1.Research of high iodine induced damage to thyroid epithelial cells through endoplasmic reticulum stress and P38MAPK signaling pathway
Xin-Na YANG ; Hui-Yao CAI ; Li-Jun CHEN ; Bo LIANG
The Chinese Journal of Clinical Pharmacology 2024;40(16):2339-2343
Objective To investigate the role of endoplasmic reticulum stress and p38 mitogen-activated protein kinase P38MAPK signaling pathway in thyroid epithelial cell injury induced by high iodine.Methods The thyroid epithelial cells Nthy-ori 3-1 were randomly divided into control group(normal culture),model group(40 mmol·L-1 potassium iodide),4-phenylbutyric acid(4-PB A)group(40 mmol·L-1 potassium iodide and 2 mmol·L-1 4-PBA)and SB203580 group(40 mmol·L-1 potassium iodide and 10 μmol·L-1 SB203580).Western blot was used to detect the expression of glucose regulated protein 78(GRP78)and p-P38/P38 of Nthy-ori 3-1 cells.MTT and colony formation experiments were used to detect the proliferation level.Flow cytometry was used to detect the apoptosis level.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of interleukin-6(IL-6).Results The expression levels of GRP78 protein in control group,model group,4-PBA group and SB203580 group were 0.15±0.03,0.61±0.07,0.27±0.03 and 0.37±0.04;the ratios of p-P38/P38 were 0.12±0.03,0.53±0.04,0.35±0.04 and 0.25±0.03;cell survival rates were(100.00±0.00)%,(53.71±6.16)%,(80.24±8.17)%and(71.29±7.36)%;the number of clones formed was 271.36±25.18,96.09±10.79,183.24±15.36 and 141.24±16.18;the apoptosis rates were(1.04±0.21)%,(9.27±1.67)%,(3.18±1.52)%and(3.82±1.09)%;IL-6 secretion levels were(0.71±0.08),(9.17±0.87),(3.26±0.29)and(4.71±0.41)nmol·L-1,respectively.For the above indicators,there was significant difference between the model group and the control group(all P<0.05);there was significant difference between the 4-PBA group,SB203580 group and the model group(all P<0.05).Conclusion High iodine can inhibit the proliferation of Nthy-ori 3-1 cells and induce apoptosis and secretion of inflammatory factors,which may be related to the activation of endoplasmic reticulum stress and P38MAPK signaling pathway by high iodine.
2.Methodological Exploration for Global Cardiovascular Academic Performance Evaluation(CAPE)System
Lu YIN ; Xueyan ZHANG ; Yeding CAO ; Wei LI ; Yan YAO ; Zhiyuan BO ; Liang WEI ; Jun CAI ; Jingang YANG ; Shengshou HU
Chinese Circulation Journal 2024;39(1):3-16,中插1-中插4
Objectives:To establish a comprehensive system of Cardiovascular Academic Performance Evaluation(CAPE)and rank global TOP100 medical institutions in the fields of cardiovascular diseases(CVD). Methods:CVD-related terms were extracted from Medical Subject Headings(MeSH),Embase thesaurus(EMtrees)and International Classification of Diseases(ICD)by CVD-related professionals,as well as by librarians and information professionals.Terminology databases(named as Fuwai Subject Headings)were established,and nine sub-disciplines were proposed,including ischemic heart diseases,hypertension,vascular diseases,arrhythmia,pulmonary vascular diseases,heart failure,congenital heart diseases,cardiomyopathy,and valvular heart diseases.The mapping patterns of sub-discipline,cardiovascular terminology and entry terms were pre-defined.The CVD-related research literature published from January 1,2016 to December 31,2022 were retrieved from Web of Science,PubMed and Scopus.Based on this,metadata were fused and duplicates were excluded.Fuwai Subject Headings were searched and matched into four respects for each literature,including subject words,titles,keywords,and abstracts,which was used to generate an information table of"Position—CVD terminology—Frequency",and to calculate CVD correlation scores and sub-discipline scores.We standardized the names of medical institutions and scholars,and make a ranking system for CAPE based on original articles with strong cardiovascular correlation(correlation