and(81.26±11.34)points,IL-23 were(122.46±11.54)and(119.49±10.26)pg·mL-1,IL-17A were(43.16±6.29)and(40.27±8.75)pg·mL-1,miR-363-5p were 1.38±0.15 and 1.36±0.15,respectively.There were statistically significant differences in the above indexes between the two groups after treatment and before treatment(all P<0.05),but no significant differences between the groups post-treatment(all P>0.05).The incidence rates of adverse drug reactions were 13.69%in the AQP-4IgG positive group and 18.51%in the negative group,with no statistically significant difference(P>0.05).Conclusion Rituximab is effective in treating NMOSD,reducing the number of annual relapses and promoting the recovery of patients'mobility.The mechanism may be related to regulating the IL-23/IL-17A axis and inhibiting the expression of miR-363-5p.
5.Expert consensus on the clinical application of efficient intelligent chromosomal karyotyping precise auxiliary diagnosis system
Hao WANG ; Juan WEN ; Yi LAI ; Lingqian WU
Chinese Journal of Medical Genetics 2024;41(9):1025-1031
Chromosomal karyotyping analysis has been considered as the gold standard for cytogenetic diagnosis and an effective measure for preventing birth defects. However, conventional karyotyping analysis relies heavily on manual recognition, which is time-consuming and labor-intensive. The application of an efficient intelligent chromosomal karyotyping precise auxiliary diagnosis system in clinical practice can significantly reduce the analysis time and greatly improve the efficiency, increase the detection rate for low-percentage mosaicisms, and promote homogenization between laboratories. This can effectively enhance the capacity and level of cytogenetic diagnosis. With the continuous application of such system in the field of karyotyping analysis, a substantial amount of clinical application data and experience has been accumulated. This consensus has been formulated after multiple rounds of discussion by experts from the clinical application collaboration group of the efficient intelligent chromosomal karyotyping precise auxiliary diagnosis system. It aims to provide a reference for peers to better utilize intelligent auxiliary diagnosis systems during chromosomal karyotyping analysis and promote the standardized development of karyotyping analysis technology.
6.Occurrence of neglected perforation of sterile surgical gloves and its effect on bacterial contamination during craniocerebral surgery
Ning NIU ; Ting-Ting LI ; Jing-Jing JI ; Wen-Juan HU ; Hao WU
Journal of Regional Anatomy and Operative Surgery 2024;33(12):1085-1088
Objective To investigate the incidence of neglected perforation of sterile surgical gloves used in craniocerebral surgery and its effect on bacterial contamination of the gloves.Methods A total of 996 sterile surgical gloves worn by surgical participants during craniocerebral surgery were selected as the study objects.Univariate and multivariate Logistic regression analysis were respectively performed to analyze the factors that might cause the neglected perforation of gloves,and the detection rate of bacteria on the outer surface of perforated and unperforated gloves were compared.Results Among 996 sterile surgical gloves used in craniocerebral surgery,84(8.43%)gloves were found to have neglected perforation,and 39(3.92%)gloves were found to have bacteria.The detection rate of bacteria on the outer surface of the neglected perforation group was significantly higher than that of the non-neglected perforation group(P<0.001).The results of univariate and multivariate Logistic analysis suggested that craniotomy,emergency surgery,surgery time≥150 minutes,the use of rotating equipment during surgery and the role of glove wearing personnel as the main surgeon were the risk factors for neglected perforation of sterile surgical gloves(OR>1,P<0.05),while the use of double-layer gloves during surgery was the protective factor for avoiding perforation(OR<1,P<0.05).Conclusion The occurrence of neglected perforation of sterile surgical gloves during craniocerebral surgery is not optimistic.The probability of neglected perforation of gloves is higher in craniotomy,emergency surgery,long surgery time,the use of rotating equipment during surgery,and gloves used by the main surgeon.For surgeries with a high incidence of neglected perforation,double-layer gloves can be worn during surgery to reduce bacterial contamination on the outer surface of sterile surgical gloves.