score≥4).When evaluating the science and technological performance for Chinese hospitals in cardiovascular diseases,National Natural Science Foundation Projects,authorized invention patents,prize achievements,research platforms,and registered data of drug clinical trials in Center for Drug Evaluation(CDE)were considered besides research papers. Results:During 2016 and 2022,1 545 103 CVD research literatures were found worldwide.After excluding meeting abstracts,books,biographies,news,videos,audio texts,retracted publications,and corrections,1 178 019 CVD research literatures were further evaluated.518 058 literatures were indexed as"strongly correlated to CVD"using Fuwai Subject Headings.Besides papers,other data sources were also collected,including 11 143 CVD-related Natural Science Foundation Projects,19 382 CVD-related effective authorized invention patents,103 CVD-related national prize achievements,24 CVD-related national research platforms,and 2 084 CDE registered data of CVD-related drug clinical trials.Research teams from nine sub-disciplines reviewed and validated research literature in respective fields,and classification rules of corresponding sub-disciplines were created and improved based on their opinions.Finally,eleven individual indexes were chosen to construct CAPE system for ranking global TOP100 medical institutions in overall CVD field and TOP30 in nine sub-disciplines.From 2016 to 2022,the number of cardiovascular disease research papers published by Chinese institutes has increased by 123.5%,with a total of approximately 76.8 thousands papers published(about 30 papers per day on average),ranked the second under the United States(approximately 114.1 thousands papers).However,the proportion of papers published by the Chinese Journal Citation Reports(JCR)and the Chinese Academy of Sciences only ranked eighth in the world.In the comprehensive academic performance of original cardiovascular research papers in global hospitals from 2020 to 2022,only two Chinese medical institutions ranked in the TOP20 as evaluated by CAPE system. Conclusions:Based on multi-source data from 2016 to 2022,CAPE initiated to establish a cardiovascular academic performance evaluation system.
3.Comparison of clinical characteristics between first-episode and relapse of major depressive disorder
Xiuyan ZHENG ; Chengxia TANG ; Zhaorui LIU ; Tingting ZHANG ; Yueqin HUANG ; Liang ZHOU ; Yuandong GONG ; Yan LIU ; Bo LIU ; Jie ZHANG ; Haiming WANG ; Zhengmin FENG ; Jun GUO ; Wenming CHEN ; Linling JIANG ; Defang CAI ; Jin LU
Chinese Mental Health Journal 2024;38(1):25-32
Objective:To describe demographic,clinical and physiological characteristics,treatment between first-episode major depressive disorder(MDD)and relapse MDD,and to explore characteristics of relapse MDD.Methods:Totally 858 patients who met the diagnostic criteria for depression of the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5),were included by using the Mini International Neuropsychiatric Interview(MINI),Clinician-Rated Dimensions of Psychosis Symptom Severity,and Hamilton Depression Scale etc.Among them,529(58.6%)were first-episode depression and 329(36.0%)were relapsed.The differences of demographic characteristics,clinical and physiological characteristics,treatment were compared byx2test and Kruskal-Wallis rank sum test.Multivariate logistic regression was used to explore the characteristics of MDD recur-rence.Results:Compared to first-episode MDD,relapse MDD had more comorbidity(OR=2.11,95%CI:1.00-4.44),more days out of role(OR=1.26,95%CI:1.01-1.56),more history of using psychiatric drug more than one month(OR=1.41,95%CI:1.02-1.97)and electroconvulsive therapy(OR=3.23,95%CI:1.42-7.36),and higher waist-hip ratio(OR=33.88,95%CI:2.88-399.32).Conclusion:Relapse MDD has positive as-sociation with comorbidity of mental disorders,out of role,and higher waist-hip ratio.
4.Application value of transanal endoscopic intersphincteric resection in sphincter preserva-tion for low rectal cancer
Gaojian CAO ; Ximo XU ; Hao ZHONG ; Zhenghao CAI ; Jun YOU ; Mingyang REN ; Liang KANG ; Bo FENG
Chinese Journal of Digestive Surgery 2024;23(6):836-844
Objective:To investigate the application value of transanal endoscopic intersphincteric resection (taE-ISR) in sphincter preservation for low rectal cancer.Methods:The pro-pensity score matching and retrospective cohort study was conducted. The clinicopathological data of 278 patients with low rectal cancer who were admitted to 5 medical centers, including Ruijin Hospital of Shanghai Jiaotong University School of Medicine et al, from January 2017 to December 2021 were collected. There were 178 males and 100 females, aged 58 (range, 49-64)years. Of 278 pati-ents, 147 cases undergoing taE-ISR were divided into the taE-ISR group, and 131 cases undergoing intersphincteric resection (ISR) were divided into the ISR group. Observation indicators:(1) propen-sity score matching and comparison of general data of patients between the two groups after matching; (2) comparison of intraoperative and postoperative conditions between the two groups; (3) long-term follow-up of the two groups; (4) analysis of risk factors affecting sphincter preservation for low rectal cancer. Propensity score matching was done by the 1∶1 nearest neighbor matching method, with a caliper value of 0.05. Propensity score matching analysis was done using the Matching package. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the Student′s t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Wilcoxon rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the Pearson chi-square test or Fisher exact probability. The Kaplan-Meier method was used to calculate survival rate and plot survival curve, and the Log-Rank test was used for survival analysis. Multivariate analysis was conducted using the Logistic regression model with the "glm2" package. The forest plot was used to show the risk factors affecting sphincter preservation for low rectal cancer. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of 278 patients, 180 cases were successfully matched, including 90 cases in the taE-ISR group and 90 cases in the ISR group, respectively. After propensity score matching, the elimination of distance between ischial tuberosities and distance from ischial tuberosity to the skin of buttocks confounding bias ensured comparability between the two groups. (2) Comparison of intraoperative and postoperative conditions between the two groups. Cases with positive distal margins, cases with specimen integrity, cases with sphincter preservation were 1, 88, 88 in the taE-ISR group and 8, 78, 74 in the ISR group, showing significant differences between the two groups ( P<0.05). (3) Long-term follow-up of the two groups. The median follow-up time was 4.3(range, 3.8-5.0)years of the taE-ISR group and 4.1(range, 3.4-4.7)years of the ISR group. The overall survival rate, disease-free survival rate and cumulative recurrence rate were 100.0%, 95.6% and 2.2% of the taE-ISR group, versus 98.9%, 87.8% and 10.0% of the ISR group, showing no significant difference in overall survival rate between the two groups ( χ2=0.97, P>0.05) and significant differences in disease-free survival rate and cumulative recurrence rate between the two groups ( χ2=4.05, 5.26, P<0.05). (4) Analysis of risk factors affecting sphincter preservation for low rectal cancer. Results of multivariate analysis showed that taE-ISR, distance from the tumor to the anus, and adjacent organ damage were independent factors affecting sphincter preservation for low rectal cancer ( odds ratio=0.86, 0.88, 1.35, 95% confidence interval as 0.79-0.93, 0.83-0.92, 1.04-1.74, P<0.05). In further analysis, there were significant differences in sphincter preservation and defecatory dysfunction between the 21 cases with neoadjuvant therapy in the taE-ISR group and the 19 cases with neoadjuvant therapy in the ISR group ( P<0.05). Conclusions:The taE-ISR is safe and feasible for patients with low rectal cancer. Compared with ISR, taE-ISR can significantly improve surgical quality, sphincter preservation rate and patient prognosis.
5.Effect of TLK2 Expression Regulated by MiR-21 on Proliferation and Apoptosis of Acute Myeloid Leukemia Cells
Bo LIANG ; Jun-Jie YIN ; Sheng-Nan ZHANG ; Chao ZHANG ; Zi-Long HU ; Yi WANG
Journal of Experimental Hematology 2024;32(3):658-662
Objective:To investigate the effect of TLK2 expression regulated by miR-21 on proliferation and apoptosis of acute myeloid leukemia cells.Methods:Seventy patients with AML admitted to our hospital from January 2019 to July 2022 were selected,while 30 patients with iron deficiency anemia were selected as the control group.Bone marrow mononuclear cells(BMMNCs)of the patients were obtained using Ficoll density gradient centrifugation.RT-qPCR was used to determine the expression levels of miR-21 and TLK2 mRNA in BMMNCs.Mimics-miR-21,mimics-NC,inhibitor-miR-21,inhibitor-NC and NC were transfected into HL-60 cells using liposome-mediated transfection technology.CCK-8 method was used to determine the activity of transfected HL-60 cells after treatment with cytarabine.The apoptosis rate of HL-60 transfected cells was determined by TUNEL method.The expression of TLK2 mRNA in HL-60 cells transfected with inhibitor-miR-21 was determined by RT-qPCR.Results:The relative expression levels of miR-21 and TLK2 mRNA in BMMNCs of AML patients were significantly higher than those of controls(both P<0.05).After HL-60 cells were treated with cytarabine,both the cell activity of inhibitor-miR-21 group and mimics-miR-21 group decreased significantly with the increase of cytarabine concentration(both P<0.05).However,at each concentration point of cytarabine,the cell activity of inhibitor-miR-21 group was lower than that of control group(P<0.05),while mimics-miR-21 group was higher than control group(P<0.05).After HL-60 cells were treated with cytarabine,the apoptosis rate of inhibitor-miR-21 group was significantly increased(P<0.05),while that of mimics-miR-21 group was significantly decreased(P<0.05).After HL-60 cells were treated with inhibitor-miR-21,the relative expression of TLK2 mRNA decreased significantly(P<0.05).Conclusion:miR-21 is highly expressed in AML patients,which may promote the apoptosis of AML cells by inhibiting the expression of TLK2.