7.Impact of different posture angles on the pain of patients with early esophageal cancer after endoscopic submucosal dissection
Tian TIAN ; Juan LI ; Zhaorong WU ; Jing WANG ; Qian WANG ; Wen LI
Chinese Journal of Practical Nursing 2024;40(16):1201-1206
Objective:To explore the effect of different posture angles on the pain and comfort in patients with early esophageal cancer after endoscopic submucosal dissection (ESD), so as to provide a basis for patients to choose the best position after ESD.Methods:This study was a randomized controlled trial. One hundred and twenty patients with early esophageal cancer who underwent ESD in the Department of Gastroenterology, Nanjing Drum Tower Hospital from March 2021 to March 2022 were selected as the study subjects, and they were randomly divided into 4 groups of 30 patients each according to the randomized numerical method. Group A was the conventional group, which was in the free position after the operation, and groups B, C, and D were the experimental groups, with group B in the head-high-feet-low 30° position, group C in the head-high-feet-low 45° position, and group D in the head-high-feet-low 60° position. The pain scores after returning to the room after the operation, at 8, 16, 24 h after the operation and comfort scores at 24 h after the operation of the patients in the four groups were evaluated by Numeric Rating Scale (NRS) and General Comfort Questionnaire (GCQ).Results:There were 17 males and 13 females in group A; there were 20 males and 10 females in group B; there were 22 males and 8 females in group C; there were 19 males and 11 females in group D. All patients were aged 30-85 years old. The time main effect, grouping main effect, and interaction effect of postoperative pain NRS score among four groups of patients were all statistically significant ( F=618.13, 12.14, 6.75, all P<0.01). There was no significant difference in the NRS scores of patients after returning to the room after the operation among the four groups ( P>0.05). The NRS scores in group D at 8 and 16 h after the operation were (1.93 ± 0.64), (0.60 ± 0.47) points, lower than the (2.87 ± 1.14), (1.97 ± 1.22) points, (2.17 ± 0.83), (1.97 ± 1.61) points, (2.30 ± 0.75), (0.80 ± 0.61) points in groups A, B, and C, the differences were statistically significant ( t values were 0.79-4.72, all P<0.05). The NRS scores in group B at 24 h after the operation was (0.23 ± 0.18) points, lower than the (1.53 ± 1.08), (0.30 ± 0.21), (0.46 ± 0.25) points in groups A, C, and D, the differences were statistically significant ( t= 5.32, 1.34, 1.37, all P<0.05). The GCQ total scores at 24 h after the operation were (96.96 ± 3.05), (99.77 ± 3.21), (93.53 ± 3.76), (92.20 ± 3.69) points in group A, B, C, D, the difference was statistically significant ( F= 29.59, P<0.05). Moreover, the GCQ total scores at 24 h after the operation in group B were higher than those in groups A, C, and D, and the differences were statistically significant ( t=3.15, 7.01, 8.52, all P<0.05). Conclusions:Targeted body position management can effectively reduce postoperative pain and improve patient comfort in early esophageal cancer patients undergoing ESD.
8.Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients (version 2024)
Yao LU ; Yang LI ; Leiying ZHANG ; Hao TANG ; Huidan JING ; Yaoli WANG ; Xiangzhi JIA ; Li BA ; Maohong BIAN ; Dan CAI ; Hui CAI ; Xiaohong CAI ; Zhanshan ZHA ; Bingyu CHEN ; Daqing CHEN ; Feng CHEN ; Guoan CHEN ; Haiming CHEN ; Jing CHEN ; Min CHEN ; Qing CHEN ; Shu CHEN ; Xi CHEN ; Jinfeng CHENG ; Xiaoling CHU ; Hongwang CUI ; Xin CUI ; Zhen DA ; Ying DAI ; Surong DENG ; Weiqun DONG ; Weimin FAN ; Ke FENG ; Danhui FU ; Yongshui FU ; Qi FU ; Xuemei FU ; Jia GAN ; Xinyu GAN ; Wei GAO ; Huaizheng GONG ; Rong GUI ; Geng GUO ; Ning HAN ; Yiwen HAO ; Wubing HE ; Qiang HONG ; Ruiqin HOU ; Wei HOU ; Jie HU ; Peiyang HU ; Xi HU ; Xiaoyu HU ; Guangbin HUANG ; Jie HUANG ; Xiangyan HUANG ; Yuanshuai HUANG ; Shouyong HUN ; Xuebing JIANG ; Ping JIN ; Dong LAI ; Aiping LE ; Hongmei LI ; Bijuan LI ; Cuiying LI ; Daihong LI ; Haihong LI ; He LI ; Hui LI ; Jianping LI ; Ning LI ; Xiying LI ; Xiangmin LI ; Xiaofei LI ; Xiaojuan LI ; Zhiqiang LI ; Zhongjun LI ; Zunyan LI ; Huaqin LIANG ; Xiaohua LIANG ; Dongfa LIAO ; Qun LIAO ; Yan LIAO ; Jiajin LIN ; Chunxia LIU ; Fenghua LIU ; Peixian LIU ; Tiemei LIU ; Xiaoxin LIU ; Zhiwei LIU ; Zhongdi LIU ; Hua LU ; Jianfeng LUAN ; Jianjun LUO ; Qun LUO ; Dingfeng LYU ; Qi LYU ; Xianping LYU ; Aijun MA ; Liqiang MA ; Shuxuan MA ; Xainjun MA ; Xiaogang MA ; Xiaoli MA ; Guoqing MAO ; Shijie MU ; Shaolin NIE ; Shujuan OUYANG ; Xilin OUYANG ; Chunqiu PAN ; Jian PAN ; Xiaohua PAN ; Lei PENG ; Tao PENG ; Baohua QIAN ; Shu QIAO ; Li QIN ; Ying REN ; Zhaoqi REN ; Ruiming RONG ; Changshan SU ; Mingwei SUN ; Wenwu SUN ; Zhenwei SUN ; Haiping TANG ; Xiaofeng TANG ; Changjiu TANG ; Cuihua TAO ; Zhibin TIAN ; Juan WANG ; Baoyan WANG ; Chunyan WANG ; Gefei WANG ; Haiyan WANG ; Hongjie WANG ; Peng WANG ; Pengli WANG ; Qiushi WANG ; Xiaoning WANG ; Xinhua WANG ; Xuefeng WANG ; Yong WANG ; Yongjun WANG ; Yuanjie WANG ; Zhihua WANG ; Shaojun WEI ; Yaming WEI ; Jianbo WEN ; Jun WEN ; Jiang WU ; Jufeng WU ; Aijun XIA ; Fei XIA ; Rong XIA ; Jue XIE ; Yanchao XING ; Yan XIONG ; Feng XU ; Yongzhu XU ; Yongan XU ; Yonghe YAN ; Beizhan YAN ; Jiang YANG ; Jiangcun YANG ; Jun YANG ; Xinwen YANG ; Yongyi YANG ; Chunyan YAO ; Mingliang YE ; Changlin YIN ; Ming YIN ; Wen YIN ; Lianling YU ; Shuhong YU ; Zebo YU ; Yigang YU ; Anyong YU ; Hong YUAN ; Yi YUAN ; Chan ZHANG ; Jinjun ZHANG ; Jun ZHANG ; Kai ZHANG ; Leibing ZHANG ; Quan ZHANG ; Rongjiang ZHANG ; Sanming ZHANG ; Shengji ZHANG ; Shuo ZHANG ; Wei ZHANG ; Weidong ZHANG ; Xi ZHANG ; Xingwen ZHANG ; Guixi ZHANG ; Xiaojun ZHANG ; Guoqing ZHAO ; Jianpeng ZHAO ; Shuming ZHAO ; Beibei ZHENG ; Shangen ZHENG ; Huayou ZHOU ; Jicheng ZHOU ; Lihong ZHOU ; Mou ZHOU ; Xiaoyu ZHOU ; Xuelian ZHOU ; Yuan ZHOU ; Zheng ZHOU ; Zuhuang ZHOU ; Haiyan ZHU ; Peiyuan ZHU ; Changju ZHU ; Lili ZHU ; Zhengguo WANG ; Jianxin JIANG ; Deqing WANG ; Jiongcai LAN ; Quanli WANG ; Yang YU ; Lianyang ZHANG ; Aiqing WEN
Chinese Journal of Trauma 2024;40(10):865-881
Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.