6.Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome (version 2024)
Junyu WANG ; Hai JIN ; Danfeng ZHANG ; Rutong YU ; Mingkun YU ; Yijie MA ; Yue MA ; Ning WANG ; Chunhong WANG ; Chunhui WANG ; Qing WANG ; Xinyu WANG ; Xinjun WANG ; Hengli TIAN ; Xinhua TIAN ; Yijun BAO ; Hua FENG ; Wa DA ; Liquan LYU ; Haijun REN ; Jinfang LIU ; Guodong LIU ; Chunhui LIU ; Junwen GUAN ; Rongcai JIANG ; Yiming LI ; Lihong LI ; Zhenxing LI ; Jinglian LI ; Jun YANG ; Chaohua YANG ; Xiao BU ; Xuehai WU ; Li BIE ; Binghui QIU ; Yongming ZHANG ; Qingjiu ZHANG ; Bo ZHANG ; Xiangtong ZHANG ; Rongbin CHEN ; Chao LIN ; Hu JIN ; Weiming ZHENG ; Mingliang ZHAO ; Liang ZHAO ; Rong HU ; Jixin DUAN ; Jiemin YAO ; Hechun XIA ; Ye GU ; Tao QIAN ; Suokai QIAN ; Tao XU ; Guoyi GAO ; Xiaoping TANG ; Qibing HUANG ; Rong FU ; Jun KANG ; Guobiao LIANG ; Kaiwei HAN ; Zhenmin HAN ; Shuo HAN ; Jun PU ; Lijun HENG ; Junji WEI ; Lijun HOU
Chinese Journal of Trauma 2024;40(5):385-396
Traumatic supraorbital fissure syndrome (TSOFS) is a symptom complex caused by nerve entrapment in the supraorbital fissure after skull base trauma. If the compressed cranial nerve in the supraorbital fissure is not decompressed surgically, ptosis, diplopia and eye movement disorder may exist for a long time and seriously affect the patients′ quality of life. Since its overall incidence is not high, it is not familiarized with the majority of neurosurgeons and some TSOFS may be complicated with skull base vascular injury. If the supraorbital fissure surgery is performed without treatment of vascular injury, it may cause massive hemorrhage, and disability and even life-threatening in severe cases. At present, there is no consensus or guideline on the diagnosis and treatment of TSOFS that can be referred to both domestically and internationally. To improve the understanding of TSOFS among clinical physicians and establish standardized diagnosis and treatment plans, the Skull Base Trauma Group of the Neurorepair Professional Committee of the Chinese Medical Doctor Association, Neurotrauma Group of the Neurosurgery Branch of the Chinese Medical Association, Neurotrauma Group of the Traumatology Branch of the Chinese Medical Association, and Editorial Committee of Chinese Journal of Trauma organized relevant experts to formulate Chinese expert consensus on the diagnosis and treatment of traumatic supraorbital fissure syndrome ( version 2024) based on evidence of evidence-based medicine and clinical experience of diagnosis and treatment. This consensus puts forward 12 recommendations on the diagnosis, classification, treatment, efficacy evaluation and follow-up of TSOFS, aiming to provide references for neurosurgeons from hospitals of all levels to standardize the diagnosis and treatment of TSOFS.