9.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.
10.The Mechanism of Brucea javanica Regulating Hela Cell Apoptosis Through TLR9-MyD88 Signaling Pathway
Juan YANG ; Weiqi WU ; Xiuyi LU ; Liuyan WEN ; Shaoping YUAN ; Yan BAI ; Qiwen WU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(6):1481-1489
Objective To investigate the molecular mechanism of Brusatol(BRU)regulating apoptosis of Hela cells through TLR9-MyD88 signaling pathway.Methods Hela cells preserved in our laboratory were treated with Brucea javanica at different concentrations(0,5.0,10.0,20.0,40.0,80.0 nmol·L-1).Hela cells were divided into Control group(normal cultured Hela cells),BRU-L group(treated with 10.0 nmol·L-1 Brucea javanica)and BRU group-H(treated with 20.0 nmol·L-1 Brucea javanica)Cells were treated with nmol·L-1 of Brucea javanica),BRU+pcDNA-NC group(transfected with pcDNA-NC+20.0 nmol·L-1 of Brucea javanica),BRU-H+pcDNA-TLR9 group(transfected with pcDNA-TLR9+20.0 nmol·L-1 of Brucea javanica).Cell proliferation was detected by CCK-8 and EdU methods.Apoptosis was detected by TUNNEL staining and flow cytometry.The protein expression levels of TLR9,MyD88,Bax,Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were detected by Western blot.Cell apoptosis and mitochondrial membrane potential detection kit(JC-1)were detected by flow cytometry,and the contents of adenosine triphosphate(ATP)and reactive oxygen species(ROS)were detected by ELISA.Results Compared with 0 nmol·L-1 group,the survival rate and IC50 value of 10 nmol·L-1 group were significantly decreased(P<0.05).After stimulation of BRU with different concentrations,the proliferation ability of cells was significantly decreased in a dose-dependent manner(P<0.05).Compared with control group,the 5-ethynyl-2'-deoxyuracil(EDU)positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein of cells in BRU-L,BRU-H and pcDNA-NC groups were significantly decreased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly increased(P<0.05).In control group,BRU-L,BRU-H group/BRU-H+pcDNA-NC,there was a continuous decreasing trend(P<0.05).Compared with the BRU-H+pcDNA-NC group,the EDU positive cell rate,TUNEL positive cell rate,apoptosis rate and Bcl-2 protein in the BRU-H+pcDNA-TLR9 group were significantly increased.The protein levels of TLR9 and MyD88,Bax,Bax/Bcl-2,Cleaved Caspase-3 and Cleaved Caspase-9 were significantly decreased(P<0.05).Compared with Control group,JC-1 level and ATP content in BRU-L,BRU-H and BRU-H+pcDNA-NC groups were significantly decreased,while ROC content and mitotracker positive cell level were significantly increased(P<0.05).Compared with BRU-L group,JC-1 level and ATP content in BRU-H group and BRU-H+pcDNA-NC group were further decreased,while ROC content and mitotracker positive cell level were further increased(P<0.05).Compared with the BRU-H+pcDNA-NC group,the levels of JC-1 and ATP in the BRU-H+pcDNA-TLR9 group were increased,while the levels of ROC,mitotracker staining positive cells were decreased(P<0.05).Conclusion Brucea javanica can produce Hela cell proliferation by regulating TLR9-MyD88 signaling pathway.

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