7.Analysis of clinical characteristics and influencing factors of adverse treatment outcomes in 238 elderly patients with multidrug-resistant pulmonary tuberculosis
LIANG Bowen ; XIAN Fuyang ; LI Bo ; LUO Jingyue ; ZHAO Liping ; AN Jun ; GAO Mengqiu ; LI Hua
China Tropical Medicine 2024;24(3):276-
Objective To analyze the clinical characteristics and factors influencing adverse treatment outcomes in elderly patients with multidrug-resistant pulmonary tuberculosis (MDR-TB) to guide the clinical diagnosis and treatment of elderly MDR-TB patients. Methods Clinical data of elderly patients with multidrug-resistant pulmonary tuberculosis initially treated at Beijing Chest Hospital from 2008 to 2023 were retrospectively collected. Complications/comorbidities, adverse drug reactions, drug resistance during initial treatment, and retreatment were statistically described. Factors influencing adverse treatment outcomes were analyzed using the chi-square test and logistic regression analysis. Results A total of 238 elderly patients with MDR-TB were collected, of which 152 (63.9%) had adverse drug reactions, 184 (77.3%) were retreated MDR-TB, 27 (11.3%) were extensively drug-resistant tuberculosis (XDR-TB), 41 were cured, 6 completed treatment, 39 failed treatment, 6 died, 107 lost to follow-up, 31 could not be evaluated, 8 did not finish treatment, and the treatment success rate was 20.4% (47/230). The adverse outcome of treatment accounted for 79.6% (183/230). Univariate analysis showed that differences in age groups, the occurrence of drug adverse reactions, and patient sources had a statistically significant impact on treatment outcomes (P<0.05). Logistic regression analysis was performed using good and adverse treatment outcomes as dependent variables for the three factors, which showed that being aged 70 and above, the occurrence of drug adverse reactions during treatment, and being a non-local patient were factors influencing adverse treatment outcomes [OR (95%CI): 2.507 (1.027-6.121), 3.253 (1.635-6.473), 2.563 (1.285-5.111), respectively]. Conclusions Elderly patients with MDR-TB exhibit a high prevalence of complications/comorbidities, a high incidence of drug adverse reactions, and unfavorable treatment outcomes. Out-of-town medical treatment, advanced age, and experiencing drug adverse reactions are risk factors for adverse treatment outcomes.
8.Current status of surgery for portal hypertension in China: a national multi-center survey analysis
Lei ZHENG ; Haiyang LI ; Jizhou WANG ; Xiao LIANG ; Jian DOU ; Jitao WANG ; Qiang FAN ; Xiong DING ; Wenlong ZHAI ; Yun JIN ; Bo LI ; Songqing HE ; Tao LI ; Jun LIU ; Kui WANG ; Zhiwei LI ; Yongyi ZENG ; Yingmei SHAO ; Yang BU ; Dong SHANG ; Yong MA ; Cheng LOU ; Xinmin YIN ; Jiefeng HE ; Haihong ZHU ; Jincai WU ; Zhidan XU ; Dunzhu BASANG ; Jianguo LU ; Liting ZHANG ; Jianguo ZHAO ; Ling LYU ; Guoyue LYU ; Nim CHOI ; To Tan CHEUNG ; Meng LUO ; Wanguang ZHANG ; Xiaolong QI ; Xiaoping CHEN
Chinese Journal of Organ Transplantation 2023;44(3):152-159
Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.
9.Clinical Comparison of the Efficacy of Systemic Thrombolysis,Catheter-Directed Thrombolysis,and AngioJet Percutaneous Mechanical Thrombectomy in Acute Lower Extremity Deep Venous Thrombosis.
Jin-Wang LI ; Jun XUE ; Fei GUO ; Lei HAN ; Ru-Bo BAN ; Xue-Liang WU
Acta Academiae Medicinae Sinicae 2023;45(3):410-415
Objective To compare the clinical effects of three treatment methods including systemic thrombolysis(ST),catheter-directed thrombolysis(CDT),and AngioJet percutaneous mechanical thrombectomy(PMT)in acute lower extremity deep venous thrombosis(LEDVT). Methods The data of 82 patients diagnosed with LEDVT in the Department of Vascular and Gland Surgery of the First Affiliated Hospital of Hebei North University from January 2017 to December 2020 were collected.The patients were assigned into a ST group(n=50),a CDT group(n=16),and a PMT group(n=16)according to different treatment methods.The efficacy and safety were compared among the three groups. Results Compared with that before treatment,the circumferential diameter difference of both lower limbs on days 1,2,and 3 of treatment in the ST,CDT,and PMT groups reduced(all P<0.001).The PMT group showed smaller circumferential diameter difference of lower limbs on days 1,2,and 3 of treatment than the ST group(all P<0.001)and smaller circumferential diameter difference of the lower patellar margin on day 1 of treatment than the CDT group(P<0.001).The PMT group showed higher diminution rate for swelling of the affected limb at the upper and lower edges of the patella than the ST group(P<0.001)and higher diminution rate for swelling at the upper edge of the patella than the CDT group(P=0.026).The incidence of complications after treatment showed no significant differences among the three groups(all P>0.05).The median of hospital stay in the PMT group was shorter than that in the ST and CDT groups(P=0.002,P=0.001).The PMT group had higher thrombus clearance rate than the ST group(P=0.002)and no significant difference in the thrombus clearance rate from the CDT group(P=0.361).The vascular recanalization rates in the PMT(all P<0.001)and CDT(P<0.001,P=0.002,P=0.009)groups 3,6,and 12 months after treatment were higher than those in ST group,and there were no significant differences between PMT and CDT groups(P=0.341,P=0.210,P=0.341). Conclusions ST,CDT,and PMT demonstrated significant efficacy in the treatment of LEDVT,and PMT was superior to ST and CDT in terms of circumferential diameter difference of the lower limbs,diminution rate for swelling of the affected limb,thrombus clearance rate,length of hospital stay,and long-term vascular recanalization.There was no obvious difference in safety among the three therapies.
Humans
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Thrombolytic Therapy/methods*
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Fibrinolytic Agents/therapeutic use*
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Treatment Outcome
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Thrombectomy/methods*
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Venous Thrombosis/drug therapy*
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Lower Extremity/blood supply*
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Catheters
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Retrospective Studies
10.Prediction of superantigen active sites and clonal expression of staphylococcal enterotoxin-like W.
Yu Hua YANG ; Xin KU ; Ya Nan GONG ; Fan Liang MENG ; Dong bo BU ; Ya Hui GUO ; Xiao Yue WEI ; Li Jin LONG ; Jia Ming FAN ; Mao Jun ZHANG ; Jian Zhong ZHANG ; Xiao Mei YAN
Chinese Journal of Epidemiology 2023;44(4):629-635
Objective: The docking and superantigen activity sites of staphylococcal enterotoxin-like W (SElW) and T cell receptor (TCR) were predicted, and its SElW was cloned, expressed and purified. Methods: AlphaFold was used to predict the 3D structure of SElW protein monomers, and the protein models were evaluated with the help of the SAVES online server from ERRAT, Ramachandran plot, and Verify_3D. The ZDOCK server simulates the docking conformation of SElW and TCR, and the amino acid sequences of SElW and other serotype enterotoxins were aligned. The primers were designed to amplify selw, and the fragment was recombined into the pMD18-T vector and sequenced. Then recombinant plasmid pMD18-T was digested with BamHⅠand Hind Ⅲ. The target fragment was recombined into the expression plasmid pET-28a(+). After identification of the recombinant plasmid, the protein expression was induced by isopropyl-beta-D- thiogalactopyranoside. The SElW expressed in the supernatant was purified by affinity chromatography and quantified by the BCA method. Results: The predicted three-dimensional structure showed that the SElW protein was composed of two domains, the amino-terminal and the carboxy-terminal. The amino-terminal domain was composed of 3 α-helices and 6 β-sheets, and the carboxy-terminal domain included 2 α-helices and 7 antiparallel β-sheets composition. The overall quality factor score of the SElW protein model was 98.08, with 93.24% of the amino acids having a Verify_3D score ≥0.2 and no amino acids located in disallowed regions. The docking conformation with the highest score (1 521.328) was selected as the analysis object, and the 19 hydrogen bonds between the corresponding amino acid residues of SElW and TCR were analyzed by PyMOL. Combined with sequence alignment and the published data, this study predicted and found five important superantigen active sites, namely Y18, N19, W55, C88, and C98. The highly purified soluble recombinant protein SElW was obtained with cloning, expression, and protein purification. Conclusions: The study found five superantigen active sites in SElW protein that need special attention and successfully constructed and expressed the SElW protein, which laid the foundation for further exploration of the immune recognition mechanism of SElW.
Humans
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Enterotoxins/genetics*
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Superantigens/genetics*
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Catalytic Domain
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Selenoprotein W/metabolism*
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Receptors, Antigen, T-Cell

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